WorldWideScience

Sample records for bioassays

  1. Bioassay Laboratory

    Data.gov (United States)

    Federal Laboratory Consortium — The Bioassay Laboratory is an accredited laboratory capable of conducting standardized and innovative environmental testing in the area of aquatic ecotoxicology. The...

  2. Bioassay guideline 2: guidelines for tritium bioassay

    International Nuclear Information System (INIS)

    1983-01-01

    This guideline is one of a series under preparation by the Federal-Provincial Working Group on Bioassay and In Vivo Monitoring Criteria. In this report tritium compounds have been grouped into four categories for the purpose of calculating Annual Limits on Intake and Investigation Levels: tritium gas, tritiated water, tritium-labelled compounds and nucleic acid precursors

  3. Bioassay guideline 1: general guidlines for bioassay programs

    International Nuclear Information System (INIS)

    1980-01-01

    This guideline is the first of a series of documents which elaborate criteria for bioassay programs, to be presented as recommendations to the Atomic Energy Control Board. It specifies which workers require routine bioassays, the accuracy and frequency of measurements, the dose levels at which specific actions must be taken, and the documentation required

  4. Bioassay programs for radiation protection

    International Nuclear Information System (INIS)

    1979-01-01

    This report discusses the rationale for the establishment of bioassay programs as a means of protection for radiation workers in the nuclear industry. The bioassay program of the Radiation Protection Bureau is described for the years 1966-1978 and plans for future changes are outlined. (auth)

  5. Fluorescence lifetime based bioassays

    Science.gov (United States)

    Meyer-Almes, Franz-Josef

    2017-12-01

    Fluorescence lifetime (FLT) is a robust intrinsic property and material constant of fluorescent matter. Measuring this important physical indicator has evolved from a laboratory curiosity to a powerful and established technique for a variety of applications in drug discovery, medical diagnostics and basic biological research. This distinct trend was mainly driven by improved and meanwhile affordable laser and detection instrumentation on the one hand, and the development of suitable FLT probes and biological assays on the other. In this process two essential working approaches emerged. The first one is primarily focused on high throughput applications employing biochemical in vitro assays with no requirement for high spatial resolution. The second even more dynamic trend is the significant expansion of assay methods combining highly time and spatially resolved fluorescence data by fluorescence lifetime imaging. The latter approach is currently pursued to enable not only the investigation of immortal tumor cell lines, but also specific tissues or even organs in living animals. This review tries to give an actual overview about the current status of FLT based bioassays and the wide range of application opportunities in biomedical and life science areas. In addition, future trends of FLT technologies will be discussed.

  6. Applied in vitro radio bioassay

    International Nuclear Information System (INIS)

    Gaburo, J.C.G.; Sordi, G.M.A.A.

    1992-11-01

    The aim of this publication is to show the concepts and in vitro bioassay techniques as well as experimental procedures related with internal contamination evaluation. The main routes of intake, metabolic behavior, and the possible types of bioassay samples that can be collected for radionuclides analysis are described. Both biological processes and the chemical and physical behavior of the radioactive material of interest are considered and the capabilities of analytical techniques to detect and quantify the radionuclides are discussed. Next, the need of quality assurance throughout procedures are considered and finally a summary of the techniques applied to the internal routine monitoring of IPEN workers is given. (author)

  7. 77 FR 14837 - Bioassay at Uranium Mills

    Science.gov (United States)

    2012-03-13

    ... NUCLEAR REGULATORY COMMISSION [NRC-2012-0057] Bioassay at Uranium Mills AGENCY: Nuclear Regulatory..., ``Bioassay at Uranium Mills.'' This guide describes a bioassay program acceptable to the NRC staff for uranium mills and applicable portions of uranium conversion facilities where the possibility of exposure...

  8. Studies on Erythropoietin Bioassay Method

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Kyoung Sam; Ro, Heung Kyu; Lee, Mun Ho [Seoul National University College of Medicine, Seoul (Korea, Republic of)

    1975-09-15

    It is the purpose of this paper to design the most preferable method of erythropoietin bioassay in Korea. Bioassay utilizing polycythemic mice are currently in general use for the indirect determination of erythropoietin. Assay animals are usually prepared either by transfusion or by exposure to reduced oxygen tension in specially constructed chamber. We prepared the polycythemic mice by the specially constructed hypobaric chamber. We observed weights and hematocrits of the mice in the hypobaric chamber, then hematocrits and 72 hours {sup 59}Fe red cell uptake ratio of the polycythemic mice induced by hypoxia after removal from the hypobaric chamber. We designed the method of erythropoietin bioassay according to the results obtained by above experiments. Then we measured the 72 hours {sup 59}Fe red cell uptake ratio of the polycythemic mice with normal saline, normal plasma and anemic plasma according to the method we designed. The results are followed:1) The hematocrits of the mice in hypobaric chamber increased to 74% in 11 days. It is preferable to maintain the pressure of the chamber to 400 mmHg for first 4 days then 300 mmHg for last 10 days to reduce the death rate and time consuming in hypobaric chamber. 2) After removal from the hypobaric chamber, the 72 hours {sup 59}Fe red cell uptake ratio decreased rapidly and maintained the lowest level from the fourth day to tenth day. 3) We design the method of erythropoietin bioassay according to the results of above experiment and to the half life of erythropoietin. 4) The Korean product {sup 59}Fe is mixture of {sup 55}Fe and {sup 59}Fe. And the {sup 59}Fe red cell uptake ratio in normal mice was far less with Korean product {sup 59}Fe than with pure {sup 59}Fe of foreign product. So it is desirable to use pure {sup 59}Fe in this method of erythropoietin bioassay. 5) Considering the cost, the technique, the time consuming and the sensitivity it is the most preferable method of erythropoietin bioassay in Korea

  9. Exposure dose assessment using bioassay

    International Nuclear Information System (INIS)

    Suga, Shinichi

    1994-01-01

    Bioassay involves following steps: sampling, pre-treatment, chemical separation and counting of radioactivity. As bioassay samples, urines are usually used, although faecal analysis may be required in some occasions for example to assess intake of non-transferable radioactive materials. Nasal smear is a useful indicator of an inhalation case. Exhalation air is used to estimate the intake of tritiated water. Sample pre-treatment includes evaporation for concentration, wet ashing, dry ashing and co-precipitation. After adding small amount of nitric acid, the sample can be concentrated by 1/10 of initial volume, which may be used to identify γ-emitters. As the pre-treatment of urine, wet ashing is used for example for analysis of Pu, and co-precipitation is used for example for analysis of Sr. Dry ashing by electric furnace is usually adopted for faecal samples. Methods of chemical separation depend on the radionuclide(s) to be analysed. The detection limit depends also on radionuclide, and for example typical detection limits are 0.4Bq / l (volume of urine sample) for 89 Sr or 90 Sr, and 0.01 Bq / l with urine and 0.01 Bq per sample with faeces for 238 Pu, 239 Pu or 241 Am. Simpler methods can be used for some radionuclides: For example, radioactivity concentration of tritium can be determined by liquid scintillation counting of urine or condensed water from exhaled air, and natural uranium in urine can be quantified by using fluorometric method. In some circumstances, gross-α or gross-β analyses are useful for quick estimation. To estimate intakes by inhalation or by ingestion from bioassay results and to assess the committed dose equivalent, commonly available bases are the relevant publications by the ICRP and domestic guides and manuals that conform to the radiation protection regulations. (author)

  10. Interpretation of thorium bioassay data

    International Nuclear Information System (INIS)

    Juliao, L.M.Q.C.; Azeredo, A.M.G.F.; Santos, M.S.; Melo, D.R.; Dantas, B.M.; Lipsztein, J.L.

    1994-01-01

    A comparison have been made between bioassay data of thorium-exposed workers from two different facilities. The first of these facilities is a monazite sand extraction plant. Isotopic equilibrium between 232 Th and 238 Th was not observed in excreta samples of these workers. The second facility is a gas mantle factory. An isotopic equilibrium between 232 Th and 228 Th was observed in extra samples. Whole body counter measurements have indicated a very low intake of thorium through inhalation. As the concentration of thorium in feces was very high it was concluded that the main pathway of entrance of the nuclide was ingestion, mainly via contamination through dirty hands. The comparison between the bioassay results of workers from the two facilities shows that the lack of Th isotopic equilibrium observed in the excretion from the workers at the monazite sand plant possibly occurred due to an additional Th intake by ingestion of contaminated fresh food. This is presumably because 228 Ra is more efficiently taken up from the soil by plants, in comparison to 228 Th or 232 Th, and subsequently, 228 Th grows in from its immediate parent, 228 Ra. (author) 5 refs.; 3 tabs

  11. Rapid bioassay for oil-contaminated soil

    Energy Technology Data Exchange (ETDEWEB)

    Ashworth, J. [ALS Environmental, Edmonton, AB (Canada); Oosterbroek, L. [HydroQual, Calgary, AB (Canada)

    2010-07-01

    This PowerPoint presentation described a study conducted to develop a rapid bioassay for soils contaminated with oil. The bioassay method was designed for a weight of evidence (WoE) approach and eco-contact guideline derivation protocol. Microtox bioassays were conducted on cyclodextrin extracts of soil quantified by solvent extraction and gas chromatography. The method was demonstrated using straight {beta}-cyclodextrin soil extracts and activated {beta}-cyclodextrin soil extracts. An analysis of the methods showed that the activation step weakens or breaks the cyclodextrin and polycyclic hydrocarbon (PHC) inclusion complex. The released PHC became toxic to the microtox organism. Results from the bioassays were then correlated with earthworm reproduction bioassay results. tabs., figs.

  12. Bioassay for uranium mill tailings

    International Nuclear Information System (INIS)

    Tschaeche, A.N.

    1986-01-01

    Uranium mill tailings are composed of fine sand that contains, among other things, some uranium (U/sup 238/ primarily), and all of the uranium daughters starting with /sup 230/Th that are left behind after the usable uranium is removed in the milling process. Millions of pounds of tailings are and continue to be generated at uranium mills around the United States. Discrete uranium mill tailings piles exist near the mills. In addition, the tailings materials were used in communities situated near mill sites for such purposes as building materials, foundations for buildings, pipe runs, sand boxes, gardens, etc. The Uranium Mill Tailings Remedial Action Project (UMTRAP) is a U.S. Department of Energy Program designed with the intention of removing or stabilizing the mill tailings piles and the tailings used to communities so that individuals are not exposed above the EPA limits established for such tailings materials. This paper discusses the bioassay programs that are established for workers who remove tailings from the communities in which they are placed

  13. Bioassay criteria for environmental restoration workers

    International Nuclear Information System (INIS)

    Carbaugh, E.H.; Bihl, D.E.

    1993-01-01

    Environmental restoration (ER) work at the U. S. Department of Energy Hanford Site posed questions concerning when to perform bioassay monitoring of workers for potential intakes of radioactivity. Application of criteria originally developed for use inside radionuclide processing facilities to ER work resulted in overly restrictive bioassay requirements. ER work typically involves site characterization or, excavating large quantities of potentially contaminated soil, rather than working with concentrated quantities of radioactivity as in a processing facility. An improved approach, tailored to ER work, provided soil contamination concentrations above which worker bioassay would be required. Soil concentrations were derived assuming acute or chronic intakes of 2% of an Annual Limit on Intake (ALI), or a potential committed effective dose equivalent of 100 mrem, and conservative dust loading of air from the work. When planning ER work, the anticipated soil concentration and corresponding need for bioassay could be estimated from work-site historical records. Once site work commenced, soil sampling and work-place surveys could be used to determine bioassay needs. This approach substantially reduced the required number of bioassay samples with corresponding reductions in analytical costs, schedules, and more flexible work-force management. (Work supported by the US Department of Energy under contract DOE-AC06-76RLO 1830.)

  14. A specific bioassay for the inhibition of flowering.

    Science.gov (United States)

    Blake, J

    1972-06-01

    A bioassay for the inhibition of flowering involving the in vitro culture of excised, partially-induced, apices of Viscaria candida is described. This bioassay has been used to detect flowering inhibition in extracts from Kalanchoe blossfeldiana.

  15. 7 Vascular Hydrophytes for Bioassay.cdr

    African Journals Online (AJOL)

    Administrator

    4 water (see Table 1). tool. The greater extension growth of macrophyte shoots in water from downstream of STWs (Fig. 1) was supported by both chemical analysis, which showed increased phosphate concentration (Table 1), and by conventional Selenastrum bioassay in which higher cell concentrations were achieved.

  16. Bioassay of naturally occurring allelochemicals for phytotoxicity.

    Science.gov (United States)

    Leather, G R; Einhellig, F A

    1988-10-01

    The bioassay has been one of the most widely used tests to demonstrate allelopathic activity. Often, claims that a particular plant species inhibits the growth of another are based entirely on the seed germination response to solvent extracts of the suspected allelopathic plant; few of these tests are of value in demonstrating allelopathy under natural conditions. The veracity of the bioassay for evaluating naturally occurring compounds for phytotoxicity depends upon the physiological and biochemical response capacity of the bioassay organism and the mechanism(s) of action of the allelochemicals. The possibility that more than one allelochemical, acting in concert at very low concentrations, may be responsible for an observed allelopathic effect makes it imperative that bioassays be extremely sensitive to chemical growth perturbation agents. Among the many measures of phytotoxicity of allelochemicals, the inhibition (or stimulation) of seed germination, radicle elongation, and/or seedling growth have been the parameters of choice for most investigations. Few of these assays have been selected with the view towards the possible mechanism of the allelopathic effect.

  17. Micro-organism distribution sampling for bioassays

    Science.gov (United States)

    Nelson, B. A.

    1975-01-01

    Purpose of sampling distribution is to characterize sample-to-sample variation so statistical tests may be applied, to estimate error due to sampling (confidence limits) and to evaluate observed differences between samples. Distribution could be used for bioassays taken in hospitals, breweries, food-processing plants, and pharmaceutical plants.

  18. The ICRP working party on bioassay interpretation

    International Nuclear Information System (INIS)

    Fry, F.A.; Lipsztein, J.L.; Birchall, A.

    2003-01-01

    In recent years there have been many developments in modelling the behaviour of radionuclides in the human body. The current generation of models are designed to be more 'realistic' than the previous generation of simple compartment models. The International Commission on Radiological Protection (ICRP) uses these models to produce dose coefficients and recognises that there is a need to give more guidance on how these models can be used to interpret bioassay data. A working party has been set up to address the issue. This paper describes some of the problems, some approaches to solving the problems and the progress of the ICRP working party. (author)

  19. Bioassay method for Uranium in urine by Delay Neutron counting

    International Nuclear Information System (INIS)

    Suratman; Purwanto; Sukarman-Aminjoyo

    1996-01-01

    A bioassay method for uranium in urine by neutron counting has been studied. The aim of this research is to obtain a bioassay method for uranium in urine which is used for the determination of internal dose of radiation workers. The bioassay was applied to the artificially uranium contaminated urine. The weight of the contaminant was varied. The uranium in the urine was irradiated in the Kartini reactor core, through pneumatic system. The delayed neutron was counted by BF3 neutron counter. Recovery of the bioassay was between 69.8-88.8 %, standard deviation was less than 10 % and the minimum detection was 0.387 μg

  20. Bioassay method for Uranium in urine by Delay Neutron counting; Metoda Bioassay Uranium dalam urin dengan pencacahan Netron Kasip

    Energy Technology Data Exchange (ETDEWEB)

    Suratman,; Purwanto,; Sukarman-Aminjoyo, [Yogyakarta Nuclear Research Centre, National Atomic Energy Agency, Yogyakarta (Indonesia)

    1996-04-15

    A bioassay method for uranium in urine by neutron counting has been studied. The aim of this research is to obtain a bioassay method for uranium in urine which is used for the determination of internal dose of radiation workers. The bioassay was applied to the artificially uranium contaminated urine. The weight of the contaminant was varied. The uranium in the urine was irradiated in the Kartini reactor core, through pneumatic system. The delayed neutron was counted by BF3 neutron counter. Recovery of the bioassay was between 69.8-88.8 %, standard deviation was less than 10 % and the minimum detection was 0.387 {mu}g.

  1. Urine sample collection protocols for bioassay samples

    Energy Technology Data Exchange (ETDEWEB)

    MacLellan, J.A.; McFadden, K.M.

    1992-11-01

    In vitro radiobioassay analyses are used to measure the amount of radioactive material excreted by personnel exposed to the potential intake of radioactive material. The analytical results are then used with various metabolic models to estimate the amount of radioactive material in the subject`s body and the original intake of radioactive material. Proper application of these metabolic models requires knowledge of the excretion period. It is normal practice to design the bioassay program based on a 24-hour excretion sample. The Hanford bioassay program simulates a total 24-hour urine excretion sample with urine collection periods lasting from one-half hour before retiring to one-half hour after rising on two consecutive days. Urine passed during the specified periods is collected in three 1-L bottles. Because the daily excretion volume given in Publication 23 of the International Commission on Radiological Protection (ICRP 1975, p. 354) for Reference Man is 1.4 L, it was proposed to use only two 1-L bottles as a cost-saving measure. This raised the broader question of what should be the design capacity of a 24-hour urine sample kit.

  2. Urine sample collection protocols for bioassay samples

    Energy Technology Data Exchange (ETDEWEB)

    MacLellan, J.A.; McFadden, K.M.

    1992-11-01

    In vitro radiobioassay analyses are used to measure the amount of radioactive material excreted by personnel exposed to the potential intake of radioactive material. The analytical results are then used with various metabolic models to estimate the amount of radioactive material in the subject's body and the original intake of radioactive material. Proper application of these metabolic models requires knowledge of the excretion period. It is normal practice to design the bioassay program based on a 24-hour excretion sample. The Hanford bioassay program simulates a total 24-hour urine excretion sample with urine collection periods lasting from one-half hour before retiring to one-half hour after rising on two consecutive days. Urine passed during the specified periods is collected in three 1-L bottles. Because the daily excretion volume given in Publication 23 of the International Commission on Radiological Protection (ICRP 1975, p. 354) for Reference Man is 1.4 L, it was proposed to use only two 1-L bottles as a cost-saving measure. This raised the broader question of what should be the design capacity of a 24-hour urine sample kit.

  3. Bioassay of procoagulant albumin in human plasma.

    Science.gov (United States)

    Grosset, A; Liu, L; Parker, C J; Rodgers, G M

    1994-09-01

    Procoagulant albumin (P-Al) is present in normal human plasma and increases monocyte and endothelial cell expression of tissue factor activity. To develop a bioassay for P-Al, we partially purified plasma from healthy volunteers and several patient groups using BaCl2 and (NH4)2SO4 precipitation. The samples were assayed for tissue factor (TF) inducing activity, expressed as a percentage increase compared to a serum-free media control. Over six months, the assay was reproducible in stored samples and in serial samples from normal volunteers. The plasma P-Al activities of 35 volunteers averaged 141 +/- 8.2% (SEM). There was no diurnal variation. There was no difference in the P-Al activity after a 12 hour fast and 2 hours after a large meal in 4 healthy volunteers. There was no increase in activity (r = 0.16) with the subject's age. The average activity from 16 poorly-controlled diabetics was 131 +/- 11% (SEM). No alteration in activity was seen with samples from patients with uremia, liver dysfunction, hemophilia, thrombotic events, or adenocarcinoma. These results indicate that P-Al activity can be bioassayed in individual patient samples; however, pathologic states associated with abnormal P-Al-induced tissue factor activity presently remain unidentified.

  4. Are bioassays useful tools to assess redox processes and biodegradation?

    DEFF Research Database (Denmark)

    Albrechtsen, Hans-Jørgen; Pedersen, Philip Grinder; Ludvigsen, L.

    2002-01-01

    sensitive hydrochemical or geochemical parameters, levels of hydrogen, and redox potential. However, all these approaches have to be evaluated against TEAP-bioassays as the most direct measure. We assessed successfully ongoing microbial-mediated redox processes by TEAP-bioassays in degradation studies...... of aromatic and chlorinated aliphatic compounds in landfill leachate plumes, and of pesticides in aquifers with various redox conditions....

  5. Circular Bioassay Platforms for Applications in Microwave-Accelerated Techniques.

    Science.gov (United States)

    Mohammed, Muzaffer; Clement, Travis C; Aslan, Kadir

    2014-12-02

    In this paper, we present the design of four different circular bioassay platforms, which are suitable for homogeneous microwave heating, using theoretical calculations (i.e., COMSOL™ multiphysics software). Circular bioassay platforms are constructed from poly(methyl methacrylate) (PMMA) for optical transparency between 400-800 nm, has multiple sample capacity (12, 16, 19 and 21 wells) and modified with silver nanoparticle films (SNFs) to be used in microwave-accelerated bioassays (MABs). In addition, a small monomode microwave cavity, which can be operated with an external microwave generator (100 W), for use with the bioassay platforms in MABs is also developed. Our design parameters for the circular bioassay platforms and monomode microwave cavity during microwave heating were: (i) temperature profiles, (ii) electric field distributions, (iii) location of the circular bioassay platforms inside the microwave cavity, and (iv) design and number of wells on the circular bioassay platforms. We have also carried out additional simulations to assess the use of circular bioassay platforms in a conventional kitchen microwave oven (e.g., 900 W). Our results show that the location of the circular bioassay platforms in the microwave cavity was predicted to have a significant effect on the homogeneous heating of these platforms. The 21-well circular bioassay platform design in our monomode microwave cavity was predicted to offer a homogeneous heating pattern, where inter-well temperature was observed to be in between 23.72-24.13°C and intra-well temperature difference was less than 0.21°C for 60 seconds of microwave heating, which was also verified experimentally.

  6. Plasmonically amplified fluorescence bioassay with microarray format

    Science.gov (United States)

    Gogalic, S.; Hageneder, S.; Ctortecka, C.; Bauch, M.; Khan, I.; Preininger, Claudia; Sauer, U.; Dostalek, J.

    2015-05-01

    Plasmonic amplification of fluorescence signal in bioassays with microarray detection format is reported. A crossed relief diffraction grating was designed to couple an excitation laser beam to surface plasmons at the wavelength overlapping with the absorption and emission bands of fluorophore Dy647 that was used as a label. The surface of periodically corrugated sensor chip was coated with surface plasmon-supporting gold layer and a thin SU8 polymer film carrying epoxy groups. These groups were employed for the covalent immobilization of capture antibodies at arrays of spots. The plasmonic amplification of fluorescence signal on the developed microarray chip was tested by using interleukin 8 sandwich immunoassay. The readout was performed ex situ after drying the chip by using a commercial scanner with high numerical aperture collecting lens. Obtained results reveal the enhancement of fluorescence signal by a factor of 5 when compared to a regular glass chip.

  7. Collection and control of tritium bioassay samples at Pantex

    International Nuclear Information System (INIS)

    Fairrow, N.L.; Ivie, W.E.

    1992-01-01

    Pantex is the final assembly/disassembly point for US nuclear weapons. The Pantex internal dosimetry section monitors radiation workers once a month for tritium exposure. In order to manage collection and control of the bioassay specimens efficiently, a bar code system for collection of samples was developed and implemented to speed up the process and decrease the number of errors probable when transferring data. In the past, all the bioassay data from samples were entered manually into a computer database. Transferring the bioassay data from the liquid scintillation counter to each individual's dosimetry record required as much as two weeks of concentrated effort

  8. Bioassays for the determination of nitrification inhibition

    Energy Technology Data Exchange (ETDEWEB)

    Grunditz, Camilla

    1999-07-01

    Requirements for nitrogen reduction in wastewater treatment plants were introduced in Sweden in the early 1990's. This was a governmental move to reduce the nitrogen discharges to the Baltic and Kattegat in order to prevent eutrophication. The nitrification process in wastewater treatment plants is performed by nitrifying bacteria. These are susceptible to inhibition and it is of great importance that the influent water does not contain toxic compounds. Therefore, there is a need for assays for the determination of nitrification inhibition. This thesis describes the development and applications of such bioassays. Pure cultures of Nitrosomonas sp. and Nitrobacter sp. were isolated from activated sludge of a wastewater treatment plant. These cultures were used as test organisms in the development of bioassays for nitrification inhibition measurements. The assays are based on two different principles; cell suspensions of the bacteria, performed in test tubes, and mediated amperometric biosensors with the bacteria immobilised. Ammonia oxidation and nitrite oxidation are studied separately without interference from other organisms, which makes it easier to interpret the results. The cell suspension assays were applied to samples of industrial and municipal wastewater. The Nitrosomonas and Nitrobacter assays showed to have different inhibition patterns. A large percentage of the Swedish municipal wastewater treatment plants were found to receive inhibitory influent water, but the inhibition level was generally low. Compared to an assay based on activated sludge, the screening method, the pure culture assays found more samples of influent water strongly inhibitory or stimulating. The highest correlation was found between the screening method and the Nitrosomonas assay. The Nitrobacter assay was found to be the most sensitive method. Assessment of toxicity of a number of chemical substances was studied using the biosensors, together with the cell suspension assays

  9. Reporter gene bioassays in environmental analysis.

    Science.gov (United States)

    Köhler, S; Belkin, S; Schmid, R D

    2000-01-01

    In parallel to the continuous development of increasingly more sophisticated physical and chemical analytical technologies for the detection of environmental pollutants, there is a progressively more urgent need also for bioassays which report not only on the presence of a chemical but also on its bioavailability and its biological effects. As a partial fulfillment of that need, there has been a rapid development of biosensors based on genetically engineered bacteria. Such microorganisms typically combine a promoter-operator, which acts as the sensing element, with reporter gene(s) coding for easily detectable proteins. These sensors have the ability to detect global parameters such as stress conditions, toxicity or DNA-damaging agents as well as specific organic and inorganic compounds. The systems described in this review, designed to detect different groups of target chemicals, vary greatly in their detection limits, specificity, response times and more. These variations reflect on their potential applicability which, for most of the constructs described, is presently rather limited. Nevertheless, present trends promise that additional improvements will make microbial biosensors an important tool for future environmental analysis.

  10. Annotating Human P-Glycoprotein Bioassay Data.

    Science.gov (United States)

    Zdrazil, Barbara; Pinto, Marta; Vasanthanathan, Poongavanam; Williams, Antony J; Balderud, Linda Zander; Engkvist, Ola; Chichester, Christine; Hersey, Anne; Overington, John P; Ecker, Gerhard F

    2012-08-01

    Huge amounts of small compound bioactivity data have been entering the public domain as a consequence of open innovation initiatives. It is now the time to carefully analyse existing bioassay data and give it a systematic structure. Our study aims to annotate prominent in vitro assays used for the determination of bioactivities of human P-glycoprotein inhibitors and substrates as they are represented in the ChEMBL and TP-search open source databases. Furthermore, the ability of data, determined in different assays, to be combined with each other is explored. As a result of this study, it is suggested that for inhibitors of human P-glycoprotein it is possible to combine data coming from the same assay type, if the cell lines used are also identical and the fluorescent or radiolabeled substrate have overlapping binding sites. In addition, it demonstrates that there is a need for larger chemical diverse datasets that have been measured in a panel of different assays. This would certainly alleviate the search for other inter-correlations between bioactivity data yielded by different assay setups.

  11. Bioassay Phantoms Using Medical Images and Computer Aided Manufacturing

    International Nuclear Information System (INIS)

    Xu, X. Geroge

    2011-01-01

    A radiation bioassay program relies on a set of standard human phantoms to calibrate and assess radioactivity levels inside a human body for radiation protection and nuclear medicine imaging purposes. However, the methodologies in the development and application of anthropomorphic phantoms, both physical and computational, had mostly remained the same for the past 40 years. We herein propose a 3-year research project to develop medical image-based physical and computational phantoms specifically for radiation bioassay applications involving internally deposited radionuclides. The broad, long-term objective of this research was to set the foundation for a systematic paradigm shift away from the anatomically crude phantoms in existence today to realistic and ultimately individual-specific bioassay methodologies. This long-term objective is expected to impact all areas of radiation bioassay involving nuclear power plants, U.S. DOE laboratories, and nuclear medicine clinics.

  12. Application of Bioassays for the Ecotoxicity Assessment of Contaminated Soils

    Science.gov (United States)

    Fernández, María D.; Babín, Mar; Tarazona, José V.

    The use of bioassays for soil characterization is receiving significant attention as a complementary tool to chemical analysis. Bioassays consist of direct toxicity assays of environmental samples that are transferred to the laboratory and analyzed for toxicity against selected organisms. Such soil samples contain the combination of the different pollutants present in situ and enable factors such as the bioavailability of contaminants or the interactions (synergic and antagonic) between them to be simultaneously studied.

  13. [Investigation on pattern and methods of quality control for Chinese materia medica based on dao-di herbs and bioassay - bioassay for Coptis chinensis].

    Science.gov (United States)

    Yan, Dan; Xiao, Xiao-he

    2011-05-01

    Establishment of bioassay methods is the technical issues to be faced with in the bioassay of Chinese materia medica. Taking the bioassay of Coptis chinensis Franch. as an example, the establishment process and application of the bioassay methods (including bio-potency and bio-activity fingerprint) were explained from the aspects of methodology, principle of selection, experimental design, method confirmation and data analysis. The common technologies were extracted and formed with the above aspects, so as to provide technical support for constructing pattern and method of the quality control for Chinese materia medica based on the dao-di herbs and bioassay.

  14. The effect of pesticide residue on caged mosquito bioassays.

    Science.gov (United States)

    Barber, J A S; Greer, Mike; Coughlin, Jamie

    2006-09-01

    Wind tunnel experiments showed that secondary pickup of insecticide residue by mosquitoes in cage bioassays had a significant effect on mortality. Cage bioassays using adult Ochlerotatus taeniorhynchus (Wiedemann) investigated the effect of exposure time to a contaminated surface. Cages were dosed in a wind tunnel using the LC50 for naled (0.124 mg a.i./ml) and an LC25 (0.0772 mg a.i./ml) for naled. Half of the bioassay mosquitoes were moved directly into clean cages with the other half remaining in the sprayed, hence contaminated, cage. Treatment mortality was assessed at 8, 15, 30, 60, 120, 240, and 1,440 min postapplication. Cage contamination had a significant effect on mosquito mortality for both the LC25 and LC50 between 15 and 30 min postapplication.

  15. Internal dosimetry performing dose assessments via bioassay measurements

    International Nuclear Information System (INIS)

    Bailey, K.M.

    1993-01-01

    The Internal Dosimetry Department at the Y-12 Plant maintains a state-of-the-art bioassay program managed under the guidance and regulations of the Department of Energy. The two major bioassay techniques currently used at Y-12 are the in vitro (urinalysis) and in vivo (lung counting) programs. Fecal analysis (as part of the in vitro program) is another alternative; however, since both urine and fecal analysis provide essentially the same capabilities for detecting exposures to uranium, the urinalysis is the main choice primarily for aesthetic reasons. The bioassay frequency is based on meeting NCRP 87 objectives which are to monitor the accumulation of radioactive material in exposed individuals, and to ensure that significant depositions are detected

  16. Bioassays for risk assessment of coal conversion products

    Energy Technology Data Exchange (ETDEWEB)

    Schacht, S.; Sinder, C.; Pfeifer, F.; Klein, J. [DMT-Gesellschaft fuer Forschung und Pruefung mbH, Essen (Germany)

    1999-07-01

    Traditional as well as biotechnological processing coal leads to complex mixtures of products. Besides chemical and physical characterization, which provides the information for product application, there is a need for bioassays to monitor properties that are probably toxic, mutagenic or cancerogenic. Investigations carried out focused on the selection, adaptation and validation of bioassays for the sensitive estimation of toxic effects. Organisms like bacteria, Daphnia magna and Scenedesmus subspicatus, representing different complexities in the biosphere, were selected as test systems for ecotoxicological and mutagenicity studies. The results obtained indicate that bioassays are, in principle, suitable tools for characterization and evaluation of coal-derived substances and bioconversion products. Using coal products, coal-relevant model compounds and bioconversion products, data for risk assessment are presented. (orig.)

  17. Manual on theory and practical aspects of bioassay

    International Nuclear Information System (INIS)

    Nuraini Hambali.

    1985-06-01

    This manual is set to provide necessary basic guidance on theory and practical aspects of bioassay specially for the newcomer in this field and the man in the laboratory. The first part is a brief information on the entry of radionuclides into the body, the metabolism and the programs of bioassay. All other factors to be considered in assessing internal contamination in man have also been brought up. In the second part, various procedures of radiochemical separations, detection and measurements are abstracted from journals and other revisions. Some methods have been attempted and to be followed where appropriate. (author)

  18. An examination of the analysis of radiostrontiums in bioassay applications

    International Nuclear Information System (INIS)

    Linauskas, S.H.; Leon, J.W.

    1993-05-01

    Radiostrontiums are among the most radiologically significant radionuclides in the nuclear reactor environment due to their relatively high fission yield, long physical half-life, volatility and mobility in the workplace, and long retention times in tissues such as bone. Effective bioassay programs include analytical processes that consider prospective monitoring requirements provided by screening measurements, as well as the retrospective monitoring requirements provided by screening measurements following an intake. Chromatography using crown ethers as well as the use of spectrometry techniques with advanced liquid-scintillation counters or solid-state surface-barrier detectors appear to have significant benefits for Sr bioassay programs. (author). 90 refs., 2 tabs., 3 figs

  19. An examination of the analysis of radiostrontiums in bioassay applications

    Energy Technology Data Exchange (ETDEWEB)

    Linauskas, S H; Leon, J W

    1993-05-01

    Radiostrontiums are among the most radiologically significant radionuclides in the nuclear reactor environment due to their relatively high fission yield, long physical half-life, volatility and mobility in the workplace, and long retention times in tissues such as bone. Effective bioassay programs include analytical processes that consider prospective monitoring requirements provided by screening measurements, as well as the retrospective monitoring requirements provided by screening measurements following an intake. Chromatography using crown ethers as well as the use of spectrometry techniques with advanced liquid-scintillation counters or solid-state surface-barrier detectors appear to have significant benefits for Sr bioassay programs. (author). 90 refs., 2 tabs., 3 figs.

  20. Initial sample extract stock concentration affects in vitro bioassay-based toxicological risk characterization

    NARCIS (Netherlands)

    Montano, M.; Loffmann, L.; Murk, A.J.; Gutleb, A.C.

    2014-01-01

    Purpose Bioassays have become an alternative for sediment risk profiling, including potential compliance with sediment quality criteria (SQC). In vitro functional bioassays have evolved through standardization and validation towards a confident toxicological hazard estimate of sediments. Sample

  1. A statistical treatment of bioassay pour fractions

    Science.gov (United States)

    Barengoltz, Jack; Hughes, David

    A bioassay is a method for estimating the number of bacterial spores on a spacecraft surface for the purpose of demonstrating compliance with planetary protection (PP) requirements (Ref. 1). The details of the process may be seen in the appropriate PP document (e.g., for NASA, Ref. 2). In general, the surface is mechanically sampled with a damp sterile swab or wipe. The completion of the process is colony formation in a growth medium in a plate (Petri dish); the colonies are counted. Consider a set of samples from randomly selected, known areas of one spacecraft surface, for simplicity. One may calculate the mean and standard deviation of the bioburden density, which is the ratio of counts to area sampled. The standard deviation represents an estimate of the variation from place to place of the true bioburden density commingled with the precision of the individual sample counts. The accuracy of individual sample results depends on the equipment used, the collection method, and the culturing method. One aspect that greatly influences the result is the pour fraction, which is the quantity of fluid added to the plates divided by the total fluid used in extracting spores from the sampling equipment. In an analysis of a single sample’s counts due to the pour fraction, one seeks to answer the question: What is the probability that if a certain number of spores are counted with a known pour fraction, that there are an additional number of spores in the part of the rinse not poured. This is given for specific values by the binomial distribution density, where detection (of culturable spores) is success and the probability of success is the pour fraction. A special summation over the binomial distribution, equivalent to adding for all possible values of the true total number of spores, is performed. This distribution when normalized will almost yield the desired quantity. It is the probability that the additional number of spores does not exceed a certain value. Of course

  2. Changes in chemical composition and bioassay assessment of ...

    African Journals Online (AJOL)

    Changes in chemical composition and bioassay assessment of nutritional potentials of almond fruit waste as an alternative feedstuff for livestock. ... AFW using day-old cockerels and considering performance parameters showed that treated AFW improved feed intake, body weight gain and feed conversion ratio even better ...

  3. Preliminary results of testing bioassay analytical performance standards

    International Nuclear Information System (INIS)

    Fisher, D.R.; Robinson, A.V.; Hadley, R.T.

    1983-08-01

    The analytical performance of both in vivo and in vitro bioassay laboratories is being studied to determine the capability of these laboratories to meet the minimum criteria for accuracy and precision specified in the draft ANSI Standard N13.30, Performance Criteria for Radiobioassay. This paper presents preliminary results of the first round of testing

  4. US Army Radiological Bioassay and Dosimetry: The RBD software package

    International Nuclear Information System (INIS)

    Eckerman, K.F.; Ward, R.C.; Maddox, L.B.

    1993-01-01

    The RBD (Radiological Bioassay and Dosimetry) software package was developed for the U. S. Army Material Command, Arlington, Virginia, to demonstrate compliance with the radiation protection guidance 10 CFR Part 20 (ref. 1). Designed to be run interactively on an IBM-compatible personal computer, RBD consists of a data base module to manage bioassay data and a computational module that incorporates algorithms for estimating radionuclide intake from either acute or chronic exposures based on measurement of the worker's rate of excretion of the radionuclide or the retained activity in the body. In estimating the intake,RBD uses a separate file for each radionuclide containing parametric representations of the retention and excretion functions. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent. For a given nuclide, if measurements exist for more than one type of assay, an auxiliary module, REPORT, estimates the intake by applying weights assigned in the nuclide file for each assay. Bioassay data and computed results (estimates of intake and committed dose equivalent) are stored in separate data bases, and the bioassay measurements used to compute a given result can be identified. The REPORT module creates a file containing committed effective dose equivalent for each individual that can be combined with the individual's external exposure

  5. Soil bioassays as tools for sludge compost quality assessment

    International Nuclear Information System (INIS)

    Domene, Xavier; Sola, Laura; Ramirez, Wilson; Alcaniz, Josep M.; Andres, Pilar

    2011-01-01

    Composting is a waste management technology that is becoming more widespread as a response to the increasing production of sewage sludge and the pressure for its reuse in soil. In this study, different bioassays (plant germination, earthworm survival, biomass and reproduction, and collembolan survival and reproduction) were assessed for their usefulness in the compost quality assessment. Compost samples, from two different composting plants, were taken along the composting process, which were characterized and submitted to bioassays (plant germination and collembolan and earthworm performance). Results from our study indicate that the noxious effects of some of the compost samples observed in bioassays are related to the low organic matter stability of composts and the enhanced release of decomposition endproducts, with the exception of earthworms, which are favored. Plant germination and collembolan reproduction inhibition was generally associated with uncomposted sludge, while earthworm total biomass and reproduction were enhanced by these materials. On the other hand, earthworm and collembolan survival were unaffected by the degree of composting of the wastes. However, this pattern was clear in one of the composting procedures assessed, but less in the other, where the release of decomposition endproducts was lower due to its higher stability, indicating the sensitivity and usefulness of bioassays for the quality assessment of composts.

  6. Assessment of acrylamide toxicity using a battery of standardised bioassays.

    Science.gov (United States)

    Zovko, Mira; Vidaković-Cifrek, Željka; Cvetković, Želimira; Bošnir, Jasna; Šikić, Sandra

    2015-12-01

    Acrylamide is a monomer widely used as an intermediate in the production of organic chemicals, e.g. polyacrylamides (PAMs). Since PAMs are low cost chemicals with applications in various industries and waste- and drinking water treatment, a certain amount of non-polymerised acrylamide is expected to end up in waterways. PAMs are non-toxic but acrylamide induces neurotoxic effects in humans and genotoxic, reproductive, and carcinogenic effects in laboratory animals. In order to evaluate the effect of acrylamide on freshwater organisms, bioassays were conducted on four species: algae Desmodesmus subspicatus and Pseudokirchneriella subcapitata, duckweed Lemna minor and water flea Daphnia magna according to ISO (International Organization for Standardisation) standardised methods. This approach ensures the evaluation of acrylamide toxicity on organisms with different levels of organisation and the comparability of results, and it examines the value of using a battery of low-cost standardised bioassays in the monitoring of pollution and contamination of aquatic ecosystems. These results showed that EC50 values were lower for Desmodesmus subspicatus and Pseudokirchneriella subcapitata than for Daphnia magna and Lemna minor, which suggests an increased sensitivity of algae to acrylamide. According to the toxic unit approach, the values estimated by the Lemna minor and Daphnia magna bioassays, classify acrylamide as slightly toxic (TU=0-1; Class 1). The results obtained from algal bioassays (Desmodesmus subspicatus and Pseudokirchneriella subcapitata) revealed the toxic effect of acrylamide (TU=1-10; Class 2) on these organisms.

  7. Microplate Bioassay for Determining Substrate Selectivity of "Candida rugosa" Lipase

    Science.gov (United States)

    Wang, Shi-zhen; Fang, Bai-shan

    2012-01-01

    Substrate selectivity of "Candida rugosa" lipase was tested using "p"-nitrophenyl esters of increasing chain length (C[subscript 1], C[subscript 7], C[subscript 15]) using the high-throughput screening method. A fast and easy 96-well microplate bioassay was developed to help students learn and practice biotechnological specificity screen. The…

  8. Soil plate bioassay: an effective method to determine ecotoxicological risks.

    Science.gov (United States)

    Boluda, R; Roca-Pérez, L; Marimón, L

    2011-06-01

    Heavy metals have become one of the most serious anthropogenic stressors for plants and other living organisms. Having efficient and feasible bioassays available to assess the ecotoxicological risks deriving from soil pollution is necessary. This work determines pollution by Cd, Co, Cr, Cu, Ni, Pb, V and Zn in two soils used for growing rice from the Albufera Natural Park in Valencia (Spain). Both were submitted to a different degree of anthropic activity, and their ecotoxicological risk was assessed by four ecotoxicity tests to compare their effectiveness: Microtox test, Zucconi test, pot bioassay (PB) and soil plate bioassay (SPB). The sensitivity of three plant species (barley, cress and lettuce) was also assessed. The results reveal a different degree of effectiveness and level of inhibition in the target organisms' growth depending on the test applied, to such an extent that the one-way analysis of variance showed significant differences only for the plate bioassay results, with considerable inhibition of root and shoot elongation in seedlings. Of the three plant species selected, lettuce was the most sensitive species to toxic effects, followed by cress and barley. Finally, the results also indicate that the SPB is an efficient, simple and economic alternative to other ecotoxicological assays to assess toxicity risks deriving from soil pollution. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. Bioassay for aquatic ecosystems review and classification; Rassegna dei principali test di ecotossicologia acquatica

    Energy Technology Data Exchange (ETDEWEB)

    Sanci, Antonella; Rosa, Silvia [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    1997-09-01

    Bioassay play a crucial role in assessing the actual or potential impacts of anthropogenic agents on the natural environment. In this technical report, literature on bioassays for aquatic ecosystems has been reviewed and classified. Problems associated with the choice and application of bioassays are discussed.

  10. Bioassay-based risk assessment of hazardous waste

    Energy Technology Data Exchange (ETDEWEB)

    Donnelly, K.C.; Brown, K.W.; He, L.Y. [Texas A and M Univ., College Station, TX (United States)

    1994-12-31

    Microbial bioassays have been used to assess the genotoxic hazard at more than 30 different hazardous waste sites. Environmental samples were extracted with dichloromethane and methanol, and the resulting residue tested using GC/MS analysis as well as the Salmonella Microsomal and E. coli Prophage Induction assays. At a munitions wastewater contaminated site, there was no correlation between mutagenicity in bacteria, and the risk as estimated from chemical analysis data of trinitrotoluene. Samples 202 and 204 from a coal gasification site contained 72 mg/kg and 9 mg/kg benzo(a)pyrene, whereas the mutagenic responses of these samples were 231 net revertants/mg and 902 revertants/mg, respectively. The data suggest that microbial bioassays provide a valuable tool for monitoring the interactions of the components of a complex mixture.

  11. Improving global laboratory capabilities for emergency radionuclide bioassay

    International Nuclear Information System (INIS)

    Li, C.; Jourdain, J.; Kramer, G. H.

    2012-01-01

    During a radiological or nuclear emergency, first-responders and the general public may be internally contaminated with the radionuclide(s) involved. A timely radionuclide bioassay provides important information about contamination, for subsequent dose assessment and medical management. Both technical and operational gaps are discussed in this paper. As many people may need to be assessed in a short period of time, any single laboratory may find its capabilities insufficient. Laboratories from other regions or other countries may be called upon for assistance. This paper proposes a road-map to improve global capabilities in emergency radionuclide bioassay, suggesting a phased approach for establishing a global laboratory network. Existing international collaboration platforms could provide the base on which to build such a network. (authors)

  12. A Rapid and Simple Bioassay Method for Herbicide Detection

    Directory of Open Access Journals (Sweden)

    Xiu-Qing Li

    2008-01-01

    Full Text Available Chlamydomonas reinhardtii, a unicellular green alga, has been used in bioassay detection of a variety of toxic compounds such as pesticides and toxic metals, but mainly using liquid culture systems. In this study, an algal lawn--agar system for semi-quantitative bioassay of herbicidal activities has been developed. Sixteen different herbicides belonging to 11 different categories were applied to paper disks and placed on green alga lawns in Petri dishes. Presence of herbicide activities was indicated by clearing zones around the paper disks on the lawn 2-3 days after application. The different groups of herbicides induced clearing zones of variable size that depended on the amount, mode of action, and chemical properties of the herbicides applied to the paper disks. This simple, paper-disk-algal system may be used to detect the presence of herbicides in water samples and act as a quick and inexpensive semi-quantitative screening for assessing herbicide contamination.

  13. The use of cultivars of Raphanus sativus for cytokinin bioassay

    Directory of Open Access Journals (Sweden)

    Dorota Kubowicz

    2013-12-01

    Full Text Available Six cultivars of radish (Raphanus sativus were tested for their usefulness in radish cytokinin bioassay by the method of Letham (1971. The best cultivar was found to be 'Sopel Lodu' which responds well to both zeatin and 2iP over a wide range of concentrations. The fresh weight of cotyledons increased at most by 71.5% (if treated with zeatin or 101.0% (if treated with 2iP compared to untreated cotyledons. This cultivar is also sensitive to the partially purified cytokinin-like fraction isolated from the pine (Pinus silvestris cambial region. The cultivar 'Sopel Lodu' is therefore proposed to be a suitable plant for cytokinin bioassays.

  14. The IMBA suite: integrated modules for bioassay analysis

    Energy Technology Data Exchange (ETDEWEB)

    Birchall, A.; Jarvis, N.S.; Peace, M.S.; Riddell, A.E.; Battersby, W.P

    1998-07-01

    The increasing complexity of models representing the biokinetic behaviour of radionuclides in the body following intake poses problems for people who are required to implement these models. The problem is exacerbated by the current paucity of suitable software. In order to remedy this situation, a collaboration between British Nuclear Fuels, Westlakes Research Institute and the National Radiological Protection Board has started with the aim of producing a suite of modules for estimating intakes and doses from bioassay measurements using the new ICRP models. Each module will have a single purpose (e.g. to calculate respiratory tract deposition) and will interface with other software using data files. The elements to be implemented initially are plutonium, uranium, caesium, iodine and tritium. It is intended to make the software available to other parties under terms yet to be decided. This paper describes the proposed suite of integrated modules for bioassay analysis, IMBA. (author)

  15. Log bioassay of residual effectiveness of insecticides against bark beetles

    Science.gov (United States)

    Richard H. Smith

    1982-01-01

    Residual effectiveness of nine insecticides applied to bark was tested against western, mountain, and Jeffrey pine beetles. Ponderosa and Jeffrey pine trees were treated and logs cut from them 2 to 13 months later, and bioassayed with the three beetles. The insecticides were sprayed at the rate of 1 gal (3.8 l) per 40- or 80-ft² (3.6 or 7.2 m²) bark surface at varying...

  16. Eubacterium brachy - Reactivity in In Vitro Bone Resorptive Bioassay,

    Science.gov (United States)

    1983-02-10

    Center Washington, D. C . 20307 If Eubacterium brachy - Reactivity in In Vitro Bone Resorptive Bioassay 1. ABSTRACT Recent studies have demonstrated an...Relative distribution of bacteria at clinically healthy and periodontally diseased sites in humans. J Clin Periodontal 5:115, 1978. 3. Evian, C ...applied foreign protein into rat gingiva. J Periodont Res 6:89, 1971. 21. Gaffer, A., Coleman, E.J., and Marcussen, H.W.: Penetration of dental plaque

  17. Activities of Jatropha curcas phorbol esters in various bioassays.

    Science.gov (United States)

    Devappa, Rakshit K; Rajesh, Sanjay K; Kumar, Vikas; Makkar, Harinder P S; Becker, Klaus

    2012-04-01

    Jatropha curcas seeds contain 30-35% oil, which can be converted to high quality biodiesel. However, Jatropha oil is toxic, ascribed to the presence of phorbol esters (PEs). In this study, isolated phorbol ester rich fraction (PEEF) was used to evaluate the activity of PEs using three aquatic species based bioassays (snail (Physa fontinalis), brine shrimp (Artemeia salina), daphnia (Daphnia magna)) and microorganisms. In all the bioassays tested, increase in concentration of PEs increased mortality with an EC(50) (48 h) of 0.33, 26.48 and 0.95 mg L(-1) PEs for snail, artemia and daphnia, respectively. The sensitivity of various microorganisms for PEs was also tested. Among the bacterial species tested, Streptococcus pyogenes and Proteus mirabilis were highly susceptible with a minimum inhibitory concentration (MIC) of 215 mg L(-1) PEs; and Pseudomonas putida were also sensitive with MIC of 251 mg L(-1) PEs. Similarly, Fusarium species of fungi exhibited EC(50) of 58 mg L(-1) PEs, while Aspergillus niger and Curvularia lunata had EC(50) of 70 mg L(-1). The snail bioassay was most sensitive with 100% snail mortality at 1 μg of PEs mL(-1). In conclusion, snail bioassay could be used to monitor PEs in Jatropha derived products such as oil, biodiesel, fatty acid distillate, kernel meal, cake, glycerol or for contamination in soil or other environmental matrices. In addition, PEs with molluscicidal/antimicrobial activities could be utilized for agricultural and pharmaceutical applications. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. BioAssay templates for the semantic web

    Directory of Open Access Journals (Sweden)

    Alex M. Clark

    2016-05-01

    Full Text Available Annotation of bioassay protocols using semantic web vocabulary is a way to make experiment descriptions machine-readable. Protocols are communicated using concise scientific English, which precludes most kinds of analysis by software algorithms. Given the availability of a sufficiently expressive ontology, some or all of the pertinent information can be captured by asserting a series of facts, expressed as semantic web triples (subject, predicate, object. With appropriate annotation, assays can be searched, clustered, tagged and evaluated in a multitude of ways, analogous to other segments of drug discovery informatics. The BioAssay Ontology (BAO has been previously designed for this express purpose, and provides a layered hierarchy of meaningful terms which can be linked to. Currently the biggest challenge is the issue of content creation: scientists cannot be expected to use the BAO effectively without having access to software tools that make it straightforward to use the vocabulary in a canonical way. We have sought to remove this barrier by: (1 defining a BioAssay Template (BAT data model; (2 creating a software tool for experts to create or modify templates to suit their needs; and (3 designing a common assay template (CAT to leverage the most value from the BAO terms. The CAT was carefully assembled by biologists in order to find a balance between the maximum amount of information captured vs. low degrees of freedom in order to keep the user experience as simple as possible. The data format that we use for describing templates and corresponding annotations is the native format of the semantic web (RDF triples, and we demonstrate some of the ways that generated content can be meaningfully queried using the SPARQL language. We have made all of these materials available as open source (http://github.com/cdd/bioassay-template, in order to encourage community input and use within diverse projects, including but not limited to our own

  19. The 10th Annual Bioassays and Bioanalytical Method Development Conference.

    Science.gov (United States)

    Ma, Mark; Tudan, Christopher; Koltchev, Dolly

    2015-01-01

    The 10th Annual Bioassays and Bioanalytical Method Development Conference was hosted in Boston, MA, USA on 20-22 October 2014. This meeting brought together scientists from the biopharmaceutical and life sciences industries, the regulatory agency and academia to share and discuss current trends in cell-based assays and bioanalysis, challenges and ideas for the future of the bioassays and bioanalytical method development. The experiences associated with new and innovative technologies were evaluated as well as their impact on the current bioassays methodologies and bioanalysis workflow, including quality, feasibility, outsourcing strategies and challenges, productivity and compliance. Several presentations were also provided by members of the US FDA, sharing both scientific and regulatory paradigms including a most recent update on the position of the FDA with specific aspects of the draft Bioanalytical Method Validation guidance following its review of the industry's responses. The meeting was jointly coincided with the 15th Annual Immunogenicity for Biotherapeutics meeting, allowing for attendees to also familiarize themselves with new and emerging approaches to overcome the effect of immunogenicity, in addition to investigative strategies.

  20. A Bioassay System Using Bioelectric Signals from Small Fish

    Science.gov (United States)

    Terawaki, Mitsuru; Soh, Zu; Hirano, Akira; Tsuji, Toshio

    Although the quality of tap water is generally examined using chemical assay, this method cannot be used for examination in real time. Against such a background, the technique of fish bioassay has attracted attention as an approach that enables constant monitoring of aquatic contamination. The respiratory rhythms of fish are considered an efficient indicator for the ongoing assessment of water quality, since they are sensitive to chemicals and can be indirectly measured from bioelectric signals generated by breathing. In order to judge aquatic contamination accurately, it is necessary to measure bioelectric signals from fish swimming freely as well as to stably discriminate measured signals, which vary between individuals. However, no bioassay system meeting the above requirements has yet been established. This paper proposes a bioassay system using bioelectric signals generated from small fish in free-swimming conditions. The system records signals using multiple electrodes to cover the extensive measurement range required in a free-swimming environment, and automatically discriminates changes in water quality from signal frequency components. This discrimination is achieved through an ensemble classification method using probability neural networks to solve the problem of differences between individual fish. The paper also reports on the results of related validation experiments, which showed that the proposed system was able to stably discriminate between water conditions before and after bleach exposure.

  1. Fluorescence-Based Bioassays for the Detection and Evaluation of Food Materials

    OpenAIRE

    Nishi, Kentaro; Isobe, Shin-Ichiro; Zhu, Yun; Kiyama, Ryoiti

    2015-01-01

    We summarize here the recent progress in fluorescence-based bioassays for the detection and evaluation of food materials by focusing on fluorescent dyes used in bioassays and applications of these assays for food safety, quality and efficacy. Fluorescent dyes have been used in various bioassays, such as biosensing, cell assay, energy transfer-based assay, probing, protein/immunological assay and microarray/biochip assay. Among the arrays used in microarray/biochip assay, fluorescence-based mi...

  2. Comparison of five bioassay techniques for assessing sediment-bound contaminants

    OpenAIRE

    Ahlf, Wolfgang; Calmano, Wolfgang; Erhard, Judith; Förstner, Ulrich

    1989-01-01

    Biological response could not be predicted based on chemical concentration of the sediment contaminants. Bioassays integrate the response of test organisms to contaminants and nutrients. Comparative results of five acute bioassays indicated that Neubauer phytoassay was the most sensitive. The mircobial biomass and algal growth tests indicated a response to the availability of contaminants and nutrients. These results suggest the usefulness of a diversity of bioassays in toxicity testing of se...

  3. Comparison of solid-phase bioassays and ecoscores to evaluate the toxicity of contaminated soils.

    OpenAIRE

    Lors , Christine; Ponge , Jean-François; Martínez Aldaya , Maite; Damidot , Denis

    2010-01-01

    International audience; Five bioassays (inhibition of lettuce germination and growth, earthworm mortality, inhibition of springtail population growth, avoidance by springtails) were compared, using four coke factory soils contaminated by PAHs and trace elements, before and after biotreatment. For each bioassay, several endpoints were combined in an 'ecoscore', a measure of test sensitivity. Ecoscores pooled over all tested bioassays revealed that most organisms were highly sensitive to the co...

  4. Fluorescent bioassays for toxic metals in milk and yoghurt

    Science.gov (United States)

    2012-01-01

    Background From a human health viewpoint, contaminated milk and its products could be a source of long-term exposure to toxic metals. Simple, inexpensive, and on-site assays would enable constant monitoring of their contents. Bioassays that can measure toxic metals in milk or yoghurt might reduce the risk. For this purpose, the green fluorescent protein (GFP)-tagged trans factors, ArsR-GFP and CadC-GFP, together with their cis elements were used to develop such bioassays. Results ArsR-GFP or CadC-GFP, which binds either toxic metal or DNA fragment including cis element, was directly mixed with cow’s milk or yoghurt within a neutral pH range. The fluorescence of GFP, which is reflected by the association/dissociation ratio between cis element and trans factor, significantly changed with increasing externally added As (III) or Cd (II) whereas smaller responses to externally added Pb (II) and Zn (II) were found. Preparation and dilution of whey fraction at low pH were essential to intrinsic zinc quantification using CadC-GFP. Using the extraction procedure and bioassay, intrinsic Zn (II) concentrations ranging from 1.4 to 4.8 mg/l for milk brands and from 1.2 to 2.9 mg/kg for yoghurt brands were determined, which correlated to those determined using inductively coupled plasma atomic emission spectroscopy. Conclusions GFP-tagged bacterial trans factors and cis elements can work in the neutralized whole composition and diluted whey fraction of milk and yoghurt. The feature of regulatory elements is advantageous for establishment of simple and rapid assays of toxic metals in dairy products. PMID:23098077

  5. Interpretation of bioassay data from nuclear fuel fabrication workers

    International Nuclear Information System (INIS)

    Melo, D.; Xavier, M.

    2005-01-01

    Full text: In nuclear fuel fabrication facilities, workers are exposed to different compounds of enriched uranium. Although in this kind of facility the main route of intake is inhalation, ingestion may occur in some situations. The interpretation of the bioassay data is very complex, since it is necessary taking into account all the different parameters, which is a big challenge. Due to the high cost of the individual monitoring programme for internal dose assessment in the routine monitoring programmes, usually only one type of measurement is assigned. In complex situations like the one described in this paper, where several parameters can compromise the accuracy of the bioassay interpretation it is need to have a combination of techniques to evaluate the internal dose. According to ICRP 78 (1997), the general order of preference in terms of accuracy of interpretation is: body activity measurement, excreta analysis and personal air sampling. Results of monitoring of working environment may provide information that assists in interpretation on particle size, chemical form and solubility, time of intake. A group of seventeen workers from controlled area of the fuel fabrication facility was selected to evaluate the internal dose using all different available techniques during a certain period. The workers were monitored for determination of uranium content in the daily urinary and faecal excretion (collected over a period of 3 consecutive days), chest counting and personal air sampling. The results have shown that at least two types of sensitivity techniques must be used, since there are some sources of uncertainties on the bioassay interpretation, like mixture of uranium compounds intake and different routes of intake. The combination of urine and faeces analysis has shown to be the more appropriate methodology for assessing internal dose in this situation. (author)

  6. Efficient algal bioassay based on short-term photosynthetic response

    International Nuclear Information System (INIS)

    Giddings, J.M.; Stewart, A.J.; O'Neill, R.V.; Gardner, R.H.

    1983-01-01

    A procedure is described for measuring the effects of toxicants on algal photosynthesis (carbon-14 bicarbonate (H 14 CO 3 )uptake) in 4-h experiments. The results for individual aromatic compounds and the water-soluble fraction (WSF) of a synthetic oil are presented as examples of applications of the bioassay. The toxicity of the WSF varied among the seven algal species tested, and the responses of some species were pH-dependent. With Selenastrum capricornutum as the test organism, the bioassay results were unaffected by variations in pH from 7.0 to 9.0, light intensity from 40 to 200 μeinsteins m -2 s -1 , culture density up to 0.5 mg chlorophyll a per litre, and agitation up to 100 rpm. The photosynthesis bioassay is simpler and faster (4 h versus 4 to 14 days), uses smaller culture volumes, and requires less space than static culture-growth tests. One person can conveniently test four materials per day, and the entire procedure, including preparation, exposure, and analysis, takes less than two days. The short incubation time reduces bottle effects such as pH changes, accumulation of metabolic products, nutrient depletion, and bacterial growth. Processes that remove or alter the test materials are also minimized. The data presented here indicate that algal photosynthesis is inhibited at toxicant concentrations similar to those that cause acute effects in aquatic animals. A model of a pelagic ecosystem is used to demonstrate that even temporary (seven-day) inhibition of algal photosynthesis can have a measurable impact on other trophic levels, particularly if the other trophic levels are also experiencing toxic effects. 25 references, 6 figures, 1 table

  7. Fluorescent bioassays for toxic metals in milk and yoghurt

    Directory of Open Access Journals (Sweden)

    Siddiki Mohammad Shohel

    2012-10-01

    Full Text Available Abstract Background From a human health viewpoint, contaminated milk and its products could be a source of long-term exposure to toxic metals. Simple, inexpensive, and on-site assays would enable constant monitoring of their contents. Bioassays that can measure toxic metals in milk or yoghurt might reduce the risk. For this purpose, the green fluorescent protein (GFP-tagged trans factors, ArsR-GFP and CadC-GFP, together with their cis elements were used to develop such bioassays. Results ArsR-GFP or CadC-GFP, which binds either toxic metal or DNA fragment including cis element, was directly mixed with cow’s milk or yoghurt within a neutral pH range. The fluorescence of GFP, which is reflected by the association/dissociation ratio between cis element and trans factor, significantly changed with increasing externally added As (III or Cd (II whereas smaller responses to externally added Pb (II and Zn (II were found. Preparation and dilution of whey fraction at low pH were essential to intrinsic zinc quantification using CadC-GFP. Using the extraction procedure and bioassay, intrinsic Zn (II concentrations ranging from 1.4 to 4.8 mg/l for milk brands and from 1.2 to 2.9 mg/kg for yoghurt brands were determined, which correlated to those determined using inductively coupled plasma atomic emission spectroscopy. Conclusions GFP-tagged bacterial trans factors and cis elements can work in the neutralized whole composition and diluted whey fraction of milk and yoghurt. The feature of regulatory elements is advantageous for establishment of simple and rapid assays of toxic metals in dairy products.

  8. Lanthanide-doped upconverting phosphors for bioassay and therapy

    Science.gov (United States)

    Guo, Huichen; Sun, Shiqi

    2012-10-01

    Lanthanide-doped fluorescent materials have gained increasing attention in recent years due to their unique luminescence properties which have led to their use in wide-ranging fields including those of biological applications. Aside from being used as agents for in vivo imaging, lanthanide-doped fluorescent materials also present many advantages for use in bioassays and therapy. In this review, we summarize the applications of lanthanide-doped up-converting phosphors (UCPs) in protein and gene detection, as well as in photodynamic and gene therapy in recent years, and outline their future potential in biological applications. The current report could serve as a reference for researchers in relevant fields.

  9. BIOSAY: a computer program to store and report bioassay data

    International Nuclear Information System (INIS)

    Williams, G.E.; Parlagreco, J.R.

    1978-12-01

    BIOSAY is a computer program designed to manipulate bioassay data. Using BIOSAY, the Dosimetry Group can generate a report suitable for an individual's dosimetry record. A second copy of this report may be mailed to the individual who provided the sample or the area health physicist for review. BIOSAY also contains a data sorting option which allows all the results for particular individuals, or groups of individuals with common attributes, to be separated from the data base. The computer code is written in a conversational tone with aids which make it usable by even casual users of the computer system

  10. Field and Bioassay Indicators for Internal Dose Intervention Therapy

    International Nuclear Information System (INIS)

    Carbaugh, Eugene H.

    2007-01-01

    Guidance is presented that is used at the U.S. Department of Energy Hanford Site to identify the potential need for medical intervention in response to intakes of radioactivity. The guidance, based on ICRP Publication 30 models and committed effective dose equivalents of 20 mSv and 200 mSv, is expressed as numerical workplace measurements and derived first-day bioassay results for large intakes. It is used by facility radiation protection staff and on-call dosimetry support staff during the first few days following an intake

  11. Herbicide impact on Hormosira banksii gametes measured by fluorescence and germination bioassays

    Energy Technology Data Exchange (ETDEWEB)

    Seery, Cliff R. [Institute for Water and Environmental Resource Management, Department of Environmental Sciences, University of Technology, Sydney, Westbourne Street, Gore Hill, 2065 NSW (Australia); Gunthorpe, Leanne [Primary Industries Research Victoria (PIRVic), VIC (Australia); Ralph, Peter J. [Institute for Water and Environmental Resource Management, Department of Environmental Sciences, University of Technology, Sydney, Westbourne Street, Gore Hill, 2065 NSW (Australia)]. E-mail: peter.ralph@uts.edu.au

    2006-03-15

    The innovative bioassay described here involves chlorophyll a fluorescence measurements of gametes from the macroalgae, Hormosira banksii, where gametes (eggs) were exposed to Diuron, Irgarol and Bromacil. Response was assessed as percent inhibition from control of effective quantum yield ({delta}F/Fm') of photosystem II, herein referred to as % PSII Inhibition. This was measured with the dual-channelled pulse amplitude modulated (PAM) fluorometer, ToxY-PAM. The fluorescence bioassay was run simultaneously with an established H. banksii germination bioassay to compare sensitivity, precision, and time-to-result. The fluorescence bioassay gave highly sensitive results evidenced by EC{sub 5}s (% PSII Inhibition) for Diuron, Irgarol and Bromacil being three, four and three orders of magnitude (respectively) lower than EC{sub 5}s generated from the germination bioassays. Precision of the fluorescence bioassay was demonstrated with low coefficient of variations (<30%) for all three toxicants. With regard to time, the fluorescence bioassay gave results within 6 h, as opposed to more than 50 h for the germination bioassay. - Chlorophyll a fluorescence measurements form the basis of a macroalgal bioassay with many advantages over germination-based methods.

  12. Herbicide impact on Hormosira banksii gametes measured by fluorescence and germination bioassays

    International Nuclear Information System (INIS)

    Seery, Cliff R.; Gunthorpe, Leanne; Ralph, Peter J.

    2006-01-01

    The innovative bioassay described here involves chlorophyll a fluorescence measurements of gametes from the macroalgae, Hormosira banksii, where gametes (eggs) were exposed to Diuron, Irgarol and Bromacil. Response was assessed as percent inhibition from control of effective quantum yield (ΔF/Fm') of photosystem II, herein referred to as % PSII Inhibition. This was measured with the dual-channelled pulse amplitude modulated (PAM) fluorometer, ToxY-PAM. The fluorescence bioassay was run simultaneously with an established H. banksii germination bioassay to compare sensitivity, precision, and time-to-result. The fluorescence bioassay gave highly sensitive results evidenced by EC 5 s (% PSII Inhibition) for Diuron, Irgarol and Bromacil being three, four and three orders of magnitude (respectively) lower than EC 5 s generated from the germination bioassays. Precision of the fluorescence bioassay was demonstrated with low coefficient of variations (<30%) for all three toxicants. With regard to time, the fluorescence bioassay gave results within 6 h, as opposed to more than 50 h for the germination bioassay. - Chlorophyll a fluorescence measurements form the basis of a macroalgal bioassay with many advantages over germination-based methods

  13. Compatibility of hydroxypropyl-β-cyclodextrin with algal toxicity bioassays

    International Nuclear Information System (INIS)

    Fai, Patricia Bi; Grant, Alastair; Reid, Brian J.

    2009-01-01

    Numerous reports have indicated that hydrophobic organic compound bioaccessibility in sediment and soil can be determined by extraction using aqueous hydroxypropyl-β-cyclodextrin (HPCD) solutions. This study establishes the compatibility of HPCD with Selenastrum capricornutum and assesses whether its presence influences the toxicity of reference toxicants. Algal growth inhibition (72 h) showed no significant (P > 0.05) difference at HPCD concentrations up to and including 20 mM. HPCD presence did not influence the toxicity of the inorganic reference toxicant (ZnSO 4 ), with IC50 values of 0.82 μM and 0.85 μM, in the presence and absence of HPCD (20 mM), respectively. However, HPCD presence (20 mM) reduced the toxicity of 2,4-dichlorophenol and the herbicides diuron and isoproturon. These reductions were attributed to inclusion complex formation between the toxicants and the HPCD cavity. Liberation of complexed toxicants, by sample manipulation prior to toxicity assessment, is proposed to provide a sensitive, high throughput, bioassay that reflects compound bioaccessibility. - Compatibility of the biomimetic HPCD extraction method with algal cell growth inhibition bioassays to assess toxicity of reference toxicants and environmental relevant herbicides

  14. Progress in herbicide determination with the thylakoid bioassay.

    Science.gov (United States)

    Trapmann, S; Etxebarria, N; Schnabl, H; Grobecker, K H

    1998-01-01

    Chloroplast thylakoids are used as biological units to determine herbicides in different kinds of water samples as well as in aqueous extracts of compost, soil or food samples. The thylakoid bioassay shows clearly inhibition of fluorescence yield in the presence of photosystem II specific herbicides. Due to this method the ecotoxicological effect of samples with unknown pollutants can be tested fast and cost effective. It has been proven that all photosynthetic active compounds are recorded at the same time because only additive interactions occur. Therefore, the contamination level can be expressed as cumulative parameter for photosystem II active substances. Application was improved clearly by the addition of the radical scavenger sodium ascorbate to the isolation media and by a higher concentration of the measuring medium. A new data evaluation method is described yielding in a lower detection limit of 0.4 microg diuron/1. The guidelines for the quality of water for human consumption with an allowable concentration of pesticides in groups is 0,5 microg/1 and can be controlled with the thylakoid bioassay without performing any preconcentration steps.

  15. Compatibility of hydroxypropyl-{beta}-cyclodextrin with algal toxicity bioassays

    Energy Technology Data Exchange (ETDEWEB)

    Fai, Patricia Bi; Grant, Alastair [School of Environmental Sciences, University of East Anglia, Norwich NR4 7TJ (United Kingdom); Reid, Brian J. [School of Environmental Sciences, University of East Anglia, Norwich NR4 7TJ (United Kingdom)], E-mail: b.reid@uea.ac.uk

    2009-01-15

    Numerous reports have indicated that hydrophobic organic compound bioaccessibility in sediment and soil can be determined by extraction using aqueous hydroxypropyl-{beta}-cyclodextrin (HPCD) solutions. This study establishes the compatibility of HPCD with Selenastrum capricornutum and assesses whether its presence influences the toxicity of reference toxicants. Algal growth inhibition (72 h) showed no significant (P > 0.05) difference at HPCD concentrations up to and including 20 mM. HPCD presence did not influence the toxicity of the inorganic reference toxicant (ZnSO{sub 4}), with IC50 values of 0.82 {mu}M and 0.85 {mu}M, in the presence and absence of HPCD (20 mM), respectively. However, HPCD presence (20 mM) reduced the toxicity of 2,4-dichlorophenol and the herbicides diuron and isoproturon. These reductions were attributed to inclusion complex formation between the toxicants and the HPCD cavity. Liberation of complexed toxicants, by sample manipulation prior to toxicity assessment, is proposed to provide a sensitive, high throughput, bioassay that reflects compound bioaccessibility. - Compatibility of the biomimetic HPCD extraction method with algal cell growth inhibition bioassays to assess toxicity of reference toxicants and environmental relevant herbicides.

  16. The use of bioassays to assess the toxicity of sediment in an acid ...

    African Journals Online (AJOL)

    Exposure of river sediment from 7 sampling sites to these bioassays provided an eco-toxicological estimation of the acute toxicity and chronic toxicity emanating from the contaminated sediments. Physico-chemical analyses revealed higher levels of sediment contamination closer to the mine. The bioassays displayed a ...

  17. Validation of a Novel Bioassay for Low-level Perchlorate Determination

    Science.gov (United States)

    2014-04-01

    was not attractive, since these reduce PMS2 , and it was thought they would interfere with the stoichiometry of NADH and perchlorate in the bioassay...these reduce PMS2 directly, and would interfere with the stoichiometry of NADH and perchlorate in the bioassay. Thus the only approach that could be

  18. Sample preparation for combined chemical analysis and bioassay application in water quality assessment

    NARCIS (Netherlands)

    Kolkman, A.; Schriks, M.; Brand, W; Bäuerlein, P.S.; van der Kooi, M.M.E.; van Doorn, R.H.; Emke, E.; Reus, A.; van der Linden, S.; de Voogt, P.; Heringa, M.B.

    2013-01-01

    The combination of in vitro bioassays and chemical screening can provide a powerful toolbox to determine biologically relevant compounds in water extracts. In this study, a sample preparation method is evaluated for the suitability for both chemical analysis and in vitro bioassays. A set of 39

  19. The CALUX bioassay: current status of its application to screening food and feed

    NARCIS (Netherlands)

    Hoogenboom, L.A.P.; Goeyens, L.; Carbonnelle, S.; Loco, van J.; Beernaert, H.; Baeyens, W.; Traag, W.A.; Bovee, T.F.H.; Jacobs, G.; Schoeters, G.

    2006-01-01

    The CALUX bioassay is at present the best screening method for dioxins and dioxin-like (dl) polychlorinated biphenyls (PCBs) in food and feed, and the only assay used in routine monitoring and during larger incidents. Furthermore, the use of bioassays in addition to chemical reference methods allows

  20. Selection of gonadotrophin surge attenuating factor phage antibodies by bioassay.

    Science.gov (United States)

    Sorsa-Leslie, Tarja; Mason, Helen D; Harris, William J; Fowler, Paul A

    2005-09-26

    We aimed to combine the generation of "artificial" antibodies with a rat pituitary bioassay as a new strategy to overcome 20 years of difficulties in the purification of gonadotrophin surge-attenuating factor (GnSAF). A synthetic single-chain antibody (Tomlinson J) phage display library was bio-panned with partially purified GnSAF produced by cultured human granulosa/luteal cells. The initial screening with a simple binding immunoassay resulted in 8 clones that were further screened using our in-vitro rat monolayer bioassay for GnSAF. Initially the antibodies were screened as pooled phage forms and subsequently as individual, soluble, single-chain antibody (scAbs) forms. Then, in order to improve the stability of the scAbs for immunopurification purposes, and to widen the range of labelled secondary antibodies available, these were engineered into full-length human immunoglobulins. The immunoglobulin with the highest affinity for GnSAF and a previously described rat anti-GnSAF polyclonal antiserum was then used to immunopurify bioactive GnSAF protein. The two purified preparations were electrophoresed on 1-D gels and on 7 cm 2-D gels (pH 4-7). The candidate GnSAF protein bands and spots were then excised for peptide mass mapping. Three of the scAbs recognised GnSAF bioactivity and subsequently one clone of the purified scAb-derived immunoglobulin demonstrated high affinity for GnSAF bioactivity, also binding the molecule in such as way as to block its bioactivity. When used for repeated immunopurification cycles and then Western blot, this antibody enabled the isolation of a GnSAF-bioactive protein band at around 66 kDa. Similar results were achieved using the rat anti-GnSAF polyclonal antiserum. The main candidate molecules identified from the immunopurified material by excision of 2-D gel protein spots was human serum albumin precursor and variants. This study demonstrates that the combination of bioassay and phage display technologies is a powerful tool in the

  1. The BIDAS: bioassay data analysis software for evaluating radionuclide intake and dose

    International Nuclear Information System (INIS)

    Lee, Tae Young; Lee, Jong Il; Chang, Si Young

    2003-01-01

    The BIDAS (BIoassay Data Analysis Software) computer code was developed for the interpretation of bioassay measurements in terms of the intake and dose using the International Commission on Radiological Protection's(ICRP's) currently recommended respiratory tract, GI-tract and biokinetic models to describe the behavior of the radioactive materials within the body. The code consists of a data base module to the manage bioassay data, a data base module containing the predicted bioassay quantities of each radionuclide, and a computational module to the estimate radionuclide intake and dose from either an acute or a chronic exposure based on the measured bioassay quantities. This paper describes the features of the code as well as the results of the BIDAS validation

  2. Plant genotoxicity: a molecular cytogenetic approach in plant bioassays.

    Science.gov (United States)

    Maluszynska, Jolanta; Juchimiuk, Jolanta

    2005-06-01

    It is important for the prevention of DNA changes caused by environment to understand the biological consequences of DNA damages and their molecular modes of action that lead to repair or alterations of the genetic material. Numerous genotoxicity assay systems have been developed to identify DNA reactive compounds. The available data show that plant bioassays are important tests in the detection of genotoxic contamination in the environment and the establishment of controlling systems. Plant system can detect a wide range of genetic damage, including gene mutations and chromosome aberrations. Recently introduced molecular cytogenetic methods allow analysis of genotoxicity, both at the chromosomal and DNA level. FISH gives a new possibility of the detection and analysis of chromosomal rearrangements in a great detail. DNA fragmentation can be estimated using the TUNEL test and the single cell gel electrophoresis (Comet assay).

  3. Neotropical electric fishes (Gymnotiformes as model organisms for bioassays

    Directory of Open Access Journals (Sweden)

    Milena Ferreira

    2015-04-01

    Full Text Available Electric fishes (Gymnotiformes inhabit Central and South America and form a relatively large group with more than 200 species. Besides a taxonomic challenge due to their still unresolved systematic, wide distribution and the variety of habitats they occupy, these fishes have been intensively studied due to their peculiar use of bioelectricity for electrolocation and communication. Conventional analysis of cells, tissues and organs have been complemented with the studies on the electric organ discharges of these fishes. This review compiles the results of 13 bioassays developed during the last 50 years, which used the quickness, low costs and functionality of the bioelectric data collection of Gymnotiformes to evaluate the effects of environmental contaminants and neuroactive drugs.

  4. Bioassay techniques for {sup 55}Fe in urine samples

    Energy Technology Data Exchange (ETDEWEB)

    Cregan, S P; Leon, J W; Linauskas, S H

    1993-11-01

    Solvent extraction, ion chromatography and several rapid screening methods were developed and evaluated for {sup 55}Fe bioassay applications. Isopropyl ether and TNOA column extractions had radiochemical recoveries exceeding 90%. These were very reproducible with a coefficient of variation less than 5%. Screening techniques investigated included direct counting of ashed urine solids, and Fe(OH){sub 3}. precipitated from urine. The sensitivities (2-50 Bq/d urine) of the screening methods were usually limited by the effective urine volume that could be counted in a liquid scintillation counter. The reference isopropyl ether and chromatography methods could easily achieve sensitivities well below the 1 Bq/d urine output target. (author). 49 refs., 3 tabs., 5 figs.

  5. Bioassay techniques for 55Fe in urine samples

    International Nuclear Information System (INIS)

    Cregan, S.P.; Leon, J.W.; Linauskas, S.H.

    1993-11-01

    Solvent extraction, ion chromatography and several rapid screening methods were developed and evaluated for 55 Fe bioassay applications. Isopropyl ether and TNOA column extractions had radiochemical recoveries exceeding 90%. These were very reproducible with a coefficient of variation less than 5%. Screening techniques investigated included direct counting of ashed urine solids, and Fe(OH) 3 . precipitated from urine. The sensitivities (2-50 Bq/d urine) of the screening methods were usually limited by the effective urine volume that could be counted in a liquid scintillation counter. The reference isopropyl ether and chromatography methods could easily achieve sensitivities well below the 1 Bq/d urine output target. (author). 49 refs., 3 tabs., 5 figs

  6. Toxicity assessment using different bioassays and microbial biosensors.

    Science.gov (United States)

    Hassan, Sedky H A; Van Ginkel, Steven W; Hussein, Mohamed A M; Abskharon, Romany; Oh, Sang-Eun

    2016-01-01

    Toxicity assessment of water streams, wastewater, and contaminated sediments, is a very important part of environmental pollution monitoring. Evaluation of biological effects using a rapid, sensitive and cost effective method can indicate specific information on ecotoxicity assessment. Recently, different biological assays for toxicity assessment based on higher and lower organisms such as fish, invertebrates, plants and algal cells, and microbial bioassays have been used. This review focuses on microbial biosensors as an analytical device for environmental, food, and biomedical applications. Different techniques which are commonly used in microbial biosensing include amperometry, potentiometry, conductometry, voltammetry, microbial fuel cells, fluorescence, bioluminescence, and colorimetry. Examples of the use of different microbial biosensors in assessing a variety of environments are summarized. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Seroprevalensi Toxoplasma gondii pada Kambing dan Bioassay Patogenitasnya pada Kucing

    Directory of Open Access Journals (Sweden)

    Ni Made Yunik Novita Dewi Dewi

    2013-11-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE The study aimed to determine seroprevalence of Toxoplasmosis in goats sloughtered at Kampung Jawa, Denpasar, Bali and to evaluate their pathogenicities through bioassay in cats.One hundred serums and meats of goats were collected. Anti-Toxoplasma gondii antibody was determined using Indirect Haemaglutination (IHA test. The pathogenicity bioassay of Toxoplasma gondii was carried out through inoculating the meats of goats which had seropositive of Toxoplasma gondii to the cats. The pathogenicity was evaluated using the intensity of oocyte sheding from the cats. The result showed that the seroprevalence of Toxoplasmosis was 46%. There was not significant difference between pathogenicity of Toxoplasma gondii in cat inoculated with meat of goat which had a high and low titer of antibody against Toxoplasma gondii. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; text-align:justify; line-height:150%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;}

  8. Evolving BioAssay Ontology (BAO): modularization, integration and applications.

    Science.gov (United States)

    Abeyruwan, Saminda; Vempati, Uma D; Küçük-McGinty, Hande; Visser, Ubbo; Koleti, Amar; Mir, Ahsan; Sakurai, Kunie; Chung, Caty; Bittker, Joshua A; Clemons, Paul A; Brudz, Steve; Siripala, Anosha; Morales, Arturo J; Romacker, Martin; Twomey, David; Bureeva, Svetlana; Lemmon, Vance; Schürer, Stephan C

    2014-01-01

    The lack of established standards to describe and annotate biological assays and screening outcomes in the domain of drug and chemical probe discovery is a severe limitation to utilize public and proprietary drug screening data to their maximum potential. We have created the BioAssay Ontology (BAO) project (http://bioassayontology.org) to develop common reference metadata terms and definitions required for describing relevant information of low-and high-throughput drug and probe screening assays and results. The main objectives of BAO are to enable effective integration, aggregation, retrieval, and analyses of drug screening data. Since we first released BAO on the BioPortal in 2010 we have considerably expanded and enhanced BAO and we have applied the ontology in several internal and external collaborative projects, for example the BioAssay Research Database (BARD). We describe the evolution of BAO with a design that enables modeling complex assays including profile and panel assays such as those in the Library of Integrated Network-based Cellular Signatures (LINCS). One of the critical questions in evolving BAO is the following: how can we provide a way to efficiently reuse and share among various research projects specific parts of our ontologies without violating the integrity of the ontology and without creating redundancies. This paper provides a comprehensive answer to this question with a description of a methodology for ontology modularization using a layered architecture. Our modularization approach defines several distinct BAO components and separates internal from external modules and domain-level from structural components. This approach facilitates the generation/extraction of derived ontologies (or perspectives) that can suit particular use cases or software applications. We describe the evolution of BAO related to its formal structures, engineering approaches, and content to enable modeling of complex assays and integration with other ontologies and

  9. 9 CFR 147.16 - Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment).

    Science.gov (United States)

    2010-01-01

    ... mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section 147.16 Animals and Animal Products... the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). This procedure has been shown... publications: (a) Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma Gallisepticum.” In: United...

  10. Comparison of solid and liquid-phase bioassays using ecoscores to assess contaminated soils

    Energy Technology Data Exchange (ETDEWEB)

    Lors, Christine [Universite Lille Nord de France, 1bis rue Georges Lefevre, 59044 Lille Cedex (France); Ecole des Mines de Douai, LGCgE-MPE-GCE, 941 rue Charles-Bourseul, 59500 Douai (France); Centre National de Recherche sur les Sites et Sols Pollues, 930 Boulevard Lahure, BP 537, 59505 Douai Cedex (France); Ponge, Jean-Francois, E-mail: ponge@mnhn.fr [Museum National d' Histoire Naturelle, Departement Ecologie et Gestion de la Biodiversite, CNRS UMR 7179, 4 Avenue du Petit-Chateau, 91800 Brunoy (France); Martinez Aldaya, Maite [Museum National d' Histoire Naturelle, Departement Ecologie et Gestion de la Biodiversite, CNRS UMR 7179, 4 Avenue du Petit-Chateau, 91800 Brunoy (France); Damidot, Denis [Universite Lille Nord de France, 1bis rue Georges Lefevre, 59044 Lille Cedex (France); Ecole des Mines de Douai, LGCgE-MPE-GCE, 941 rue Charles-Bourseul, 59500 Douai (France)

    2011-10-15

    Bioassays on aqueous and solid phases of contaminated soils were compared, belonging to a wide array of trophic and response levels and using ecoscores for evaluating ecotoxicological and genotoxicological endpoints. The method was applied to four coke factory soils contaminated mainly with PAHs, but also to a lesser extent by heavy metals and cyanides. Aquatic bioassays do not differ from terrestrial bioassays when scaling soils according to toxicity but they are complementary from the viewpoint of ecological relevance. Both aquatic and terrestrial endpoints are strongly correlated with concentrations of 3-ring PAHs. This evaluation procedure allows us to propose a cost-effective battery which embraces a wide array of test organisms and response levels: it includes two rapid bioassays (Microtox) and springtail avoidance), a micronucleus test and three bioassays of a longer duration (algal growth, lettuce germination and springtail reproduction). This battery can be recommended for a cost-effective assessment of polluted/remediated soils. - Highlights: > Comparison of liquid- and solid-phase bioassays on contaminated soils, using ecoscores. > Complementarity of liquid- and solid-phase bioassays for the evaluation of environmental hazards. > Proposal for a restricted battery of 5 most sensitive tests. > Use of this restricted battery for a cost-effective assessment of polluted/remediated soils. - Aqueous and solid phases of contaminated soils give similar results in terms of toxicity but are complementary for the evaluation of environmental hazards by ecoscores.

  11. Comparison of solid and liquid-phase bioassays using ecoscores to assess contaminated soils

    International Nuclear Information System (INIS)

    Lors, Christine; Ponge, Jean-Francois; Martinez Aldaya, Maite; Damidot, Denis

    2011-01-01

    Bioassays on aqueous and solid phases of contaminated soils were compared, belonging to a wide array of trophic and response levels and using ecoscores for evaluating ecotoxicological and genotoxicological endpoints. The method was applied to four coke factory soils contaminated mainly with PAHs, but also to a lesser extent by heavy metals and cyanides. Aquatic bioassays do not differ from terrestrial bioassays when scaling soils according to toxicity but they are complementary from the viewpoint of ecological relevance. Both aquatic and terrestrial endpoints are strongly correlated with concentrations of 3-ring PAHs. This evaluation procedure allows us to propose a cost-effective battery which embraces a wide array of test organisms and response levels: it includes two rapid bioassays (Microtox) and springtail avoidance), a micronucleus test and three bioassays of a longer duration (algal growth, lettuce germination and springtail reproduction). This battery can be recommended for a cost-effective assessment of polluted/remediated soils. - Highlights: → Comparison of liquid- and solid-phase bioassays on contaminated soils, using ecoscores. → Complementarity of liquid- and solid-phase bioassays for the evaluation of environmental hazards. → Proposal for a restricted battery of 5 most sensitive tests. → Use of this restricted battery for a cost-effective assessment of polluted/remediated soils. - Aqueous and solid phases of contaminated soils give similar results in terms of toxicity but are complementary for the evaluation of environmental hazards by ecoscores.

  12. Establishment of a bioassay for the toxicity evaluation and quality control of Aconitum herbs

    International Nuclear Information System (INIS)

    Qin, Yi; Wang, Jia-bo; Zhao, Yan-ling; Shan, Li-mei; Li, Bao-cai; Fang, Fang; Jin, Cheng; Xiao, Xiao-he

    2012-01-01

    Highlights: ► A new bioassay was optimized to evaluate the toxicity of Aconitum herbs. ► Characterizing total toxicity is its unique advantage over chemical analysis methods. ► The application of this bioassay promotes the safe use of Aconitum herbs in clinic. - Abstract: Currently, no bioassay is available for evaluating the toxicity of Aconitum herbs, which are well known for their lethal cardiotoxicity and neurotoxicity. In this study, we established a bioassay to evaluate the toxicity of Aconitum herbs. Test sample and standard solutions were administered to rats by intravenous infusion to determine their minimum lethal doses (MLD). Toxic potency was calculated by comparing the MLD. The experimental conditions of the method were optimized and standardized to ensure the precision and reliability of the bioassay. The application of the standardized bioassay was then tested by analyzing 18 samples of Aconitum herbs. Additionally, three major toxic alkaloids (aconitine, mesaconitine, and hypaconitine) in Aconitum herbs were analyzed using a liquid chromatographic method, which is the current method of choice for evaluating the toxicity of Aconitum herbs. We found that for all Aconitum herbs, the total toxicity of the extract was greater than the toxicity of the three alkaloids. Therefore, these three alkaloids failed to account for the total toxicity of Aconitum herbs. Compared with individual chemical analysis methods, the chief advantage of the bioassay is that it characterizes the total toxicity of Aconitum herbs. An incorrect toxicity evaluation caused by quantitative analysis of the three alkaloids might be effectively avoided by performing this bioassay. This study revealed that the bioassay is a powerful method for the safety assessment of Aconitum herbs.

  13. [Application of bioassay in quality control of Chinese materia medica-taking Radix Isatidis as an example].

    Science.gov (United States)

    Yan, Dan; Ren, Yongshen; Luo, Jiaoyang; Li, Hanbing; Feng, Xue; Xiao, Xiaohe

    2010-10-01

    Bioassay, which construct the characteristics consistents with Chinese medical science, is the core mode and methods for the quality control of Chinese materia medica. Taking the bioassay of Radix Isatidis as an example, the contribution, status and application of bioassay in the quality control of Chinese materia medica were introduced in this article, and two key issue (the selection of reference and measurement methods) in the process of establishing bioassay were also explained. This article expects to provide a reference for the development and improvement of the bioassay of Chinese materia medica in a practical manipulation level.

  14. Review of literature on bioassay methods for estimating radionuclides in urine

    International Nuclear Information System (INIS)

    Prasad, M.V.R.; Surya Narayana, D.S.; Jeevanram, R.K.; Sundarajan, A.R.

    1991-01-01

    Bioassay methods of certain important radionuclides encountered in the nuclear fuel cycle operations, viz., thorium, uranium, sup(239)Pu, sup(241)Am, sup(90)Sr, sup(99)Tc, sup(106)Ru, sup(137)Cs are reviewed, with special emphasis on urinalysis. Since the preconcentration is an important prerequisite for bioassay, various preconcentration methods are also discussed. Brief account of various instruments both nuclear and analytical used in the bioassay programme is included. The sensitivities of the methods cited in the literature vis-a-vis the derived recording levels indicated in ICRP recommendations are compared. Literature surveyed up to 1990 is tabulated. (author). 96 refs., 1 fig ., 3 tabs

  15. Bioassay of Phenol and its Intermediate Products Using Daphnia magna

    Directory of Open Access Journals (Sweden)

    Afshin Maleki

    2008-06-01

    Full Text Available Phenol is one of the most common compounds found in many industrial effluents such as petroleum refining and petrochemicals, pharmaceuticals, pesticides, paint and dye industries, organic chemicals manufacturing, etc. The contamination of bodies of water with phenol is a serious problem in terms of environmental considerations due to its high toxicity. In this study, toxicity of phenol and its degradation mixtures by sonochemical, photochemical, and photosonochemical processes were investigated. Toxicity assay tests were carried out using Daphnia magna as a bio-indicator. The sonochemical and photochemical experiments were carried out using a bath sonicator (500 W working at 35 and 130 kHz frequencies and with a 400 W medium pressure mercury lamp, respectively. Experiments were performed at initial concentrations of 100 mg L-1. Bioassay tests showed that phenol was toxic to D.magna and so resulted in quite low LC50 values. Comparison of toxicity units (TU between phenol and effluent toxicity showed that TU value for photosonochemical effluent was lower than that obtained for phenol, photochemical effluent, and sonochemical effluent. It was found that the toxicity unit of photochemical effluent was lower than that obtained for sonochemical effluent. According to the D.magna acute toxicity test, it is concluded that photosonolysis and photolysis are capable of decreasing the toxicity of by-products formed during the degradation of phenol aqueous solutions. Photosonic and photolytic processes can, therefore, be recommended as a potential approach to the treatment of phenolic wastewater.

  16. Bioassay-based risk assessment of complex mixtures

    Energy Technology Data Exchange (ETDEWEB)

    Donnelly, K.C.; Safe, S.H. [Texas A& M Univ., Houston, TX (United States); Randerath, K.; Randerath, E. [College Station and Baylor College of Medicine, Houston, TX (United States)

    1994-12-31

    To compare the standard chemical-based risk assessment with in vitro genotoxicity assays, two complex environmental mixtures from a wood preserving site were analyzed in the Salmonella/microsome and E. coli prophage induction assays. Using GC/MS, sample 003 was found to contain relatively low levels of polycyclic aromatic hydrocarbons (PNAs) and elevated levels of polychlorinated dibenzo-p-dioxins (PCDDs), while sample 005 had higher levels of PNAs and relatively low levels of PCDDs. The complex mixtures were sequentially extracted with methylene chloride and methanol for analysis in Salmonella, or extracted with 1:1 hexane: acetone mixture for analysis in the prophage induction assay. At a dose of 1.0 mg/plate in Salmonella strain TA98 with metabolic activation, the methanol extract of sample 003 induced 197 net revertants, while sample 005 induced 436 net revertants. In the prophage induction assay, with activation, the hexane:acetone extract of sample 003 induced a fold increase that was slightly lower than that observed with sample 005. The estimated incremental carcinogenic risk for dermal adsorption and ingestion was 1.5E-3 for sample 003, while for sample 005 the estimated risk was 1.5E-2. Thus, the sample which induced the maximum response in both bioassays also had the highest estimated cancer risk. However, the frequency of PNA-DNA adducts in both skin and liver tissues was appreciably higher with sample 005 than with sample 003.

  17. Bioassay-based risk assessment of complex mixtures

    International Nuclear Information System (INIS)

    Donnelly, K.C.; Huebner, H.J.

    1996-01-01

    The baseline risk assessment often plays an integral role in various decision-making processes at Superfund sites. The present study reports on risk characterizations prepared for seven complex mixtures using biological and chemical analysis. Three of the samples (A, B, and C) were complex mixtures of polycyclic aromatic hydrocarbons (PAHs) extracted from coal tar; while four samples extracted from munitions-contaminated soil contained primarily nitroaromatic hydrocarbons. The chemical-based risk assessment ranked sample C as least toxic, while the risk associated with samples A and B was approximately equal. The microbial bioassay was in general agreement for the coal tar samples. The weighted activity of the coal tar extracts in Salmonella was 4,960 for sample C, and 162,000 and 206,000 for samples A and B, respectively. The bacterial mutagenicity of 2,4,6-trinitrotoluene contaminated soils exhibited an indirect correlation with chemical-based risk assessment. The aqueous extract of sample 004 induced 1,292 net revertants in Salmonella, while the estimated risk to ingestion and dermal adsorption was 2E-9. The data indicate that the chemical-based risk assessment accurately predicted the genotoxicity of the PAHs, while the accuracy of the risk assessment for munitions contaminated soils was limited due to the presence of metabolites of TNT degradation. The biological tests used in this research provide a valuable compliment to chemical analysis for characterizing the genotoxic risk of complex mixtures

  18. Detection of organic compounds with whole-cell bioluminescent bioassays.

    Science.gov (United States)

    Xu, Tingting; Close, Dan; Smartt, Abby; Ripp, Steven; Sayler, Gary

    2014-01-01

    Natural and manmade organic chemicals are widely deposited across a diverse range of ecosystems including air, surface water, groundwater, wastewater, soil, sediment, and marine environments. Some organic compounds, despite their industrial values, are toxic to living organisms and pose significant health risks to humans and wildlife. Detection and monitoring of these organic pollutants in environmental matrices therefore is of great interest and need for remediation and health risk assessment. Although these detections have traditionally been performed using analytical chemical approaches that offer highly sensitive and specific identification of target compounds, these methods require specialized equipment and trained operators, and fail to describe potential bioavailable effects on living organisms. Alternatively, the integration of bioluminescent systems into whole-cell bioreporters presents a new capacity for organic compound detection. These bioreporters are constructed by incorporating reporter genes into catabolic or signaling pathways that are present within living cells and emit a bioluminescent signal that can be detected upon exposure to target chemicals. Although relatively less specific compared to analytical methods, bioluminescent bioassays are more cost-effective, more rapid, can be scaled to higher throughput, and can be designed to report not only the presence but also the bioavailability of target substances. This chapter reviews available bacterial and eukaryotic whole-cell bioreporters for sensing organic pollutants and their applications in a variety of sample matrices.

  19. Luminescent Lanthanide Reporters for High-Sensitivity Novel Bioassays

    Energy Technology Data Exchange (ETDEWEB)

    Anstey, Mitchell R. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Fruetel, Julia A. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Foster, Michael E. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Hayden, Carl C. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Buckley, Heather L. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Arnold, John [Sandia National Lab. (SNL-CA), Livermore, CA (United States)

    2013-09-01

    Biological imaging and assay technologies rely on fluorescent organic dyes as reporters for a number of interesting targets and processes. However, limitations of organic dyes such as small Stokes shifts, spectral overlap of emission signals with native biological fluorescence background, and photobleaching have all inhibited the development of highly sensitive assays. To overcome the limitations of organic dyes for bioassays, we propose to develop lanthanide-based luminescent dyes and demonstrate them for molecular reporting applications. This relatively new family of dyes was selected for their attractive spectral and chemical properties. Luminescence is imparted by the lanthanide atom and allows for relatively simple chemical structures that can be tailored to the application. The photophysical properties offer unique features such as narrow and non-overlapping emission bands, long luminescent lifetimes, and long wavelength emission, which enable significant sensitivity improvements over organic dyes through spectral and temporal gating of the luminescent signal.Growth in this field has been hindered due to the necessary advanced synthetic chemistry techniques and access to experts in biological assay development. Our strategy for the development of a new lanthanide-based fluorescent reporter system is based on chelation of the lanthanide metal center using absorbing chromophores. Our first strategy involves "Click" chemistry to develop 3-fold symmetric chelators and the other involves use of a new class of tetrapyrrole ligands called corroles. This two-pronged approach is geared towards the optimization of chromophores to enhance light output.

  20. Analyzing bioassay data using Bayesian methods-A primer

    International Nuclear Information System (INIS)

    Miller, G.; Inkret, W.C.; Schillaci, M.E.

    1997-01-01

    The classical statistics approach used in health physics for the interpretation of measurements is deficient in that it does not allow for the consideration of needle in a haystack effects, where events that are rare in a population are being detected. In fact, this is often the case in health physics measurements, and the false positive fraction is often very large using the prescriptions of classical statistics. Bayesian statistics provides an objective methodology to ensure acceptably small false positive fractions. The authors present the basic methodology and a heuristic discussion. Examples are given using numerically generated and real bioassay data (Tritium). Various analytical models are used to fit the prior probability distribution, in order to test the sensitivity to choice of model. Parametric studies show that the normalized Bayesian decision level k α -L c /σ 0 , where σ 0 is the measurement uncertainty for zero true amount, is usually in the range from 3 to 5 depending on the true positive rate. Four times σ 0 rather than approximately two times σ 0 , as in classical statistics, would often seem a better choice for the decision level

  1. Analyzing bioassay data using Bayesian methods -- A primer

    Energy Technology Data Exchange (ETDEWEB)

    Miller, G.; Inkret, W.C.; Schillaci, M.E.; Martz, H.F.; Little, T.T.

    2000-06-01

    The classical statistics approach used in health physics for the interpretation of measurements is deficient in that it does not take into account needle in a haystack effects, that is, correct identification of events that are rare in a population. This is often the case in health physics measurements, and the false positive fraction (the fraction of results measuring positive that are actually zero) is often very large using the prescriptions of classical statistics. Bayesian statistics provides a methodology to minimize the number of incorrect decisions (wrong calls): false positives and false negatives. The authors present the basic method and a heuristic discussion. Examples are given using numerically generated and real bioassay data for tritium. Various analytical models are used to fit the prior probability distribution in order to test the sensitivity to choice of model. Parametric studies show that for typical situations involving rare events the normalized Bayesian decision level k{sub {alpha}} = L{sub c}/{sigma}{sub 0}, where {sigma}{sub 0} is the measurement uncertainty for zero true amount, is in the range of 3 to 5 depending on the true positive rate. Four times {sigma}{sub 0} rather than approximately two times {sigma}{sub 0}, as in classical statistics, would seem a better choice for the decision level in these situations.

  2. Comparison of solid-phase bioassays and ecoscores to evaluate the toxicity of contaminated soils.

    Science.gov (United States)

    Lors, Christine; Ponge, Jean-François; Martínez Aldaya, Maite; Damidot, Denis

    2010-08-01

    Five bioassays (inhibition of lettuce germination and growth, earthworm mortality, inhibition of springtail population growth, avoidance by springtails) were compared, using four coke factory soils contaminated by PAHs and trace elements, before and after biotreatment. For each bioassay, several endpoints were combined in an 'ecoscore', a measure of test sensitivity. Ecoscores pooled over all tested bioassays revealed that most organisms were highly sensitive to the concentration of 3-ring PAHs. When four soils were combined, behavioural tests using the springtail Folsomia candida showed higher ecoscores, i.e. they were most sensitive to soil contamination. However, despite overall higher sensitivity of behavioural tests, which could be used for cheap and rapid assessment of soil toxicity, especially at low levels of contamination, some test endpoints were more sensitive than others, and this may differ from a soil to another, pointing to the need for a battery of bioassays when more itemized results are expected. Copyright 2010 Elsevier Ltd. All rights reserved.

  3. Biomonitoring of cyanotoxins in two tropical reservoirs by cladoceran toxicity bioassays.

    Science.gov (United States)

    da S Ferrão-Filho, Aloysio; Soares, Maria Carolina S; de Freitas Magalhães, Valeria; Azevedo, Sandra M F O

    2009-02-01

    This study evaluates the potential for the use of cladocerans in biomonitoring of cyanobacterial toxins. Two zooplankton species (Daphnia gessneri and Moina micrura) were cultivated in the laboratory for use in acute (48 h) and chronic (10 days) bioassays. Water samples were collected from two reservoirs and diluted in mineral water at four concentrations. Survivorship in the acute bioassays was used to calculate LC50, and survivorship and fecundity in chronic bioassays were used to calculate the intrinsic population growth rate (r) and the EC50. Analysis of phytoplankton in the water samples from one reservoir revealed that cyanobacteria were the dominant group, represented by the genera Anabaena, Cylindrospermopsis, and Microcystis. Results of bioassays showed adverse effects including death, paralysis, and reduced population growth rate, generally proportional to the reservoir water concentration. These effects may be related to the presence of cyanobacteria toxins (microcystins or saxitoxins) in the water.

  4. Comparative susceptibility of bemisia tabaci to imidacloprid in field- and laboratory-based bioassays

    Science.gov (United States)

    Bemisia tabaci biotype B is a resistance-prone pest of protected and open agriculture. Systemic uptake bioassays used in resistance monitoring programs have provided important information on susceptibility to neonicotinoid insecticides, but have remained decoupled from field performance. Simultaneou...

  5. Phototoxicity activity of Psoralea drupacea L. using Atremia salina bioassay system

    Directory of Open Access Journals (Sweden)

    Mohammad Ramezani

    2011-07-01

    Conclusion: The result showed that P. drupacea methanolic extract and chloroform fraction have phototoxicity in A. salina bioassay system and their toxic effect is related to phototoxic constituents such as psoralen.

  6. Integration of laboratory bioassays into the risk-based corrective action process

    International Nuclear Information System (INIS)

    Edwards, D.; Messina, F.; Clark, J.

    1995-01-01

    Recent data generated by the Gas Research Institute (GRI) and others indicate that residual hydrocarbon may be bound/sequestered in soil such that it is unavailable for microbial degradation, and thus possibly not bioavailable to human/ecological receptors. A reduction in bioavailability would directly equate to reduced exposure and, therefore, potentially less-conservative risk-based cleanup soil goals. Laboratory bioassays which measure bioavailability/toxicity can be cost-effectively integrated into the risk-based corrective action process. However, in order to maximize the cost-effective application of bioassays several site-specific parameters should be addressed up front. This paper discusses (1) the evaluation of parameters impacting the application of bioassays to soils contaminated with metals and/or petroleum hydrocarbons and (2) the cost-effective integration of bioassays into a tiered ASTM type framework for risk-based corrective action

  7. Hierarchical responses to organic contaminants in aquatic ecotoxicological bioassays: from microcystins to biodegradation

    OpenAIRE

    Montenegro, Katia

    2008-01-01

    In this thesis I explore the ecotoxicological responses of aquatic organisms at different hierarchical levels to organic contaminants by means of bioassays. The bioassays use novel endpoints or approaches to elucidate the effects of exposure to contaminants and attempt to give mechanistic explanations that could be used to interpret effects at higher hierarchical scales. The sensitivity of population growth rate in the cyanobacteria species Microcystis aeruginosa to the herbicide glyp...

  8. Issues in weighting bioassay data for use in regressions for internal dose assessments

    International Nuclear Information System (INIS)

    Strom, D.J.

    1992-11-01

    For use of bioassay data in internal dose assessment, research should be done to clarify the goal desired, the choice of method to achieve the goal, the selection of adjustable parameters, and on the ensemble of information that is available. Understanding of these issues should determine choices of weighting factors for bioassay data used in regression models. This paper provides an assessment of the relative importance of the various factors

  9. Development and validation of microbial bioassay for quantification of Levofloxacin in pharmaceutical preparations

    Directory of Open Access Journals (Sweden)

    Nishant A. Dafale

    2015-02-01

    Full Text Available The aim of this study was to develop and validate a simple, sensitive, precise and cost-effective one-level agar diffusion (5+1 bioassay for estimation of potency and bioactivity of Levofloxacin in pharmaceutical preparation which has not yet been reported in any pharmacopoeia. Among 16 microbial strains, Bacillus pumilus ATCC-14884 was selected as the most significant strain against Levofloxacin. Bioassay was optimized by investigating several factors such as buffer pH, inoculums concentration and reference standard concentration. Identification of Levofloxacin in commercial sample Levoflox tablet was done by FTIR spectroscopy. Mean potency recovery value for Levofloxacin in Levoflox tablet was estimated as 100.90%. A validated bioassay method showed linearity (r2=0.988, precision (Interday RSD=1.05%, between analyst RSD=1.02% and accuracy (101.23%, RSD=0.72%. Bioassay was correlated with HPLC using same sample and estimated potencies were 100.90% and 99.37%, respectively. Results show that bioassay is a suitable method for estimation of potency and bioactivity of Levofloxacin pharmaceutical preparations. Keywords: Levofloxacin, Antibiotic resistance, Microbiological bioassay, HPLC, Pharmacopoeia

  10. A rapid bioassay for detecting saxitoxins using a Daphnia acute toxicity test

    Energy Technology Data Exchange (ETDEWEB)

    Ferrao-Filho, Aloysio da S., E-mail: aloysio@ioc.fiocruz.b [Laboratorio de Avaliacao e Promocao da Saude Ambiental, Departamento de Biologia, Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil 4365, Manguinhos, Rio de Janeiro, RJ 21045-900 (Brazil); Soares, Maria Carolina S., E-mail: mcarolsoares@gmail.co [Departamento de Engenharia Sanitaria e Ambiental Faculdade de Engenharia, Universidade Federal de Juiz de Fora, Juiz de Fora, MG 36036-900 (Brazil); Freitas de Magalhaes, Valeria, E-mail: valeria@biof.ufrj.b [Laboratorio de Ecofisiologia e Toxicologia de Cianobacterias, Instituto de Biofisica Carlos Chagas Filho, CCS, Universidade Federal do Rio de Janeiro, Ilha do Fundao, Rio de Janeiro, RJ 21949-900 (Brazil); Azevedo, Sandra M.F.O., E-mail: sazevedo@biof.ufrj.b [Laboratorio de Ecofisiologia e Toxicologia de Cianobacterias, Instituto de Biofisica Carlos Chagas Filho, CCS, Universidade Federal do Rio de Janeiro, Ilha do Fundao, Rio de Janeiro, RJ 21949-900 (Brazil)

    2010-06-15

    Bioassays using Daphnia pulex and Moina micrura were designed to detect cyanobacterial neurotoxins in raw water samples. Phytoplankton and cyanotoxins from seston were analyzed during 15 months in a eutrophic reservoir. Effective time to immobilize 50% of the exposed individuals (ET{sub 50}) was adopted as the endpoint. Paralysis of swimming movements was observed between approx0.5-3 h of exposure to lake water containing toxic cyanobacteria, followed by an almost complete recovery of the swimming activity within 24 h after being placed in control water. The same effects were observed in bioassays with a saxitoxin-producer strain of Cylindrospermopsis raciborskii isolated from the reservoir. Regression analysis showed significant relationships between ET{sub 50}vs. cell density, biomass and saxitoxins content, suggesting that the paralysis of Daphnia in lake water samples was caused by saxitoxins found in C. raciborskii. Daphnia bioassay was found to be a sensitive method for detecting fast-acting neurotoxins in natural samples, with important advantages over mouse bioassays. - A new Daphnia bioassay, as an alternative to the mouse bioassay, is able to detect effects of fast-acting, potent neurotoxins in raw water.

  11. A rapid bioassay for detecting saxitoxins using a Daphnia acute toxicity test

    International Nuclear Information System (INIS)

    Ferrao-Filho, Aloysio da S.; Soares, Maria Carolina S.; Freitas de Magalhaes, Valeria; Azevedo, Sandra M.F.O.

    2010-01-01

    Bioassays using Daphnia pulex and Moina micrura were designed to detect cyanobacterial neurotoxins in raw water samples. Phytoplankton and cyanotoxins from seston were analyzed during 15 months in a eutrophic reservoir. Effective time to immobilize 50% of the exposed individuals (ET 50 ) was adopted as the endpoint. Paralysis of swimming movements was observed between ∼0.5-3 h of exposure to lake water containing toxic cyanobacteria, followed by an almost complete recovery of the swimming activity within 24 h after being placed in control water. The same effects were observed in bioassays with a saxitoxin-producer strain of Cylindrospermopsis raciborskii isolated from the reservoir. Regression analysis showed significant relationships between ET 50 vs. cell density, biomass and saxitoxins content, suggesting that the paralysis of Daphnia in lake water samples was caused by saxitoxins found in C. raciborskii. Daphnia bioassay was found to be a sensitive method for detecting fast-acting neurotoxins in natural samples, with important advantages over mouse bioassays. - A new Daphnia bioassay, as an alternative to the mouse bioassay, is able to detect effects of fast-acting, potent neurotoxins in raw water.

  12. An ion quencher operated lamp for multiplexed fluorescent bioassays.

    Science.gov (United States)

    Qing, Taiping; Sun, Huanhuan; He, Xiaoxiao; Huang, Xiaoqin; He, Dinggeng; Bu, Hongchang; Qiao, Zhenzhen; Wang, Kemin

    2018-02-01

    A novel and adjustable lamp based on competitive interaction among dsDNA-SYBR Green I (SGI), ion quencher, and analyte was designed for bioanalysis. The "filament" and switch of the lamp could be customized by employing different dsDNA and ion quencher. The poly(AT/TA) dsDNA was successfully screened as the most effective filament of the lamp. Two common ions, Hg 2+ and Fe 3+ , were selected as the model switch, and the corresponding ligand molecules cysteine (Cys) and pyrophosphate ions (PPi) were selected as the targets. When the fluorescence-quenched dsDNA/SGI-ion complex was introduced into a target-containing system, ions could be bound by competitive molecules and separate from the complex, thereby lighting the lamp. However, no light was observed if the biomolecule could not snatch the metal ions from the complex. Under the optimal conditions, sensitive and selective detection of Cys and PPi was achieved by the lamp, with practical applications in fetal bovine serum and human urine. This ion quencher regulated lamp for fluorescent bioassays is simple in design, fast in operation, and is more convenient than other methods. Significantly, as many molecules could form stable complexes with metal ions selectively, this ion quencher operated lamp has potential for the detection of a wide spectrum of analytes. Graphical abstract A novel and adjustable lamp on the basis of competitive interaction among dsDNA-SYBR Green I, ions quencher and analyte was designed for bioanalysis. The filament and switch of lamp could be customized by employing different dsDNA and ions quencher.

  13. Development of Androgen- and Estrogen-Responsive bio-assays, members of a panel of human cell line-based highly selective steroid-responsive bioassays

    NARCIS (Netherlands)

    Sonneveld, E.; Jansen, H.J.

    2004-01-01

    We have established highly sensitive and specific androgen and estrogen reporter cell lines which we have named AR (androgen receptor) and ERα (estrogen receptor alpha) CALUX® (Chemically Activated LUciferase eXpression), respectively. Both bioassays are member of a panel of CALUX reporter cell

  14. Development of androgen-and estrogen-responsive bioassays, members of a panel of human cell line-based highly selective steroid-responsive bioassays

    NARCIS (Netherlands)

    Sonneveld, E.; Jansen, H.J..; Riteco, J.A.C.; Brouwer, A.

    2005-01-01

    We have established highly sensitive and specific androgen and estrogen reporter cell lines which we have named AR (androgen receptor) and ERα (estrogen receptor alpha) CALUX® (Chemically Activated LUciferase eXpression), respectively. Both bioassays are member of a panel of CALUX reporter cell

  15. Evaluation of the toxicity of two soils from Jales Mine (Portugal) using aquatic bioassays.

    Science.gov (United States)

    Loureiro, Susana; Ferreira, Abel L G; Soares, Amadeu M V M; Nogueira, António J A

    2005-10-01

    Soil contamination can be one path for streams and groundwater contamination. As a complement of chemical analysis and total contaminants determination, bioassays can provide information on the bioavailable fraction of chemical compounds, focusing on the retention and habitat function of soils. In this study the evaluation of the toxicity of two soils from the abandoned Jales Mine (Portugal) regarded both functions. The buffer capacity of soils was tested with bioassays carried out using the cladoceran Daphnia magna and the marine bacteria Vibrio fischeri. The habitat function of soils was evaluated with the reproduction bioassay with the collembolan Folsomia candida. The Microtox solid-phase test was performed with V. fischeri using soil as test medium, and soil elutriates were extracted to perform the Microtox basic test, and an immobilization and reproduction bioassay with D. magna. The marine bacteria showed high sensitivity to the soil with low heavy metal content (JNC soil) and to JNC soil elutriates, while the soil with highest heavy metal content (JC soil) or soil elutriates exposure did not cause any toxic effect. In the bioassays with D. magna, organisms showed sensitivity to JNC and also to JC soil elutriates. Both mobilization and reproduction features were inhibited. The bioassay with F. candida did not reflect any influence of the contaminants on their reproduction. Although JNC soil presented lower heavy metal contents, elutriates showed different patterns of contamination when compared to JC soil and elutriates, which indicates different retention and buffer capacities between soils. Results obtained in this study underlined the sensitivity and importance of soil elutriate bioassays with aquatic organisms in the evaluation strategy in soil ERA processes.

  16. The limits of two-year bioassay exposure regimens for identifying chemical carcinogens.

    Science.gov (United States)

    Huff, James; Jacobson, Michael F; Davis, Devra Lee

    2008-11-01

    Chemical carcinogenesis bioassays in animals have long been recognized and accepted as valid predictors of potential cancer hazards to humans. Most rodent bioassays begin several weeks after birth and expose animals to chemicals or other substances, including workplace and environmental pollutants, for 2 years. New findings indicate the need to extend the timing and duration of exposures used in the rodent bioassay. In this Commentary, we propose that the sensitivity of chemical carcinogenesis bio-assays would be enhanced by exposing rodents beginning in utero and continuing for 30 months (130 weeks) or until their natural deaths at up to about 3 years. Studies of three chemicals of different structures and uses-aspartame, cadmium, and toluene-suggest that exposing experimental animals in utero and continuing exposure for 30 months or until their natural deaths increase the sensitivity of bioassays, avoid false-negative results, and strengthen the value and validity of results for regulatory agencies. Government agencies, drug companies, and the chemical industry should conduct and compare the results of 2-year bioassays of known carcinogens or chemicals for which there is equivocal evidence of carcinogenicity with longer-term studies, with and without in utero exposure. If studies longer than 2 years and/or with in utero exposure are found to better identify potential human carcinogens, then regulatory agencies should promptly revise their testing guidelines, which were established in the 1960s and early 1970s. Changing the timing and dosing of the animal bioassay would enhance protection of workers and consumers who are exposed to potentially dangerous workplace or home contaminants, pollutants, drugs, food additives, and other chemicals throughout their lives.

  17. Benchmarking organic micropollutants in wastewater, recycled water and drinking water with in vitro bioassays.

    Science.gov (United States)

    Escher, Beate I; Allinson, Mayumi; Altenburger, Rolf; Bain, Peter A; Balaguer, Patrick; Busch, Wibke; Crago, Jordan; Denslow, Nancy D; Dopp, Elke; Hilscherova, Klara; Humpage, Andrew R; Kumar, Anu; Grimaldi, Marina; Jayasinghe, B Sumith; Jarosova, Barbora; Jia, Ai; Makarov, Sergei; Maruya, Keith A; Medvedev, Alex; Mehinto, Alvine C; Mendez, Jamie E; Poulsen, Anita; Prochazka, Erik; Richard, Jessica; Schifferli, Andrea; Schlenk, Daniel; Scholz, Stefan; Shiraishi, Fujio; Snyder, Shane; Su, Guanyong; Tang, Janet Y M; van der Burg, Bart; van der Linden, Sander C; Werner, Inge; Westerheide, Sandy D; Wong, Chris K C; Yang, Min; Yeung, Bonnie H Y; Zhang, Xiaowei; Leusch, Frederic D L

    2014-01-01

    Thousands of organic micropollutants and their transformation products occur in water. Although often present at low concentrations, individual compounds contribute to mixture effects. Cell-based bioassays that target health-relevant biological endpoints may therefore complement chemical analysis for water quality assessment. The objective of this study was to evaluate cell-based bioassays for their suitability to benchmark water quality and to assess efficacy of water treatment processes. The selected bioassays cover relevant steps in the toxicity pathways including induction of xenobiotic metabolism, specific and reactive modes of toxic action, activation of adaptive stress response pathways and system responses. Twenty laboratories applied 103 unique in vitro bioassays to a common set of 10 water samples collected in Australia, including wastewater treatment plant effluent, two types of recycled water (reverse osmosis and ozonation/activated carbon filtration), stormwater, surface water, and drinking water. Sixty-five bioassays (63%) showed positive results in at least one sample, typically in wastewater treatment plant effluent, and only five (5%) were positive in the control (ultrapure water). Each water type had a characteristic bioanalytical profile with particular groups of toxicity pathways either consistently responsive or not responsive across test systems. The most responsive health-relevant endpoints were related to xenobiotic metabolism (pregnane X and aryl hydrocarbon receptors), hormone-mediated modes of action (mainly related to the estrogen, glucocorticoid, and antiandrogen activities), reactive modes of action (genotoxicity) and adaptive stress response pathway (oxidative stress response). This study has demonstrated that selected cell-based bioassays are suitable to benchmark water quality and it is recommended to use a purpose-tailored panel of bioassays for routine monitoring.

  18. Assessing arsenic bioavailability through the use of bioassays

    Science.gov (United States)

    Diesel, E.; Nadimpalli, M.; Hull, M.; Schreiber, M. E.; Vikesland, P.

    2009-12-01

    Various methods have been used to characterize the bioavailability of a contaminant, including chemical extractions from soils, toxicity tests, bioaccumulation measurements, estimation from soil properties, in vitro/in vivo tests, and microbial biossays. Unfortunately, these tests are all unique (i.e. they measure bioavailability through different mechanisms) and it is difficult to compare measurements collected using one method to those collected from another. Additionally, there are fundamental aspects of bioavailability research that require further study. In particular, changes in bioavailability over time are not well understood, as well as what the geochemical controls are on changes in bioavailability. In addition, there are no studies aimed at the integration of bioavailability measurements and potential geochemical controls. This research project seeks to find a standard set of assays and sensors that can be used to assess arsenic bioavailability at any field site, as well as to use these tools and techniques to better understand changes in, and controls on, arsenic bioavailability. The bioassays to be utilized in this research are a bioluminescent E. coli assay and a Corbicula fluminea (Asian clam) assay. Preliminary experiments to determine the suitability of the E. coli and C. fluminea assays have been completed. The E. coli assay can be utilized to analyze As(III) and As(V) with a linear standard curve between 5 and 200 ppb for As(III) and 100 ppb and 5 ppm for As(V); no bioluminescent response above background was elicited in the presence of Roxarsone, an organoarsenical. The C. fluminea assay is capable of bioaccumulating As(III), As(V), Roxarsone, and MSMA, with As(III) being the most readily accumulated, followed by As(V), Roxarsone and MSMA, respectively. Additional research will include assessing bioavailability of various arsenic species adsorbed to natural colloidal materials (i.e. clays, iron oxides, NOM) to the E. coli and C. fluminea assays

  19. Structuring a risk-based bioassay program for uranium usage in university laboratories

    Science.gov (United States)

    Dawson, Johnne Talia

    Bioassay programs are integral in a radiation safety program. They are used as a method of determining whether individuals working with radioactive material have been exposed and have received a resulting dose. For radionuclides that are not found in nature, determining an exposure is straightforward. However, for a naturally occurring radionuclide like uranium, it is not as straightforward to determine whether a dose is the result of an occupational exposure. The purpose of this project is to address this issue within the University of Nevada, Las Vegas's (UNLV) bioassay program. This project consisted of two components that studied the effectiveness of a bioassay program in determining the dose for an acute inhalation of uranium. The first component of the plan addresses the creation of excretion curves, utilizing MATLAB that would allow UNLV to be able to determine at what time an inhalation dose can be attributed to. The excretion curves were based on the ICRP 30 lung model, as well as the Annual Limit Intake (ALI) values located in the Nuclear Regulatory Commission's 10CFR20 which is based on ICRP 30 (International Commission on Radiological Protection). The excretion curves would allow UNLV to be able to conduct in-house investigations of inhalation doses without solely depending on outside investigations and sources. The second component of the project focused on the creation of a risk based bioassay program to be utilized by UNLV that would take into account bioassay frequency that depended on the individual. Determining the risk based bioassay program required the use of baseline variance in order to minimize the investigation of false positives among those individuals who undergo bioassays for uranium work. The proposed program was compared against an evaluation limit of 10 mrem per quarter, an investigational limit of 125 mrem per quarter, and the federal/state requirement of 1.25 rem per quarter. It was determined that a bioassay program whose bioassay

  20. Comparison of solid-phase bioassays and ecoscores to evaluate the toxicity of contaminated soils

    International Nuclear Information System (INIS)

    Lors, Christine; Ponge, Jean-Francois; Martinez Aldaya, Maite; Damidot, Denis

    2010-01-01

    Five bioassays (inhibition of lettuce germination and growth, earthworm mortality, inhibition of springtail population growth, avoidance by springtails) were compared, using four coke factory soils contaminated by PAHs and trace elements, before and after biotreatment. For each bioassay, several endpoints were combined in an 'ecoscore', a measure of test sensitivity. Ecoscores pooled over all tested bioassays revealed that most organisms were highly sensitive to the concentration of 3-ring PAHs. When four soils were combined, behavioural tests using the springtail Folsomia candida showed higher ecoscores, i.e. they were most sensitive to soil contamination. However, despite overall higher sensitivity of behavioural tests, which could be used for cheap and rapid assessment of soil toxicity, especially at low levels of contamination, some test endpoints were more sensitive than others, and this may differ from a soil to another, pointing to the need for a battery of bioassays when more itemized results are expected. - The avoidance test using the soil springtail Folsomia candida is globally more sensitive to PAH contamination than acute and chronic toxicity bioassays using plants and animals but a battery of tests could reveal better in detail.

  1. Comparison of solid-phase bioassays and ecoscores to evaluate the toxicity of contaminated soils

    Energy Technology Data Exchange (ETDEWEB)

    Lors, Christine [Universite Lille Nord de France, 1bis rue Georges Lefevre, 59044 Lille Cedex (France); Ecole des Mines de Douai, MPE-GCE, 941 rue Charles-Bourseul, 59500 Douai (France); Centre National de Recherche sur les Sites et Sols Pollues, 930 Boulevard Lahure, BP 537, 59505 Douai Cedex (France); Ponge, Jean-Francois, E-mail: ponge@mnhn.f [Museum National d' Histoire Naturelle, CNRS UMR 7179, 4 Avenue du Petit-Chateau, 91800 Brunoy (France); Martinez Aldaya, Maite [Museum National d' Histoire Naturelle, CNRS UMR 7179, 4 Avenue du Petit-Chateau, 91800 Brunoy (France); Damidot, Denis [Universite Lille Nord de France, 1bis rue Georges Lefevre, 59044 Lille Cedex (France); Ecole des Mines de Douai, MPE-GCE, 941 rue Charles-Bourseul, 59500 Douai (France)

    2010-08-15

    Five bioassays (inhibition of lettuce germination and growth, earthworm mortality, inhibition of springtail population growth, avoidance by springtails) were compared, using four coke factory soils contaminated by PAHs and trace elements, before and after biotreatment. For each bioassay, several endpoints were combined in an 'ecoscore', a measure of test sensitivity. Ecoscores pooled over all tested bioassays revealed that most organisms were highly sensitive to the concentration of 3-ring PAHs. When four soils were combined, behavioural tests using the springtail Folsomia candida showed higher ecoscores, i.e. they were most sensitive to soil contamination. However, despite overall higher sensitivity of behavioural tests, which could be used for cheap and rapid assessment of soil toxicity, especially at low levels of contamination, some test endpoints were more sensitive than others, and this may differ from a soil to another, pointing to the need for a battery of bioassays when more itemized results are expected. - The avoidance test using the soil springtail Folsomia candida is globally more sensitive to PAH contamination than acute and chronic toxicity bioassays using plants and animals but a battery of tests could reveal better in detail.

  2. Natural products phytotoxicity A bioassay suitable for small quantities of slightly water-soluble compounds.

    Science.gov (United States)

    Dornbos, D L; Spencer, G F

    1990-02-01

    A large variety of secondary metabolites that can inhibit germination and/or seedling growth are produced by plants in low quantities. The objective of this study was to develop a bioassay capable of reliably assessing reductions in germination percentage and seedling length of small-seeded plant species caused by exposure to minute quantities of these compounds. The germination and growth of alfalfa (Medicago saliva), annual ryegrass (Lolium multiflorum), and velvetleaf (Abutilon theophrasti) were evaluated against six known phytotoxins from five chemical classes; cinmethylin (a herbicidal cineole derivative) was selected as a comparison standard. Each phytotoxin, dissolved in a suitable organic solvent, was placed on water-agar in small tissue culture wells. After the solvent evaporated, imbibed seeds were placed on the agar; after three days, germination percentages and seedling lengths were measured. Compared to a commonly used filter paper procedure, this modified agar bioassay required smaller quantities of compound per seed for comparable bioassay results. This bioassay also readily permitted the measurement of seedling length, a more sensitive indicator of phytotoxicity than germination. Seedling length decreased sigmoidally as the toxin concentration increased logarithmically. Phytotoxicity was a function of both compound and plant species. Cinmethylin, a grass herbicide, reduced the length of annual ryegrass seedlings by 90-100%, whereas that of alfalfa and velvetleaf was inhibited slightly. The agar bioassay facilitated the rapid and reliable testing of slightly water-soluble compounds, requiring only minute quantities of each compound to give reproducible results.

  3. Development by flow cytometry of bioassays based on chlorella for environmental monitoring

    Directory of Open Access Journals (Sweden)

    Petrescu C-M,

    2016-05-01

    Full Text Available In ecotoxicological assessments, bioassays (ecotoxicity tests or biotests are one of the main tools, defined as methods which use living cells, tissues, organism or communities to assess exposure-related effects of chemicals. The increasing complexity of environmental degradation requires an increase in the capacity of scientific approach in monitoring and notification as early as possible risks. Our own objective concerns the detection of aquatic environment pollution in Romania and particularly in the Danube basin. For assessing aquatic environment pollution degree or for assessing cytotoxicity or ecotoxicity of pollutants (heavy metals, nanoparticles, pesticides, etc. we developed news experimental bioassays based on the use of viability and apoptosis biomarkers of Chlorella cells by flow cytometry. Our proposed bioassays could be rapid and very sensitive tests for in laboratory aquatic risk assessment and biomonitoring.

  4. Methodology for estimation of 32P in bioassay samples by Cerenkov counting

    International Nuclear Information System (INIS)

    Wankhede, Sonal; Sawant, Pramilla D.; Yadav, R.K.B.; Rao, D.D.

    2016-01-01

    Radioactive phosphorus ( 32 P) as phosphate is used to effectively reduce bone pain in terminal cancer patients. Several hospitals in India carry out this palliative care procedure on a regular basis. Thus, production as well as synthesis of 32 P compounds has increased over the years to meet this requirement. Monitoring of radiation workers handling 32 P compounds is important for further strengthening of radiological protection program at processing facility. 32 P being a pure beta emitter (β max = 1.71 MeV, t 1/2 = 14.3 d), bioassay is the preferred individual monitoring technique. Method standardized at Bioassay Lab, Trombay, includes estimation of 32 P in urine by co-precipitation with ammonium phosphomolybdate (AMP) followed by gross beta counting. In the present study, feasibility of Cerenkov counting for detection of 32 P in bioassay samples was explored and the results obtained were compared with the gross beta counting technique

  5. Comparison of liquid chromatographic and bioassay procedures for determining depletion of intramuscularly injected tylosin.

    Science.gov (United States)

    Moats, W A; Harris, E W; Steele, N C

    1985-01-01

    Crossbred pigs weighing 80-110 kg were injected intramuscularly in the ham with 8.8 mg/kg tylosin. Animals were slaughtered in groups of 3 at intervals of 4 h, and 1, 2, 4, and 8 days after injection, and samples of blood, injected muscle, uninjected muscle, liver, and kidney were analyzed by liquid chromatography (LC) and by bioassay using Sarcina lutea as the test organism. The LC method was far more sensitive with a detection limit of less than 0.1 ppm, while the detection limit by bioassay was about 0.5 ppm in tissue. Results by bioassay and LC sometimes differed considerably for tissue samples. Residues in all tissues were below the tolerance limit of 0.2 ppm at 24 h, except in the injected muscle in one animal. Residues were not detected in any tissue of any animal at 48 h after treatment.

  6. Requirements for radiation emergency urine bioassay techniques for the public and first responders.

    Science.gov (United States)

    Li, Chunsheng; Vlahovich, Slavica; Dai, Xiongxin; Richardson, Richard B; Daka, Joseph N; Kramer, Gary H

    2010-11-01

    Following a radiation emergency, the affected public and the first responders may need to be quickly assessed for internal contamination by the radionuclides involved. Urine bioassay is one of the most commonly used methods for assessing radionuclide intake and radiation dose. This paper attempts to derive the sensitivity requirements (from inhalation exposure) for the urine bioassay techniques for the top 10 high-risk radionuclides that might be used in a terrorist attack. The requirements are based on a proposed reference dose to adults of 0.1 Sv (CED, committed effective dose). In addition, requirements related to sample turnaround time and field deployability of the assay techniques are also discussed. A review of currently available assay techniques summarized in this paper reveals that method development for ²⁴¹Am, ²²⁶Ra, ²³⁸Pu, and ⁹⁰Sr urine bioassay is needed.

  7. A rapid and inexpensive bioassay to evaluate the decontamination of organophosphates.

    Science.gov (United States)

    Claborn, David M; Martin-Brown, Skylar A; Sagar, Sanjay Gupta; Durham, Paul

    2012-01-01

    An inexpensive and rapid bioassay using adult red flour beetles was developed for use in assessing the decontamination of environments containing organophosphates and related chemicals. A decontamination protocol was developed which demonstrated that 2 to 3 applications of 5% bleach solution were required to obtain nearly complete decontamination of malathion. The bioassay was also used to screen common household cleaners as potential decontaminating agents, but only 5% bleach was effective at improving survival of insects on steel plates treated with 25% malathion. A toxic degradation product (malaoxon) was detected using gas chromatography/mass spectrophotometry; this toxin affected the decontamination efficacy and resulted in continued toxicity to the beetles until subsequent decontaminations. The bioassay provides evidence to support the use of red flour beetles as a sensitive, less expensive method for determining safety levels of environments contaminated with malathion and other toxins, and may have application in the study of chemical warfare agents.

  8. Sampling method, storage and pretreatment of sediment affect AVS concentrations with consequences for bioassay responses.

    Science.gov (United States)

    De Lange, H J; Van Griethuysen, C; Koelmans, A A

    2008-01-01

    Sediment treatment and sediment storage may alter sediment toxicity, and consequently biotic response. Purpose of our study was to combine these three aspects (treatment-toxicity-biotic response) in one integrated approach. We used Acid Volatile Sulfide (AVS) concentrations as a proxy of the disturbance of the sediment. AVS and Simultaneously Extracted Metal (SEM) concentrations were compared to bioassay responses with the freshwater benthic macroinvertebrate Asellus aquaticus. Storage conditions and sediment treatment affected AVS but not SEM levels. AVS can be used as a proxy for sediment disturbance. The best way to pretreat the sediment for use in a bioassay in order to maintain initial AVS conditions was to sample the sediment with an Ekman grab, immediately store it in a jar without headspace, and freeze it as soon as possible. In a survey using seven different sediments, bioassay responses of A. aquaticus were correlated with SEM and AVS characteristics.

  9. In vitro bioassays to evaluate complex chemical mixtures in recycled water

    Science.gov (United States)

    Jia, Ai; Escher, Beate I.; Leusch, Frederic D.L.; Tang, Janet Y.M.; Prochazka, Erik; Dong, Bingfeng; Snyder, Erin M.; Snyder, Shane A.

    2016-01-01

    With burgeoning population and diminishing availability of freshwater resources, the world continues to expand the use of alternative water resources for drinking, and the quality of these sources has been a great concern for the public as well as public health professionals. In vitro bioassays are increasingly being used to enable rapid, relatively inexpensive toxicity screening that can be used in conjunction with analytical chemistry data to evaluate water quality and the effectiveness of water treatment. In this study, a comprehensive bioassay battery consisting of 36 bioassays covering 18 biological endpoints was applied to screen the bioactivity of waters of varying qualities with parallel treatments. Samples include wastewater effluent, ultraviolet light (UV) and/or ozone advanced oxidation processed (AOP) recycled water, and infiltrated recycled groundwater. Based on assay sensitivity and detection frequency in the samples, several endpoints were highlighted in the battery, including assays for genotoxicity, mutagenicity, estrogenic activity, glucocorticoid activity, aryl hydrocarbon receptor activity, oxidative stress response, and cytotoxicity. Attenuation of bioactivity was found to be dependent on the treatment process and bioassay endpoint. For instance, ozone technology significantly removed oxidative stress activity, while UV based technologies were most efficient for the attenuation of glucocorticoid activity. Chlorination partially attenuated genotoxicity and greatly decreased herbicidal activity, while groundwater infiltration efficiently attenuated most of the evaluated bioactivity with the exception of genotoxicity. In some cases, bioactivity (e.g., mutagenicity, genotoxicity, and arylhydrocarbon receptor) increased following water treatment, indicating that transformation products of water treatment may be a concern. Furthermore, several types of bioassays with the same endpoint were compared in this study, which could help guide the selection

  10. In vitro bioassays to evaluate complex chemical mixtures in recycled water.

    Science.gov (United States)

    Jia, Ai; Escher, Beate I; Leusch, Frederic D L; Tang, Janet Y M; Prochazka, Erik; Dong, Bingfeng; Snyder, Erin M; Snyder, Shane A

    2015-09-01

    With burgeoning population and diminishing availability of freshwater resources, the world continues to expand the use of alternative water resources for drinking, and the quality of these sources has been a great concern for the public as well as public health professionals. In vitro bioassays are increasingly being used to enable rapid, relatively inexpensive toxicity screening that can be used in conjunction with analytical chemistry data to evaluate water quality and the effectiveness of water treatment. In this study, a comprehensive bioassay battery consisting of 36 bioassays covering 18 biological endpoints was applied to screen the bioactivity of waters of varying qualities with parallel treatments. Samples include wastewater effluent, ultraviolet light (UV) and/or ozone advanced oxidation processed (AOP) recycled water, and infiltrated recycled groundwater. Based on assay sensitivity and detection frequency in the samples, several endpoints were highlighted in the battery, including assays for genotoxicity, mutagenicity, estrogenic activity, glucocorticoid activity, arylhydrocarbon receptor activity, oxidative stress response, and cytotoxicity. Attenuation of bioactivity was found to be dependent on the treatment process and bioassay endpoint. For instance, ozone technology significantly removed oxidative stress activity, while UV based technologies were most efficient for the attenuation of glucocorticoid activity. Chlorination partially attenuated genotoxicity and greatly decreased herbicidal activity, while groundwater infiltration efficiently attenuated most of the evaluated bioactivity with the exception of genotoxicity. In some cases, bioactivity (e.g., mutagenicity, genotoxicity, and arylhydrocarbon receptor) increased following water treatment, indicating that transformation products of water treatment may be a concern. Furthermore, several types of bioassays with the same endpoint were compared in this study, which could help guide the selection

  11. Implementation of bioassay methods to improve assessment of incorporated radionuclides

    International Nuclear Information System (INIS)

    Oeh, U.; Hoellriegl, V.; Li, W.B.; Roth, P.; Wahl, W.; Andrasi, A.; Zombori, P.; Bouvier, C.; Carlan de, L.; Franck, D.; Ritt, J.; Fischer, H.; Schmitzer, C.

    2005-01-01

    Full text: Internal exposure to uranium and thorium can principally be assessed from external radiation measurements, exhalation measurements, or the assay of these elements excreted in urine or feces. Since both 232 Th and 238 U emit only photon radiations of low energy and with low emission probabilities, their detection limits by in vivo counting are of the order of kBq even when sophisticated devices are used. Consequently, usually bioassay methods are used for the incorporation monitoring of workers. Alpha spectrometry is the commonly applied technique, usually employed to measure 232 Th and 238 U in urine or fecel samples. For accurate analysis of body contents, 24 hours collections of urine or feces are usually used. The fecal activity, however, resembles predominantly the intake by ingestion of these nuclides during the last few days whereas the urinary excretion is more closely related to the body content of the nuclides. However, urinary excretion is also varying with the actual intake of 232 Th and/or 238 U. The measurement of these nuclides in urine by alpha-spectrometry requires tedious and time-consuming chemical work-up to prepare the samples for spectrometric analysis. Therefore, the number of analyses, which can be carried out is quite low and the results are available only after a time lag of several days. Additionally, under certain conditions the alpha-spectrometry is not sensitive enough. Other methods that have been developed may be confined to the availability of certain devices being difficult to access (e.g. nuclear reactors for radiochemical neutron activation analysis). Much better suitable as routine method is the application of inductively coupled plasma mass spectrometry (ICP-MS) for measurements of 232 Th and 238 U concentrations in urine. For elemental analyses, ICP-MS can already be considered as commonly used method. The present work which was carried out in the framework of an EU project (IDEA: Internal Dosimetry - Enhancements in

  12. Review of Bioassays for Monitoring Fate and Transport ofEstrogenic Endocrine Disrupting Compounds in Water

    Energy Technology Data Exchange (ETDEWEB)

    CGCampbell@lbl.gov

    2004-01-30

    Endocrine disrupting compounds (EDCs) are recognizedcontaminants threatening water quality. Despite efforts in sourceidentification, few strategies exist for characterization or treatment ofthis environmental pollution. Given that there are numerous EDCs that cannegatively affect humans and wildlife, general screening techniques likebioassays and biosensors provide an essential rapid and intensiveanalysis capacity. Commonly applied bioassays include the ELISA and YESassays, but promising technologies include ER-CALUXa, ELRA, Endotecta,RIANA, and IR-bioamplification. Two biosensors, Endotecta and RIANA, arefield portable using non-cellular biological detection strategies.Environmental management of EDCs in water requires integration ofbiosensors and bioassays for monitoring and assessment.

  13. Resolutions of ICRP models with BIOKMOD: Application for the bioassays evaluation

    International Nuclear Information System (INIS)

    Sanchez, G.

    2005-01-01

    Biokmod is a tool box developed using Mathematic for solving compartmental modes. It gives analytic and numeric solutions. Biokmod solves the current ICRP models including Acute, constant, continuous variable, multi-inputs and random intakes. All parameters (deposition factors, rate transfer coefficients, fractional rate of absorption, etc.) can be modified by users. It can be also applied for evaluating unknown intakes fitting bioassay experimental data and for evacuating uncertainties in the ICRP models. There is a web version (BiokmodWeb) at http://www3.enusa.es//webMathematica/public/biokmode.html. In this article we describe the application of Biokmod for evaluating Bioassays. (Author) 8 refs

  14. A versatile electrowetting-based digital microfluidic platform for quantitative homogeneous and heterogeneous bio-assays

    Science.gov (United States)

    Vergauwe, Nicolas; Witters, Daan; Ceyssens, Frederik; Vermeir, Steven; Verbruggen, Bert; Puers, Robert; Lammertyn, Jeroen

    2011-05-01

    Electrowetting-on-dielectric (EWOD) lab-on-a-chip systems have already proven their potential within a broad range of bio-assays. Nevertheless, research on the analytical performance of those systems is limited, yet crucial for a further breakthrough in the diagnostic field. Therefore, this paper presents the intrinsic possibilities of an EWOD lab-on-a-chip as a versatile platform for homogeneous and heterogeneous bio-assays with high analytical performance. Both droplet dispensing and splitting cause variations in droplet size, thereby directly influencing the assay's performance. The extent to which they influence the performance is assessed by a theoretical sensitivity analysis, which allows the definition of a basic framework for the reduction of droplet size variability. Taking advantage of the optimized droplet manipulations, both homogeneous and heterogeneous bio-assays are implemented in the EWOD lab-on-a-chip to demonstrate the analytical capabilities and versatility of the device. A fully on-chip enzymatic assay is realized with high analytical performance. It demonstrates the promising capabilities of an EWOD lab-on-a-chip in food-related and medical applications, such as nutritional and blood analyses. Further, a magnetic bio-assay for IgE detection using superparamagnetic nanoparticles is presented whereby the nanoparticles are used as solid carriers during the bio-assay. Crucial elements are the precise manipulation of the superparamagnetic nanoparticles with respect to dispensing and separation. Although the principle of using nano-carriers is demonstrated for protein detection, it can be easily extended to a broader range of bio-related applications like DNA sensing. In heterogeneous bio-assays the chip surface is actively involved during the execution of the bio-assay. Through immobilization of specific biological compounds like DNA, proteins and cells a reactive chip surface is realized, which enhances the bio-assay performance. To demonstrate

  15. Standardization of a fluconazole bioassay and correlation of results with those obtained by high-pressure liquid chromatography.

    Science.gov (United States)

    Rex, J H; Hanson, L H; Amantea, M A; Stevens, D A; Bennett, J E

    1991-01-01

    An improved bioassay for fluconazole was developed. This assay is sensitive in the clinically relevant range (2 to 40 micrograms/ml) and analyzes plasma, serum, and cerebrospinal fluid specimens; bioassay results correlate with results obtained by high-pressure liquid chromatography (HPLC). Bioassay and HPLC analyses of spiked plasma, serum, and cerebrospinal fluid samples (run as unknowns) gave good agreement with expected values. Analysis of specimens from patients gave equivalent results by both HPLC and bioassay. HPLC had a lower within-run coefficient of variation (less than 2.5% for HPLC versus less than 11% for bioassay) and a lower between-run coefficient of variation (less than 5% versus less than 12% for bioassay) and was more sensitive (lower limit of detection, 0.1 micrograms/ml [versus 2 micrograms/ml for bioassay]). The bioassay is, however, sufficiently accurate and sensitive for clinical specimens, and its relative simplicity, low sample volume requirement, and low equipment cost should make it the technique of choice for analysis of routine clinical specimens. PMID:1854166

  16. High-throughput mosquito and fly bioassay system for natural and artificial substrates treated with residual insecticides.

    Science.gov (United States)

    Aldridge, Robert L; Wynn, W Wayne; Britch, Seth C; Allan, Sandra A; Walker, Todd W; Geden, Christopher J; Hogsette, Jerome A; Linthicum, Kenneth J

    2013-03-01

    A high-throughput bioassay system to evaluate the efficacy of residual pesticides against mosquitoes and muscid flies with minimal insect handling was developed. The system consisted of 4 components made of readily available materials: 1) a CO2 anaesthetizing chamber, 2) a specialized aspirator, 3) a cylindrical flat-bottomed glass bioassay chamber assembly, and 4) a customized rack.

  17. UTILITY OF A FULL LIFE-CYCLE COPEPOD BIOASSAY APPROACH FOR ASSESSMENT OF SEDIMENT-ASSOCIATED CONTAMINANT MIXTURES. (R825279)

    Science.gov (United States)

    AbstractWe compared a 21 day full life-cycle bioassay with an existing 14 day partial life-cycle bioassay for two species of meiobenthic copepods, Microarthridion littorale and Amphiascus tenuiremis. We hypothesized that full life-cycle tests would bette...

  18. Comparison of mouse and swine bioassays for determination of soil arsenic relative bioavailability

    Science.gov (United States)

    Evaluation of soil arsenic (As) relative bioavailability (RBA) is essential to accurately assess human exposure to As contaminated soils via the incidental ingestion pathway. A variety of animal bioassays have been developed to estimate As RBA in contaminated soils and dusts, wit...

  19. Development of K-bioassay for the efficient potassium fertilization of citrus tree

    Energy Technology Data Exchange (ETDEWEB)

    U, Jang Kual [Cheju National University, Cheju (Korea, Republic of); Han, Hae Ryong [Cheju National University, Cheju (Korea, Republic of); Moon, Duk Young; Kim, Chang Myung; Lim, Han Cheol; Moon, Do Kyung [Cheju Citrus Research Institute, Cheju (Korea, Republic of); Song, Sung Jun [Cheju National Univerisity, Cheju (Korea, Republic of)

    1994-12-31

    a Similar to the {sup 42} K uptake, {sup 86} Rb uptake by the roots of Hordeum distichum grown in the hydroponic culture was negatively correlated with the concentration of K supplied previously, showing that {sup 86} Rb can be used for the K-bioassay. {sup 86} Rb having longer half life(18.86 day) than {sup 42} K(12.36 hr) allowed the use of larger number of root samples. {sup 86} Rb uptake of 3 years old Citrus unshiu Marc. grown in water culture decreased drastically with the increase of K concentration of the culture solution, thus demonstrating that the nutrition status of K for citrus trees can be diagnosed by K-bioassay using {sup 86} Rb tracer. {sup 86} Rb uptake by the excised roots of Hordeum distichum correlated with the exchangeable K in soil. The amount of exchangeable K in soil for the optimal plant growth can be determined by its relationship. {sup 42} K- and {sup 86} Rb-uptake by the Hordeum distichum roots were markedly inhibited by 5 x 10{sup -3} M KCN in the bioassay solution, indicating that uptake is metabolically controlled. There was no significant relationship between K content in citrus leaves and K concentration in the water-culture medium. It is concluded that K-bioassay is a potentially useful tool for determining of K requirement in citrus trees. (author)

  20. Sensitivity and Specificity of Bioassay of Estrogenicity on Mammary Gland and Uterus of Female Mice

    Czech Academy of Sciences Publication Activity Database

    Škarda, Josef

    2002-01-01

    Roč. 51, - (2002), s. 407-412 ISSN 0862-8408 R&D Projects: GA ČR GA523/99/0843; GA AV ČR KSK5020115 Institutional research plan: CEZ:AV0Z5045916 Keywords : Bioassay * Estrogenicity * Mammary gland Subject RIV: ED - Physiology Impact factor: 0.984, year: 2002

  1. Bioassays for Evaluating Water Quality: Screening for total bioactivity to assess water safety

    Science.gov (United States)

    Bioassays are a potential solution for assessing complex samples since they screen for total bioactivity for a given pathway or mode of action (MOA), such as estrogen receptor activation, in the samples. Overall, they can account for the three challenges listed above, and can sim...

  2. Ecotoxicological assessment of metal-polluted urban soils using bioassays with three soil invertebrates.

    NARCIS (Netherlands)

    Santarufo, L.; van Gestel, C.A.M.; Maisto, G.

    2012-01-01

    This study aimed at assessing the quality of urban soils by integrating chemical and ecotoxicological approaches. Soils from five sites in downtown Naples, Italy, were sampled and characterized for physical-chemical properties and total and water-extractable metal concentrations. Bioassays with

  3. Evaluation of soil bioassays for use at Washington state hazardous waste sites: A pilot study

    International Nuclear Information System (INIS)

    Blakley, N.; Norton, D.; Stinson, M.; Boyer, R.

    1994-01-01

    The Washington State Department of Ecology (Ecology) is developing guidelines to assess soil toxicity at hazardous waste sites being investigated under the Washington Model Toxics Control Act Cleanup Regulation. To evaluate soil toxicity, Ecology selected five bioassay protocols -- Daphnia, Earthworm, Seedling, Fathead Minnow, and Frog Embryo Teratogenesis Assay Xenopus (FETAX) -- for use as screening level assessment tools at six State hazardous waste sites. Sites contained a variety of contaminants including metals, creosote, pesticides, and petroleum products (leaking underground storage tanks). Three locations, representing high, medium, and low levels of contamination, were samples at each site. In general, the high contaminant samples resulted in the highest toxic response in all bioassays. The order of site toxicity, as assessed by overall toxic response, is creosote, petroleum products, metals, and pesticides. Results indicate that human health standards, especially for metals, may not adequately protect some of the species tested. The FETAX bioassay had the greatest overall number of toxic responses and lowest variance. The seedling and Daphnia bioassays had lower and similar overall toxic response results, followed by the earthworm and fathead minnow. Variability was markedly highest for the seedling. The Daphnia and fathead minnow variability were similar to the FETAX level, while the earthworm variability was slightly higher

  4. APPLICATION OF PLANT AND EARTHWORM BIOASSAYS TO EVALUATE REMEDIATION OF A LEAD-CONTAMINATED SOIL

    Science.gov (United States)

    Earthworm acute toxicity, plant seed germination/root elongation (SG/RE) and plant genotoxicity bioassays were employed to evaluate the remediation of a lead-contaminated soil. The remediation involved removal of heavy metals by a soil washing/soil leaching treatment process. A p...

  5. Strategies for Transferring Mixtures of Organic Contaminants from Aquatic Environments into Bioassays

    DEFF Research Database (Denmark)

    Jahnke, Annika; Mayer, Philipp; Schäfer, Sabine

    2016-01-01

    and monitoring of such mixtures, a variety of cell-based in vitro and low-complexity in vivo bioassays based on algae, daphnids or fish embryos are available. A very important and sometimes unrecognized challenge is how to combine sampling, extraction and dosing to transfer the mixtures from the environment...

  6. BIOASSAY STUDIES OF METAL(II) COMPLEXES OF 2,2'-(ETHANE ...

    African Journals Online (AJOL)

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    diyldiimino)diacetic acid (EDDA) were prepared and characterized. Coordination complexes of the EDDA ... corresponding amines with alkyl halide to bear diammines of the same class with different substituents. ... Bioassay studies of metal(II) complexes of 2,2'-(ethane-1,2-diyldiimino)diacetic acid. Bull. Chem. Soc. Ethiop.

  7. Bioassay-guided studies on the cytotoxic and in vitro trypanocidal ...

    African Journals Online (AJOL)

    This paper reports a bioassay-guided study to search for possible biological activity (cytotoxic and trypanocidal) in two Ugandan medicinal plants. The methodology adopted was the so-called ping-pong approach, involving phytochemical purification (column chromatography and preparative thin layer chromatography), ...

  8. Bioassay directed identification of natural aryl hydrocarbon-receptor agonists in marmalade

    NARCIS (Netherlands)

    Ede, van K.I.; Li, A.; Antunes Fernandes, E.C.; Mulder, P.P.J.; Peijnenburg, A.A.C.M.; Hoogenboom, L.A.P.

    2008-01-01

    Citrus fruit and citrus fruit products, like grapefruit, lemon and marmalade were shown to contain aryl hydrocarbon receptor (AhR) agonists, as detected with the DR CALUX® bioassay. This is of interest regarding the role of the Ah-receptor pathway in the adverse effects of dioxins, PCBs and other

  9. Experience with NQA-1 quality assurance standards applied to in vitro bioassay

    International Nuclear Information System (INIS)

    Bihl, D.E.; MacLellan, J.A.

    1991-10-01

    On June 1, 1990, the large (about 4000 samples per year) excreta bioassay program at the Hanford Site ceased abruptly when the contract with the bioassay laboratory was terminated. An intense, high-priority effort was begun to replace the services on an interim basis until a new contract could be procured. Despite the urgency to get the excreta bioassay program going again, the Hanford Internal Dosimetry Program was constrained to use only labs that could meet stringent quality assurance (QA) requirements, even during the interim period. The QA requirements were based on NQA-1 with selected additions from the Environmental Protection Agency's QAMS 005/80 (EPA 1983) and the American Society for Testing and Materials' C 1009-83 (ASTM 1984). This constraint was driven both by legal reasons and by the Hanford Site contractors and workers not wanting the quality of the data to be sacrificed. Finding labs that could (1) handle the large throughput, (2) meet the technical requirements, and (3) pass the QA audit proved more difficult than first anticipated. This presentation focuses on the QA requirements that the labs had to meet and how those very broad requirements were applied specifically to excreta bioassay. 5 refs

  10. The Intersection of CMOS Microsystems and Upconversion Nanoparticles for Luminescence Bioimaging and Bioassays

    Directory of Open Access Journals (Sweden)

    Liping Wei

    2014-09-01

    Full Text Available Organic fluorophores and quantum dots are ubiquitous as contrast agents for bio-imaging and as labels in bioassays to enable the detection of biological targets and processes. Upconversion nanoparticles (UCNPs offer a different set of opportunities as labels in bioassays and for bioimaging. UCNPs are excited at near-infrared (NIR wavelengths where biological molecules are optically transparent, and their luminesce in the visible and ultraviolet (UV wavelength range is suitable for detection using complementary metal-oxide-semiconductor (CMOS technology. These nanoparticles provide multiple sharp emission bands, long lifetimes, tunable emission, high photostability, and low cytotoxicity, which render them particularly useful for bio-imaging applications and multiplexed bioassays. This paper surveys several key concepts surrounding upconversion nanoparticles and the systems that detect and process the corresponding luminescence signals. The principle of photon upconversion, tuning of emission wavelengths, UCNP bioassays, and UCNP time-resolved techniques are described. Electronic readout systems for signal detection and processing suitable for UCNP luminescence using CMOS technology are discussed. This includes recent progress in miniaturized detectors, integrated spectral sensing, and high-precision time-domain circuits. Emphasis is placed on the physical attributes of UCNPs that map strongly to the technical features that CMOS devices excel in delivering, exploring the interoperability between the two technologies.

  11. Effect of the Changes of Respiratory Tract Model on the Uranium Bioassay Data

    Energy Technology Data Exchange (ETDEWEB)

    Kwon, Taeeun; Noh, Siwan; Kim, Meeryeong; Lee, Jaiki [Hanyang Univ., Seoul (Korea, Republic of); Lee, Jongil; Kim, Jang Lyul [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2014-05-15

    The HRTM, however, was revised based on the recent experimental data in OIR (Occupational Intakes of Radionuclides) draft report of ICRP. The changes of respiratory tract model are predicted to directly affect bioassay data like retention and excretion functions. Lung retention function is especially important to internal exposure assessment for workers related to fuel manufacturing because the place could be contaminated by uranium. In addition, faecel samples are recommended to be used for in-vitro bioassay of uranium because of very slow excretion via urine. More reliable assessments for the workers in fuel manufacturing could be achieved by recalculation of bioassay data for uranium and the comparing study using original and revised HRTM. In this study, therefore, the lung retention and faecal excretion functions for inhalation of UO{sub 2} and U{sub 3}O{sub 8} were recalculated using revised HRTM and the results were compared with those of original HRTM. In this study the lung retention and faecal excretion functions for inhalation of UO{sub 2} and U{sub 3}O{sub 8} were calculated based on original and revised HRTM. The results show that the revised HRTM increases lung retention and uptakes to alimentary tract which cause the more faecal excretion. The results in this study confirm the effect of the changes of respiratory tract model on the uranium bioassay data although the more study is needed to apply to practical fields.

  12. Kinetic microplate bioassays for relative potency of antibiotics improved by partial Least Square (PLS) regression.

    Science.gov (United States)

    Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Almeida, Túlia de Souza Botelho; Lourenço, Felipe Rebello

    2016-05-01

    Microbiological assays are widely used to estimate the relative potencies of antibiotics in order to guarantee the efficacy, safety, and quality of drug products. Despite of the advantages of turbidimetric bioassays when compared to other methods, it has limitations concerning the linearity and range of the dose-response curve determination. Here, we proposed to use partial least squares (PLS) regression to solve these limitations and to improve the prediction of relative potencies of antibiotics. Kinetic-reading microplate turbidimetric bioassays for apramacyin and vancomycin were performed using Escherichia coli (ATCC 8739) and Bacillus subtilis (ATCC 6633), respectively. Microbial growths were measured as absorbance up to 180 and 300min for apramycin and vancomycin turbidimetric bioassays, respectively. Conventional dose-response curves (absorbances or area under the microbial growth curve vs. log of antibiotic concentration) showed significant regression, however there were significant deviation of linearity. Thus, they could not be used for relative potency estimations. PLS regression allowed us to construct a predictive model for estimating the relative potencies of apramycin and vancomycin without over-fitting and it improved the linear range of turbidimetric bioassay. In addition, PLS regression provided predictions of relative potencies equivalent to those obtained from agar diffusion official methods. Therefore, we conclude that PLS regression may be used to estimate the relative potencies of antibiotics with significant advantages when compared to conventional dose-response curve determination. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. 'Dose per unit content' functions: A robust tool for the interpretation of bioassay data

    International Nuclear Information System (INIS)

    Berkovski, V.; Bonchuk, Y.; Ratia, G.

    2003-01-01

    The purposes of this study were to investigate the influence of the consequences of the lack of primary bioassay information and to elaborate approaches which could improve the reliability of dose assessments. The aggregated time-dependent functions 'dose per unit organ (excretion) content' z(t) have been proposed in this study as a convenient and reliable tool for bioassay. The analysis of the variation of z with changes of AMAD has demonstrated the existence of areas of the relative invariance of z, which permits the selection of one (reference) function z for the whole area of stability. Within the framework of such an approach an arbitrary set of bioassay data can be approximated by the linear combination F(t) S i E/ i z(t-t i ), whereI> F(t) function of time t, which approximates the observed bioassay time trend; t i = time shift of the acute intake i; E i effective dose, associated with the acute intake i (the two last parameters are results of the approximation procedure). (author)

  14. Experience with NQA-1 quality assurance standards applied to in vitro bioassay

    International Nuclear Information System (INIS)

    Bihl, D.E.; MacLellan, J.A.

    1991-01-01

    On June 1, 1990, the large (about 4,000 samples per year) excreta bioassay program at the Hanford Site ceased abruptly when the contract with the bioassay laboratory was terminated. An intense, high-priority effort was begun to replace the services on an interim basis until a new contract could be procured. Despite the urgency to get the excreta bioassay program going again, the Hanford Internal Dosimetry Program was constrained to use only labs that could meet stringent quality assurance (QA) requirements, even during the interim period. The QA requirements were based on NQA-1 with selected additions from the Environmental Protection Agency's QAMS 005/80 (EPA 1983) and the American Society for Testing and Materials' C 1009-83 (ASTM 1984). This constraint was driven both by legal reasons and by the Hanford Site contractors and workers not wanting the quality of the data to be sacrificed. Finding labs that could (1) handle the large throughput, (2) meet the technical requirements, and (3) pass the QA audit proved more difficult than first anticipated. This presentation focuses on the QA requirements that the labs had to meet and how those very broad requirements were applied specifically to excreta bioassay

  15. Androgen Bioassay for the Detection of Nonlabeled Androgenic Compounds in Nutritional Supplements.

    Science.gov (United States)

    Cooper, Elliot R; McGrath, Kristine C Y; Li, XiaoHong; Heather, Alison K

    2018-01-01

    Both athletes and the general population use nutritional supplements. Athletes often turn to supplements hoping that consuming the supplement will help them be more competitive and healthy, while the general population hopes to improve body image or vitality. While many supplements contain ingredients that may have useful properties, there are supplements that are contaminated with compounds that are banned for use in sport or have been deliberately adulterated to fortify a supplement with an ingredient that will produce the advertised effect. In the present study, we have used yeast cell and mammalian cell androgen bioassays to characterize the androgenic bioactivity of 112 sports supplements available from the Australian market, either over the counter or via the Internet. All 112 products did not declare an androgen on the label as an included ingredient. Our findings show that six out of 112 supplements had strong androgenic bioactivity in the yeast cell bioassay, indicating products spiked or contaminated with androgens. The mammalian cell bioassay confirmed the strong androgenic bioactivity of five out of six positive supplements. Supplement 6 was metabolized to weaker androgenic bioactivity in the mammalian cells. Further to this, Supplement 6 was positive in a yeast cell progestin bioassay. Together, these findings highlight that nutritional supplements, taken without medical supervision, could expose or predispose users to the adverse consequences of androgen abuse. The findings reinforce the need to increase awareness of the dangers of nutritional supplements and highlight the challenges that clinicians face in the fast-growing market of nutritional supplements.

  16. New in vitro reporter gene bioassays for screening of hormonal active compounds in the environment

    Czech Academy of Sciences Publication Activity Database

    Svobodová, Kateřina; Cajthaml, Tomáš

    2010-01-01

    Roč. 88, č. 4 (2010), s. 839-847 ISSN 0175-7598 R&D Projects: GA ČR GAP503/10/0408 Institutional research plan: CEZ:AV0Z50200510 Keywords : endocrine disruptors * in vitro bioassays * reporter gene assays Subject RIV: EE - Microbiology, Virology Impact factor: 3.280, year: 2010

  17. Development and application of bioassays for a site-specific risk assessment of contaminated soil

    NARCIS (Netherlands)

    Rila, J.-P.

    2008-01-01

    Soil risk assessment based on generic approaches is accompanied by a large number of uncertainties. In site-specific risk assessment aimed at identifying the actual effects on the ecosystem by using e.g. bioassays in soil elutriates and taking into account land-use these uncertainties can be largely

  18. A rapid bioassay for detecting saxitoxins using a Daphnia acute toxicity test.

    Science.gov (United States)

    Ferrão-Filho, Aloysio da S; Soares, Maria Carolina S; de Magalhães, Valéria Freitas; Azevedo, Sandra M F O

    2010-06-01

    Bioassays using Daphnia pulex and Moina micrura were designed to detect cyanobacterial neurotoxins in raw water samples. Phytoplankton and cyanotoxins from seston were analyzed during 15 months in a eutrophic reservoir. Effective time to immobilize 50% of the exposed individuals (ET50) was adopted as the endpoint. Paralysis of swimming movements was observed between approximately 0.5-3 h of exposure to lake water containing toxic cyanobacteria, followed by an almost complete recovery of the swimming activity within 24 h after being placed in control water. The same effects were observed in bioassays with a saxitoxin-producer strain of Cylindrospermopsis raciborskii isolated from the reservoir. Regression analysis showed significant relationships between ET50 vs. cell density, biomass and saxitoxins content, suggesting that the paralysis of Daphnia in lake water samples was caused by saxitoxins found in C. raciborskii. Daphnia bioassay was found to be a sensitive method for detecting fast-acting neurotoxins in natural samples, with important advantages over mouse bioassays. Copyright 2010 Elsevier Ltd. All rights reserved.

  19. Determination of Biochemical Oxygen Demand of Area Waters: A Bioassay Procedure for Environmental Monitoring

    Science.gov (United States)

    Riehl, Matthew

    2012-01-01

    A graphical method for determining the 5-day biochemical oxygen demand (BOD5) for a body of water is described. In this bioassay, students collect a sample of water from a designated site, transport it to the laboratory, and evaluate the amount of oxygen consumed by naturally occurring bacteria during a 5-day incubation period. An accuracy check,…

  20. Olfactoryresponse of the predatory mite Typhlodromus pyri (Acari: Phytoseiidae) to methyl salicylate in laboratory bioassays

    Science.gov (United States)

    The response of Typhlodromus pyri, a key predator of grapevine rust mite (Calepitrimerus vitis), to MeSA was tested using a Y-tube olfactometer in laboratory bioassays. Six doses ranging from 200 to 0.002 µg of diluted MeSA were tested. Significantly higher proportions of T. pyri preferred MeSA at ...

  1. Worldwide bioassay data resources for plutonium/americium internal dosimetry studies

    International Nuclear Information System (INIS)

    Miller, G.; Bertelli, L.; Little, T.; Guilmette, R.; Riddell, T.; Filipy, R.

    2005-01-01

    Full text: Biokinetic models are the scientific underpinning of internal dosimetry. These models describe how materials of interest taken into the body by various routes (for example inhalation) are transported through the body, allowing the modelling of bioassay measurements and the estimation of radiation dose. The International Commission on Radiation Protection (ICRP) publishes biokinetic models for use in internal dosimetry. These models represent the consensus judgement of a committee of experts, based on human and animal data. Nonetheless, it is important to validate biokinetic models using directly applicable data, in a scientifically transparent manner, especially for internal dosimetry research purposes (as opposed to radiation protection), as in epidemiology studies. Two major goals would be to determine individual variations of model parameters for the purpose of assessing this source of uncertainty in internal dose calculations, and to determine values of workplace specific parameters (such as particle solubility in lung fluids) for different representative workplaces. Furthermore, data on the observed frequency of intakes under various conditions can be used in the interpretation of bioassay data. All of the above may be couched in the terminology of Bayesian statistical analysis and amount to the determination of the Bayesian prior probability distributions needed in a Bayesian interpretation of bioassay data. The authors have direct knowledge of several significant databases of plutonium/americium bioassay data (including autopsy data). The purpose of this paper is to acquaint the worldwide community with these resources and to invite others who may know of other such databases to participate with us in a publication that would document the content, form, and the procedures for seeking access to these databases. These databases represent a tremendous scientific resource in this field. Examples of databases known to the authors include: the

  2. Paper-based chromatic toxicity bioassay by analysis of bacterial ferricyanide reduction.

    Science.gov (United States)

    Pujol-Vila, F; Vigués, N; Guerrero-Navarro, A; Jiménez, S; Gómez, D; Fernández, M; Bori, J; Vallès, B; Riva, M C; Muñoz-Berbel, X; Mas, J

    2016-03-03

    Water quality assessment requires a continuous and strict analysis of samples to guarantee compliance with established standards. Nowadays, the increasing number of pollutants and their synergistic effects lead to the development general toxicity bioassays capable to analyse water pollution as a whole. Current general toxicity methods, e.g. Microtox(®), rely on long operation protocols, the use of complex and expensive instrumentation and sample pre-treatment, which should be transported to the laboratory for analysis. These requirements delay sample analysis and hence, the response to avoid an environmental catastrophe. In an attempt to solve it, a fast (15 min) and low-cost toxicity bioassay based on the chromatic changes associated to bacterial ferricyanide reduction is here presented. E. coli cells (used as model bacteria) were stably trapped on low-cost paper matrices (cellulose-based paper discs, PDs) and remained viable for long times (1 month at -20 °C). Apart from bacterial carrier, paper matrices also acted as a fluidic element, allowing fluid management without the need of external pumps. Bioassay evaluation was performed using copper as model toxic agent. Chromatic changes associated to bacterial ferricyanide reduction were determined by three different transduction methods, i.e. (i) optical reflectometry (as reference method), (ii) image analysis and (iii) visual inspection. In all cases, bioassay results (in terms of half maximal effective concentrations, EC50) were in agreement with already reported data, confirming the good performance of the bioassay. The validation of the bioassay was performed by analysis of real samples from natural sources, which were analysed and compared with a reference method (i.e. Microtox). Obtained results showed agreement for about 70% of toxic samples and 80% of non-toxic samples, which may validate the use of this simple and quick protocol in the determination of general toxicity. The minimum instrumentation

  3. Responses of lone star tick (acari: ixodidae) nymphs to the repellent deet applied in acetone and ethanol solutions in vitro bioassays

    Science.gov (United States)

    Behavioral bioassays remain a standard tool in the discovery, development, and registration of repellents. Although tick repellent bioassays tend to be rather uncomplicated, several factors can influence their outcomes. Typically repellent bioassays use a solvent, such as acetone or ethanol, to disp...

  4. Analysing traces of autoinducer-2 requires standardization of the Vibrio harveyi bioassay.

    Science.gov (United States)

    Vilchez, Ramiro; Lemme, André; Thiel, Verena; Schulz, Stefan; Sztajer, Helena; Wagner-Döbler, Irene

    2007-01-01

    Autoinducer-2 (furanosyl borate diester) is a biologically active compound whose role as a universal bacterial signalling molecule is currently under intense investigation. Because of its instability and the low concentrations of it found in biological samples, its detection relies at present on a bioassay that measures the difference in the timing of the luminescence of the Vibrio harveyi BB170 sensor strain with and without externally added AI-2. Here we systematically investigated which parameters affected the fold induction values of luminescence obtained in the bioassay and developed a modified protocol. Our experiments showed that growth and luminescence of V. harveyi BB170 are strongly influenced by trace elements. In particular, addition of Fe(3+) within a certain concentration range to the growth medium of the preinoculum culture improved the reproducibility and reduced the variance of the bioassay. In contrast, trace elements and vitamins introduced directly into the bioassay caused inhibitory effects. The initial density and luminescence of the sensor strain are very important and the values required for these parameters were defined. Borate interferes with the detection of AI-2 by giving false positive results. The response of V. harveyi BB170 to chemically synthesized AI-2 in the bioassay is nonlinear except over a very small concentration range; it is maximum over three orders of magnitude and shows inhibition above 35 microM. Based on the modified protocol, we were able to detect AI-2 in the absence of inhibitors with maximum fold induction values for the positive control (chemically synthesized AI-2) of >120 with a standard deviation of approximately 30% in a reliable and reproducible way.

  5. Impact of Spodoptera frugiperda neonate pretreatment conditions on Vip3Aa19 insecticidal protein activity and laboratory bioassay variation.

    Science.gov (United States)

    Da Silva, Karen F; Spencer, Terence A; Camargo Gil, Carolina; Siegfried, Blair D; Walters, Frederick S

    2016-04-01

    Variation in response to insecticidal proteins is common upon repetition of insect bioassays. Understanding this variation is a prerequisite to detecting biologically important differences. We tracked neonate Spodoptera frugiperda (J.E. Smith) susceptibility to Vip3Aa19 over 17 generations using standardized bioassay methods. Five larval pretreatment conditions and one bioassay condition were tested to determine whether susceptibility was affected. These included: storage time; prefeeding; storage at reduced temperature; storage at reduced humidity; colony introgression of field-collected individuals. Extremes of photoperiod during the bioassay itself were also examined. LC50 values for two strains of S. frugiperda varied 6.6-fold or 8.8-fold over 17 generations. Storage time and humidity had no impact on Vip3Aa19 susceptibility, whereas prefeeding significantly reduced subsequent mortality (by 27%). Storage at reduced temperature increased mortality for one colony (from 45.6 to 73.0%) but not for the other. Introgression of field-collected individuals affected susceptibility at the first generation but not for subsequent generations. A 24 h bioassay photophase significantly reduced susceptibility (by 26%) for both colonies. Certain pretreatment and bioassay conditions were identified that can affect S. frugiperda Vip3Aa19 susceptibility, but innate larval heterogeneity was also present. Our observations should help to increase the consistency of insecticidal protein bioassay results. © 2015 Syngenta Crop Protection, LLC. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

  6. [Investigation on pattern of quality control for Chinese materia medica based on famous-region drug and bioassay--the work reference].

    Science.gov (United States)

    Yan, Dan; Xiao, Xiaohe

    2011-05-01

    Selection and standardization of the work reference are the technical issues to be faced with in the bioassay of Chinese materia medica. Taking the bioassay of Coptis chinensis. as an example, the manufacture process of the famous-region drugs extraction was explained from the aspects of original identification, routine examination, component analysis and bioassay. The common technologies were extracted, and the selection and standardization procedures of the work reference for the bioassay of Chinese materia medica were drawn up, so as to provide technical support for constructing a new mode and method of the quality control of Chinese materia medica based on the famous-region drugs and bioassay.

  7. Fathead minnows (Pimephales promelas) and goldfish (Carassius auratus) as standard fish in bioassays and their reaction to potential reference toxicants

    Energy Technology Data Exchange (ETDEWEB)

    Adelman, I.R.; Smith, L.L. Jr.

    1976-02-01

    Fathead minnows (Pimephales promelas) and goldfish (Carassius auratus) were compared for their suitability as standard bioassay fish. Both species showed the same variability of bioassay results when tested with four toxicants. Fathead minnows are recommended on the basis of their small size and on their capability for use in complete life cycle tests. On the basis of minimum variability of bioassay results, sodium chloride was superior for use as a reference toxicant. Both sodium chloride and pentachlorophenol seemed capable of detecting abnormal fish. On the basis of seven listed criteria either sodium chloride or pentachlorophenol would be acceptable as a reference toxicant.

  8. Evaluating the efficacy of biological and conventional insecticides with the new 'MCD bottle' bioassay.

    Science.gov (United States)

    Sternberg, Eleanore D; Waite, Jessica L; Thomas, Matthew B

    2014-12-16

    Control of mosquitoes requires the ability to evaluate new insecticides and to monitor resistance to existing insecticides. Monitoring tools should be flexible and low cost so that they can be deployed in remote, resource poor areas. Ideally, a bioassay should be able to simulate transient contact between mosquitoes and insecticides, and it should allow for excito-repellency and avoidance behaviour in mosquitoes. Presented here is a new bioassay, which has been designed to meet these criteria. This bioassay was developed as part of the Mosquito Contamination Device (MCD) project and, therefore, is referred to as the MCD bottle bioassay. Presented here are two experiments that serve as a proof-of-concept for the MCD bottle bioassay. The experiments used four insecticide products, ranging from fast-acting, permethrin-treated, long-lasting insecticide nets (LLINs) that are already widely used for malaria vector control, to the slower acting entomopathogenic fungus, Beauveria bassiana, that is currently being evaluated as a prospective biological insecticide. The first experiment used the MCD bottle to test the effect of four different insecticides on Anopheles stephensi with a range of exposure times (1 minute, 3 minutes, 1 hour). The second experiment is a direct comparison of the MCD bottle and World Health Organization (WHO) cone bioassay that tests a subset of the insecticides (a piece of LLIN and a piece of netting coated with B. bassiana spores) and a further reduced exposure time (5 seconds) against both An. stephensi and Anopheles gambiae. Immediate knockdown and mortality after 24 hours were assessed using logistic regression and daily survival was assessed using Cox proportional hazards models. Across both experiments, fungus performed much more consistently than the chemical insecticides but measuring the effect of fungus required monitoring of mosquito mortality over several days to a week. Qualitatively, the MCD bottle and WHO cone performed comparably

  9. Brine shrimp bioassay: importance of correct taxonomic identification of Artemia (Anostraca) species.

    Science.gov (United States)

    Ruebhart, David R; Cock, Ian E; Shaw, Glen R

    2008-08-01

    Despite the common use of the brine shrimp bioassay in toxicology, there is confusion in the literature regarding citation of the correct taxonomic identity of the Artemia species used. The genus Artemia, once thought to be represented by a single species Artemia salina, is now known to be composed of several bisexual species as well as parthenogenetic populations. Artemia franciscana is the best studied of the Artemia species and is considered to represent the vast majority of studies in which Artemia is used as an experimental test organism. We found that in studies referring to the use of A. salina, the zoogeography of the cyst harvest site indicated that the species used was actually A. franciscana. Those performing bioassays with Artemia need to exercise diligence in assigning correct species identification, as the identity of the test organism is an important parameter in assuring the validity of the results of the assay.

  10. Genotoxic evaluation of an industrial effluent from an oil refinery using plant and animal bioassays

    Directory of Open Access Journals (Sweden)

    Fernando Postalli Rodrigues

    2010-01-01

    Full Text Available Polycyclic aromatic hydrocarbons (PAHs are genotoxic chemicals commonly found in effluents from oil refineries. Bioassays using plants and cells cultures can be employed for assessing environmental safety and potential genotoxicity. In this study, the genotoxic potential of an oil refinery effluent was analyzed by means of micronucleus (MN testing of Alium cepa, which revealed no effect after 24 h of treatment. On the other hand, primary lesions in the DNA of rat (Rattus norvegicus hepatoma cells (HTC were observed through comet assaying after only 2 h of exposure. On considering the capacity to detect DNA damage of a different nature and of these cells to metabolize xenobiotics, we suggest the association of the two bioassays with these cell types, plant (Allium cepa and mammal (HTC cells, for more accurately assessing genotoxicity in environmental samples.

  11. Genotoxic evaluation of an industrial effluent from an oil refinery using plant and animal bioassays.

    Science.gov (United States)

    Rodrigues, Fernando Postalli; Angeli, José Pedro Friedmann; Mantovani, Mário Sérgio; Guedes, Carmen Luisa Barbosa; Jordão, Berenice Quinzani

    2010-01-01

    Polycyclic aromatic hydrocarbons (PAHs) are genotoxic chemicals commonly found in effluents from oil refineries. Bioassays using plants and cells cultures can be employed for assessing environmental safety and potential genotoxicity. In this study, the genotoxic potential of an oil refinery effluent was analyzed by means of micronucleus (MN) testing of Alium cepa, which revealed no effect after 24 h of treatment. On the other hand, primary lesions in the DNA of rat (Rattus norvegicus) hepatoma cells (HTC) were observed through comet assaying after only 2 h of exposure. On considering the capacity to detect DNA damage of a different nature and of these cells to metabolize xenobiotics, we suggest the association of the two bioassays with these cell types, plant (Allium cepa) and mammal (HTC) cells, for more accurately assessing genotoxicity in environmental samples.

  12. Methods for conducting bioassays using embryos and larvae of Pacific herring, Clupea pallasi.

    Science.gov (United States)

    Dinnel, Paul A; Middaugh, Douglas P; Schwarck, Nathan T; Farren, Heather M; Haley, Richard K; Hoover, Richard A; Elphick, James; Tobiason, Karen; Marshall, Randall R

    2011-02-01

    The rapid decrease of several stocks of Pacific herring, Clupea pallasi, in Puget Sound, Washington, has led to concerns about the effects of industrial and nonpoint source contamination on the embryo and larval stages of this and related forage fish species. To address these concerns, the state of Washington and several industries have funded efforts to develop embryo and larval bioassay protocols that can be used by commercial laboratories for routine effluent testing. This article presents the results of research to develop herring embryo and larval bioassay protocols. Factors evaluated during protocol development included temperature, salinity, dissolved oxygen (DO), light intensity, photoperiod, larval feeding regimes, use of brine and artificial sea salts, gonad sources, collection methods, and egg quality.

  13. Biomagnification of bioassay derived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents

    Science.gov (United States)

    Jones, P.D.; Ankley, G.T.; Best, D. A.; Crawford, R.; DeGalan, N.; Giesy, J.P.; Kubiak, T.J.; Ludwig, J. P.; Newsted, J.L.; Tillitt, D. E.; Verbrugge, D.A.

    1993-01-01

    In recent years contamination of the Great Lakes ecosystem with planar chlorinated hydrocarbons (PCHs) has attracted considerable concern due to their known reproductive and teratogenic effects. The H4IIE bioassay has been standardized as a means of measuring the biological potency of a PCH mixture as 2,3,7,8-tetrachloro-p-dibenzodioxin equivalents (TCDD-EQ). Using this bioassay we have investigated the biomagnification of TCDD-EQ in a semi-closed ecosystem. The biomagnification of TCDD-EQ is demonstrated and results indicate that the food chain is the major pathway for TCDD-EQ through this ecosystem. The H4IIE assay system is demonstrated to be a viable integrative measure of the total concentration of TCDD-EQ in different trophic levels.

  14. Isolation of Fungi from Heterodera glycines and in vitro Bioassays for Their Antagonism to Eggs.

    Science.gov (United States)

    Meyer, S L; Huettel, R N; Sayre, R M

    1990-10-01

    Twenty fungi were assayed in vitro for antagonism to eggs of Heterodera glycines. Eight of the fungi were isolated from cysts or eggs of H. glycines during the current study, one was isolated from Panagrellus redivivus, and eleven were obtained from other researchers or collections. The bioassays were conducted on eggs from nematodes that had been grown monoxenically on excised root tips. Phoma chrysanthemicola, one strain of Verticillium chlamydosporium, and one strain of V. lecanii caused a decrease (P Trichoderma polysporum infected live eggs but enhanced (P Fusarium sp., Neocosmospora vasinfecta, Scytalidium fulvum, Trichoderma harzianum (two strains), V. chlamydosporium (one strain), V. lecanii (three strains), and an unidentified fungus did not measurably affect egg viability, even though hyphae of five of these fungi were seen in live eggs. The bioassay provides a useful step in the selection of a biological control agent for this major nematode pest.

  15. Rapid bioassay method for estimation of 90Sr in urine samples by liquid scintillation counting

    International Nuclear Information System (INIS)

    Wankhede, Sonal; Chaudhary, Seema; Sawant, Pramilla D.

    2018-01-01

    Radiostrontium (Sr) is a by-product of the nuclear fission of uranium and plutonium in nuclear reactors and is an important radionuclide in spent nuclear fuel and radioactive waste. Rapid bioassay methods are required for estimating Sr in urine following internal contamination. Decision regarding medical intervention, if any can be based upon the results of urinalysis. The present method used at Bioassay Laboratory, Trombay is by Solid Extraction Chromatography (SEC) technique. The Sr separated from urine sample is precipitated as SrCO 3 and analyzed gravimetrically. However, gravimetric procedure is time consuming and therefore, in the present study, feasibility of Liquid Scintillation Counting for direct detection of radiostrontium in effluent was explored. The results obtained in the present study were compared with those obtained using gravimetric method

  16. High-throughput tri-colour flow cytometry technique to assess Plasmodium falciparum parasitaemia in bioassays

    DEFF Research Database (Denmark)

    Tiendrebeogo, Regis W; Adu, Bright; Singh, Susheel K

    2014-01-01

    BACKGROUND: Unbiased flow cytometry-based methods have become the technique of choice in many laboratories for high-throughput, accurate assessments of malaria parasites in bioassays. A method to quantify live parasites based on mitotracker red CMXRos was recently described but consistent...... distinction of early ring stages of Plasmodium falciparum from uninfected red blood cells (uRBC) remains a challenge. METHODS: Here, a high-throughput, three-parameter (tri-colour) flow cytometry technique based on mitotracker red dye, the nucleic acid dye coriphosphine O (CPO) and the leucocyte marker CD45...... for enumerating live parasites in bioassays was developed. The technique was applied to estimate the specific growth inhibition index (SGI) in the antibody-dependent cellular inhibition (ADCI) assay and compared to parasite quantification by microscopy and mitotracker red staining. The Bland-Altman analysis...

  17. Intake retention functions and their applications to bioassay and the estimation of internal radiation doses

    International Nuclear Information System (INIS)

    Skrable, K.W.; Chabot, G.E.; French, C.S.; La Bone, T.R.

    1988-01-01

    This paper describes a way of obtaining and gives applications of intake retention functions. These functions give the fraction of an intake of radioactive material expected to be present in a specified bioassay compartment at any time after a single acute exposure or after onset of a continuous exposure. The intake retention functions are derived from a multicompartmental model and a recursive catenary kinetics equation that completely describe the metabolism of radioelements from intake to excretion, accounting for the delay in uptake from compartments in the respiratory and gastrointestinal tracts and the recycling of radioelements between systemic compartments. This approach, which treats excretion as the 'last' compartment of all catenary metabolic pathways, avoids the use of convolution integrals and provides algebraic solutions that can be programmed on hand held calculators or personal computers. The estimation of intakes and internal radiation doses and the use of intake retention functions in the design of bioassay programs are discussed along with several examples

  18. The application of bioassays as indicators of petroleum-contaminated soil remediation.

    Science.gov (United States)

    Płaza, Grazyna; Nałecz-Jawecki, Grzegorz; Ulfig, Krzysztof; Brigmon, Robin L

    2005-04-01

    Bioremediation has proven successful in numerous applications to petroleum contaminated soils. However, questions remain as to the efficiency of bioremediation in lowering long-term soil toxicity. In the present study, the bioassays Spirotox, Microtox, Ostracodtoxkit F, umu-test with S-9 activation, and plant assays were applied, and compared to evaluate bioremediation processes in heavily petroleum contaminated soils. Six higher plant species (Secale cereale L., Lactuca sativa L., Zea mays L., Lepidium sativum L., Triticum vulgare L., Brassica oleracea L.) were used for bioassay tests based on seed germination and root elongation. The ecotoxicological analyses were made in DMSO/H2O and DCM/DMSO soil extracts. Soils were tested from two biopiles at the Czechowice oil refinery, Poland, that have been subjected to different bioremediation applications. In biopile 1 the active or engineered bioremediation process lasted four years, while biopile 2 was treated passively or non-engineered for eight months. The test species demonstrated varying sensitivity to soils from both biopiles. The effects on test organisms exposed to biopile 2 soils were several times higher compared to those in biopile 1 soils, which correlated with the soil contaminants concentration. Soil hydrocarbon concentrations indeed decreased an average of 81% in biopile 1, whereas in biopile 2 TPH/TPOC concentrations only decreased by 30% after eight months of bioremediation. The bioassays were presented to be sensitive indicators of soil quality and can be used to evaluate the quality of bioremediated soil. The study encourages the need to combine the bioassays with chemical monitoring for evaluation of the bioremediation effectiveness and assessing of the contaminated/remediated soils.

  19. Mouse bioassay to assess oestrogenic and anti-oestrogenic compounds: Hydroxytamoxifen, Diethylstilbestrol and Genistein

    Czech Academy of Sciences Publication Activity Database

    Köhlerová, Eva; Škarda, Josef

    2004-01-01

    Roč. 51, č. 5 (2004), s. 209-217 ISSN 0931-184X R&D Projects: GA ČR GA524/02/0406; GA AV ČR IBS5045302; GA AV ČR KSK5020115 Institutional research plan: CEZ:AV0Z5045916 Keywords : bioassay * anti-oestrogens * oestrogenicity Subject RIV: FB - Endocrinology, Diabetology, Metabolism, Nutrition Impact factor: 0.471, year: 2004

  20. Seasonally and regionally determined indication potential of bioassays in contaminated river sediments.

    Science.gov (United States)

    Hilscherová, Klára; Dusek, Ladislav; Sídlová, Tereza; Jálová, Veronika; Cupr, Pavel; Giesy, John P; Nehyba, Slavomír; Jarkovský, Jirí; Klánová, Jana; Holoubek, Ivan

    2010-03-01

    River sediments are a dynamic system, especially in areas where floods occur frequently. In the present study, an integrative approach is used to investigate the seasonal and spatial dynamics of contamination of sediments from a regularly flooded industrial area in the Czech Republic, which presents a suitable model ecosystem for pollutant distribution research at a regional level. Surface sediments were sampled repeatedly to represent two different hydrological situations: spring (after the peak of high flow) and autumn (after longer period of low flow). Samples were characterized for abiotic parameters and concentrations of priority organic pollutants. Toxicity was assessed by Microtox test; genotoxicity by SOS-chromotest and green fluorescent protein (GFP)-yeast test; and the presence of compounds with specific mode of action by in vitro bioassays for dioxin-like activity, anti-/androgenicity, and anti-/estrogenicity. Distribution of organic contaminants varied among regions and seasonally. Although the results of Microtox and genotoxicity tests were relatively inconclusive, all other specific bioassays led to statistically significant regional and seasonal differences in profiles and allowed clear separation of upstream and downstream regions. The outcomes of these bioassays indicated an association with concentrations of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) as master variables. There were significant interrelations among dioxin-like activity, antiandrogenicity and content of organic carbon, clay, and concentration of PAHs and PCBs, which documents the significance of abiotic factors in accumulation of pollutants. The study demonstrates the strength of the specific bioassays in indicating the changes in contamination and emphasizes the crucial role of a well-designed sampling plan, in which both spatial and temporal dynamics should be taken into account, for the correct interpretations of information in risk assessments.

  1. Polycyclic aromatic hydrocarbons bioavailability in industrial and agricultural soils: Linking SPME and Tenax extraction with bioassays.

    Science.gov (United States)

    Guo, Meixia; Gong, Zongqiang; Li, Xiaojun; Allinson, Graeme; Rookes, James; Cahill, David

    2017-06-01

    The aims of this study were to evaluate the bioavailability of polycyclic aromatic hydrocarbons (PAHs) in industrial and agricultural soils using chemical methods and a bioassay, and to study the relationships between the methods. This was conducted by comparing the quantities of PAHs extracted from two manufactured gas plant (MGP) soils and an agricultural soil with low level contamination by solid-phase micro-extraction (SPME) and Tenax-TA extraction with the quantities taken up by the earthworm (Eisenia fetida). In addition, a biodegradation experiment was conducted on one MGP soil (MGP-A) to clarify the relationship between PAH removal by biodegradation and the variation in PAH concentrations in soil pore water. Results demonstrated that the earthworm bioassay could not be used to examine PAH bioavailability in the tested MGP soils; which was the case even in the diluted MGP-A soils after biodegradation. However, the bioassay was successfully applied to the agricultural soil. These results suggest that earthworms can only be used for bioassays in soils with low toxicity. In general, rapidly desorbing concentrations extracted by Tenax-TA could predict PAH concentrations accumulated in earthworms (R 2 =0.66), while SPME underestimated earthworm concentrations by a factor of 2.5. Both SPME and Tenax extraction can provide a useful tool to predict PAH bioavailability for earthworms, but Tenax-TA extraction was proven to be a more sensitive and precise method than SPME for the prediction of earthworm exposure in the agricultural soil. Copyright © 2017. Published by Elsevier Inc.

  2. Bioassays with terrestrial and aquatic species as monitoring tools of hydrocarbon degradation.

    Science.gov (United States)

    Bori, Jaume; Vallès, Bettina; Ortega, Lina; Riva, Maria Carme

    2016-09-01

    In this study chemical analyses and ecotoxicity tests were applied for the assessment of a heavily hydrocarbon-contaminated soil prior and after the application of a remediation procedure that consisted in the stimulation of soil autochthonous populations of hydrocarbon degraders in static-ventilated biopiles. Terrestrial bioassays were applied in mixtures of test soils and artificial control soil and studied the survival and reproduction of Eisenia fetida and the avoidance response of E. fetida and Folsomia candida. Effects on aquatic organisms were studied by means of acute tests with Vibrio fischeri, Raphidocelis subcapitata, and Daphnia magna performed on aqueous elutriates from test soils. The bioremediation procedure led to a significant reduction in the concentration of hydrocarbons (from 34264 to 3074 mg kg(-1), i.e., 91 % decrease) and toxicity although bioassays were not able to report a percentage decrease of toxicity as high as the percentage reduction. Sublethal tests proved the most sensitive terrestrial bioassays and avoidance tests with earthworms and springtails showed potential as monitoring tools of hydrocarbon remediation due to their high sensitivity and short duration. The concentrations of hydrocarbons in water extracts from test soils were 130 and 100 μg L(-1) before and after remediation, respectively. Similarly to terrestrial tests, most aquatic bioassays detected a significant reduction in toxicity, which was almost negligible at the end of the treatment. D. magna survival was the most affected by soil elutriates although toxicity to the crustacean was associated to the salinity of the samples rather than to the concentration of hydrocarbons. Ecotoxicity tests with aqueous soil elutriates proved less relevant in the assessment of hydrocarbon-contaminated soils due to the low hydrosolubility of hydrocarbons and the influence of the physicochemical parameters of the aquatic medium.

  3. Cryopreserved semen in ecotoxicological bioassays: sensitivity and reliability of cryopreserved Sparus aurata spermatozoa.

    Science.gov (United States)

    Fabbrocini, Adele; D'Adamo, Raffaele; Del Prete, Francesco; Langellotti, Antonio Luca; Rinna, Francesca; Silvestri, Fausto; Sorrenti, Gerarda; Vitiello, Valentina; Sansone, Giovanni

    2012-10-01

    The aim of this study was to evaluate the feasibility of using cryopreserved S. aurata semen in spermiotoxicity tests. Cryopreservation is a biotechnology that can provide viable gametes and embryos on demand, rather than only in the spawning season, thus overcoming a limitation that has hindered the use of some species in ecotoxicological bioassays. Firstly, the sperm motility pattern of cryopreserved semen was evaluated after thawing by means of both visual and computer-assisted analyses. Motility parameters in the cryopreserved semen did not change significantly in the first hour after thawing, meaning that they were maintained for long enough to enable their use in spermiotoxicity tests. In the second phase of the research, bioassays were performed, using cadmium as the reference toxicant, in order to evaluate the sensitivity of cryopreserved S. aurata semen to ecotoxicological contamination. The sensitivity of the sperm motility parameters used as endpoints (motility percentages and velocities) proved to be comparable to what has been recorded for the fresh semen of other aquatic species (LOECs from 0.02 to 0.03 mg L(-1)). The test showed good reliability and was found to be rapid and easy to perform, requiring only a small volume of the sample. Moreover, cryopreserved semen is easy to store and transfer and makes it possible to perform bioassays in different sites or at different times with the same batch of semen. The proposed bioassay is therefore a promising starting point for the development of toxicity tests that are increasingly tailored to the needs of ecotoxicology and environmental quality evaluation strategies. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. Functional diagnostics for thyrotropin hormone receptor autoantibodies: bioassays prevail over binding assays.

    Science.gov (United States)

    Lytton, Simon David; Schluter, Anke; Banga, Paul J

    2018-06-01

    Autoantibodies to the thyrotropin hormone receptor (TSH-R) are directly responsible for the hyperthyroidism in Graves' disease and mediate orbital manifestations in Graves' orbitopathy (otherwise known as thyroid eye disease). These autoantibodies are heterogeneous in their function and collectively referred to as TRAbs. Measurement of TRAbs is clinically important for diagnosis of a variety of conditions and different commercial assays with high sensitivity and specificity are available for diagnostic purposes. This review provides overwhelming evidence that the TRAbs detected in binding assays by mainly the automated electrochemical luminescence immunoassays (ECLIA) do not distinguish TRAbs that stimulate the TSH-R (called TSIs or TSAbs) and TRAbs that just inhibit the binding of TSH without stimulating the TSH-R (called TBAbs). However, TSAbs and TBAbs have divergent pathogenic roles, and depending which fraction predominates cause different clinical symptoms and engender different therapeutic regimen. Therefore, diagnostic distinction of TSAbs and TBAbs is of paramount clinical importance. To date, only bioassays such as the Mc4 TSH-R bioassay (Thyretain TM , Quidel) and the Bridge assay (Immulite 2000, Siemens) can measure TSAbs, with only the former being able to distinguish between TSAbs and TBAbs. On this note, it is strongly recommended to only use the term TSI or TSAb when reporting the results of bioassays, whereas the results of automated TRAb binding assays should be reported as TRAbs (of undetermined functional significance). This review aims to present a technical and analytical account of leading commercial diagnostic methods of anti-TSH-R antibodies, a metaanalysis of their clinical performance and a perspective for the use of cell based TSH-R bioassays in the clinical diagnostics of Graves' disease.

  5. Bioassay method for toxicity studies of insecticide formulations to Tuta absoluta (meyrick, 1917)

    OpenAIRE

    Galdino, Tarcísio Visintin da Silva; Picanço, Marcelo Coutinho; Morais, Elisangela Gomes Fidelis de; Silva, Nilson Rodrigues; Silva, Geverson Aelton Rezende da; Lopes, Mayara Cristina

    2011-01-01

    Chemical control is the main method for controlling the tomato leafminer, Tuta absoluta (Meyrick, 1917) (Lepidoptera: Gelechiidae). Reported techniques for the evaluation of insecticide toxicity to the tomato leafminer are not in agreement with field conditions and do not allow us to verify whether doses used in the field are efficient for control. Thus, the objective of this work was to develop a bioassay methodology to study the toxicity of insecticide formulations to T. absoluta that repre...

  6. Bioassay method for toxicity studies of insecticide formulations to tuta absoluta (meyrick, 1917).

    OpenAIRE

    GALDINO, T. V. da S.; PICANÇO, M. C.; MORAIS, E. G. F. de; SILVA, N. R.; SILVA, G. A. R da; LOPES, M. C.

    2014-01-01

    Chemical control is the main method for controlling the tomato leafminer, Tuta absoluta (Meyrick, 1917) (Lepidoptera: Gelechiidae). Reported techniques for the evaluation of insecticide toxicity to the tomato leafminer are not in agreement with field conditions and do not allow us to verify whether doses used in the field are efficient for control. Thus, the objective of this work was to develop a bioassay methodology to study the toxicity of insecticide formulations to T. absoluta that repre...

  7. Bioassay using the water soluble fraction of a Nigerian Light Crude ...

    African Journals Online (AJOL)

    Summary: A 96-hour bioassay was conducted using the water soluble fraction of a Nigerian light crude oil sample on Clarias gariepinus fingerlings. 0, 2.5, 5.0, 7.5 and 10 mls of water soluble fractions (WSF) of the oil were added to 1000 litres of de-chlorinated tap water to form 0, 25, 50 , 75 and 100 parts per million ...

  8. Genotoxicity monitoring of industrial wastes using plant bioassays and management through vermitechnology: A review

    Directory of Open Access Journals (Sweden)

    Sartaj Ahmad Bhat

    2017-10-01

    Full Text Available The main objective of this review was to summarize and present a comprehensive account of the cytotoxic, genotoxic and mutagenic potential of various industrial wastes/sludges using some well-known plant bioassays followed by their bioremediation using vermitechnology. Industries are the main origin of discharges of various types of chemical wastes and are the main causes of environmental degradation. The direct application of industrial sludges could also harm the local biota. The genotoxicity of industrial sludges is assessed using various plant bioassays (for example Allium cepa, Vicia faba and these bioassays are comparatively more sensitive and cost-effective compared to other in-vitro genotoxicity bioassays. In addition, the materials used for toxicity evaluation are easily available and are being routinely used for the monitoring of environmental pollution. In most studies, the increases in root length and mitotic index, as well as the decrease in chromosomal aberrations in post vermicomposted sludges/wastes indicate that earthworms have the ability to reduce the ecotoxicogenetic effects of sludges/wastes. Post vermicompost is considered an excellent material of a homogenous nature as it has reduced levels of contaminants and holds more nutrients over a longer time without affecting the environment. The biotransformation potential of earthworms and their ability to detoxify most of the heavy metals in industrial sludges is because of their strong metabolic system and the involvement of diverse intestinal microflora and chloragocytic cells that reduce toxic forms to nontoxic forms. This unique ability of earthworms confirms the effectiveness of vermitechnology in reducing the toxicity of industrial wastes. Keywords: Allium cepa, Earthworm, Industrial sludge, Toxicity, Vermicomposting

  9. Development of a chick bioassay for determination of infectivity of viral pathogens in poultry litter.

    Science.gov (United States)

    Islam, A F M F; Walkden-Brown, S W; Groves, P J; Wells, B

    2013-01-01

    To develop a chicken bioassay to detect infective viral pathogens in poultry litter and to determine the effects of type of chicken and age of exposure, as well as the effect of simulated litter transportation, on the level of viral infectivity detected. A 5 × 2 × 2 factorial design, plus negative controls. Five chicken litters, including two with deliberate contamination (one transported and one not), two chicken types (specific-pathogen-free (SPF) Leghorns and Cobb broilers) and two ages at initial exposure (days 1 and 8). Two replicates of each treatment combination. The 10 chickens in each of 22 isolators were either exposed (20 isolators) or not (2 isolators) to 8 L of previously used or deliberately contaminated poultry litter in two deep scratch trays. At day 35 post-exposure, sera were assayed for antibodies against chicken anaemia virus (CAV), infectious bronchitis virus (IBV), infectious bursal disease virus (IBDV), Newcastle disease virus (NDV) and fowl adenovirus (FAV). Spleen samples were tested for Marek's disease virus (MDV) using real-time polymerase chain reaction. The bioassay detected CAV, IBDV and FAV, but not NDV, IBV or MDV, in chickens exposed to infected litters. Infection in SPF chickens was detected with greater sensitivity than in the broiler chickens. Sensitivity increased with age at exposure in broiler but not SPF chickens. Simulated transportation for 24 h had little effect on pathogen detection. A bioassay based on the exposure of day-old SPF chickens to poultry litter and measurement of seroconversion at day 35 post-exposure is a useful semi-quantitative assay for viral infectivity in poultry litter, with overnight transportation of litter having little effect on the level of viral infectivity detected. This bioassay has applications in research on litter treatment protocols. © 2013 The Authors. Australian Veterinary Journal © 2013 Australian Veterinary Association.

  10. Estimation of uranium in bioassay samples of occupational workers by laser fluorimetry

    International Nuclear Information System (INIS)

    Suja, A.; Prabhu, S.P.; Sawant, P.D.; Sarkar, P.K.; Tiwari, A.K.; Sharma, R.

    2010-01-01

    A newly established uranium processing facility has been commissioned at BARC, Trombay. Monitoring of occupational workers at regulars intervals is essential to assess intake of uranium by the workers in this facility. The design and engineering safety features of the plant are such that there is very low probability of uranium getting air borne during normal operations. However, the leakages from the system during routine maintenance of the plant may result in intake of uranium by workers. As per the new biokinetic model for uranium, 63% of uranium entering the blood stream gets directly excreted in urine. Therefore, bioassay monitoring (urinalysis) was recommended for these workers. A group of 21 workers was selected for bioassay monitoring to assess the existing urinary excretion levels of uranium before the commencement of actual work. For this purpose, sample collection kit along with an instruction slip was provided to the workers. Bioassay samples received were wet ashed with conc. nitric acid and hydrogen peroxide to break down the metabolized complexes of uranium and it was co-precipitated with calcium phosphate. Separation of uranium from the matrix was done using ion exchange technique and final activity quantification in these samples was done using laser fluorimeter (Quantalase, Model No. NFL/02). Calibration of the laser fluorimeter is done using 10 ppb uranium standard (WHO, France Ref. No. 180000). Verification of the system performance is done by measuring concentration of uranium in the standards (1 ppb to 100 ppb). Standard addition method was followed for estimation of uranium concentration in the samples. Uranyl ions present in the sample get excited by pulsed nitrogen laser at 337.1 nm, and on de-excitation emit fluorescence light (540 nm) intensity which is measured by the PMT. To estimate the uranium in the bioassay samples, a known aliquot of the sample was mixed with 5% sodium pyrophosphate and fluorescence intensity was measured

  11. Recombinant cell bioassays for the detection of (gluco) corticosteroids and endocrine-disrupting potencies of several enviromental PCB contaminants

    NARCIS (Netherlands)

    Bovee, T.F.H.; Helsdingen, J.R.; Hamers, A.R.M.; Brouwer, B.A.; Nielen, M.W.F.

    2011-01-01

    Sensitive and robust bioassays for glucocorticoids are very useful for the pharmaceutical industry, environmental scientists and veterinary control. Here, a recombinant yeast cell was constructed that expresses the human glucocorticoid receptor alpha and a green fluorescent reporter protein in

  12. Continuous flow bioassay method to evaluate the effects of outboard motor exhausts and selected aromatic toxicants on fish. [Carassius auratus

    Energy Technology Data Exchange (ETDEWEB)

    Brenniman, G. (Univ. of Illinois, Chicago); Hartung, R.; Weber, W.J. Jr.

    1976-01-01

    A continuous flow bioassay system was designed to measure the effects of outboard motor exhaust (OME) emissions and selected volatile and evaporative aromatic toxicants on goldfish (Carassius auratus). Continuous flow bioassays were run for 24, 48, 72, 96, and 720 h to determine lethal concentrations for 50 percent of individuals (LC 50's) for leaded OME, non-leaded OME, toluene, xylene, and 1,3,5 trimethylbenzene, the three individual compounds having been identified as significant aromatic components of OME. The 96 h LC-50's for these substances were found to be 171, 168, 23, 17, and 13 ppm, respectively. The values of 171 and 168 ppm for the two OME's are given in terms of gallons of fuel burned per million gallons of water. The continuous flow bioassay method was demonstrated to be a more reliable indicator of the effects of OME pollutants on aquatic organisms than is the static bioassay method.

  13. In Vitro Androgen Bioassays as a Detection Method for Designer Androgens

    Directory of Open Access Journals (Sweden)

    Alison K. Heather

    2013-02-01

    Full Text Available Androgens are the class of sex steroids responsible for male sexual characteristics, including increased muscle mass and decreased fat mass. Illicit use of androgen doping can be an attractive option for those looking to enhance sporting performance and/or physical appearance. The use of in vitro bioassays to detect androgens, especially designer or proandrogens, is becoming increasingly important in combating androgen doping associated with nutritional supplements. The nutritional sports supplement market has grown rapidly throughout the past decade. Many of these supplements contain androgens, designer androgens or proandrogens. Many designer or proandrogens cannot be detected by the standard highly-sensitive screening methods such as gas chromatography-mass spectrometry because their chemical structure is unknown. However, in vitro androgen bioassays can detect designer and proandrogens as these assays are not reliant on knowing the chemical structure but instead are based on androgen receptor activation. For these reasons, it may be advantageous to use routine androgen bioassay screening of nutraceutical samples to help curb the increasing problem of androgen doping.

  14. Application of the CALUX bioassay for epidemiological study. Analyses of Belgian human plasma

    Energy Technology Data Exchange (ETDEWEB)

    Wouwe, N. van; Debacker, N.; Sasse, A. [Scientific Institute of Public Health, Brussels (BE)] (and others)

    2004-09-15

    The CALUX bioassay is a promising screening method for the detection of dioxin-like compounds. The observed good sensitivity, low number of false negative results as well as the good correlations with the GC-HRMS TEQ-values in case of feed and food analyses allow this method to climb in the first assessment methods' scale. The low amount of sample needed in addition to those latest advantages suggest that the CALUX bioassay could be a good screening method for epidemiological studies. The Belgian epidemiological study concerning the possible effect of the dioxin incident on the body burden of the Belgian population was an opportunity to test this method in comparison to the gold reference one: the GC-HRMS. The first part of this abstract presents epidemiological parameters (sensibility, specificity,) of the CALUX bioassay using CALUX TEQ-values as estimators of the TEQ-values of the 17 PCDD/Fs. The second part examines epidemiological determinants observed for CALUX and GCHRMS TEQ-values.

  15. Intercomparison programs - a tool for the implementation of a quality assurance program in bioassay

    International Nuclear Information System (INIS)

    Mesquita, Sueli A. de; Sousa, Wanderson O.; Juliao, Ligia M.Q.C.; Santos, Maristela S.; Fernandes, Paulo C.P.

    2009-01-01

    In vitro bioassay laboratories need to have means to demonstrate that they are technically competent, operate an effective quality system, and are able to generate technically valid calibration and test results. The reliability of the results of measurements has a high influence on the reliability of the dose assessment. Inter-laboratory tests are one of the tools for assessing the analytical consistency of in vitro bioassay laboratories. The intercomparison exercises provide an opportunity to compare radiochemistry techniques for in vitro analysis of biological samples. The in vitro Laboratory of the Instituto de Radioprotecao e Dosimetria has therefore participated in the intercomparison exercises sponsored by PROCORAD, ARCAL and IAEA since 1998. The intercomparison exercises comprise measurements of gamma and beta emitters in urine samples and alpha emitters in urine and faecal samples. This paper presents the performance of the IRD in vitro bioassay laboratory in the past intercomparisons. The results demonstrate that in vitro laboratory is able to generate technically valid results, which also guarantee the support for a quality assurance program and accreditation by competent organism in Brazil. (author)

  16. Development of bacteria-based bioassays for arsenic detection in natural waters.

    Science.gov (United States)

    Diesel, Elizabeth; Schreiber, Madeline; van der Meer, Jan Roelof

    2009-06-01

    Arsenic contamination of natural waters is a worldwide concern, as the drinking water supplies for large populations can have high concentrations of arsenic. Traditional techniques to detect arsenic in natural water samples can be costly and time-consuming; therefore, robust and inexpensive methods to detect arsenic in water are highly desirable. Additionally, methods for detecting arsenic in the field have been greatly sought after. This article focuses on the use of bacteria-based assays as an emerging method that is both robust and inexpensive for the detection of arsenic in groundwater both in the field and in the laboratory. The arsenic detection elements in bacteria-based bioassays are biosensor-reporter strains; genetically modified strains of, e.g., Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Rhodopseudomonas palustris. In response to the presence of arsenic, such bacteria produce a reporter protein, the amount or activity of which is measured in the bioassay. Some of these bacterial biosensor-reporters have been successfully utilized for comparative in-field analyses through the use of simple solution-based assays, but future methods may concentrate on miniaturization using fiberoptics or microfluidics platforms. Additionally, there are other potential emerging bioassays for the detection of arsenic in natural waters including nematodes and clams.

  17. Fluorescence-based bioassays for the detection and evaluation of food materials.

    Science.gov (United States)

    Nishi, Kentaro; Isobe, Shin-Ichiro; Zhu, Yun; Kiyama, Ryoiti

    2015-10-13

    We summarize here the recent progress in fluorescence-based bioassays for the detection and evaluation of food materials by focusing on fluorescent dyes used in bioassays and applications of these assays for food safety, quality and efficacy. Fluorescent dyes have been used in various bioassays, such as biosensing, cell assay, energy transfer-based assay, probing, protein/immunological assay and microarray/biochip assay. Among the arrays used in microarray/biochip assay, fluorescence-based microarrays/biochips, such as antibody/protein microarrays, bead/suspension arrays, capillary/sensor arrays, DNA microarrays/polymerase chain reaction (PCR)-based arrays, glycan/lectin arrays, immunoassay/enzyme-linked immunosorbent assay (ELISA)-based arrays, microfluidic chips and tissue arrays, have been developed and used for the assessment of allergy/poisoning/toxicity, contamination and efficacy/mechanism, and quality control/safety. DNA microarray assays have been used widely for food safety and quality as well as searches for active components. DNA microarray-based gene expression profiling may be useful for such purposes due to its advantages in the evaluation of pathway-based intracellular signaling in response to food materials.

  18. Fluorescence-Based Bioassays for the Detection and Evaluation of Food Materials

    Directory of Open Access Journals (Sweden)

    Kentaro Nishi

    2015-10-01

    Full Text Available We summarize here the recent progress in fluorescence-based bioassays for the detection and evaluation of food materials by focusing on fluorescent dyes used in bioassays and applications of these assays for food safety, quality and efficacy. Fluorescent dyes have been used in various bioassays, such as biosensing, cell assay, energy transfer-based assay, probing, protein/immunological assay and microarray/biochip assay. Among the arrays used in microarray/biochip assay, fluorescence-based microarrays/biochips, such as antibody/protein microarrays, bead/suspension arrays, capillary/sensor arrays, DNA microarrays/polymerase chain reaction (PCR-based arrays, glycan/lectin arrays, immunoassay/enzyme-linked immunosorbent assay (ELISA-based arrays, microfluidic chips and tissue arrays, have been developed and used for the assessment of allergy/poisoning/toxicity, contamination and efficacy/mechanism, and quality control/safety. DNA microarray assays have been used widely for food safety and quality as well as searches for active components. DNA microarray-based gene expression profiling may be useful for such purposes due to its advantages in the evaluation of pathway-based intracellular signaling in response to food materials.

  19. A new approach for bioassays based on frequency- and time-domain measurements of magnetic nanoparticles.

    Science.gov (United States)

    Oisjöen, Fredrik; Schneiderman, Justin F; Astalan, Andrea Prieto; Kalabukhov, Alexey; Johansson, Christer; Winkler, Dag

    2010-01-15

    We demonstrate a one-step wash-free bioassay measurement system capable of tracking biochemical binding events. Our approach combines the high resolution of frequency- and high speed of time-domain measurements in a single device in combination with a fast one-step bioassay. The one-step nature of our magnetic nanoparticle (MNP) based assay reduces the time between sample extraction and quantitative results while mitigating the risks of contamination related to washing steps. Our method also enables tracking of binding events, providing the possibility of, for example, investigation of how chemical/biological environments affect the rate of a binding process or study of the action of certain drugs. We detect specific biological binding events occurring on the surfaces of fluid-suspended MNPs that modify their magnetic relaxation behavior. Herein, we extrapolate a modest sensitivity to analyte of 100 ng/ml with the present setup using our rapid one-step bioassay. More importantly, we determine the size-distributions of the MNP systems with theoretical fits to our data obtained from the two complementary measurement modalities and demonstrate quantitative agreement between them. Copyright 2009 Elsevier B.V. All rights reserved.

  20. Investigation of animal and algal bioassays for reliable saxitoxin ecotoxicity and cytotoxicity risk evaluation.

    Science.gov (United States)

    Perreault, François; Matias, Marcelo Seleme; Melegari, Silvia Pedroso; Pinto, Catia Regina Silva de Carvalho; Creppy, Edmond Ekué; Popovic, Radovan; Matias, William Gerson

    2011-05-01

    Contamination of water bodies by saxitoxin can result in various toxic effects in aquatic organisms. Saxitoxin contamination has also been shown to be a threat to human health in several reported cases, even resulting in death. In this study, we evaluated the sensitivity of animal (Neuro-2A) and algal (Chlamydomonas reinhardtii) bioassays to saxitoxin effect. Neuro-2A cells were found to be sensitive to saxitoxin, as shown by a 24 h EC50 value of 1.5 nM, which was obtained using a cell viability assay. Conversely, no saxitoxin effect was found in any of the algal biomarkers evaluated, for the concentration range tested (2-128 nM). These results indicate that saxitoxin may induce toxic effects in animal and human populations at concentrations where phytoplankton communities are not affected. Therefore, when evaluating STX risk of toxicity, algal bioassays do not appear to be reliable indicators and should always be conducted in combination with animal bioassays. Copyright © 2011 Elsevier Inc. All rights reserved.

  1. Development of acute and chronic sediment bioassays with the harpacticoid copepod Quinquelaophonte sp.

    Science.gov (United States)

    Stringer, Tristan J; Glover, Chris N; Keesing, Vaughan; Northcott, Grant L; Gaw, Sally; Tremblay, Louis A

    2014-01-01

    Reliable environmentally realistic bioassay methodologies are increasingly needed to assess the effects of environmental pollution. This study describes two estuarine sediment bioassays, one acute (96 h) and one chronic (14 d), with the New Zealand harpacticoid copepod Quinquelaophonte sp. utilising behavioural and reproductive endpoints. Spiked sediments were used to expose Quinquelaophonte sp. to three reference compounds representing important categories of estuarine chemical stressors: zinc (a metal), atrazine (a pesticide), and phenanthrene (a polycyclic aromatic hydrocarbon). Acute-to-chronic ratios (ACR) were used to further characterise species responses. Acute sediment (sandy and low total organic content) 96 h EC50 values for the sublethal inhibition of mobility for zinc, atrazine and phenanthrene were 137, 5.4, and 2.6 µg/g, respectively. The chronic EC50 values for inhibition of reproduction (total offspring) were 54.5, 0.0083, and 0.067 µg/g for zinc, atrazine, and phenanthrene, respectively. For phenanthrene, a potentially novel mode of action was identified on reproduction. Quinquelaophonte sp. was found to be more sensitive than several other estuarine species indicating choice of test organism is important to characterising the effects of environmentally relevant levels of contamination. The bioassay sediment results demonstrate the sensitivity and suitability of Quinquelaophonte sp. as a tool for the assessment use of estuarine health. © 2013 Published by Elsevier Inc.

  2. Determination of an Environmental Background Level of Sr-90 in Urine for the Hanford Bioassay Program

    International Nuclear Information System (INIS)

    Antonio, Cheryl L.; Rivard, James W.

    2009-01-01

    During the decommissioning and maintenance of some of the facilities at the U.S. Department of Energy Hanford Site in Washington State, workers have potential for a 90Sr intake. However, because of worldwide radioactive fallout, 90Sr is present in our environment, and can be detectable in routine urine bioassay samples. It is important for the Hanford Site bioassay program to discern an occupational intake from a non-occupational environmental one. A detailed study of the background 90Sr in the urine of unexposed Hanford workers was performed. A survey of the Hanford Site bioassay database found 128 Hanford workers who were hired between 1997 and 2002 and who had a very low potential for an occupational exposure prior to the baseline strontium urinalysis. Each urinalysis sample represented excretion during an approximate 24-hr period. The arithmetic mean value for the 128 pre-exposure baselines was 3.6 ± 5.1 mBq d-1. The 90Sr activities in urine varied from -12 to 20 mBq. The 99th percentile result was 16.4 mBqd-1, which was interpreted to mean that 1% of Hanford workers not occupationally exposed to strontium might exceed 16.4 mBq d-1.

  3. Assessing the genotoxicity of urban air pollutants using two in situ plant bioassays

    Energy Technology Data Exchange (ETDEWEB)

    Villarini, M.; Fatigoni, C.; Dominici, L.; Maestri, S. [Department of Medical-Surgical Specialties and Public Health, University of Perugia, I-06126 (Italy); Ederli, L.; Pasqualini, S. [Department of Applied Biology, University of Perugia, I-06121 (Italy); Monarca, S. [Department of Medical-Surgical Specialties and Public Health, University of Perugia, I-06126 (Italy); Moretti, M., E-mail: massimo.moretti@unipg.i [Department of Medical-Surgical Specialties and Public Health, University of Perugia, I-06126 (Italy)

    2009-12-15

    Genotoxicity of urban air has been analysed almost exclusively in airborne particulates. We monitored the genotoxic effects of airborne pollutants in the urban air of Perugia (Central Italy). Two plant bioindicators with different genetic endpoints were used: micronuclei in meiotic pollen mother cells using Tradescantia-micronucleus bioassay (Trad-MCN) and DNA damage in nuclei of Nicotiana tabacum leaves using comet assay (Nicotiana-comet). Buds of Tradescantia clone no. 4430 and young N. tabacum cv. Xanthi plants were exposed for 24 h at three sites with different pollution levels. One control site (indoor control) was also used. The two bioassays showed different sensitivities toward urban pollutants: Trad-MCN assay was the most sensitive, but DNA damage in N. tabacum showed a better correlation with the pollutant concentrations. In situ biomonitoring of airborne genotoxins using higher plants combined with chemical analysis is thus recommended for characterizing genotoxicity of urban air. - Plant bioassays used to explore in situ the correlation between air pollution and genotoxicity.

  4. Development of bacteria-based bioassays for arsenic detection in natural waters

    Energy Technology Data Exchange (ETDEWEB)

    Diesel, Elizabeth; Schreiber, Madeline [Virginia Tech, Department of Geosciences, Blacksburg, VA (United States); Meer, Jan Roelof van der [University of Lausanne, Department of Fundamental Microbiology, Lausanne (Switzerland)

    2009-06-15

    Arsenic contamination of natural waters is a worldwide concern, as the drinking water supplies for large populations can have high concentrations of arsenic. Traditional techniques to detect arsenic in natural water samples can be costly and time-consuming; therefore, robust and inexpensive methods to detect arsenic in water are highly desirable. Additionally, methods for detecting arsenic in the field have been greatly sought after. This article focuses on the use of bacteria-based assays as an emerging method that is both robust and inexpensive for the detection of arsenic in groundwater both in the field and in the laboratory. The arsenic detection elements in bacteria-based bioassays are biosensor-reporter strains; genetically modified strains of, e.g., Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Rhodopseudomonas palustris. In response to the presence of arsenic, such bacteria produce a reporter protein, the amount or activity of which is measured in the bioassay. Some of these bacterial biosensor-reporters have been successfully utilized for comparative in-field analyses through the use of simple solution-based assays, but future methods may concentrate on miniaturization using fiberoptics or microfluidics platforms. Additionally, there are other potential emerging bioassays for the detection of arsenic in natural waters including nematodes and clams. (orig.)

  5. Noninvasive quantitation of human liver steatosis using magnetic resonance and bioassay methods

    Energy Technology Data Exchange (ETDEWEB)

    D' Assignies, Gaspard; Ruel, Martin; Khiat, Abdesslem; Lepanto, Luigi; Kauffmann, Claude; Tang, An [Centre Hospitalier de l' Universite de Montreal (CHUM), Departement de Radiologie, Montreal, Quebec (Canada); Chagnon, Miguel [Universite de Montreal (UDEM), Departement de Mathematiques et de Statistique, Montreal, Quebec (Canada); Gaboury, Louis [Centre Hospitalier de l' Universite de Montreal (CHUM), Departement d' Anatomo-Pathologie, Montreal, Quebec (Canada); Boulanger, Yvan [Centre Hospitalier de l' Universite de Montreal (CHUM), Departement de Radiologie, Montreal, Quebec (Canada); Hopital Saint-Luc du CHUM, Departement de Radiologie, Montreal, Quebec (Canada)

    2009-08-15

    The purpose was to evaluate the ability of three magnetic resonance (MR) techniques to detect liver steatosis and to determine which noninvasive technique (MR, bioassays) or combination of techniques is optimal for the quantification of hepatic fat using histopathology as a reference. Twenty patients with histopathologically proven steatosis and 24 control subjects underwent single-voxel proton MR spectroscopy (MRS; 3 voxels), dual-echo in phase/out of phase MR imaging (DEI) and diffusion-weighted MR imaging (DWI) examinations of the liver. Blood or urine bioassays were also performed for steatosis patients. Both MRS and DEI data allowed to detect steatosis with a high sensitivity (0.95 for MRS; 1 for DEI) and specificity (1 for MRS; 0.875 for DEI) but not DWI. Strong correlations were found between fat fraction (FF) measured by MRS, DEI and histopathology segmentation as well as with low density lipoprotein (LDL) and cholesterol concentrations. A Bland-Altman analysis showed a good agreement between the FF measured by MRS and DEI. Partial correlation analyses failed to improve the correlation with segmentation FF when MRS or DEI data were combined with bioassay results. Therefore, FF from MRS or DEI appear to be the best parameters to both detect steatosis and accurately quantify fat liver noninvasively. (orig.)

  6. Experimental and Computational Characterization of Biological Liquid Crystals: A Review of Single-Molecule Bioassays

    Directory of Open Access Journals (Sweden)

    Sungsoo Na

    2009-09-01

    Full Text Available Quantitative understanding of the mechanical behavior of biological liquid crystals such as proteins is essential for gaining insight into their biological functions, since some proteins perform notable mechanical functions. Recently, single-molecule experiments have allowed not only the quantitative characterization of the mechanical behavior of proteins such as protein unfolding mechanics, but also the exploration of the free energy landscape for protein folding. In this work, we have reviewed the current state-of-art in single-molecule bioassays that enable quantitative studies on protein unfolding mechanics and/or various molecular interactions. Specifically, single-molecule pulling experiments based on atomic force microscopy (AFM have been overviewed. In addition, the computational simulations on single-molecule pulling experiments have been reviewed. We have also reviewed the AFM cantilever-based bioassay that provides insight into various molecular interactions. Our review highlights the AFM-based single-molecule bioassay for quantitative characterization of biological liquid crystals such as proteins.

  7. Sampling method, storage and pretreatment of sediment affect AVS concentrations with consequences for bioassay responses

    Energy Technology Data Exchange (ETDEWEB)

    Lange, H.J. de [Aquatic Ecology and Water Quality Management Group, Wageningen University, Wageningen University and Research Centre, P.O. Box 8080, 6700 DD, Wageningen (Netherlands); Centre for Ecosystem Studies, Alterra, Wageningen University and Research Centre, P.O. Box 47, 6700 AA, Wageningen (Netherlands)], E-mail: marieke.delange@wur.nl; Griethuysen, C. van; Koelmans, A.A. [Aquatic Ecology and Water Quality Management Group, Wageningen University, Wageningen University and Research Centre, P.O. Box 8080, 6700 DD, Wageningen (Netherlands)

    2008-01-15

    Sediment treatment and sediment storage may alter sediment toxicity, and consequently biotic response. Purpose of our study was to combine these three aspects (treatment-toxicity-biotic response) in one integrated approach. We used Acid Volatile Sulfide (AVS) concentrations as a proxy of the disturbance of the sediment. AVS and Simultaneously Extracted Metal (SEM) concentrations were compared to bioassay responses with the freshwater benthic macroinvertebrate Asellus aquaticus. Storage conditions and sediment treatment affected AVS but not SEM levels. AVS can be used as a proxy for sediment disturbance. The best way to pretreat the sediment for use in a bioassay in order to maintain initial AVS conditions was to sample the sediment with an Ekman grab, immediately store it in a jar without headspace, and freeze it as soon as possible. In a survey using seven different sediments, bioassay responses of A. aquaticus were correlated with SEM and AVS characteristics. - Change in AVS is a good proxy for sediment disturbance and combined with SEM it can be used as a suitable predictor for biotic effects of sediment contamination.

  8. Assessing the genotoxicity of urban air pollutants using two in situ plant bioassays

    International Nuclear Information System (INIS)

    Villarini, M.; Fatigoni, C.; Dominici, L.; Maestri, S.; Ederli, L.; Pasqualini, S.; Monarca, S.; Moretti, M.

    2009-01-01

    Genotoxicity of urban air has been analysed almost exclusively in airborne particulates. We monitored the genotoxic effects of airborne pollutants in the urban air of Perugia (Central Italy). Two plant bioindicators with different genetic endpoints were used: micronuclei in meiotic pollen mother cells using Tradescantia-micronucleus bioassay (Trad-MCN) and DNA damage in nuclei of Nicotiana tabacum leaves using comet assay (Nicotiana-comet). Buds of Tradescantia clone no. 4430 and young N. tabacum cv. Xanthi plants were exposed for 24 h at three sites with different pollution levels. One control site (indoor control) was also used. The two bioassays showed different sensitivities toward urban pollutants: Trad-MCN assay was the most sensitive, but DNA damage in N. tabacum showed a better correlation with the pollutant concentrations. In situ biomonitoring of airborne genotoxins using higher plants combined with chemical analysis is thus recommended for characterizing genotoxicity of urban air. - Plant bioassays used to explore in situ the correlation between air pollution and genotoxicity.

  9. Building bio-assays with magnetic particles on a digital microfluidic platform.

    Science.gov (United States)

    Kokalj, Tadej; Pérez-Ruiz, Elena; Lammertyn, Jeroen

    2015-09-25

    Digital microfluidics (DMF) has emerged as a promising liquid handling technology for a variety of applications, demonstrating great potential both in terms of miniaturization and automation. DMF is based on the manipulation of discrete, independently controllable liquid droplets, which makes it highly reconfigurable and reprogrammable. One of its most exclusive advantages, compared to microchannel-based microfluidics, is its ability to precisely handle solid nano- and microsized objects, such as magnetic particles. Magnetic particles have become very popular in the last decade, since their high surface-to-volume ratio and the possibility to magnetically separate them from the matrix make them perfect suitable as a solid support for bio-assay development. The potential of magnetic particles in DMF-based bio-assays has been demonstrated for various applications. In this review we discuss the latest developments of magnetic particle-based DMF bio-assays with the aim to present, identify and analyze the trends in the field. We also discuss the state-of-the art of device integration, current status of commercialization and issues that still need to be addressed. With this paper we intend to stimulate researchers to exploit and unveil the potential of these exciting tools, which will shape the future of modern biochemistry, microbiology and biomedical diagnostics. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Use of a modified microplate bioassay method to investigate antibacterial activity in the Peruvian medicinal plant Peperomia galioides.

    Science.gov (United States)

    Langfield, Richard D; Scarano, Frank J; Heitzman, Mary E; Kondo, Miwako; Hammond, Gerald B; Neto, Catherine C

    2004-10-01

    A versatile microplate bioassay for quick and sensitive determination of antibacterial activity was developed for use in screening medicinal plants and identification of their active principles. This assay can be used to determine minimum inhibitory concentrations for small quantities of organic or water-soluble plant extracts. Bioassay-guided fractionation of the stem and leaves of Peperomia galioides using this method found fractions containing grifolin and grifolic acid, which inhibited growth of Staphylococcus aureus and Staphylococcus epidermidis.

  11. In vitro bioassays for detecting dioxin-like activity--application potentials and limits of detection, a review.

    Science.gov (United States)

    Eichbaum, Kathrin; Brinkmann, Markus; Buchinger, Sebastian; Reifferscheid, Georg; Hecker, Markus; Giesy, John P; Engwall, Magnus; van Bavel, Bert; Hollert, Henner

    2014-07-15

    Use of in vitro assays as screening tool to characterize contamination of a variety of environmental matrices has become an increasingly popular and powerful toolbox in the field of environmental toxicology. While bioassays cannot entirely substitute analytical methods such as gas chromatography-mass spectrometry (GC-MS), the increasing improvement of cell lines and standardization of bioassay procedures enhance their utility as bioanalytical pre-screening tests prior to more targeted chemical analytical investigations. Dioxin-receptor-based assays provide a holistic characterization of exposure to dioxin-like compounds (DLCs) by integrating their overall toxic potential, including potentials of unknown DLCs not detectable via e.g. GC-MS. Hence, they provide important additional information with respect to environmental risk assessment of DLCs. This review summarizes different in vitro bioassay applications for detection of DLCs and considers the comparability of bioassay and chemical analytically derived toxicity equivalents (TEQs) of different approaches and various matrices. These range from complex samples such as sediments through single reference to compound mixtures. A summary of bioassay derived detection limits (LODs) showed a number of current bioassays to be equally sensitive as chemical methodologies, but moreover revealed that most of the bioanalytical studies conducted to date did not report their LODs, which represents a limitation with regard to low potency samples. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Toxicity of hydrogen sulfide to goldfish (Carassius auratus L. ) as influenced by temperature, oxygen, and bioassay techniques

    Energy Technology Data Exchange (ETDEWEB)

    Adelman, I.R.; Smith, L.L. Jr.

    1972-01-01

    Bioassays were conducted to test the effect of temperature and oxygen on H/sub 2/S toxicity to goldfish (Carassius auratus L.) and to investigate some factors that influence bioassay results. Relation of H/sub 2/S toxicity to temperature is negatively logarithmic over the range of 6.5-25 C. The mean 96-hr TL50 at 6 C was 530 ..mu..g/liter and at 25 C was 4 ..mu..g/liter. At temperatures of 14, 20, and 26 C, most acute mortality from H/sub 2/S ended by 11 days and the 11-day TL50's at these temperatures were significantly different. In bioassays with and without prior oxygen acclimation, decreasing oxygen concentrations increased H/sub 2/S toxicity. In the former, mean TL50's were 62 and 48 ..mu..g/liter H/sub 2/S at oxygen concentrations of 6 and 1.5 mg/liter, respectively, and in the latter, 71 and 53 ..mu..g/liter H/sub 2/S at the same oxygen concentrations. Variability in bioassay results was not affected by test temperatures of 14, 20, and 26 C, and in most cases 1 week of temperature acclimation was adequate. Stocks of fish responded differently after 11 days of bioassay, although differences were not detected after 4 days of bioassay. 27 references, 4 figures, 3 tables.

  13. A novel method for standardized application of fungal spore coatings for mosquito exposure bioassays.

    Science.gov (United States)

    Farenhorst, Marit; Knols, Bart G J

    2010-01-20

    Interest in the use of fungal entomopathogens against malaria vectors is growing. Fungal spores infect insects via the cuticle and can be applied directly on the insect to evaluate infectivity. For flying insects such as mosquitoes, however, application of fungal suspensions on resting surfaces is more realistic and representative of field settings. For this type of exposure, it is essential to apply specific amounts of fungal spores homogeneously over a surface for testing the effects of fungal dose and exposure time. Contemporary methods such as spraying or brushing spore suspensions onto substrates do not produce the uniformity and consistency that standardized laboratory assays require. Two novel fungus application methods using equipment developed in the paint industry are presented and compared. Wired, stainless steel K-bars were tested and optimized for coating fungal spore suspensions onto paper substrates. Different solvents and substrates were evaluated. Two types of coating techniques were compared, i.e. manual and automated coating. A standardized bioassay set-up was designed for testing coated spores against malaria mosquitoes. K-bar coating provided consistent applications of spore layers onto paper substrates. Viscous Ondina oil formulations were not suitable and significantly reduced spore infectivity. Evaporative Shellsol T solvent dried quickly and resulted in high spore infectivity to mosquitoes. Smooth proofing papers were the most effective substrate and showed higher infectivity than cardboard substrates. Manually and mechanically applied spore coatings showed similar and reproducible effects on mosquito survival. The standardized mosquito exposure bioassay was effective and consistent in measuring effects of fungal dose and exposure time. K-bar coating is a simple and consistent method for applying fungal spore suspensions onto paper substrates and can produce coating layers with accurate effective spore concentrations. The mosquito bioassay

  14. A novel method for standardized application of fungal spore coatings for mosquito exposure bioassays

    Directory of Open Access Journals (Sweden)

    Knols Bart GJ

    2010-01-01

    Full Text Available Abstract Background Interest in the use of fungal entomopathogens against malaria vectors is growing. Fungal spores infect insects via the cuticle and can be applied directly on the insect to evaluate infectivity. For flying insects such as mosquitoes, however, application of fungal suspensions on resting surfaces is more realistic and representative of field settings. For this type of exposure, it is essential to apply specific amounts of fungal spores homogeneously over a surface for testing the effects of fungal dose and exposure time. Contemporary methods such as spraying or brushing spore suspensions onto substrates do not produce the uniformity and consistency that standardized laboratory assays require. Two novel fungus application methods using equipment developed in the paint industry are presented and compared. Methods Wired, stainless steel K-bars were tested and optimized for coating fungal spore suspensions onto paper substrates. Different solvents and substrates were evaluated. Two types of coating techniques were compared, i.e. manual and automated coating. A standardized bioassay set-up was designed for testing coated spores against malaria mosquitoes. Results K-bar coating provided consistent applications of spore layers onto paper substrates. Viscous Ondina oil formulations were not suitable and significantly reduced spore infectivity. Evaporative Shellsol T solvent dried quickly and resulted in high spore infectivity to mosquitoes. Smooth proofing papers were the most effective substrate and showed higher infectivity than cardboard substrates. Manually and mechanically applied spore coatings showed similar and reproducible effects on mosquito survival. The standardized mosquito exposure bioassay was effective and consistent in measuring effects of fungal dose and exposure time. Conclusions K-bar coating is a simple and consistent method for applying fungal spore suspensions onto paper substrates and can produce coating layers

  15. Using a macroalgal δ15N bioassay to detect cruise ship waste water effluent inputs

    International Nuclear Information System (INIS)

    Kaldy, James

    2011-01-01

    Highlights: → Green macroalgae exposed to nutrient solutions exhibited changes in tissue 15 N signatures. → Macroalgae exhibited no fractionation with NO 3 and slight fractionation with NH 4 . → Algae exposed to cruise ship waste water had increased tissue δ 15 N indicating a heavy N source. → Field bioassays exhibited decreased δ 15 N indicating isotopically light riverine δ 15 N-NO 3 was likely the dominant N source. → Algal bioassays could not detect a δ 15 N cruise ship waste water signal in this system. - Abstract: Green macroalgae bioassays were used to determine if the δ 15 N signature of cruise ship waste water effluent (CSWWE) could be detected in a small harbor. Opportunistic green macroalgae (Ulva spp.) were collected, cultured under nutrient depleted conditions and characterized with regard to N content and δ 15 N. Samples of algae were used in controlled incubations to evaluate the direction of isotope shift from exposure to CSWWE. Algae samples exposed to CSWWE exhibited an increase of 1-2.5 per mille in δ 15 N values indicating that the CSWWE had an enriched isotope signature. In contrast, algae samples exposed to field conditions exhibited a significant decrease in the observed δ 15 N indicating that a light N source was used. Isotopically light, riverine nitrogen derived from N 2 -fixing trees in the watershed may be a N source utilized by algae. These experiments indicate that the δ 15 N CSWWE signature was not detectable under the CSWWE loading conditions of this experiment.

  16. Bioassay requirements for 125I and 131I in medical, teaching and research institutions

    International Nuclear Information System (INIS)

    1983-09-01

    The more widespread use of radioactive isotopes of iodine (collectively referred to as radioiodines) as a research tool, coupled with their diagnostic and therapeutic uses in nuclear medicine, has resulted in an increased number of personnel who are exposed to these radioisotopes and who therefore should be monitored for internal radioiodine contamination. This document describes the minimum acceptable features of a bioassay programme which the Atomic Energy Control Board (AECB) requires to be available in institutions holding a prescribed substance licence authorising the use of significant quantities of 125 I or 131 I or both. A licensee may submit details of his own proposed bioassay programme to the AECB for approval. If such a programme fails to be approved, the programme described below shall be adhered to. This document does not deal with individuals who are likely to maintain a significant chronic thyroid burden of radioiodine. It is assumed that the radioiodine taken into the body is in a soluble, inorganic form (I 2 , iodide or iodate) or in an organic form (e.g. methyl iodide) which is metabolised in the body with a resultant release of iodide. Radioiodinated organic compounds which are not catabolised to iodide in the body to any significant degree are not the subject of this document, since the metabolism of the radioiodine will be dictated by the metabolism of the compound. This means that individuals whose only exposure to radioiodine is in the form of prepared radioiodinated compounds such as antigens and antibodies (e.g. individuals using radio immuno assay kits in which the antigen or antibody is supplied as radioiodinated material) are not required to participate in this bioassay programme for radioiodine

  17. Detection of anabolic androgenic steroid abuse in doping control using mammalian reporter gene bioassays.

    Science.gov (United States)

    Houtman, Corine J; Sterk, Saskia S; van de Heijning, Monique P M; Brouwer, Abraham; Stephany, Rainer W; van der Burg, Bart; Sonneveld, Edwin

    2009-04-01

    Anabolic androgenic steroids (AAS) are a class of steroid hormones related to the male hormone testosterone. They are frequently detected as drugs in sport doping control. Being similar to or derived from natural male hormones, AAS share the activation of the androgen receptor (AR) as common mechanism of action. The mammalian androgen responsive reporter gene assay (AR CALUX bioassay), measuring compounds interacting with the AR can be used for the analysis of AAS without the necessity of knowing their chemical structure beforehand, whereas current chemical-analytical approaches may have difficulty in detecting compounds with unknown structures, such as designer steroids. This study demonstrated that AAS prohibited in sports and potential designer AAS can be detected with this AR reporter gene assay, but that also additional steroid activities of AAS could be found using additional mammalian bioassays for other types of steroid hormones. Mixtures of AAS were found to behave additively in the AR reporter gene assay showing that it is possible to use this method for complex mixtures as are found in doping control samples, including mixtures that are a result of multi drug use. To test if mammalian reporter gene assays could be used for the detection of AAS in urine samples, background steroidal activities were measured. AAS-spiked urine samples, mimicking doping positive samples, showed significantly higher androgenic activities than unspiked samples. GC-MS analysis of endogenous androgens and AR reporter gene assay analysis of urine samples showed how a combined chemical-analytical and bioassay approach can be used to identify samples containing AAS. The results indicate that the AR reporter gene assay, in addition to chemical-analytical methods, can be a valuable tool for the analysis of AAS for doping control purposes.

  18. Evaluation of internal exposure of nuclear medicine staff through in vivo and in vitro bioassay techniques

    Energy Technology Data Exchange (ETDEWEB)

    Lucena, E.A.; Araujo, F.; Sousa, W.O.; Dantas, A.L.A.; Dantas, B.M. [Instituto de Radioprotecao e Dosimetria, CNEN, Av. Salvador Allende s/n, CEP 22780-160 Rio de Janeiro (Brazil); Rebelo, A.M.O.; Corbo, R. [Hospital Universitario Clementino Fraga Filho, HU-UFRJ, Av. Brigadeiro Trompowsky, s/n, ILHA do Fundao, CEP 21945-560, Rio de Janeiro, RJ (Brazil)

    2007-07-01

    The manipulation of a wide variety of unsealed sources in Nuclear Medicine results in a significant risk of internal exposure of the workers. {sup 131}I should be highlighted among the most frequently used radionuclides because of its large application for diagnosis and therapy of thyroid diseases. The increasing use of radionuclides for medical purposes creates a demand for feasible methodologies to perform occupational control of internal contamination. Currently in Brazil, there are {approx}300 nuclear medicine centres in operation but individual monitoring is still restricted to the control of external exposure. This work presents the development of in vivo and in vitro bioassay techniques aimed to quantify incorporation of radionuclides used in Nuclear Medicine. It is also presented the results of a preliminary survey of internal exposure of a group of workers involved in the preparation of therapeutic doses of {sup 131}I. Workers were monitored with a gamma camera available in the Nuclear Medicine Service of the University Hospital of Rio de Janeiro and at the Institute of Radiation Protection and Dosimetry Whole-Body Counter (IRDWBC). The in vivo detection systems were calibrated with a neck-thyroid phantom developed in IRD. Urine samples from radiopharmacy workers were collected after preparation and administration of therapeutic doses (10-250 mCi) of {sup 131}I and measured with a HPGe detection system available in the Bioassay Laboratory of IRD. The results show that the bioassay methods developed in this work present enough sensitivity for routine monitoring of nuclear medicine workers. All workers monitored in this survey presented positive results for {sup 131}I in urine samples and two workers presented detectable activities in thyroid when measured at the IRD-WBC. The highest committed effective dose per preparation was estimated to be 17 {mu}Sv. (authors)

  19. Redox-flexible NADH oxidase biosensor: A platform for various dehydrogenase bioassays and biosensors

    International Nuclear Information System (INIS)

    Serban, Simona; El Murr, Nabil

    2006-01-01

    A generic amperometric bioassay based on the enzymatic oxidation catalysed by the stable NADH oxidase (NAox) from Thermus thermophilus has been developed for NADH measurements. The NAox uses O 2 as its natural electron acceptor and produces H 2 O 2 in a two-electron process. Electrochemical and spectrophotometric experiments showed that the NAox used in this work, presents a very good activity towards its substrate and, in contrary to previously mentioned NADH oxidases, does not require the addition of any exogenous flavin cofactor neither to promote nor to maintain its activity. In addition, the NAox used also works with artificial electron acceptors like ferrocene derivatives. O 2 was successfully replaced by redox mediators such as hydroxymethyl ferrocene (FcCH 2 OH) for the regeneration of the active enzyme. Combining the NAox with the mediator and the horseradish peroxidase we developed an original, high sensitive 'redox-flexible' NADH amperometric bioassay working in a large window of applied potentials in both oxidation and reduction modes. The biosensor has a continuous and complementary linearity range permitting to measure NADH concentrations starting from 5 x 10 -6 M in reduction until 2 x 10 3 M in oxidation. This redox-flexibility allows choosing the applied potential in order to avoid electrochemical interferences. The association of the 'redox-flexible' concept with NADH dependent enzymes opens a novel strategy for dehydrogenases based bioassays and biosensors. The great number of dehydrogenases available makes the concept applicable for numerous substrates to analyse. Moreover it allows the development of a wide range of biosensors on the basis of a generic platform. This gives several advantages over the previous manufacturing techniques and offers a general and flexible scheme for the fabrication of biosensors presenting high sensitivities, wide calibration ranges and less affected by electrochemical interferences

  20. Guidance document for prepermit bioassay testing of low-level radioactive waste

    International Nuclear Information System (INIS)

    Anderson, S.L.; Harrison, F.L.

    1990-11-01

    In response to the mandate of Public Law 92-532, the Marine Protection, Research, and Sanctuaries Act (MPRSA) of 1972, as amended, the Environmental Protection Agency (EPA) has developed a program to promulgate regulations and criteria to control the ocean disposal of radioactive wastes. The EPA seeks to understand the mechanisms for biological response of marine organisms to the low levels of radioactivity that may arise from the release of these wastes as a result of ocean-disposal practices. Such information will play an important role in determining the adequacy of environmental assessments provided to the EPA in support of any disposal permit application. Although the EPA requires packaging of low-level radioactive waste to prevent release during radiodecay of the materials, some release of radioactive material into the deep-sea environment may occur when a package deteriorates. Therefore, methods for evaluating the impact on biota are being evaluated. Mortality and phenotypic responses are not anticipated at the expected low environmental levels that might occur if radioactive materials were released from the low-level waste packages. Therefore, traditional bioassay systems are unsuitable for assessing sublethal effects on biota in the marine environment. The EPA Office of Radiation Programs (ORP) has had an ongoing program to examine sublethal responses to radiation at the cellular level, using cytogenetic end points. This technical guidance report represents prepermit bioassay procedures that potentially may be applicable to the assessment of effects from a mixture of radionuclides that could be released from a point source at the ocean bottom. Methodologies along with rationale and a discussion of uncertainty are presented for the sediment benthic bioassay protocols identified in this report

  1. GHSI emergency radionuclide bioassay laboratory network - summary of the second exercise

    International Nuclear Information System (INIS)

    Li, Chunsheng; Ko, Raymond; Quayle, Debora; Sadi, Baki; Bartizel, Christine; Battisti, Paolo; Boettger, Axel; Bouvier, Celine; Paquet, Francois; CapoteCuellar, Antonio; Carr, Zhanat; Hammond, Derek; Hartmann, Martina; Heikkinen, Tarja; Jones, Robert L.; Kim, Eunjoo; Koga, Roberto; Kukhta, Boris; Mitchell, Lorna; Morhard, Ryan; Rulik, Petr; Sergei, Aleksanin; Sierra, Inmaculada; Oliveira Sousa, Wandersonde; Szabo, Gyula

    2017-01-01

    The Global Health Security Initiative (GHSI) established a laboratory network within the GHSI community to develop collective surge capacity for radionuclide bioassay in response to a radiological or nuclear emergency as a means of enhancing response capability, health outcomes and community resilience. GHSI partners conducted an exercise in collaboration with the WHO Radiation Emergency Medical Preparedness and Assistance Network and the IAEA Response and Assistance Network, to test the participating laboratories (18) for their capabilities in in vitro assay of biological samples, using a urine sample spiked with multiple high-risk radionuclides ( 90 Sr, 106 Ru, 137 Cs, and 239 Pu). Laboratories were required to submit their reports within 72 h following receipt of the sample, using a pre-formatted template, on the procedures, methods and techniques used to identify and quantify the radionuclides in the sample, as well as the bioassay results with a 95% confidence interval. All of the participating laboratories identified and measured all or some of the radionuclides in the sample. However, gaps were identified in both the procedures used to assay multiple radionuclides in one sample, as well as in the methods or techniques used to assay specific radionuclides in urine. Two-third of the participating laboratories had difficulties in determining all the radionuclides in the sample. Results from this exercise indicate that challenges remain with respect to ensuring that results are delivered in a timely, consistent and reliable manner to support medical interventions. Laboratories within the networks are encouraged to work together to develop and maintain collective capabilities and capacity for emergency bioassay, which is an important component of radiation emergency response. (authors)

  2. Guidance document for prepermit bioassay testing of low-level radioactive waste

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, S.L.; Harrison, F.L.

    1990-11-01

    In response to the mandate of Public Law 92-532, the Marine Protection, Research, and Sanctuaries Act (MPRSA) of 1972, as amended, the Environmental Protection Agency (EPA) has developed a program to promulgate regulations and criteria to control the ocean disposal of radioactive wastes. The EPA seeks to understand the mechanisms for biological response of marine organisms to the low levels of radioactivity that may arise from the release of these wastes as a result of ocean-disposal practices. Such information will play an important role in determining the adequacy of environmental assessments provided to the EPA in support of any disposal permit application. Although the EPA requires packaging of low-level radioactive waste to prevent release during radiodecay of the materials, some release of radioactive material into the deep-sea environment may occur when a package deteriorates. Therefore, methods for evaluating the impact on biota are being evaluated. Mortality and phenotypic responses are not anticipated at the expected low environmental levels that might occur if radioactive materials were released from the low-level waste packages. Therefore, traditional bioassay systems are unsuitable for assessing sublethal effects on biota in the marine environment. The EPA Office of Radiation Programs (ORP) has had an ongoing program to examine sublethal responses to radiation at the cellular level, using cytogenetic end points. This technical guidance report represents prepermit bioassay procedures that potentially may be applicable to the assessment of effects from a mixture of radionuclides that could be released from a point source at the ocean bottom. Methodologies along with rationale and a discussion of uncertainty are presented for the sediment benthic bioassay protocols identified in this report.

  3. Fast and accurate semantic annotation of bioassays exploiting a hybrid of machine learning and user confirmation.

    Science.gov (United States)

    Clark, Alex M; Bunin, Barry A; Litterman, Nadia K; Schürer, Stephan C; Visser, Ubbo

    2014-01-01

    Bioinformatics and computer aided drug design rely on the curation of a large number of protocols for biological assays that measure the ability of potential drugs to achieve a therapeutic effect. These assay protocols are generally published by scientists in the form of plain text, which needs to be more precisely annotated in order to be useful to software methods. We have developed a pragmatic approach to describing assays according to the semantic definitions of the BioAssay Ontology (BAO) project, using a hybrid of machine learning based on natural language processing, and a simplified user interface designed to help scientists curate their data with minimum effort. We have carried out this work based on the premise that pure machine learning is insufficiently accurate, and that expecting scientists to find the time to annotate their protocols manually is unrealistic. By combining these approaches, we have created an effective prototype for which annotation of bioassay text within the domain of the training set can be accomplished very quickly. Well-trained annotations require single-click user approval, while annotations from outside the training set domain can be identified using the search feature of a well-designed user interface, and subsequently used to improve the underlying models. By drastically reducing the time required for scientists to annotate their assays, we can realistically advocate for semantic annotation to become a standard part of the publication process. Once even a small proportion of the public body of bioassay data is marked up, bioinformatics researchers can begin to construct sophisticated and useful searching and analysis algorithms that will provide a diverse and powerful set of tools for drug discovery researchers.

  4. Fast and accurate semantic annotation of bioassays exploiting a hybrid of machine learning and user confirmation

    Directory of Open Access Journals (Sweden)

    Alex M. Clark

    2014-08-01

    Full Text Available Bioinformatics and computer aided drug design rely on the curation of a large number of protocols for biological assays that measure the ability of potential drugs to achieve a therapeutic effect. These assay protocols are generally published by scientists in the form of plain text, which needs to be more precisely annotated in order to be useful to software methods. We have developed a pragmatic approach to describing assays according to the semantic definitions of the BioAssay Ontology (BAO project, using a hybrid of machine learning based on natural language processing, and a simplified user interface designed to help scientists curate their data with minimum effort. We have carried out this work based on the premise that pure machine learning is insufficiently accurate, and that expecting scientists to find the time to annotate their protocols manually is unrealistic. By combining these approaches, we have created an effective prototype for which annotation of bioassay text within the domain of the training set can be accomplished very quickly. Well-trained annotations require single-click user approval, while annotations from outside the training set domain can be identified using the search feature of a well-designed user interface, and subsequently used to improve the underlying models. By drastically reducing the time required for scientists to annotate their assays, we can realistically advocate for semantic annotation to become a standard part of the publication process. Once even a small proportion of the public body of bioassay data is marked up, bioinformatics researchers can begin to construct sophisticated and useful searching and analysis algorithms that will provide a diverse and powerful set of tools for drug discovery researchers.

  5. Bioassay of circulating luteinizing hormone in the rhesus monkey: comparison with radioimmunoassay during physiological changes

    International Nuclear Information System (INIS)

    Dufau, M.L.; Hodgen, G.D.; Goodman, A.L.; Catt, K.J.

    1977-01-01

    The concentration of biologically active LH in Rhesus monkey (Macaca mulatta) serum was measured by a highly sensitive bioassay based upon testosterone production by dispersed rat interstitial cells. The sensitivity of the in vitro bioassay was equal to or higher than that of radioimmunoassay, with detection limits of 0.1 mIU of human menopausal gonadotropin (hMG) or 10 ng of a Rhesus pituitary gonadotropin preparation (LER-1909-2). Parallel dose-response curves were obtained for hMG and Rhesus monkey pituitary gonadotropin. The method permits bioassay of LH in 20--100 μl of serum from adult male monkeys, and from female monkeys during the follicular and luteal phases of the menstrual cycle. Bioactive LH concentrations could be assayed in 0.25 to 5 μl of serum from mid-cycle, postmenopausal, and castrated female monkeys. Serum LH was undetectable in two hypophysectomized adult female monkeys and six intact immature animals, and was 13 +- 6 (SD) mIU/ml in adult male monkeys. In adult females, follicular phase LH levels ranged from 17 to 169 mIU/ml, with a mean of 76 +- 52 mIU/ml. The midcycle LH peak was 1738 +- 742 mIU/ml and the luteal phase values ranged from 6-47 mIU/ml, with a mean of 35 +- 5 mIU/ml. Serum LH concentrations ranged from 100 to 900 mIU/ml in two menopausal females, and from 590--1480 mIU/ml in castrated females. Treatment of castrated female monkeys with estrogen plus progesterone produced an initial two-fold rise in sepum LH within 3 days, followed by a gradual decline to one-fourth to one-tenth of the initial levels after 10 days of treatment. Serum LH was suppressed to undetectable levels during the third week, and remained so for the duration of the 60-day treatment period

  6. Toxicity of Single and Mixed Contaminants in Seawater Measured with Acute Toxicity Bioassays

    Directory of Open Access Journals (Sweden)

    A.R. Fernandez-Alba

    2002-01-01

    Full Text Available Different types of organic pollutants commonly detected in seawater have been evaluated by acute toxicity bioassays. Vibrio fischeri, Daphnia magna, and Selenastrum capricornotum were selected to test toxic effects of individual compounds and mixtures of these compounds, obtaining EC50 values in the range of 0.001 to 28.9 mg/l. In the case of mixtures, synergistic toxic responses were seen for a clear majority of the cases (>60%. Mixtures containing methyl-tertiary-butyl ether (MTBE exhibit accelerated processes that result in a change in concentration required to produce a toxic effect; for example, in the case of mixtures containing MTBE and Diuron and Dichlofluanid.

  7. 'In-vivo' and bioassay results from two contrasting cases of plutonium-239 inhalation

    International Nuclear Information System (INIS)

    Ramsden, D.; Bains, M.E.D.; Fraser, D.C.

    1969-06-01

    'In-vivo' and bioassay measurements following two incidents involving plutonium-239 inhalation are described and contrasted. Incident 1, involving the inhalation of insoluble plutonium oxide, resulted in a lung content of about 20 nCi after the initial clearance. Urine excretion was negligible and the estimation of exposure was based on 'in-vivo' data and faecal excretion. Incident,2, involving the inhalation of soluble plutonium, proved negligible and the estimation of exposure, based on urinary excretion, was 0.6 nCi. (author)

  8. A Bayesian approach to the analysis of quantal bioassay studies using nonparametric mixture models.

    Science.gov (United States)

    Fronczyk, Kassandra; Kottas, Athanasios

    2014-03-01

    We develop a Bayesian nonparametric mixture modeling framework for quantal bioassay settings. The approach is built upon modeling dose-dependent response distributions. We adopt a structured nonparametric prior mixture model, which induces a monotonicity restriction for the dose-response curve. Particular emphasis is placed on the key risk assessment goal of calibration for the dose level that corresponds to a specified response. The proposed methodology yields flexible inference for the dose-response relationship as well as for other inferential objectives, as illustrated with two data sets from the literature. © 2013, The International Biometric Society.

  9. Analysis of polonium-210 in food products and bioassay samples by isotope-dilution alpha spectrometry

    International Nuclear Information System (INIS)

    Lin Zhichao; Wu Zhongyu

    2009-01-01

    A rapid and reliable radiochemical method coupled with a simple and compact plating apparatus was developed, validated, and applied for the analysis of 210 Po in variety of food products and bioassay samples. The method performance characteristics, including accuracy, precision, robustness, and specificity, were evaluated along with a detailed measurement uncertainty analysis. With high Po recovery, improved energy resolution, and effective removal of interfering elements by chromatographic extraction, the overall method accuracy was determined to be better than 5% with measurement precision of 10%, at 95% confidence level.

  10. Analysis of polonium-210 in food products and bioassay samples by isotope-dilution alpha spectrometry.

    Science.gov (United States)

    Lin, Zhichao; Wu, Zhongyu

    2009-05-01

    A rapid and reliable radiochemical method coupled with a simple and compact plating apparatus was developed, validated, and applied for the analysis of (210)Po in variety of food products and bioassay samples. The method performance characteristics, including accuracy, precision, robustness, and specificity, were evaluated along with a detailed measurement uncertainty analysis. With high Po recovery, improved energy resolution, and effective removal of interfering elements by chromatographic extraction, the overall method accuracy was determined to be better than 5% with measurement precision of 10%, at 95% confidence level.

  11. Use of 236Pu and 242Pu as a radiochemical tracer for estimation of Pu in bioassay samples by fission track analysis

    International Nuclear Information System (INIS)

    Sawant, Pramilla D.; Prabhu, Supreetha P.; Kalsi, P.C.

    2008-01-01

    236 Pu and 242 Pu are routinely used as radiochemical yield monitors in India for bioassay monitoring of occupational workers by alpha spectrometry. Fission Track Analysis (FTA) is also being standardized for trace level determination of Pu in bioassay samples. The present study, reports the utility of 236 Pu and 242 Pu as radiochemical tracers in estimation of Pu in bioassay samples by FTA technique. The advantages of using 236 Pu tracer in FTA over 242 Pu as well as the interference caused due to presence of 241 Pu in the bioassay samples of occupational workers handling power reactor grade Pu is discussed. (author)

  12. Single-core magnetic markers in rotating magnetic field based homogeneous bioassays and the law of mass action

    Energy Technology Data Exchange (ETDEWEB)

    Dieckhoff, Jan, E-mail: j.dieckhoff@tu-bs.de [Institut fuer Elektrische Messtechnik und Grundlagen der Elektrotechnik, TU Braunschweig, Braunschweig (Germany); Schrittwieser, Stefan; Schotter, Joerg [Molecular Diagnostics, AIT Austrian Institute of Technology, Vienna (Austria); Remmer, Hilke; Schilling, Meinhard; Ludwig, Frank [Institut fuer Elektrische Messtechnik und Grundlagen der Elektrotechnik, TU Braunschweig, Braunschweig (Germany)

    2015-04-15

    In this work, we report on the effect of the magnetic nanoparticle (MNP) concentration on the quantitative detection of proteins in solution with a rotating magnetic field (RMF) based homogeneous bioassay. Here, the phase lag between 30 nm iron oxide single-core particles and the RMF is analyzed with a fluxgate-based measurement system. As a test analyte anti-human IgG is applied which binds to the protein G functionalized MNP shell and causes a change of the phase lag. The measured phase lag changes for a fixed MNP and a varying analyte concentration are modeled with logistic functions. A change of the MNP concentration results in a nonlinear shift of the logistic function with the analyte concentration. This effect results from the law of mass action. Furthermore, the bioassay results are used to determine the association constant of the binding reaction. - Highlights: • A rotating magnetic field based homogeneous bioassay concept was presented. • Here, single-core iron oxide nanoparticles are applied as markers. • The impact of the particle concentration on the bioassay results is investigated. • The relation between particle concentration and bioassay sensitivity is nonlinear. • This finding can be reasonably explained by the law of mass action.

  13. Establishing principal soil quality parameters influencing earthworms in urban soils using bioassays

    International Nuclear Information System (INIS)

    Hankard, Peter K.; Bundy, Jacob G.; Spurgeon, David J.; Weeks, Jason M.; Wright, Julian; Weinberg, Claire; Svendsen, Claus

    2005-01-01

    Potential contamination at ex-industrial sites means that, prior to change of use, it will be necessary to quantify the extent of risks to potential receptors. To assess ecological hazards, it is often suggested to use biological assessment to augment chemical analyses. Here we investigate the potential of a commonly recommended bioassay, the earthworm reproduction test, to assess the status of urban contaminated soils. Sample points at all study sites had contaminant concentrations above the Dutch soil criteria Target Values. In some cases, the relevant Intervention Values were exceeded. Earthworm survival at most points was high, but reproduction differed significantly in soil from separate patches on the same site. When the interrelationships between soil parameters and reproduction were studied, it was not possible to create a good model of site soil toxicity based on single or even multiple chemical measurements of the soils. We thus conclude that chemical analysis alone is not sufficient to characterize soil quality and confirms the value of biological assays for risk assessment of potentially contaminated soils. - Bioassays must be applied for the risk assessment complexly-polluted sites to complement chemical analysis of soils

  14. Estimation of uranium in bioassay samples of occupational workers by laser fluorimetry

    International Nuclear Information System (INIS)

    Suja, A.; Prabhu, S.P.; Sawant, P.D.; Sarkar, P.K.; Tiwari, A.K.; Sharma, R.

    2012-01-01

    A newly established uranium processing facility has been commissioned at BARC, Trombay. Monitoring of occupational workers is essential to assess intake of uranium in this facility. A group of 21 workers was selected for bioassay monitoring to assess the existing urinary excretion levels of uranium before the commencement of actual work. Bioassay samples collected from these workers were analyzed by ion-exchange technique followed by laser fluorimetry. Standard addition method was followed for estimation of uranium concentration in the samples. The minimum detectable activity by this technique is about 0.2 ng. The range of uranium observed in these samples varies from 19 to 132 ng/L. Few of these samples were also analyzed by fission track analysis technique and the results were found to be comparable to those obtained by laser fluorimetry. The urinary excretion rate observed for the individual can be regarded as a 'personal baseline' and will be treated as the existing level of uranium in urine for these workers at the facility. (author)

  15. Screening the Toxicity of Selected Personal Care Products Using Embryo Bioassays: 4-MBC, Propylparaben and Triclocarban

    Directory of Open Access Journals (Sweden)

    Tiago Torres

    2016-10-01

    Full Text Available Recently, several emerging pollutants, including Personal Care Products (PCPs, have been detected in aquatic ecosystems, in the ng/L or µg/L range. Available toxicological data is limited, and, for certain PCPs, evidence indicates a potential risk for the environment. Hence, there is an urgent need to gather ecotoxicological data on PCPs as a proxy to improve risk assessment. Here, the toxicity of three different PCPs (4-Methylbenzylidene Camphor (4-MBC, propylparaben and triclocarban was tested using embryo bioassays with Danio rerio (zebrafish and Paracentrotus lividus (sea urchin. The No Observed Effect Concentration (NOEC for triclocarban was 0.256 µg/L for sea urchin and 100 µg/L for zebrafish, whereas NOEC for 4-MBC was 0.32 µg/L for sea urchin and 50 µg/L for zebrafish. Both PCPs impacted embryo development at environmentally relevant concentrations. In comparison with triclocarban and 4-MBC, propylparaben was less toxic for both sea urchin (NOEC = 160 µg/L and zebrafish (NOEC = 1000 µg/L. Overall, this study further demonstrates the sensitivity of embryo bioassays as a high-throughput approach for testing the toxicity of emerging pollutants.

  16. Efficiency Calibration of Phantom Family for Use in Direct Bioassay of Radionuclide in the Body

    International Nuclear Information System (INIS)

    Kim, Ji Seok; Ha, Wi Ho; Kim, Hyun Ki; Park, Gyung Deok; Lee, Jai Ki

    2008-01-01

    A major source of uncertainties of in vivo bioassay using a whole body counter calibrated against a body phantom containing known radioactivities is variation of counting geometry caused by the differences in body size of the subject from that of the phantom. Phantoms such as the BOMAB phantom are based on the body size of the reference man and usually single phantom is used in usual calibration of the counter. This is because it is difficult to apply a set of phantoms having different sizes. In order to reduce the potential errors due to variation of counting geometry, use of a set of phantoms having different body-shapes have been attempted. The efficiency files are stored in the computer analyzing the measurement data and a suitable one is retrieved for the specific subject. Experimental or computational approach can be employed in generation of the efficiency files. Carlan et al. demonstrated that Monte Carlo simulations can provide acceptable efficiencies by use of the IGOR phantom family. The body size of the individual subject undergoing in vivo bioassay should be determined by an appropriate method

  17. Comparative study of Graves' ophthalmopathy by ultrasonography, computed tomography, and fish bioassay

    International Nuclear Information System (INIS)

    Mann, K.; Schoener, W.; Juengst, D.; Karl, H.J.; Maier-Hauff, K.; Rothe, R.

    1979-01-01

    In 35 patients with Graves' ophthalmopathy (GO) thyroid function was tested by T 3 -RIA, T 4 -RIA, TBI, TRH-test, thyroid scanning, and determination of thyroid autoantibodies. Additional ultrasonography (A-scan), computed tomography (CT) of the orbit, and the determination of an exophthalmogenic serum activity in fish bioassay was performed. Typical alterations for GO were observed in 26 cases with ultrasonography. CT showed an enlargement of medial and/or lateral rectus muscles in 24 of 33 patients, and in 17 cases a region of high density in the apex of the muscle cone. The density of retrobulbar fat after i.v. injection of contrast medium did not differ significantly from that observed in normal men. Characteristic signs of GO were not detected in only 2 cases using both methods together. Exophthalmogenic serum activity was found in the IgG fraction of serum protein. The incidence rate was high (69%), but for diagnostic purpose the fish bioassay cannot be recommended. (orig.) 891 AJ/orig. 892 BRE [de

  18. [Quality evaluation of rhubarb dispensing granules based on multi-component simultaneous quantitative analysis and bioassay].

    Science.gov (United States)

    Tan, Peng; Zhang, Hai-Zhu; Zhang, Ding-Kun; Wu, Shan-Na; Niu, Ming; Wang, Jia-Bo; Xiao, Xiao-He

    2017-07-01

    This study attempts to evaluate the quality of Chinese formula granules by combined use of multi-component simultaneous quantitative analysis and bioassay. The rhubarb dispensing granules were used as the model drug for demonstrative study. The ultra-high performance liquid chromatography (UPLC) method was adopted for simultaneously quantitative determination of the 10 anthraquinone derivatives (such as aloe emodin-8-O-β-D-glucoside) in rhubarb dispensing granules; purgative biopotency of different batches of rhubarb dispensing granules was determined based on compound diphenoxylate tablets-induced mouse constipation model; blood activating biopotency of different batches of rhubarb dispensing granules was determined based on in vitro rat antiplatelet aggregation model; SPSS 22.0 statistical software was used for correlation analysis between 10 anthraquinone derivatives and purgative biopotency, blood activating biopotency. The results of multi-components simultaneous quantitative analysisshowed that there was a great difference in chemical characterizationand certain differences inpurgative biopotency and blood activating biopotency among 10 batches of rhubarb dispensing granules. The correlation analysis showed that the intensity of purgative biopotency was significantly correlated with the content of conjugated anthraquinone glycosides (Panalysis and bioassay can achieve objective quantification and more comprehensive reflection on overall quality difference among different batches of rhubarb dispensing granules. Copyright© by the Chinese Pharmaceutical Association.

  19. Bioassay-Guided Isolated Compounds from Morinda officinalis Inhibit Alzheimer’s Disease Pathologies

    Directory of Open Access Journals (Sweden)

    Yoon Kyoung Lee

    2017-09-01

    Full Text Available Due to the side effects of synthetic drugs, the therapeutic potential of natural products for Alzheimer’s disease (AD has gained interest. Morinda officinalis has demonstrated inhibitory effects on geriatric diseases, such as bone loss and osteoporosis. However, although AD is a geriatric disease, M. officinalis has not been evaluated in an AD bioassay. Therefore, M. officinalis extracts and fractions were tested for AD-related activity, including inhibition of acetylcholinesterase (AChE, butyrylcholinesterase (BChE, β-site amyloid precursor protein cleaving enzyme 1 (BACE1, and advanced glycation end-product (AGE formation. A bioassay-guided approach led to isolation of 10 active compounds, eight anthraquinones (1–8, one coumarin (9, and one phytosterol (10, from n-hexane and ethyl acetate fractions of M. officinalis. The five anthraquinones (4–8 were stronger inhibitors of AChE than were other compounds. Compounds 3 and 9 were good inhibitors of BChE, and compounds 3 and 8 were good inhibitors of BACE1. Compounds 1–5 and 7–9 were more active than the positive control in inhibiting AGE formation. In addition, we first suggested a structure-activity relationship by which anthraquinones inhibit AChE and BACE1. Our findings demonstrate the preventive and therapeutic efficacy of M. officinalis for AD and its potential use as a natural alternative medicine.

  20. Evaluation and modeling of the parameters affecting fluoride toxicity level in aquatic environments by bioassay method

    Directory of Open Access Journals (Sweden)

    Hamid Reza Shamsollahi

    2014-04-01

    Full Text Available Background: Fluoride exists in various forms in nature and water resources. , The rising level of fluoride in water resources due to discharge of industrial effluents can cause toxicity in aquatic organisms. To prevent toxicity, it is necessary to determine maximum fluoride toxicity as well as effluent discharge limits. The aim of this study was to determine the maximum fluoride toxicity and the factors affecting fluoride toxicity to provide a model in order to determine the effluent discharge limits. Methods: Daphnia magna bioassay in the absence of confounding factors was used to determine the maximum level of fluoride toxicity. Then, bioassay was repeated in the presence of the confounding factors (hardness, temperature and exposure time to determine their effects. Results: In the absence of intervening factors, fluoride LC50 levels determined after 24, 48 and 72 hours exposure were 4.9, 46.5 and 38.7 mg/l, respectively.. Also, the influence of confounding factors on LC50 values was reported significant by Minitab software. Conclusion: Increasing the water hardness reduced fluoride toxicity, and increasing the water temperature and exposure time increased fluoride toxicity in aquatic environments. Therefore, while determining the wastewater discharge limit in terms of fluoride concentration, it is essential to take the effect of confounding factors on fluoride toxicity into account in order to prevent toxicity in the open water resources.

  1. DOSEXPRT: A bioassay dosimetry code for Martin Marietta Energy Systems, Inc.

    Energy Technology Data Exchange (ETDEWEB)

    Ward, R.C.; Eckerman, K.F.

    1992-04-01

    The bioassay code DOSEXPRT was developed for Martin Marietta Energy Systems, Inc., to provide compliance with Department of Energy (DOE) Order 5480, Chapter 11. DOSEXPRT computes the intake of a radionuclide in any year (considering both acute and chronic intakes) from in vivo measurements of the retained activity and/or measurements of the activity in excreta. The committed effective and organ doses for the intake are computed as well as the effective and organ doses expected to be received in each calendar year out to 50 years beyond the year of intake. The bioassay records used as input for DOSEXPRT are extracted from the Martin Marietta Energy Systems Occupational Health Information System (OHIS). DOSEXPRT implements a set of algorithms with parameters governing the translocation, retention, and excretion of the nuclide contained in data files specific to the nuclide. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent for the intakes in the year. Annual organ and effective doses are computed using additional dose-rate files that contain data on the dose rate at various times following a unit intake. If measurements are presented for more than one assay for a given nuclide, DOSEXPRT estimates the intake by applying weights assigned in the nuclide file for each assay. DOSEXPRT is accessed off the OHIS MENU No. 4 and designed to be run as a batch processor, but can also be run interactively for testing purposes.

  2. DOSEXPRT: A bioassay dosimetry code for Martin Marietta Energy Systems, Inc

    Energy Technology Data Exchange (ETDEWEB)

    Ward, R.C.; Eckerman, K.F.

    1992-04-01

    The bioassay code DOSEXPRT was developed for Martin Marietta Energy Systems, Inc., to provide compliance with Department of Energy (DOE) Order 5480, Chapter 11. DOSEXPRT computes the intake of a radionuclide in any year (considering both acute and chronic intakes) from in vivo measurements of the retained activity and/or measurements of the activity in excreta. The committed effective and organ doses for the intake are computed as well as the effective and organ doses expected to be received in each calendar year out to 50 years beyond the year of intake. The bioassay records used as input for DOSEXPRT are extracted from the Martin Marietta Energy Systems Occupational Health Information System (OHIS). DOSEXPRT implements a set of algorithms with parameters governing the translocation, retention, and excretion of the nuclide contained in data files specific to the nuclide. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent for the intakes in the year. Annual organ and effective doses are computed using additional dose-rate files that contain data on the dose rate at various times following a unit intake. If measurements are presented for more than one assay for a given nuclide, DOSEXPRT estimates the intake by applying weights assigned in the nuclide file for each assay. DOSEXPRT is accessed off the OHIS MENU No. 4 and designed to be run as a batch processor, but can also be run interactively for testing purposes.

  3. Bioassay standardization for the detection of allelopathic compounds and environmental toxicants using lettuce

    Directory of Open Access Journals (Sweden)

    Mateus Salomão Simões

    2013-09-01

    Full Text Available The purpose of this study was to assess different experimental conditions to determine a protocol for bioassays based on seed germination and early seedling growth using lettuce (Lactuca sativa L. cv. Grand Rapids as indicator species. This protocol aims to provide support for the standardization of assays of various chemicals such as allelochemicals and environmental toxicants. The following tests were performed: time of germination, temperature, light, solution volume and Petri dish size. For each test (except for time of germination, the influence of the conditions investigated was determined by the endpoints germination percentage, germination speed index, root length, seedling fresh weight and total dry weight. The results showed that variations in the methods altered the results. It is recommended that bioassays using L. sativa L. cv. Grand Rapids be carried out for a minimum period of four days for assessments of both germination and initial growth and that the experimental conditions include a temperature of 20°C, 90-mm Petri dishes or larger, 0.1 mL cypsela solution, and continuous light or 12-hour photoperiod.

  4. Bioassay and characterization of soil microorganisms involved in the biodegradation of the fungicide, metalaxyl

    International Nuclear Information System (INIS)

    Bailey, A.M.

    1985-01-01

    A sensitive bioassay was developed to detect low concentrations of metalaxyl in soils. The quantitative estimation of metalaxyl in soils was based on a significant positive relationship between the radial growth of Phytophthora boehmeriae and the log concentration of the fungicide in the agar. The isolate of P. boehmeriae was chosen for its sensitivity to metalaxyl as manifested in a linear growth response on cornmeal agar over a range of 2 to 30 ng/ml. The sensitivity and quantitative nature of the bioassay was confirmed by comparison with data obtained by using 14 C-metalaxyl. Metabolism of metalaxyl was detected in three of five avocado soils that had repeated applications of the fungicide over 2-5 yr. The average disappearance of metalaxyl was 28 days, and in the most active soils was 14 days. The composition and level of the microbial populations of soils, either active or inactive in the breakdown of metalaxyl, did not differ. Fungal and bacterial microflora recovered from these two soils by use of either selective media or filtration techniques were capable of metabolizing metalaxyl over a 45-day period

  5. Antiplasmodial Properties and Bioassay-Guided Fractionation of Ethyl Acetate Extracts from Carica papaya Leaves

    Science.gov (United States)

    Melariri, Paula; Campbell, William; Etusim, Paschal; Smith, Peter

    2011-01-01

    We investigated the antiplasmodial properties of crude extracts from Carica papaya leaves to trace the activity through bioassay-guided fractionation. The greatest antiplasmodial activity was observed in the ethyl acetate crude extract. C. papaya showed a high selectivity for P. falciparum against CHO cells with a selectivity index of 249.25 and 185.37 in the chloroquine-sensitive D10 and chloroquine-resistant DD2 strains, respectively. Carica papaya ethyl acetate extract was subjected to bioassay-guided fractionation to ascertain the most active fraction, which was purified and identified using high-pressure liquid chromatography (HPLC) and GC-MS (Gas chromatography-Mass spectrometry) methods. Linoleic and linolenic acids identified from the ethyl acetate fraction showed IC50 of 6.88 μg/ml and 3.58 μg/ml, respectively. The study demonstrated greater antiplasmodial activity of the crude ethyl acetate extract of Carica papaya leaves with an IC50 of 2.96 ± 0.14 μg/ml when compared to the activity of the fractions and isolated compounds. PMID:22174990

  6. Identification and characterization of antifungal compounds using a Saccharomyces cerevisiae reporter bioassay.

    Directory of Open Access Journals (Sweden)

    Brad Tebbets

    Full Text Available New antifungal drugs are urgently needed due to the currently limited selection, the emergence of drug resistance, and the toxicity of several commonly used drugs. To identify drug leads, we screened small molecules using a Saccharomyces cerevisiae reporter bioassay in which S. cerevisiae heterologously expresses Hik1, a group III hybrid histidine kinase (HHK from Magnaporthe grisea. Group III HHKs are integral in fungal cell physiology, and highly conserved throughout this kingdom; they are absent in mammals, making them an attractive drug target. Our screen identified compounds 13 and 33, which showed robust activity against numerous fungal genera including Candida spp., Cryptococcus spp. and molds such as Aspergillus fumigatus and Rhizopus oryzae. Drug-resistant Candida albicans from patients were also highly susceptible to compounds 13 and 33. While the compounds do not act directly on HHKs, microarray analysis showed that compound 13 induced transcripts associated with oxidative stress, and compound 33, transcripts linked with heavy metal stress. Both compounds were highly active against C. albicans biofilm, in vitro and in vivo, and exerted synergy with fluconazole, which was inactive alone. Thus, we identified potent, broad-spectrum antifungal drug leads from a small molecule screen using a high-throughput, S. cerevisiae reporter bioassay.

  7. In vitro bioassays reveal that additives are significant contributors to the toxicity of commercial household pesticides.

    Science.gov (United States)

    van de Merwe, Jason P; Neale, Peta A; Melvin, Steven D; Leusch, Frederic D L

    2018-06-01

    Pesticides commonly used around households can contain additives of unknown concentrations and toxicity. Given the likelihood of these chemicals washing into urban waterways, it is important to understand the effects that these additives may have on aquatic organisms. The aim of this study was to compare the toxicity of commercially available household pesticides to that of the active ingredient(s) alone. The toxicity of five household pesticides (three herbicides and two insecticides) was investigated using a bacterial cytotoxicity bioassay and an algal photosynthesis bioassay. The commercial products were up to an order of magnitude more toxic than the active ingredient(s) alone. In addition, two commercial products with the same listed active ingredients in the same ratio had a 600× difference in potency. These results clearly demonstrate that additives in commercial formulations are significant contributors to the toxicity of household pesticides. The toxicity of pesticides in aquatic systems is therefore likely underestimated by conventional chemical monitoring and risk assessment when only the active ingredients are considered. Regulators and customers should require more clarity from pesticide manufacturers about the nature and concentrations of not only the active ingredients, but also additives used in commercial formulations. In addition, monitoring programmes and chemical risk assessments schemes should develop a structured approach to assessing the toxic effects of commercial formulations, including additives, rather than simply those of the listed active ingredients. Copyright © 2018. Published by Elsevier B.V.

  8. Use of a germination bioassay to test compost maturity in Tekelan Village

    Science.gov (United States)

    Oktiawan, Wiharyanto; Zaman, Badrus; Purwono

    2018-02-01

    Livestock waste from cattle farms in Tekelan village, Getasan Subdistrict, Semarang Regency can be grouped into three types, namely solid waste, slurry and waste water. Solid waste (cow dung) was processed into compost, while slurry and waste water were used to make liquid fertilizer. This compost was used as a component of planting media in horticultural crops and potted plants production. We evaluated the toxicity (phytochemical and ecotoxicological) test of compost by using germination index (GI). Vigna radiata seeds are sown on filter paper dampened with compost extract for different times. GI was calculated by relative germination (G) and relative radical length (L). The germination index (GI) = G / G0 x L / L0 x 100, where G0 and L0 are values obtained by distilled water as a control. The results showed that germination bioassay and radical length using aquades and groundwater in Tekelan village did not affect the radical length of Vigna radiata . Technically, groundwater in Tekelan village can be used as a germination bioassay control. The cow dung compost substrate appears to have a major influence on compost toxicity. Mature compost was produced on day 14 with a GI of 104.03.

  9. Verifying Identities of Plant-Based Multivitamins Using Phytochemical Fingerprinting in Combination with Multiple Bioassays.

    Science.gov (United States)

    Lim, Yeni; Ahn, Yoon Hee; Yoo, Jae Keun; Park, Kyoung Sik; Kwon, Oran

    2017-09-01

    Sales of multivitamins have been growing rapidly and the concept of natural multivitamin, plant-based multivitamin, or both has been introduced in the market, leading consumers to anticipate additional health benefits from phytochemicals that accompany the vitamins. However, the lack of labeling requirements might lead to fraudulent claims. Therefore, the objective of this study was to develop a strategy to verify identity of plant-based multivitamins. Phytochemical fingerprinting was used to discriminate identities. In addition, multiple bioassays were performed to determine total antioxidant capacity. A statistical computation model was then used to measure contributions of phytochemicals and vitamins to antioxidant activities. Fifteen multivitamins were purchased from the local markets in Seoul, Korea and classified into three groups according to the number of plant ingredients. Pearson correlation analysis among antioxidant capacities, amount phenols, and number of plant ingredients revealed that ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picryhydrazyl (DPPH) assay results had the highest correlation with total phenol content. This suggests that FRAP and DPPH assays are useful for characterizing plant-derived multivitamins. Furthermore, net effect linear regression analysis confirmed that the contribution of phytochemicals to total antioxidant capacities was always relatively higher than that of vitamins. Taken together, the results suggest that phytochemical fingerprinting in combination with multiple bioassays could be used as a strategy to determine whether plant-derived multivitamins could provide additional health benefits beyond their nutritional value.

  10. Evaluating the performance of the ORTECR DetectiveTM for emergency urine bioassay

    International Nuclear Information System (INIS)

    Li, C.; Ko, R.; Moodie, G.; Kramer, G. H.

    2011-01-01

    The performance of the ORTEC R Detective TM as a field deployable tool for emergency urine bioassay of 137 Cs, 60 Co, 192 Ir, 169 Yb and 75 Se was evaluated against ANSI N13.30. The tested activity levels represent 10 % RL (reference level) and 1 % RL defined by [Li C., Vlahovich S., Dai X., Richardson R. B., Daka J. N. and Kramer G. H. Requirements for radiation emergency urine bioassay techniques for the public and first responders. Health Phys (in press, 99(5), 702-707 (2010)]. The tests were conducted for both single radionuclide and mixed radionuclides at two geometries, one conventional geometry (CG) and one improved geometry (IG) which improved the MDAs (minimum detectable amounts) by a factor of 1.6-2.7. The most challenging radionuclide was 169 Yb. The measurement of the mixture radionuclides for 169 Yb at the CG did not satisfy the ANSI N13.30 requirements even at 10 % RL. At 1 % RL, 169 Yb and 192 Ir were not detectable at either geometry, while the measurement of 60 Co in the mixed radionuclides satisfied the ANSI N13.30 requirements only at the IG. (authors)

  11. Relative proportions of polycyclic aromatic hydrocarbons differ between accumulation bioassays and chemical methods to predict bioavailability

    Energy Technology Data Exchange (ETDEWEB)

    Gomez-Eyles, Jose L., E-mail: j.l.gomezeyles@reading.ac.u [University of Reading, School of Human and Environmental Sciences, Department of Soil Science, Reading RG6 6DW, Berkshire (United Kingdom); Collins, Chris D.; Hodson, Mark E. [University of Reading, School of Human and Environmental Sciences, Department of Soil Science, Reading RG6 6DW, Berkshire (United Kingdom)

    2010-01-15

    Chemical methods to predict the bioavailable fraction of organic contaminants are usually validated in the literature by comparison with established bioassays. A soil spiked with polycyclic aromatic hydrocarbons (PAHs) was aged over six months and subjected to butanol, cyclodextrin and tenax extractions as well as an exhaustive extraction to determine total PAH concentrations at several time points. Earthworm (Eisenia fetida) and rye grass root (Lolium multiflorum) accumulation bioassays were conducted in parallel. Butanol extractions gave the best relationship with earthworm accumulation (r{sup 2} <= 0.54, p <= 0.01); cyclodextrin, butanol and acetone-hexane extractions all gave good predictions of accumulation in rye grass roots (r{sup 2} <= 0.86, p <= 0.01). However, the profile of the PAHs extracted by the different chemical methods was significantly different (p < 0.01) to that accumulated in the organisms. Biota accumulated a higher proportion of the heavier 4-ringed PAHs. It is concluded that bioaccumulation is a complex process that cannot be predicted by measuring the bioavailable fraction alone. - The ability of chemical methods to predict PAH accumulation in Eisenia fetida and Lolium multiflorum was hindered by the varied metabolic fate of the different PAHs within the organisms.

  12. A bioassay for the detection of benzimidazoles reveals their presence in a range of environmental samples

    Directory of Open Access Journals (Sweden)

    Terence S Crofts

    2014-11-01

    Full Text Available Cobamides are a family of enzyme cofactors that include vitamin B12 (cobalamin and are produced solely by prokaryotes. Structural variability in the lower axial ligand has been observed in cobamides produced by diverse organisms. Of the three classes of lower ligands, the benzimidazoles are uniquely found in cobamides, whereas the purine and phenolic bases have additional biological functions. Many organisms acquire cobamides by salvaging and remodeling cobamides or their precursors from the environment. These processes require free benzimidazoles for incorporation as lower ligands, though the presence of benzimidazoles in the environment has not been previously investigated. Here, we report a new purification method and bioassay to measure the total free benzimidazole content of samples from microbial communities and laboratory media components. The bioassay relies on the calcofluor-bright phenotype of a bluB mutant of the model cobalamin-producing bacterium Sinorhizobium meliloti. The concentrations of individual benzimidazoles in these samples were measured by liquid chromatography-tandem mass spectrometry. Several benzimidazoles were detected in subpicomolar to subnanomolar concentrations in host-associated and environmental samples. In addition, benzimidazoles were found to be common contaminants of laboratory media components. These results suggest that benzimidazoles present in the environment and in laboratory media have the potential to influence microbial metabolic activities.

  13. Mimicking Daphnia magna bioassay performance by an electronic tongue for urban water quality control

    Energy Technology Data Exchange (ETDEWEB)

    Kirsanov, Dmitry, E-mail: d.kirsanov@gmail.com [Laboratory of Chemical Sensors, St. Petersburg State University, St. Petersburg (Russian Federation); Laboratory of Artificial Sensor Systems, ITMO University, St. Petersburg (Russian Federation); Legin, Evgeny [Laboratory of Artificial Sensor Systems, ITMO University, St. Petersburg (Russian Federation); Sensor Systems LLC, St. Petersburg (Russian Federation); Zagrebin, Anatoly; Ignatieva, Natalia; Rybakin, Vladimir [Institute of Limnology, Russian Academy of Sciences, St. Petersburg (Russian Federation); Legin, Andrey [Laboratory of Chemical Sensors, St. Petersburg State University, St. Petersburg (Russian Federation); Laboratory of Artificial Sensor Systems, ITMO University, St. Petersburg (Russian Federation)

    2014-05-01

    Highlights: • -Daphnia magna bioassay can be simulated with multisensor system. • Urban water toxicity can be predicted from potentiometric ET data. • Independent test set validation confirms statistical significance of the results. - Abstract: Toxicity is one of the key parameters of water quality in environmental monitoring. However, being evaluated as a response of living beings (as their mobility, fertility, death rate, etc.) to water quality, toxicity can only be assessed with the help of these living beings. This imposes certain restrictions on toxicity bioassay as an analytical method: biotest organisms must be properly bred, fed and kept under strictly regulated conditions and duration of tests can be quite long (up to several days), thus making the whole procedure the prerogative of the limited number of highly specialized laboratories. This report describes an original application of potentiometric multisensor system (electronic tongue) when the set of electrochemical sensors was calibrated against Daphnia magna death rate in order to perform toxicity assessment of urban waters without immediate involvement of living creatures. PRM (partial robust M) and PLS (projections on latent structures) regression models based on the data from this multisensor system allowed for prediction of toxicity of unknown water samples in terms of biotests but in the fast and simple instrumental way. Typical errors of water toxicity predictions were below 20% in terms of Daphnia death rate which can be considered as a good result taking into account the complexity of the task.

  14. High performance wash-free magnetic bioassays through microfluidically enhanced particle specificity.

    Science.gov (United States)

    Bechstein, Daniel J B; Lee, Jung-Rok; Ooi, Chin Chun; Gani, Adi W; Kim, Kyunglok; Wilson, Robert J; Wang, Shan X

    2015-06-30

    Magnetic biosensors have emerged as a sensitive and versatile platform for high performance medical diagnostics. These magnetic biosensors require well-tailored magnetic particles as detection probes, which need to give rise to a large and specific biological signal while showing very low nonspecific binding. This is especially important in wash-free bioassay protocols, which do not require removal of particles before measurement, often a necessity in point of care diagnostics. Here we show that magnetic interactions between magnetic particles and magnetized sensors dramatically impact particle transport and magnetic adhesion to the sensor surfaces. We investigate the dynamics of magnetic particles' biomolecular binding and magnetic adhesion to the sensor surface using microfluidic experiments. We elucidate how flow forces can inhibit magnetic adhesion, greatly diminishing or even eliminating nonspecific signals in wash-free magnetic bioassays, and enhancing signal to noise ratios by several orders of magnitude. Our method is useful for selecting and optimizing magnetic particles for a wide range of magnetic sensor platforms.

  15. Bioassay of Lake Onego bottom sediments toxicity based on their chemical composition and deepwater macrozoobenthos state

    Directory of Open Access Journals (Sweden)

    Kalinkina Nataliya Michailovna

    2017-03-01

    Full Text Available The bioassay of the toxicity of bottom sediments sampled in different areas of Lake Onega was carried out by crustaceans biotesting (Ceriodaphnia affinis Lillijeborg. It was shown that in the most areas of Lake Onega there are non-toxic bottom sediments. Toxic bottom sediments were found in Kondopogskaya Bay, intensively polluted with pulp-and-paper mill wastewaters. For the first time in the deep central part of Lake Onega the area was revealed where the toxic bottom sediments contain a high content of iron, manganese and other trace elements typical for the central areas of the lake. The mapping of the bottom of Lake Onega was accomplished, and three zones were identified based on the analysis of the data concerning the chemical composition of bottom sediments, bioassay toxicity data and the results of the deepwater macrozoobenthos assessment. For each zone the parameters of the main groups of benthos (Amphipoda, Oligochaeta, Chironomidae were defined. The first zone is located in the area of intensive anthropogenic influence (Kondopogskaya Bay, Petrozavodskaya Bay, Povenets Bay, Kizhi Skerries. The second zone is located mostly in the deep part of Petrozavodskaya Bay, where the most intensive development of amphipods is observed. The third area is identified for the first time: it is located in the central deep part of Lake Onega, where the communities of macrozoobenthos are limited by a natural toxic factor.

  16. Intemational collaborative study on the preparation of 1st international standard for rhTSH for bioassay

    International Nuclear Information System (INIS)

    Huang Ying; Shen Hongzheng; Yu Ting; Xu Ligen

    2007-01-01

    The history of the international collaborative studies on the preparation of standards of TSH for bioassay and immunoassay was reviewed. The result of collaborative study on the 1st international standard for thyroid-stimulating hormone, recombinant, human, for bioassay was reported in detail in this article. Based on the results of this collaborative study, it is proposed that the candidate standard be established as the international standard for rhTSH for bioassay, and be assigned an activity of 9.5 IU per ampoule. The national standard preparation of TSH for immunoassay was also reassayed, revealing the potency to be 0.557 mIU/ampoule, i.e. 92. 8% of the labelled value of 0.600mIU/ampoule, a reasonable consistency. (authors)

  17. Optimization and field use of a bioassay to monitor sea lice Lepeophtheirus salmonis sensitivity to emamectin benzoate.

    Science.gov (United States)

    Westcott, Jillian D; Stryhn, Henrik; Burka, John F; Hammell, K Larry

    2008-04-01

    A bioassay for sea lice Lepeophtheirus salmonis sensitivity towards emamectin benzoate (EMB) was validated for field use. A probit regression model with natural responsiveness was used for the number of affected (moribund or dead) sea lice in bioassays involving different concentrations of EMB. Bioassay optimization included an evaluation of the inter-rater reliability of sea lice responsiveness to EMB and an evaluation of gender-related differences in susceptibility. Adoption of a set of bioassay response criteria improved the concordance (evaluated using the concordance correlation coefficient) between raters' assessments and the model estimation of EC50 values (the 'effective concentration' leading to a response of 50% of the lice not prone to natural response). An evaluation of gender-related differences in EMB susceptibility indicated that preadult stage female sea lice exhibited a significantly larger sensitivity towards EMB in 12 of 19 bioassays compared to preadult males. In order to evaluate sea lice sensitivity to EMB in eastern Canada, the intensive salmon farming area in the Bay of Fundy in southwestern New Brunswick was divided into 4 distinct regions based on industry health management practices and hydrographics. A total of 38 bioassays were completed from 2002 to 2005 using populations of preadult stage sea lice collected from Atlantic salmon Salmo salar farms within the 4 described regions. There was no significant overall effect of region or year on EC50 values; however, analysis of variance indicated a significant effect of time of year on EC50 values in 2002 and a potential effect in 2004 to 2005. Although the range of EC50 values obtained in this 3 yr study did not appear sufficient to affect current clinical success in the control of sea lice, the results suggest a seasonal- or temperature-associated variation in sensitivity to EMB. This will need to be considered if changes in EMB efficacy occur in the future.

  18. Contaminated sediments and bioassay responses of three macroinvertebrates, the midge larva Chironomus riparius, the water louse Asellus aquaticus and the mayfly nymph Ephoron virgo

    NARCIS (Netherlands)

    Lange, de H.J.; Haas, de E.M.; Maas, H.; Peeters, E.T.H.M.

    2005-01-01

    Bioassays are widely used to estimate ecological risks of contaminated sediments. We compared the results of three whole sediment bioassays, using the midge larva Chironomus riparius, the water louse Asellus aquaticus, and the mayfly nymph Ephoron virgo. We used sediments from sixteen locations in

  19. In vivo genotoxicity of furan in F344 rats at cancer bioassay doses

    International Nuclear Information System (INIS)

    Ding, Wei; Petibone, Dayton M.; Latendresse, John R.; Pearce, Mason G.; Muskhelishvili, Levan; White, Gene A.; Chang, Ching-Wei; Mittelstaedt, Roberta A.; Shaddock, Joseph G.; McDaniel, Lea P.; Doerge, Daniel R.; Morris, Suzanne M.; Bishop, Michelle E.; Manjanatha, Mugimane G.; Aidoo, Anane; Heflich, Robert H.

    2012-01-01

    Furan, a potent rodent liver carcinogen, is found in many cooked food items and thus represents a human cancer risk. Mechanisms for furan carcinogenicity were investigated in male F344 rats using the in vivo Comet and micronucleus assays, combined with analysis of histopathological and gene expression changes. In addition, formamidopyrimidine DNA glycosylase (Fpg) and endonuclease III (EndoIII)-sensitive DNA damage was monitored as a measure of oxidative DNA damage. Rats were treated by gavage on four consecutive days with 2, 4, and 8 mg/kg bw furan, doses that were tumorigenic in 2-year cancer bioassays, and with two higher doses, 12 and 16 mg/kg. Rats were killed 3 h after the last dose, a time established as producing maximum levels of DNA damage in livers of furan-treated rats. Liver Comet assays indicated that both DNA strand breaks and oxidized purines and pyrimidines increased in a near-linear dose-responsive fashion, with statistically significant increases detected at cancer bioassay doses. No DNA damage was detected in bone marrow, a non-target tissue for cancer, and peripheral blood micronucleus assays were negative. Histopathological evaluation of liver from furan-exposed animals produced evidence of inflammation, single-cell necrosis, apoptosis, and cell proliferation. In addition, genes related to apoptosis, cell-cycle checkpoints, and DNA-repair were expressed at a slightly lower level in the furan-treated livers. Although a mixed mode of action involving direct DNA binding cannot be ruled out, the data suggest that furan induces cancer in rat livers mainly through a secondary genotoxic mechanism involving oxidative stress, accompanied by inflammation, cell proliferation, and toxicity. -- Highlights: ► Furan is a potent rodent liver carcinogen and represents a human cancer risk. ► Furan induces DNA damage in rat liver at cancer bioassay doses. ► Furan induces oxidative stress, inflammation and cell proliferation in rat liver. ► Expression of

  20. Human sperm bioassay has potential in evaluating the quality of cumulus-oocyte complexes.

    Science.gov (United States)

    Hossain, A M; Rizk, B; Huff, C; Helvacioglu, A; Thorneycroft, I H

    1996-01-01

    Human sperm bioassay is routinely used as a quality control check for the culture media. This is one of the three bioassays chosen by the College of American Pathologists (CAP) for interlaboratory proficiency testing to assess the standards of in vitro fertilization (IVF) and andrology laboratories. This study utilized sperm bioassay to assess the quality of cumulus-oocyte complexes (COCs) retrieved in IVF procedures COCs, harvested from the female partner of IVF couples, undergoing identical ovarian stimulation protocols, were individually inseminated with the sperm of the corresponding male partner. Sperm motility in sperm-COC cocultures were compared. Cocultures were established by inseminating the 103 COCs, retrieved from 18 IVF couples with 1 x 10(5) to 2 x 10(5) sperm of the corresponding male partners of the couples. In all 18 cases, the sperm were prepared identically using the Percoll wash method. The cocultures were maintained for 48 h but the oocytes were removed immediately after the fertilization check (approximately 16 h). The motility of sperm in the cocultures and in the insemination stocks were noted and 17 of 18 sperm stocks used for insemination had similar high preinsemination motility (90.2 +/- 5.0%). At 48 h the sperm motility had significantly decreased in the cocultures compared to the insemination stocks; 52.7 +/- 19.9% versus 67.2 +/- 10.4%. There was no difference in the motility among the small, medium, and large COCs (56.4 +/- 24.6%, 52.5 +/- 17.9%, and 50.8 +/- 20.9%, respectively). In 45% of IVF cases, the motility in cocultures varied widely, falling below as well as above that of their corresponding insemination stocks. Furthermore, the sperm motility varied among the cocultures in both pregnant and nonpregnant patients but the extent of variation appears to be greater in the latter. The inter-COC coculture sperm motility variation most likely is due to the differences in the quality of cumulus-oocyte complexes.

  1. Coupling of In Vitro Bioassays with Planar Chromatography in Effect-Directed Analysis.

    Science.gov (United States)

    Weiss, Stefan C; Egetenmeyer, Nicole; Schulz, Wolfgang

    Modern analytical test methods increasingly detect anthropogenic organic substances and their transformation products in water samples and in the environment. The presence of these compounds might pose a risk to the aquatic environment. To determine a possible (eco)toxicological risk, aquatic samples are tested using various bioassays, including sub-organismic assays such as the luminescent bacteria inhibition test, the acetylcholinesterase inhibition test, and the umu-test. The effect-directed analysis (EDA) combines physicochemical separation methods with biological (in vitro) tests. High-performance thin-layer chromatography (HPTLC) has proved to be particularly well suited for the separation of organic compounds and the subsequent analysis of effects by the application of the biotests directly on the surface of the HPTLC plate. The advantage of using HPTLC in comparison to high-performance liquid chromatography (HPLC) for EDA is that the solvent which is used as a mobile phase during chromatography is completely evaporated after the separation and therefore can no longer influence the applied bioassays.A prioritization during the complex identification process can be achieved when observed effects are associated with the separated zones in HPTLC. This increases the probability of identifying the substance responsible for an adverse effect from the multitude of organic trace substances in environmental samples. Furthermore, by comparing the pattern of biological effects of a separated sample, it is possible to track and assess changes in biological activity over time, over space, or in the course of a process, even without identifying the substance. HPTLC has already been coupled with various bioassays.Because HPTLC is a very flexible system, various detection techniques can be used and combined. In addition to the UV/Vis absorption and fluorescence measurements, TLC can also be coupled with a mass spectrometer (MS) for compound identification. In addition

  2. Symposium on Short-Term Genetic Bioassays in the Evaluation of Complex Environmental Mixtures

    CERN Document Server

    Sandhu, Shahbeg; Lewtas, Joellen; Claxton, Larry; Strauss, Gary; Nesnow, Stephen

    1985-01-01

    With this proceedings of the fourth symposium on complex mixtures, we continue to revise and extend our knowledge of genetic methods for the evaluation of chemical mixtures in the environment. The early chapters of this volume are devoted to new bioassay techniques that are directly applicable to the monitoring of environments contaminated with genotoxic chemicals. Microbiological methods have been further refined to meet the special needs of atmospheric monitoring so that very small samples may now be efficiently tested. New in situ methods utilizing green plants actually avoid many of the usual difficulties of sample collection and preparation and offer special advantages in monitoring wastewater, sludges, and hazardous wastes. Insects also are being employed very effectively in the evaluation of gaseous air pollutants in controlled laboratory investigations. Increased emphasis has been placed on a comprehensive assessment of the potential of complex mixtures t9 cause various kinds of genetic damage. New as...

  3. High throughput octal alpha/gamma spectrometer for low level bioassay estimations

    International Nuclear Information System (INIS)

    Bhasin, B.D.; Shirke, S.H.; Suri, M.M.; Vaidya, P.P.; Ghodgaonkar, M.D.

    1995-01-01

    The present paper describes the development of a high throughput octal alpha spectrometry system specially developed for the estimation of low levels of actinides in bioassay and environmental samples. The system processes simultaneously the outputs coming from eight independent detectors. It can be configured to simultaneously record low level alpha and gamma spectra. The high throughput is achieved by using a prioritised multiplexer router. The prioritised multiplexing and routing coupled with fast 8K ADC (conversion time 20 μsec) allow simultaneous acquisition of multiple spectra without any significant loss in counts. The dual (8K, 24bit) port memory facilitates easy online viewing of spectrum buildup. A menu driven user friendly software makes the operating system convenient to use. A specially developed software provides built-in routines for processing the spectra and estimating the isotopic activity. The interactive mode of software provides easy identification of isotopes compatible with the separation chemistry of different actinides. (author). 6 refs., 2 figs

  4. Mixture effects in samples of multiple contaminants - An inter-laboratory study with manifold bioassays.

    Science.gov (United States)

    Altenburger, Rolf; Scholze, Martin; Busch, Wibke; Escher, Beate I; Jakobs, Gianina; Krauss, Martin; Krüger, Janet; Neale, Peta A; Ait-Aissa, Selim; Almeida, Ana Catarina; Seiler, Thomas-Benjamin; Brion, François; Hilscherová, Klára; Hollert, Henner; Novák, Jiří; Schlichting, Rita; Serra, Hélène; Shao, Ying; Tindall, Andrew; Tolefsen, Knut-Erik; Umbuzeiro, Gisela; Williams, Tim D; Kortenkamp, Andreas

    2018-05-01

    Chemicals in the environment occur in mixtures rather than as individual entities. Environmental quality monitoring thus faces the challenge to comprehensively assess a multitude of contaminants and potential adverse effects. Effect-based methods have been suggested as complements to chemical analytical characterisation of complex pollution patterns. The regularly observed discrepancy between chemical and biological assessments of adverse effects due to contaminants in the field may be either due to unidentified contaminants or result from interactions of compounds in mixtures. Here, we present an interlaboratory study where individual compounds and their mixtures were investigated by extensive concentration-effect analysis using 19 different bioassays. The assay panel consisted of 5 whole organism assays measuring apical effects and 14 cell- and organism-based bioassays with more specific effect observations. Twelve organic water pollutants of diverse structure and unique known modes of action were studied individually and as mixtures mirroring exposure scenarios in freshwaters. We compared the observed mixture effects against component-based mixture effect predictions derived from additivity expectations (assumption of non-interaction). Most of the assays detected the mixture response of the active components as predicted even against a background of other inactive contaminants. When none of the mixture components showed any activity by themselves then the mixture also was without effects. The mixture effects observed using apical endpoints fell in the middle of a prediction window defined by the additivity predictions for concentration addition and independent action, reflecting well the diversity of the anticipated modes of action. In one case, an unexpectedly reduced solubility of one of the mixture components led to mixture responses that fell short of the predictions of both additivity mixture models. The majority of the specific cell- and organism

  5. Estimation of 244Cm intake by bioassay measurements following a contamination incident

    International Nuclear Information System (INIS)

    Thein, M.; Bogard, J.S.; Eckerman, K.F.

    1990-01-01

    An employee was contaminated with radioactive material consisting primarily of 244 Cm and 246 Cm as a consequence of handling a curium nitrate solution at a reprocessing facility. In vivo gamma analysis and in vitro (urine and fecal) bioassay measurements were performed. A sample of the curium solution from the workplace was obtained to confirm that the nitrate was the chemical form and to identify the isotopes of curium present. The mass ratio of 244 Cm/ 246 Cm was determined to be 91 to 7. Observed excretion rates were consistent with available information on curium. The results of the in vivo and in vitro measurements are presented and intake estimates for the incident are developed. (author) 11 refs.; 3 figs.; 2 tabs

  6. Ecotoxicological assessment of metal-polluted urban soils using bioassays with three soil invertebrates.

    Science.gov (United States)

    Santorufo, Lucia; Van Gestel, Cornelis A M; Maisto, Giulia

    2012-07-01

    This study aimed at assessing the quality of urban soils by integrating chemical and ecotoxicological approaches. Soils from five sites in downtown Naples, Italy, were sampled and characterized for physical-chemical properties and total and water-extractable metal concentrations. Bioassays with Eisenia andrei, Enchytraeus crypticus and Folsomia candida were performed to assess toxicity of the soils, using survival, reproduction and growth as the endpoints. Metal bioaccumulation in the animals was also measured. The properties and metal concentrations of the soils strongly differed. Metal bioaccumulation was related with total metal concentrations in soil and was highest in E. crypticus, which was more sensitive than E. andrei and F. candida. Responses of the three species to the investigated soils seemed due to both metal contamination and soil properties. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Luciferase-Specific Coelenterazine Analogues for Optical Contamination-Free Bioassays.

    Science.gov (United States)

    Nishihara, Ryo; Abe, Masahiro; Nishiyama, Shigeru; Citterio, Daniel; Suzuki, Koji; Kim, Sung Bae

    2017-04-19

    Spectral overlaps among the multiple optical readouts commonly cause optical contamination in fluorescence and bioluminescence. To tackle this issue, we created five-different lineages of coelenterazine (CTZ) analogues designed to selectively illuminate a specific luciferase with unique luciferase selectivity. In the attempt, we found that CTZ analogues with ethynyl or styryl groups display dramatically biased bioluminescence to specific luciferases and pHs by modifying the functional groups at the C-2 and C-6 positions of the imidazopyradinone backbone of CTZ. The optical contamination-free feature was exemplified with the luciferase-specific CTZ analogues, which illuminated anti-estrogenic and rapamycin activities in a mixture of optical probes. This unique bioluminescence platform has great potential for specific and high throughput imaging of multiple optical readouts in bioassays without optical contamination.

  8. Early-stage bioassay for monitoring radioactive contamination in living livestock.

    Science.gov (United States)

    Yamaguchi, Toshiro; Sawano, Kaita; Kishimoto, Miori; Furuhama, Kazuhisa; Yamada, Kazutaka

    2012-12-01

    Soil samples from the ground surface and feces and blood from a mixed-breed male pig were collected on April 10, 2011 at a farm within 20 km of the Fukushima Daiichi nuclear power plant. The radioactivity of each sample was measured using a Ge semiconductor detector. Despite the fact that the pig had been fed non-contaminated imported feed, (131)I, (134)Cs and (137)Cs were detected in the feces, and (134)Cs and (137)Cs were detected in the blood clots. Because it is considerably difficult to measure radioactive contamination in the edible muscle of living livestock, bioassays are an option for the screening of radioactive contamination in living livestock to ensure food safety.

  9. Guide to the bioassay of uranium at uranium mine-mill facilities

    International Nuclear Information System (INIS)

    1981-01-01

    As a result of occupational exposure, uranium may be taken into the body by inhalation, ingestion or absorption through skin wounds. The organs at risk are the lung, kidney, and bones. Analysis of urine samples for uranium is recommended on a regular monthly basis, before and after a rest period, and it is suggested that a worker be removed from a working area if a level above 300 μg/l is found before a rest period, or 150 μg/l after a rest period. Background information on the development of a bioassay program is given, and a recommended program for uranium mine and mill facilities is included. (L.L.)

  10. Theobroma cacao: Review of the Extraction, Isolation, and Bioassay of Its Potential Anti-cancer Compounds

    Science.gov (United States)

    Baharum, Zainal; Akim, Abdah Md; Hin, Taufiq Yap Yun; Hamid, Roslida Abdul; Kasran, Rosmin

    2016-01-01

    Plants have been a good source of therapeutic agents for thousands of years; an impressive number of modern drugs used for treating human diseases are derived from natural sources. The Theobroma cacao tree, or cocoa, has recently garnered increasing attention and become the subject of research due to its antioxidant properties, which are related to potential anti-cancer effects. In the past few years, identifying and developing active compounds or extracts from the cocoa bean that might exert anti-cancer effects have become an important area of health- and biomedicine-related research. This review provides an updated overview of T. cacao in terms of its potential anti-cancer compounds and their extraction, in vitro bioassay, purification, and identification. This article also discusses the advantages and disadvantages of the techniques described and reviews the processes for future perspectives of analytical methods from the viewpoint of anti-cancer compound discovery. PMID:27019680

  11. A new bioassay for the inspection and identification of TBT-containing antifouling paint.

    Science.gov (United States)

    Gueuné, Hervé; Thouand, Gérald; Durand, Marie-José

    2009-11-01

    Since the 1960s tributyl (TBT)-based antifouling paints are widely applied to protect ship's hulls from biofouling. Due to its high toxicity to aquatic ecosystem most of the countries (28 nations in 2008) signed the AFS convention to control the use of harmful antifouling systems on ships. Nevertheless there is currently no simple method to control the presence of organotin in paint. In this study, we propose a bioassay based on the use of a recombinant bioluminescent bacteria to detect directly in paint the presence of TBT. We also propose a simple device as an inspection system to control the absence of organotin in the ship's hull paint. The presence of organotin could be revealed in less than three hours.

  12. Use of a cytochemical bioassay for determination of thyroid stimulating hormone (TSH) in clinical investigation

    International Nuclear Information System (INIS)

    Doehler, K.D.; Hashimoto, T.; Zur Muehlen, A. von

    1977-01-01

    Recently a highly sensitive cytochemical bioassay (CBA) for the determination of human TSH has been developed. We could show that this assay is specific for TSH and measurements done on plasma of normal euthyroid persons agree well with radioimmunological findings. Due to the extreme sensitivity of the CBA we were able to detect low but measurable TSH levels in patients with primary hyperthyroidism, which were not increased by TRH treatment before therapeutic treatment. After therapeutic treatment, TRH application was able to stimulate additional biologically active TSH release which, however, barely reached the lowest limit of detection by RIA. In certain pathological cases we were able to detect elevated plasma TSH levels, which were active immunologically but inactive biologically. (orig.) [de

  13. Isolation of bioactive allelochemicals from sunflower (variety Suncross-42) through fractionation-guided bioassays.

    Science.gov (United States)

    Anjum, Tehmina; Bajwa, Rukhsana

    2010-11-01

    Plants are rich source of biologically active allelochemicals. However, natural product discovery is not an easy task. Many problems encountered during this laborious practice can be overcome through the modification of preliminary trials. Bioassay-directed isolation of active plant compounds can increase efficiency by eliminating many of the problems encountered. This strategy avoids unnecessary compounds, concentrating on potential components and thus reducing the cost and time required. In this study, a crude aqueous extract of sunflower leaves was fractionated through high performance liquid chromatography. The isolated fractions were checked against Chenopodium album and Rumex dentatus. The fraction found active against two selected weeds was re-fractionated, and the active components were checked for their composition. Thin layer chromatography isolated a range of phenolics, whereas the presence of bioactive terpenoids was confirmed through mass spectroscopy and nuclear magnetic resonance spectroscopy.

  14. Thyroid Stimulating Immunoglobulin Bioassay Using Cultured Human Thyroid Cells; A Simplified Micromethod

    International Nuclear Information System (INIS)

    Lee, Myung Chul; Chung, June Key; Cho, Bo Youn; Koh, Chang Soon; Lee, Moon Ho; Ahn, Il Min; Ahn, Hee Kwon

    1985-01-01

    The activation of adenylate cyclase of human thymocytes in primary cell culture and the release of c-AMP into the medium are used to detect b-TSH and TSAb in sera of patients with autoimmune thyroid disease. Sera of patients are used directly as a part of cell culture without immunoglobulin precipitation. In the above TSI bioassay, TSAb pooled serum show c-AMP concentration between that of 1 mU/ml and 10 mU/ml b-TSH but normal control pooled serum doesn't show any detectable c-AMP response. Ninety five percent of untreated Graves' patients shows TSAb activity above normal range, 20% of Hashimoto's and 363/0 of euthyroid Graves' patients show detectable TSAb activity.

  15. Digitized molecular diagnostics: reading disk-based bioassays with standard computer drives.

    Science.gov (United States)

    Li, Yunchao; Ou, Lily M L; Yu, Hua-Zhong

    2008-11-01

    We report herein a digital signal readout protocol for screening disk-based bioassays with standard optical drives of ordinary desktop/notebook computers. Three different types of biochemical recognition reactions (biotin-streptavidin binding, DNA hybridization, and protein-protein interaction) were performed directly on a compact disk in a line array format with the help of microfluidic channel plates. Being well-correlated with the optical darkness of the binding sites (after signal enhancement by gold nanoparticle-promoted autometallography), the reading error levels of prerecorded audio files can serve as a quantitative measure of biochemical interaction. This novel readout protocol is about 1 order of magnitude more sensitive than fluorescence labeling/scanning and has the capability of examining multiplex microassays on the same disk. Because no modification to either hardware or software is needed, it promises a platform technology for rapid, low-cost, and high-throughput point-of-care biomedical diagnostics.

  16. Enumeration of Antibacterial Activity of Few Medicinal Plants by Bioassay Method

    Directory of Open Access Journals (Sweden)

    B. Uma Reddy

    2010-01-01

    Full Text Available The present study was aimed to investigate the antibacterial activity of some common locally available plants, in order to estimate the biological potential of these herbs. The alcoholic extract of Tagetes erecta L (Asteraceae, Argemone mexicana L (Papavaraceae, Datura stramonium L. (Solanaceae and Tylophora indica (Burm.f. Merr. (Asclepiadaceae were evaluated for antibacterial activity using broth dilution bioassay method. It is clear from the results that, the extracts of these plants acts as a good source of antibiotics against various bacterial pathogens tested and exhibited broad spectrum of antibacterial activity. These plant extracts were shown to be moderate to maximum inhibitory effect against different bacterial forms such as Salmonella typhii, Proteus vulgaris, Pseudomonas aeruginosa and Escherichia coli, where as, mild to moderate activity against Klebsiella pneumoniae and Staphylococcus aureus. The results of these studies revealed most valuable information and also support the continued sustainable use of these plants in traditional systems of medicine.

  17. An algorithm for robust non-linear analysis of radioimmunoassays and other bioassays

    International Nuclear Information System (INIS)

    Normolle, D.P.

    1993-01-01

    The four-parameter logistic function is an appropriate model for many types of bioassays that have continuous response variables, such as radioimmunoassays. By modelling the variance of replicates in an assay, one can modify the usual parameter estimation techniques (for example, Gauss-Newton or Marquardt-Levenberg) to produce parameter estimates for the standard curve that are robust against outlying observations. This article describes the computation of robust (M-) estimates for the parameters of the four-parameter logistic function. It describes techniques for modelling the variance structure of the replicates, modifications to the usual iterative algorithms for parameter estimation in non-linear models, and a formula for inverse confidence intervals. To demonstrate the algorithm, the article presents examples where the robustly estimated four-parameter logistic model is compared with the logit-log and four-parameter logistic models with least-squares estimates. (author)

  18. Correlation and comparison of Nb2 lymphoma cell bioassay with radioimmunoassay for human prolactin

    International Nuclear Information System (INIS)

    Subramanian, M.G.; Spirtos, N.J.; Moghissi, K.S.; Magyar, D.M.; Hayes, M.F.; Gala, R.R.

    1984-01-01

    Serum samples from groups of men and women with normal and elevated prolactin (PRL) levels were assayed by radioimmunoassay (RIA) and by Nb 2 lymphoma cell bioassay (BA) for the presence of PRL. Because the Nb 2 lymphoma cells respond to both PRL and growth hormone, BA for PRL activity was carried out before and after neutralization of growth hormone in the serum samples. There were excellent correlations between RIA and BA both in euprolactinemic and hyperprolactinemic subjects. On an absolute basis, RIA and BA values were similar in the euprolactinemic group (6.6 +/- 0.8 versus 6.2 +/- 1.0), whereas in the hyperprolactinemic group, RIA values were significantly higher than the BA results. The two assay systems also appeared to correlate better in women who were hyperprolactinemic, with obvious menstrual cycle disturbances, than in hyperprolactinemic women without menstrual cycle disturbances

  19. The Canadian National Calibration Reference Center for Bioassay and in-vivo Monitoring: A program summary

    International Nuclear Information System (INIS)

    Kramer, G.H.; Zamora, M.L.

    1994-01-01

    The Canadian National Calibration Reference Center for Bioassay and in-vivo Monitoring is part of the Radiation Protection Bureau, Department of Health. The Reference Center operates a variety of different intercomparison programs that are designed to confirm that workplace monitoring results are accurate and provide the necessary external verification required by the Canadian regulators. The programs administered by the Reference Center currently include urinalysis intercomparisons for tritium, natural uranium, and 14 C, and in-vivo programs for whole-body, thorax, and thyroid monitoring. The benefits of the intercomparison programs to the participants are discussed by example. Future programs that are planned include dual spiked urine sample which contain both tritium and 14 C and the in-vivo measurement of 99m Tc. 18 refs., 1 fig., 2 tabs

  20. Evaluation of oxidation techniques for preparing bioassay and environmental samples for liquid scintillation counting

    International Nuclear Information System (INIS)

    Miller, H.H.

    1979-10-01

    In environmental and biological monitoring for carbon-14 and tritium, the presence of color and chemical quenching agents in the samples can degrade the efficiency of liquid scintillation counting. A series of experiments was performed to evaluate the usefulness, under routine conditions, of first oxidizing the samples to improve the counting by removing the color and quenching agents. The scintillation counter was calibrated for the effects of quenching agents on its counting efficiency. Oxidizing apparatus was tested for its ability to accurately recover the 14 C and 3 H in the samples. Scintillation counting efficiences were compared for a variety of oxidized and unoxidized environmental and bioassay samples. The overall conclusion was that, for routine counting, oxidation of such samples is advantageous when they are highly quenched or in solid form

  1. Rapid Bioassay-Guided Isolation of Antibacterial Clerodane Type Diterpenoid from Dodonaea viscosa (L. Jaeq.

    Directory of Open Access Journals (Sweden)

    Muhammad Khurram

    2015-08-01

    Full Text Available Plant extracts are complex matrices and, although crude extracts are widely in use, purified compounds are pivotal in drug discovery. This study describes the application of automated preparative-HPLC combined with a rapid off-line bacterial bioassay, using reduction of a tetrazolium salt as an indicator of bacterial metabolism. This approach enabled the identification of fractions from Dodonaea viscosa that were active against Staphylococcus aureus and Escherichia coli, which, ultimately, resulted in the identification of a clerodane type diterpenoid, 6β-hydroxy-15,16-epoxy-5β, 8β, 9β, 10α-cleroda-3, 13(16, 14-trien-18-oic acid, showing bacteriostatic activity (minimum inhibitory concentration (MIC = 64–128 µg/mL against test bacteria. To the best of our knowledge, this is the first report on antibacterial activity of this metabolite from D. viscosa.

  2. Bioassay-Guided Isolation of Cytotoxic Isocryptoporic Acids from Cryptoporus volvatus

    Directory of Open Access Journals (Sweden)

    Ling-Yun Zhou

    2016-12-01

    Full Text Available The present work constitutes a contribution to the phytochemical investigation of Cryptoporus volvatus aiming to search for effective cytotoxic constituents against tumor cell lines in vivo. Bioassay-guided separation of the ethylacetate extract of C. volvatus afforded four new isocryptoporic acid (ICA derivatives, ICA-B trimethyl ester (1, ICA-E (2, ICA-E pentamethyl ester (3, and ICA-G (4, together with nine known cryptoporic acids. These isocryptoporic acids are isomers of the cryptoporic acids with drimenol instead of albicanol as the terpenoid fragment; their structures were elucidated on the basis of spectroscopic evidences (UV, IR, HRMS, and NMR and comparison with literature values. All isolates show certain cytotoxic activities against five tumor cell lines. Among them, compound 4 showed an comparable activity to that of the positive control cis-platin, while other compounds exhibited weak cytotoxic activities.

  3. Analysis, separation, and bioassay of pyrrolizidine alkaloids from comfrey (Symphytum officinale).

    Science.gov (United States)

    Couet, C E; Crews, C; Hanley, A B

    1996-01-01

    Pyrrolizidine alkaloids have been linked to liver and lung cancers and a range of other deleterious effects. As with many natural toxicants, major problems arise in determining the effects of the different members of the class and the importance of various forms of ingestion. In this study we have investigated the levels of pyrrolizidine alkaloids in comfrey (Symphytum officinale), determined the levels in different parts of the plant and in herbal remedies, separated the alkaloids into two main groups--the principal parent alkaloids and the corresponding N-oxides--and, finally, carried out a simple bioassay based upon the mutagenic capability of the separated compounds in a human cell line. We conclude that the part of the plant ingested is important in terms of alkaloid challenge and that the effect of two of the major groups of alkaloids individually is different from that of alkaloids in the whole plant extract.

  4. Comparison of ICRP Publication 30 lung model-based predictions with measured bioassay data for airborne natural UO2 exposure

    International Nuclear Information System (INIS)

    Thind, K.S.

    1987-01-01

    In this paper a comparison is made between the build-up of U thorax burdens and the predicted total lung (lung and lymph) burden, based on the lung model provided in ICRP Publication 30 for a group of 29 atomic radiation workers at a Canadian fuel fabrication facility. A similar comparison is made between the predicted ratio of the total lung burden to urinary excretion and the ratio obtained from bioassay data. The study period for the comparison is 5 y. The inhalation input for the lung model calculations was derived from air-sampling data and the choice of particle size activity median aerodynamic diameter (AMAD) was guided by particle size measurements made at representative work locations. The pulmonary clearance half-times studied were 100, 250 and 500 d. For the purpose of this comparison, averaged exposure and averaged bioassay data for the group were used. This comparison indicates that for the conditions of this facility, the assumption of a 500-d pulmonary clearance half-time and a particle size of 1 micron (AMAD) may be too conservative. It is suggested that measurements of air concentrations and particle size used as input parameters for the ICRP Publication 30 lung model may be used to calculate bioassay parameters which may then be tested against bioassay data obtained as part of an operational health physics program, thereby giving a useful step towards defining a derived air concentration value for U in the workplace

  5. Responses in sediment bioassays used in the Netherlands: can observed toxicity be explained by routinely monitored priority pollutants?

    NARCIS (Netherlands)

    Lahr, J.; Maas-Diepeveen, J.L.; Stuijfzand, S.C.; Leonards, P.E.G.; Drueke, J.M.; Luecker, S.; Espeldoorn, A.

    2003-01-01

    In order to identify the cause of toxicity in sediments and suspended matter, a large number of samples with different degrees of contamination was taken at various locations in The Netherlands. Standard acute bioassays were carried out with the bacterium Vibrio fischeri, the rotifer Brachionus

  6. Sediment toxicity assessment in the Lagoon of Venice (Italy) using Paracentrotus lividus (Echinodermata: Echinoidea) fertilization and embryo bioassays.

    Science.gov (United States)

    Volpi Ghirardini, A; Arizzi Novelli, A; Tagliapietra, D

    2005-09-01

    The capacity of two toxicity bioassays (fertilization and embryo toxicity tests) to discriminate sediment toxicity using the sea urchin Paracentrotus lividus was tested in five stations with different levels of pollution in the Lagoon of Venice. Two stations were located in estuarine sites, two in the industrial zone, and one in a site at the top of our quality gradient (reference). Elutriate was chosen as sediment matrix to assess the potential effects of bioavailable pollutants in the water column as a consequence of sediment resuspension (dredging and dumping, fishing gear, etc.). An experimental design based on Quality Assurance/Quality Control procedures (QA/QC) was adopted in order to set the methodological basis for an effective use of these bioassays in monitoring programs. Results revealed both higher embriotoxicity than spermiotoxicity in all stations and the efficacy of combined use of both toxicity bioassays in discriminating differing pollution/bioavailability between stations and periods. The good representativeness of the integrated sampling scheme and the standardization of all experimental phases yielded high precision of results. Clear Toxicity Fingerprints were evidenced for the investigated sites through the combined use of both bioassays. A good fit between ecotoxicological data and chemical contamination levels was found, except for unnatural sediment texture.

  7. Translating bioassay results to field population responses using a Leslie-matrix model for the marine amphipod Corophium volutator

    NARCIS (Netherlands)

    Smit, M.G.D.; Kater, B.J.; Jak, R.G.; Heuvel-Greve, van den M.J.

    2006-01-01

    Bioassays can be used for the assessment of sediment contamination. The response is classified based on a statistical scale indicating a certain effect percentage being significantly different from the controls (e.g. mortality classes of 0¿10%, 10¿20% etc.). The ecological relevance of this

  8. Real-time PCR for the early detection and quantification of Coxiella burnetii as an alternative to the murine bioassay.

    Science.gov (United States)

    Howe, Gerald B; Loveless, Bonnie M; Norwood, David; Craw, Philip; Waag, David; England, Marilyn; Lowe, John R; Courtney, Bernard C; Pitt, M Louise; Kulesh, David A

    2009-01-01

    Real-time PCR was used to analyze archived blood from non-human primates (NHP) and fluid samples originating from a well-controlled Q fever vaccine efficacy trial. The PCR targets were the IS1111 element and the com1 gene of Coxiella burnetii. Data from that previous study were used to evaluate real-time PCR as an alternative to the use of sero-conversion by mouse bioassay for both quantification and early detection of C. burnetii bacteria. Real-time PCR and the mouse bioassay exhibited no statistical difference in quantifying the number of microorganisms delivered in the aerosol challenge dose. The presence of C. burnetii in peripheral blood of non-human primates was detected by real-time PCR as early after exposure as the mouse bioassay with results available within hours instead of weeks. This study demonstrates that real-time PCR has the ability to replace the mouse bioassay to measure dosage and monitor infection of C. burnetii in a non-human primate model.

  9. Evaluation of the toxic effects of arsenite, chromate, cadmium, and copper using a battery of four bioassays

    Energy Technology Data Exchange (ETDEWEB)

    Ko, Kyung-Seok; Lee, Pyeong-Koo [Korea Institute of Geoscience and Mineral Resources (KIGAM), Daejeon (Korea, Republic of). Geologic Environment Div.; Kong, In Chul [Yeungnam Univ., Kyungbuk (Korea, Republic of). Dept. of Environmental Engineering

    2012-09-15

    The sensitivities of four different kinds of bioassays to the toxicities of arsenite, chromate, cadmium, and copper were compared. The different bioassays exhibited different sensitivities, i.e., they responded to different levels of toxicity of each of the different metals. However, with the exception of the {alpha}-glucosidase enzyme activity, arsenite was the most toxic compound towards all the tested organisms, exhibiting the highest toxic effect on the seeds of Lactuca, with an EC{sub 50} value of 0.63 mg/L. The sensitivities of Lactuca and Raphanus were greater than the sensitivities of two other kinds of seeds tested. Therefore, these were the seeds appropriate for use in a seed germination assay. A high revertant mutagenic ratio (5:1) of Salmonella typhimurium was observed with an arsenite concentration of 0.1 {mu}g/plate, indicative of a high possibility of mutagenicity. These different results suggested that a battery of bioassays, rather than one bioassay alone, is needed as a more accurate and better tool for the bioassessment of environmental pollutants. (orig.)

  10. A fish-feeding laboratory bioassay to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms.

    Science.gov (United States)

    Marty, Micah J; Pawlik, Joseph R

    2015-01-11

    Marine chemical ecology is a young discipline, having emerged from the collaboration of natural products chemists and marine ecologists in the 1980s with the goal of examining the ecological functions of secondary metabolites from the tissues of marine organisms. The result has been a progression of protocols that have increasingly refined the ecological relevance of the experimental approach. Here we present the most up-to-date version of a fish-feeding laboratory bioassay that enables investigators to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms. Organic metabolites of all polarities are exhaustively extracted from the tissue of the target organism and reconstituted at natural concentrations in a nutritionally appropriate food matrix. Experimental food pellets are presented to a generalist predator in laboratory feeding assays to assess the antipredatory activity of the extract. The procedure described herein uses the bluehead, Thalassoma bifasciatum, to test the palatability of Caribbean marine invertebrates; however, the design may be readily adapted to other systems. Results obtained using this laboratory assay are an important prelude to field experiments that rely on the feeding responses of a full complement of potential predators. Additionally, this bioassay can be used to direct the isolation of feeding-deterrent metabolites through bioassay-guided fractionation. This feeding bioassay has advanced our understanding of the factors that control the distribution and abundance of marine invertebrates on Caribbean coral reefs and may inform investigations in diverse fields of inquiry, including pharmacology, biotechnology, and evolutionary ecology.

  11. Sensitivity and specificity of the bioassay of estrogenicity in mammary gland and seminal vesicles of male mice

    Czech Academy of Sciences Publication Activity Database

    Škarda, Josef

    2002-01-01

    Roč. 51, č. 3 (2002), s. 267-276 ISSN 0862-8408 R&D Projects: GA ČR GA523/99/0843; GA AV ČR KSK5020115 Keywords : bioassay * estrogenicity * mammary gland Subject RIV: ED - Physiology Impact factor: 0.984, year: 2002

  12. Cleavable DNA-protein hybrid molecular beacon: A novel efficient signal translator for sensitive fluorescence anisotropy bioassay.

    Science.gov (United States)

    Hu, Pan; Yang, Bin

    2016-01-15

    Due to its unique features such as high sensitivity, homogeneous format, and independence on fluorescent intensity, fluorescence anisotropy (FA) assay has become a hotspot of study in oligonucleotide-based bioassays. However, until now most FA probes require carefully customized structure designs, and thus are neither generalizable for different sensing systems nor effective to obtain sufficient signal response. To address this issue, a cleavable DNA-protein hybrid molecular beacon was successfully engineered for signal amplified FA bioassay, via combining the unique stable structure of molecular beacon and the large molecular mass of streptavidin. Compared with single DNA strand probe or conventional molecular beacon, the DNA-protein hybrid molecular beacon exhibited a much higher FA value, which was potential to obtain high signal-background ratio in sensing process. As proof-of-principle, this novel DNA-protein hybrid molecular beacon was further applied for FA bioassay using DNAzyme-Pb(2+) as a model sensing system. This FA assay approach could selectively detect as low as 0.5nM Pb(2+) in buffer solution, and also be successful for real samples analysis with good recovery values. Compatible with most of oligonucleotide probes' designs and enzyme-based signal amplification strategies, the molecular beacon can serve as a novel signal translator to expand the application prospect of FA technology in various bioassays. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds

    Science.gov (United States)

    ZHAO, Bin; BASTON, David S.; KHAN, Elaine; SORRENTINO, Claudio; DENISON, Michael S.

    2011-01-01

    Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications. Over the past 10 years, reporter gene assays have been widely accepted and used for analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like compounds in various types of matrices, such as biological, environmental, food and feed samples, given that high-resolution instrumental analysis techniques are impractical for large-scale screening analysis. The most sensitive cell-based reporter gene bioassay systems developed are the mechanism-based CALUX (Chemically Activated Luciferase Expression) and CAFLUX (Chemically Activated Fluorescent Expression) bioassays, which utilize recombinant cell lines containing stably transfected dioxin (AhR)-responsive firefly luciferase or enhanced green fluorescent protein (EGFP) reporter genes, respectively. While the current CALUX and CAFLUX bioassays are very sensitive, increasing their lower limit of sensitivity, magnitude of response and dynamic range for chemical detection would significantly increase their utility, particularly for those samples that contain low levels of dioxin-like HAHs (i.e., serum). In this study, we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays. PMID:21394221

  14. Proof of concept for a novel insecticide bioassay based on sugar feeding by adult Aedes aegypti (Stegomyia aegypti).

    Science.gov (United States)

    Stell, F M; Roe, R M; Arellano, C; Kennedy, L; Thornton, H; Saavedra-Rodriguez, K; Wesson, D M; Black, W C; Apperson, C S

    2013-09-01

    Aedes aegypti L. (Stegomyia aegypti) (Diptera: Culicidae) is the principal vector of dengue and yellow fever viruses in tropical and subtropical regions of the world. Disease management is largely based on mosquito control achieved by insecticides applied to interior resting surfaces and through space sprays. Population monitoring to detect insecticide resistance is a significant component of integrated disease management programmes. We developed a bioassay method for assessing insecticide susceptibility based on the feeding activity of mosquitoes on plant sugars. Our prototype sugar-insecticide feeding bioassay system was composed of inexpensive, disposable components, contained minimal volumes of insecticide, and was compact and highly transportable. Individual mosquitoes were assayed in a plastic cup that contained a sucrose-permethrin solution. Trypan blue dye was added to create a visual marker in the mosquito's abdomen for ingested sucrose-permethrin solution. Blue faecal spots provided further evidence of solution ingestion. With the sugar-insecticide feeding bioassay, the permethrin susceptibility of Ae. aegypti females from two field-collected strains was characterized by probit analysis of dosage-response data. The field strains were also tested by forced contact of females with permethrin residues on filter paper. Dosage-response patterns were similar, indicating that the sugar-insecticide feeding bioassay had appropriately characterized the permethrin susceptibility of the two strains. © 2012 The Royal Entomological Society.

  15. A miniature bioassay for testing the acute phytotoxicity of photosystem II herbicides on seagrass.

    Directory of Open Access Journals (Sweden)

    Adam D Wilkinson

    Full Text Available Photosystem II (PSII herbicides have been detected in nearshore tropical waters such as those of the Great Barrier Reef and may add to the pressure posed by runoff containing sediments and nutrients to threatened seagrass habitats. There is a growing number of studies into the potential effects of herbicides on seagrass, generally using large experimental setups with potted plants. Here we describe the successful development of an acute 12-well plate phytotoxicity assay for the PSII herbicide Diuron using isolated Halophila ovalis leaves. Fluorescence images demonstrated Diuron affected the entire leaf surface evenly and responses were not influenced by isolating leaves from the plant. The optimum exposure duration was 24 h, by which time the inhibition of effective quantum yield of PSII (∆F/F(m' was highest and no deterioration of photosystems was evident in control leaves. The inhibition of ∆F/F(m' by Diuron in isolated H. ovalis leaves was identical to both potted and hydroponically grown plants (with leaves remaining attached to rhizomes, indicating similar reductions in photosynthetic activity in these acute well-plate assays. The sensitivity of the assay was not influenced by irradiance (range tested 40 to 400 μmol photons m(-2 s(-1. High irradiance, however, caused photo-oxidative stress in H. ovalis and this generally impacted in an additive or sub-additive way with Diuron to damage PSII. The bioassay using isolated leaves is more rapid, uses far less biological material and does not rely on specialised aquarium facilities in comparison with assays using potted plants. The development and validation of this sensitive bioassay will be useful to reliably screen and monitor the phytotoxicity of existing and emerging PSII herbicides and contribute to risk assessments and water quality guideline development in the future.

  16. A Miniature Bioassay for Testing the Acute Phytotoxicity of Photosystem II Herbicides on Seagrass

    Science.gov (United States)

    Wilkinson, Adam D.; Collier, Catherine J.; Flores, Florita; Mercurio, Phil; O’Brien, Jake; Ralph, Peter J.; Negri, Andrew P.

    2015-01-01

    Photosystem II (PSII) herbicides have been detected in nearshore tropical waters such as those of the Great Barrier Reef and may add to the pressure posed by runoff containing sediments and nutrients to threatened seagrass habitats. There is a growing number of studies into the potential effects of herbicides on seagrass, generally using large experimental setups with potted plants. Here we describe the successful development of an acute 12-well plate phytotoxicity assay for the PSII herbicide Diuron using isolated Halophila ovalis leaves. Fluorescence images demonstrated Diuron affected the entire leaf surface evenly and responses were not influenced by isolating leaves from the plant. The optimum exposure duration was 24 h, by which time the inhibition of effective quantum yield of PSII (∆F/Fm’) was highest and no deterioration of photosystems was evident in control leaves. The inhibition of ∆F/Fm’ by Diuron in isolated H. ovalis leaves was identical to both potted and hydroponically grown plants (with leaves remaining attached to rhizomes), indicating similar reductions in photosynthetic activity in these acute well-plate assays. The sensitivity of the assay was not influenced by irradiance (range tested 40 to 400 μmol photons m-2 s-1). High irradiance, however, caused photo-oxidative stress in H. ovalis and this generally impacted in an additive or sub-additive way with Diuron to damage PSII. The bioassay using isolated leaves is more rapid, uses far less biological material and does not rely on specialised aquarium facilities in comparison with assays using potted plants. The development and validation of this sensitive bioassay will be useful to reliably screen and monitor the phytotoxicity of existing and emerging PSII herbicides and contribute to risk assessments and water quality guideline development in the future. PMID:25674791

  17. Effects of sporophyll storage on giant kelp Macrocystis pyrifera (Agardh) bioassay

    Energy Technology Data Exchange (ETDEWEB)

    Gully, J.R.; Bottomley, J.P.; Baird, R.B.

    1999-07-01

    The giant kelp Macrocystis pyrifera (Agardh) is a US Environmental Protection Agency (US EPA)-approved west coast marine species for chronic toxicity monitoring and compliance in the National Pollution Discharge Elimination System (NPDES). The protocol allows field-collected sporophylls to be stored for up to 24 h at 9 to 12 C prior to use. However, the effects of sporophyll storage on the bioassay results have not been fully investigated, particularly with kelp collected from beds south of Point Conception, CA, USA. Therefore, 13 matched-pair reference toxicant bioassays using fresh and stored sporophylls collected from a subtidal kelp bed near Laguna Beach, CA, USA, were performed and compared. The results indicate that a lower percentage of spores germinate and the germ tube lengths are reduced when stored sporophylls are used. The intratest precision of the germination endpoint decreased as evidenced by significant increases in the percent minimum significant difference (%MSD) statistic. The intertest precision also decreased in the germination endpoint as demonstrated by significant increases in the coefficient of variation (CV) values at four effect levels. Conversely, a significant reduction in the CVs was observed in the germ tube length data, possibly as a consequence of the decrease in germ tube length associated with storage. Finally, significant decreases in mean effect concentrations in the germination endpoint in tests using stored sporophylls indicated that storage increased the sensitivity of the spores to the toxic effects of CuCl{sub 2}. The toxicological sensitivity and intratest precision of the germ tube length endpoint were not significantly affected by storage of the sporophylls. The effects of sporophyll storage resulted in a high frequency of invalid tests, lower statistical power, less effective quality assurance standards, and apparent bias in the observed toxicity of CuCl{sub 2}.

  18. A novel bioassay for the activity determination of therapeutic human brain natriuretic peptide (BNP.

    Directory of Open Access Journals (Sweden)

    Lei Yu

    Full Text Available BACKGROUND: Recombinant human brain natriuretic peptide (rhBNP is an important peptide-based therapeutic drug indicated for the treatment of acute heart failure. Accurate determination of the potency of therapeutic rhBNP is crucial for the safety and efficacy of the drug. The current bioassay involves use of rabbit aortic strips, with experiments being complicated and time-consuming and markedly variable in results. Animal-less methods with better precision and accuracy should be explored. We have therefore developed an alternative cell-based assay, which relies on the ability of BNP to induce cGMP production in HEK293 cells expressing BNP receptor guanylyl cyclase-A. METHODOLOGY/PRINCIPAL FINDINGS: An alternative assay based on the measurement of BNP-induced cGMP production was developed. Specifically, the bioassay employs cells engineered to express BNP receptor guanylyl cyclase-A (GCA. Upon rhBNP stimulation, the levels of the second messager cGMP in these cells drastically increased and subsequently secreted into culture supernatants. The quantity of cGMP, which corresponds to the rhBNP activity, was determined using a competitive ELISA developed by us. Compared with the traditional assay, the novel cell-based assay demonstrated better reproducibility and precision. CONCLUSION/SIGNIFICANCE: The optimized cell-based assay is much simpler, more rapid and precise compared with the traditional assay using animal tissues. To our knowledge, this is the first report on a novel and viable alternative assay for rhBNP potency analysis.

  19. Bioassay-guided supercritical fluid extraction of cyclooxygenase-2 inhibiting substances in Plantago major L.

    Science.gov (United States)

    Stenholm, A; Göransson, U; Bohlin, L

    2013-02-01

    Selective extraction of plant materials is advantageous for obtaining extracts enriched with desired constituents, thereby reducing the need for subsequent chromatography purification. Such compounds include three cyclooxygenase-2 (COX-2) inhibitory substances in Plantago major L. targeted in this investigation: α-linolenic acid (α-LNA) (18:3 ω-3) and the triterpenic acids ursolic acid and oleanolic acid. To investigate the scope for tuning the selectivity of supercritical fluid extraction (SFE) using bioassay guidance, and Soxhlet extraction with dichloromethane as solvent as a reference technique, to optimise yields of these substances. Extraction parameters were varied to optimise extracts' COX-2/COX-1 inhibitory effect ratios. The crude extracts were purified initially using a solid phase extraction (SPE) clean-up procedure and the target compounds were identified with GC-MS, LC-ESI-MS and LC-ESI-MS² using GC-FID for quantification. α-LNA was preferentially extracted in dynamic mode using unmodified carbon dioxide at 40°C and 172 bar, at a 0.04% (w/w) yield with a COX-2/COX-1 inhibitory effect ratio of 1.5. Ursolic and oleanolic acids were dynamically extracted at 0.25% and 0.06% yields, respectively, with no traces of (α-LNA) and a COX-2/COX-1-inhibitory effect ratio of 1.1 using 10% (v/v) ethanol as polar modifier at 75°C and 483 bar. The Soxhlet extracts had ursolic acid, oleanolic acid and αLNA yields up to 1.36%, 0.34% and 0.15%, respectively, with a COX-2/COX-1 inhibitory effect ratio of 1.2. The target substances can be extracted selectively by bioassay guided optimisation of SFE conditions. Copyright © 2012 John Wiley & Sons, Ltd.

  20. Developing a quick and inexpensive in vitro (non-animal) bioassay for mascara irritation.

    Science.gov (United States)

    Thomason, H; Montagnes, D J S

    2014-04-01

    Mascara is a mild irritant that causes a range of medical problems. Animal models to predict ocular irritation have, however, been questioned at a number of levels, and there is a continued need to develop in vitro testing methods. We assess changes in an easily quantifiable attribute, ciliated protozoan growth rate, as a sensitive, sublethal measure. Specifically, we test six, randomly chosen, commercial mascara products against a control (as treatments) and reveal through ANOVA (n = 6, α = 0.05) significant differences in the specific growth rate to treatments (for both protozoa). We provide evidence that two easily cultured protozoa (Paramecium caudatum, Blepharisma japonicum) should be considered as models to assess ocular irritancy (and possibly cosmetics in general) and establish the groundwork for such studies to be applied at a more commercial level. We do this by developing a bioassay for mascara toxicity and indicate the low cost (after equipment is purchased, on the order of $100s) and the ease of performing such tests (able to be conducted by undergraduate students), as a consideration for their future commercial application. We first examined dose dependence of responses, revealing that there was a need to conduct preliminary work to determine appropriate levels for sublethal responses. We then show that some products resulted in mortality at high concentrations, others decreased growth rate by >50% (compared with the control), whereas others had no significant effect, compared with the control. We have provided a novel, quick and inexpensive means to assess mascara; the next step is to validate these ciliate bioassays by comparison with animal testing and epidemiological studies, which is beyond the scope of this fundamental 'proof-of-concept' study. © 2013 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  1. In situ and laboratory bioassays to evaluate the impact of effluent discharges on receiving aquatic ecosystems

    International Nuclear Information System (INIS)

    Smolders, R.; Bervoets, L.; Blust, R.

    2004-01-01

    Effluents are a main source of direct and often continuous input of pollutants into aquatic ecosystems with long-term implications on ecosystem functioning. Therefore, the study of the effects of effluent exposure on organisms, populations or communities within the framework of impact assessment has a high ecological relevance. The aim of this study was to assess the toxicological impact of two effluents, one household wastewater treatment effluent (Effluent 1) and one industrial effluent (Effluent 2), on the receiving aquatic ecosystem using two test species under both in situ and laboratory conditions. Zebra mussel (Dreissena polymorpha) and common carp (Cyprinus carpio) were exposed under laboratory conditions in an online monitoring flow-through system (receiving different concentrations of Effluent 2) and under in situ conditions along the pollution gradient established by these two effluent discharges. Bioassays focussed on growth and condition related endpoints (i.e. condition, growth, lipid budget), since these are key functional processes within organisms and populations. Under laboratory conditions, increasing concentrations of the industrial effluent (Effluent 2) had a negative effect on both zebra mussel and carp energy reserves and condition. Under in situ conditions, the same negative impact of Effluent 2 was observed for zebra mussels, while Effluent 1 had no apparent effect on exposed zebra mussels. Carp growth and condition, on the other hand, were significantly increased at the discharge sites of both effluents when compared to the reference site, probably due to differences in food availability. The results indicate that a combination of in situ and laboratory exposures can illustrate how ecological processes influence bioassay studies. The incorporation of indirect, ecological effects, like changes in food availability, provides considerable benefit in understanding and predicting effects of effluents on selected species under realistic exposure

  2. In situ and laboratory bioassays to evaluate the impact of effluent discharges on receiving aquatic ecosystems

    Energy Technology Data Exchange (ETDEWEB)

    Smolders, R. [Laboratory for Ecophysiology, Biochemistry and Toxicology, University of Antwerp (RUCA), Groenenborgerlaan 171, B-2020 Antwerp (Belgium)]. E-mail: roel.smolders@ua.ac.be; Bervoets, L. [Laboratory for Ecophysiology, Biochemistry and Toxicology, University of Antwerp (RUCA), Groenenborgerlaan 171, B-2020 Antwerp (Belgium); Blust, R. [Laboratory for Ecophysiology, Biochemistry and Toxicology, University of Antwerp (RUCA), Groenenborgerlaan 171, B-2020 Antwerp (Belgium)

    2004-11-01

    Effluents are a main source of direct and often continuous input of pollutants into aquatic ecosystems with long-term implications on ecosystem functioning. Therefore, the study of the effects of effluent exposure on organisms, populations or communities within the framework of impact assessment has a high ecological relevance. The aim of this study was to assess the toxicological impact of two effluents, one household wastewater treatment effluent (Effluent 1) and one industrial effluent (Effluent 2), on the receiving aquatic ecosystem using two test species under both in situ and laboratory conditions. Zebra mussel (Dreissena polymorpha) and common carp (Cyprinus carpio) were exposed under laboratory conditions in an online monitoring flow-through system (receiving different concentrations of Effluent 2) and under in situ conditions along the pollution gradient established by these two effluent discharges. Bioassays focussed on growth and condition related endpoints (i.e. condition, growth, lipid budget), since these are key functional processes within organisms and populations. Under laboratory conditions, increasing concentrations of the industrial effluent (Effluent 2) had a negative effect on both zebra mussel and carp energy reserves and condition. Under in situ conditions, the same negative impact of Effluent 2 was observed for zebra mussels, while Effluent 1 had no apparent effect on exposed zebra mussels. Carp growth and condition, on the other hand, were significantly increased at the discharge sites of both effluents when compared to the reference site, probably due to differences in food availability. The results indicate that a combination of in situ and laboratory exposures can illustrate how ecological processes influence bioassay studies. The incorporation of indirect, ecological effects, like changes in food availability, provides considerable benefit in understanding and predicting effects of effluents on selected species under realistic exposure

  3. Assessment of N and P in organic fertilizer using the missing element technique and a microbial bioassay

    International Nuclear Information System (INIS)

    Salas, E.; Ramirez, C.

    2002-01-01

    Assessment of N and P in organic fertilizers using the missing element technique and a microbial bioassay.Through a greenhouse bioassay, using sorghum (Sorghum vulgare) as a test plant, and a microbial assay the availability of N and P in 6 substrates was determined, namely: soil alone and in combination with several organic amendments 10% W/W of chicken manure (CM), compost (C), bocashi (B), vermicompost (V) and coffe hulls (Br). In the microbial assay a complete randomized design with 6 replicates was used; the microbial biomass (BM) was determined 2 days after the glucose amendment of each treatment. In both bioassays a 2 X 2 factorial (N and P fertilization) was establish and the following combinations resulted: +N, +P, +P+N and -P-N (control). For the greenhouse experiment, a complete randomized design with 4 replicates was used. Above-ground plant material of sorghum was harvested 34 days after showing to determine plant dry weight (PS) and content of N and P. Both assays showed a response to the soil amendment with N and P. Soil treatments with CM, C and B showed the highest values of PS and BM. Soil treatment with CM amended with N, P or both did not showed a response in PS or BM, in C and B there was a response to N addition but not to P. In treatments with V and Br, the lowest values for PS and BM were obtained, and there was a growth response to N and P. Both bioassays were able to pinpoint N and P defficiencies in the soil as well in some of the mixtures of soil with organic amendments. A high correlation was encountered between the greenhouse assay and the microbial bioassay (r= 0.86, P=0.0001). Therefore, the microbial bioassay can be a cheaper alternative to the plant bioassay not only to evaluate the nutritional quality of compost but also to identify nutrient deficiencies in soils as well as in substrates amended with organic fertilizers. (Author) [es

  4. Integrating bioassays and analytical chemistry as an improved approach to support safety assessment of food contact materials.

    Science.gov (United States)

    Veyrand, Julien; Marin-Kuan, Maricel; Bezencon, Claudine; Frank, Nancy; Guérin, Violaine; Koster, Sander; Latado, Hélia; Mollergues, Julie; Patin, Amaury; Piguet, Dominique; Serrant, Patrick; Varela, Jesus; Schilter, Benoît

    2017-10-01

    Food contact materials (FCM) contain chemicals which can migrate into food and result in human exposure. Although it is mandatory to ensure that migration does not endanger human health, there is still no consensus on how to pragmatically assess the safety of FCM since traditional approaches would require extensive toxicological and analytical testing which are expensive and time consuming. Recently, the combination of bioassays, analytical chemistry and risk assessment has been promoted as a new paradigm to identify toxicologically relevant molecules and address safety issues. However, there has been debate on the actual value of bioassays in that framework. In the present work, a FCM anticipated to release the endocrine active chemical 4-nonyphenol (4NP) was used as a model. In a migration study, the leaching of 4NP was confirmed by LC-MS/MS and GC-MS. This was correlated with an increase in both estrogenic and anti-androgenic activities as measured with bioassays. A standard risk assessment indicated that according to the food intake scenario applied, the level of 4NP measured was lower, close or slightly above the acceptable daily intake. Altogether these results show that bioassays could reveal the presence of an endocrine active chemical in a real-case FCM migration study. The levels reported were relevant for safety assessment. In addition, this work also highlighted that bioactivity measured in migrate does not necessarily represent a safety issue. In conclusion, together with analytics, bioassays contribute to identify toxicologically relevant molecules leaching from FCM and enable improved safety assessment.

  5. Investigation of independence in inter-animal tumor-type occurrences within the NTP rodent-bioassay database

    Energy Technology Data Exchange (ETDEWEB)

    Bogen, K.T. [Lawrence Livermore National Lab., CA (United States); Seilkop, S. [Analytical Sciences, Inc., Durham, NC (United States)

    1993-05-01

    Statistically significant elevation in tumor incidence at multiple histologically distinct sites is occasionally observed among rodent bioassays of chemically induced carcinogenesis. If such data are to be relied on (as they have, e.g., by the US EPA) for quantitative cancer potency assessment, their proper analysis requires a knowledge of the extent to which multiple tumor-type occurrences are independent or uncorrelated within individual bioassay animals. Although difficult to assess in a statistically rigorous fashion, a few significant associations among tumor-type occurrences in rodent bioassays have been reported. However, no comprehensive studies of animal-specific tumor-type occurrences at death or sacrifice have been conducted using the extensive set of available NTP rodent-bioassay data, on which most cancer-potency assessment for environmental chemicals is currently based. This report presents the results of such an analysis conducted on behalf of the National Research Council`s Committee on Risk Assessment for Hazardous Air Pollutants. Tumor-type associations among individual animals were examined for {approximately}2500 to 3000 control and {approximately}200 to 600 treated animals using pathology data from 62 B6C3F1 mouse studies and 61 F/344N rat studies obtained from a readily available subset of the NTP carcinogenesis bioassay database. No evidence was found for any large correlation in either the onset probability or the prevalence-at-death or sacrifice of any tumor-type pair investigated in control and treated rats and niece, although a few of the small correlations present were statistically significant. Tumor-type occurrences were in most cases nearly independent, and departures from independence, where they did occur, were small. This finding is qualified in that tumor-type onset correlations were measured only indirectly, given the limited nature of the data analyzed.

  6. A review of metal (Pb and Zn) sensitive and pH tolerant bioassay organisms for risk screening of metal-contaminated acidic soils

    International Nuclear Information System (INIS)

    Chapman, E.Emily V.; Dave, Göran; Murimboh, John D.

    2013-01-01

    To improve risk estimates at the screening stage of Ecological Risk Assessment (ERA), short duration bioassays tailored to undisturbed soil cores from the contaminated site could be useful. However, existing standardized bioassays use disturbed soil samples and often pH sensitive organisms. This is a problem as naturally acidic soils are widespread. Changing soil properties to suit the test organism may change metal bioavailability, leading to erroneous risk estimates. For bioassays in undisturbed soil cores to be effective, species able to withstand natural soil properties must be identified. This review presents a critical examination of bioassay species' tolerance of acidic soils and sensitivity to metal contaminants such as Pb and Zn. Promising organisms include; Dendrobaena octaedra, Folsomia candida, Caenorhabditis elegans, Oppia nitens, Brassica rapa, Trifolium pratense, Allium cepa, Quercus rubra and Acer rubrum. The MetSTICK test and the Bait lamina test were also identified as suitable microorganism tests. -- Highlights: •Risk screening of metal contaminated soils should consider metal bioavailability. •Metal bioavailability is dependent on soil properties such as pH. •Many standardized bioassay organisms are sensitive to acidic soils. •This review identifies acid tolerant and metal sensitive bioassays and species. •The identified tests can improve risk screening of acidic metal contaminated soil. -- This review identifies bioassay species able to withstand naturally acidic soils while being sensitive to metal contaminants

  7. Effect-based trigger values for in vitro bioassays: Reading across from existing water quality guideline values.

    Science.gov (United States)

    Escher, Beate I; Neale, Peta A; Leusch, Frederic D L

    2015-09-15

    Cell-based bioassays are becoming increasingly popular in water quality assessment. The new generations of reporter-gene assays are very sensitive and effects are often detected in very clean water types such as drinking water and recycled water. For monitoring applications it is therefore imperative to derive trigger values that differentiate between acceptable and unacceptable effect levels. In this proof-of-concept paper, we propose a statistical method to read directly across from chemical guideline values to trigger values without the need to perform in vitro to in vivo extrapolations. The derivation is based on matching effect concentrations with existing chemical guideline values and filtering out appropriate chemicals that are responsive in the given bioassays at concentrations in the range of the guideline values. To account for the mixture effects of many chemicals acting together in a complex water sample, we propose bioanalytical equivalents that integrate the effects of groups of chemicals with the same mode of action that act in a concentration-additive manner. Statistical distribution methods are proposed to derive a specific effect-based trigger bioanalytical equivalent concentration (EBT-BEQ) for each bioassay of environmental interest that targets receptor-mediated toxicity. Even bioassays that are indicative of the same mode of action have slightly different numeric trigger values due to differences in their inherent sensitivity. The algorithm was applied to 18 cell-based bioassays and 11 provisional effect-based trigger bioanalytical equivalents were derived as an illustrative example using the 349 chemical guideline values protective for human health of the Australian Guidelines for Water Recycling. We illustrate the applicability using the example of a diverse set of water samples including recycled water. Most recycled water samples were compliant with the proposed triggers while wastewater effluent would not have been compliant with a few

  8. Low-Level Plutonium Bioassay Measurements at the Lawrence Livermore National Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Hamilton, T; Brown, T; Hickman, D; Marchetti, A; Williams, R; Kehl, S

    2007-06-18

    Plutonium-239 ({sup 239}Pu) and plutonium-240 ({sup 240}Pu) are important alpha emitting radionuclides contained in radioactive debris from nuclear weapons testing. {sup 239}Pu and {sup 240}Pu are long-lived radionuclides with half-lives of 24,400 years and 6580 years, respectively. Concerns over human exposure to plutonium stem from knowledge about the persistence of plutonium isotopes in the environment and the high relative effectiveness of alpha-radiation to cause potential harm to cells once incorporated into the human body. In vitro bioassay tests have been developed to assess uptakes of plutonium based on measured urinary excretion patterns and modeled metabolic behaviors of the absorbed radionuclides. Systemic plutonium absorbed by the deep lung or from the gastrointestinal tract after ingestion is either excreted or distributed to other organs, primarily to the liver and skeleton, where it is retained for biological half-times of around 20 and 50 years, respectively. Dose assessment and atoll rehabilitation programs in the Marshall Islands have historically given special consideration to residual concentrations of plutonium in the environment even though the predicted dose from inhalation and/or ingestion of plutonium accounts for less than 5% of the annual effective dose from exposure to fallout contamination. Scientists from the Lawrence Livermore National Laboratory (LLNL) have developed a state-of-the-art bioassay test to assess urinary excretion rates of plutonium from Marshallese populations. This new heavy-isotope measurement system is based on Accelerator Mass Spectrometry (AMS). The AMS system at LLNL far exceeds the standard measurement requirements established under the latest United States Department of Energy (DOE) regulation, 10CFR 835, for occupational monitoring of plutonium, and offers several advantages over classical as well as competing new technologies for low-level detection and measurement of plutonium isotopes. The United States

  9. [Application study of human sperm motility bioassay in IVF laboratory quality control].

    Science.gov (United States)

    Cai, Xia; Pomeroy, Kimball O; Mattox, John H

    2006-07-01

    To investigate the sensitivity of human sperm survival bioassay to using known concentrations of potential toxin of formalin and to elevate the application value of human sperm motility assay as a quality control method in detecting the components used in IVF program. Fresh semen was obtained from healthy males at andrology laboratory by masturbation. Sperm was processed on a gradient column of isolate medium and PBS medium. In experiment 1, the medium with 0.25%, 0.75% concentration of formalin and control medium were added to the Falcon culture tubes containing HTF medium with or without 0.3% bovine albumin serum and with or without light mineral oil. In experiment 2, in 3 types of culture tubes containing HTF medium with or without 0.3% bovine albumin serum and with or without light mineral oil, the sperm was exposed to each culture tube and cultured for 24 and 48 hrs at room temperature, and the motile sperms were counted under the microscope. The average sperm motility index in the HTF medium with 0.25% formalin at 24 hrs was 0.594 +/- 0.331, significantly higher than in the HTF medium with 0.75% formalin (0.450 +/- 0.284) (P average sperm survival indexes were 0.683 +/- 0.334 and 0.527 +/- 0.345, respectively, higher than without bovine albumin serum and light mineral oil (0.394 +/- 0.311 and 0.424 +/- 0.311). The average sperm index of 7 ml tissue culture tube made in Denmark was 0.677 +/- 0.335, higher than the other two types of culture tubes made in the USA (0.551 +/- 0.317 and 0.596 +/- 0.327) (P sperm cultured in the medium with 0.3% bovine albumin serum and light mineral oil, the average sperm survival indexes were 0.821 +/- 0.259 and 0.645 +/- 0.335, respectively, higher than without bovine albumin serum or light mineral oil (0.571 +/- 0.321 and 0.395 +/- 0.245) (P sperm survival bioassay is a sensitivity quality control method to detect the components in the IVF laboratory. The 7 ml tissue culture tube made in Denmark is most suitable for culturing

  10. Assessment of mobility and bioavailability of contaminants in MSW incineration ash with aquatic and terrestrial bioassays.

    Science.gov (United States)

    Ribé, V; Nehrenheim, E; Odlare, M

    2014-10-01

    Incineration of municipal solid waste (MSW) is a waste treatment method which can be sustainable in terms of waste volume reduction as well as a source of renewable energy. In the process fly and bottom ash is generated as a waste material. The ash residue may vary greatly in composition depending on the type of waste incinerated and it can contain elevated levels of harmful contaminants such as heavy metals. In this study, the ecotoxicity of a weathered, untreated incineration bottom ash was characterized as defined by the H14 criterion of the EU Waste Framework Directive by means of an elemental analysis, leaching tests followed by a chemical analysis and a combination of aquatic and solid-phase bioassays. The experiments were conducted to assess the mobility and bioavailability of ash contaminants. A combination of aquatic and terrestrial bioassays was used to determine potentially adverse acute effects of exposure to the solid ash and aqueous ash leachates. The results from the study showed that the bottom ash from a municipal waste incineration plant in mid-Sweden contained levels of metals such as Cu, Pb and Zn, which exceeded the Swedish EPA limit values for inert wastes. The chemical analysis of the ash leachates showed high concentrations of particularly Cr. The leachate concentration of Cr exceeded the limit value for L/S 10 leaching for inert wastes. Filtration of leachates prior to analysis may have underestimated the leachability of complex-forming metals such as Cu and Pb. The germination test of solid ash and ash leachates using T. repens showed a higher inhibition of seedling emergence of seeds exposed to the solid ash than the seeds exposed to ash leachates. This indicated a relatively low mobility of toxicants from the solid ash into the leachates, although some metals exceeded the L/S 10 leaching limit values for inert wastes. The Microtox® toxicity test showed only a very low toxic response to the ash leachate exposure, while the D. magna

  11. Could some procedures commonly used in bioassays with the copepod Acartia tonsa Dana 1849 distort results?

    Science.gov (United States)

    Lopes, Laís Fernanda de Palma; Agostini, Vanessa Ochi; Muxagata, Erik

    2018-04-15

    Many organizations have suggested the use of the Calanoid copepod Acartia tonsa in protocols for acute toxicity tests. Nevertheless, these protocols present some problems, such as using 60-180µm meshes to separate specific stages of A. tonsa or carrying out the tests using small volumes that reflect high densities of A. tonsa that do not occur in nature, which could lead to distorted results. In addition, ecotoxicological studies may use statistical approaches that are inadequate for the type of data being analysed. For these reasons, some methodological approaches for bioassays using A. tonsa need to be clarified and revised. In this study, we present information about (i) the retention of copepodite stages of A. tonsa on 180, 330 and 500µm net meshes; (ii) tested storage volumes of 1 organism per 5, 10 or 20mL in each test container (TC); and (iii) considerations about the statistics employed. The results demonstrated that a net mesh of 180µm is capable of retaining all copepodite stages (CI to CVI), contrasting with the recommendation of using a 180µm mesh to separate out adults only. Coarser meshes (330 and 500µm) can also retain different proportions of all copepodite stages, but cannot separate out one developmental stage only. Twenty-five millilitres of medium in an open TC, commonly employed in bioassays simulating densities of 1 organism 5mL -1 , completely evaporated, and the results showed that the TCs need to be covered (e.g., PVC film) and filled with a minimum of 100mL of culture medium (simulating densities of 1 organism 20mL -1 ) to avoid evaporation and increases in salinity. The current use of ANOVA in ecotoxicological studies with proportions of surviving organisms should also be reconsidered since the data are discrete and have a binomial distribution; general linear models (GLMs) are considered more adequate. The information presented here suggests some adjustments that hopefully will enable the improvement of the procedures and methods

  12. Bioassay measurements of individuals living near the US Department of Energy's Hanford Site in Washington State, Fall 1985

    International Nuclear Information System (INIS)

    Sula, M.J.; Bihl, D.E.

    1986-05-01

    The purpose of the bioassay measurements was to provide individuals, living within a specific area near the Hanford Site, information on the current levels of radionuclides in their bodies. The measurements included whole body counter (in vivo) examinations and urine sample analyses for detecting the presence of major radionuclides related to current and historical operations at Hanford. Notifications of the special measurements were sent by letter to 515 residences in north Franklin County. Eighty-nine individuals from 52 of the 515 residences requested and received whole body counts. Of these, 32 also provided urine samples. The measurements gave no evidence of unusual levels of radioactivity in any individual. The ability of bioassay measurements to detect the presence of radioactivity in an individual following an exposure is dependent on the quality of the measurement and the nature of the exposure. This report includes a discussion of the capability, under various circumstances, of the measurements that were provided

  13. Feasibility studies to assess the use of 236Pu as a radiochemical yield monitor in bioassay samples

    International Nuclear Information System (INIS)

    Sawant, P.D.; Kalsi, P.C.

    2007-01-01

    Various plutonium compounds are handled in nuclear facilities of BARC. Hence, there is a possibility of occupational workers getting exposed to Pu. In vitro bioassay monitoring in which Pu is separated by chemical procedures from excreta samples and estimated by alpha-spectrometry, is the method of choice for the evaluation of internal dose to the occupational workers handling Pu. However, this method requires a suitable Pu tracer for reducing the uncertainties due to chemical yield in the separation, electro-deposition and counting efficiency. 242 Pu is commonly used as a tracer but due to its non-availability, efforts were made earlier to indigenously synthesis 236 Pu by proton irradiation of 237 Np in BARC-TIFR pelletron facility. The present study, reports the feasibility of using 236 Pu as a radiochemical yield monitor (tracer) in bioassay samples. (author)

  14. Application of bioassays to evaluate a copper contaminated soil before and after a pilot-scale electrokinetic remediation

    Energy Technology Data Exchange (ETDEWEB)

    Wang Quanying [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Zhou Dongmei [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China)], E-mail: dmzhou@issas.ac.cn; Cang Long [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Sun Tianran [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China)

    2009-02-15

    Remediation programmes are considered to be complete when human risk-based criteria are met. However, these targets are often unsatisfied with the ecological parameters that may be important with regard to future soil use. Five soil subsamples, collecting along a pilot-scale soil column after electrokinetic treatment, were studied, from which about 42.0%-93.3% soil Cu had been successfully removed. A series of biological assays including soil microbial biomass carbon, basal soil respiration, soil urease activity, earthworm assays, and seed assays were used to evaluate their ecological risks. The results showed that the bioassay data from the treatment variants did not supposedly reflecting the decreased soil Cu concentrations after the electrokinetic treatment, but were highly correlated with some soil physicochemical characteristics. It suggests that bioassays are necessary to assess the ecotoxicity of soil after electrokinetic treatment. - There has been a motivation towards using biological indicators for risk assessment of contaminated soil after electrokinetic remediation.

  15. Addressing the recovery of feeding rates in post-exposure feeding bioassays: Cyathura carinata as a case study

    Energy Technology Data Exchange (ETDEWEB)

    Pais-Costa, Antonia Juliana [IMAR—Institute of Marine Research, MARE—Marine and Environmental Sciences Centre, Faculty of Sciences and Technology, University of Coimbra, 3004-517 Coimbra (Portugal); Acevedo, Pelayo [SaBio IREC, Instituto de Investigación en Recursos Cinegéticos (UCLM-CSIC-JCCM), Ciudad Real 13005 (Spain); Marques, João Carlos [IMAR—Institute of Marine Research, MARE—Marine and Environmental Sciences Centre, Faculty of Sciences and Technology, University of Coimbra, 3004-517 Coimbra (Portugal); Martinez-Haro, Mónica, E-mail: monica.martinezharo@gmail.com [IMAR—Institute of Marine Research, MARE—Marine and Environmental Sciences Centre, Faculty of Sciences and Technology, University of Coimbra, 3004-517 Coimbra (Portugal)

    2015-02-15

    Post-exposure bioassays are used in environmental assessment as a cost-effective tool, but the effects of organism's recovery after exposure to pollutant has not yet been addressed in detail. The recoveries of post-exposure feeding rates after being exposed to two sublethal concentrations of cadmium during two different exposure periods (48 h and 96 h) were evaluated under laboratory conditions using the estuarine isopod Cyathura carinata. Results showed that feeding depression was a stable endpoint up to 24 h after cadmium exposure, which is useful for ecotoxicological bioassays. - Highlights: • We studied recovery of post-exposure feeding rates 48–96 h after cadmium exposure. • The assay is based on the isopod Cyathura carinata. • Post-exposure feeding inhibition is a stable sublethal endpoint.

  16. Application of bioassays to evaluate a copper contaminated soil before and after a pilot-scale electrokinetic remediation

    International Nuclear Information System (INIS)

    Wang Quanying; Zhou Dongmei; Cang Long; Sun Tianran

    2009-01-01

    Remediation programmes are considered to be complete when human risk-based criteria are met. However, these targets are often unsatisfied with the ecological parameters that may be important with regard to future soil use. Five soil subsamples, collecting along a pilot-scale soil column after electrokinetic treatment, were studied, from which about 42.0%-93.3% soil Cu had been successfully removed. A series of biological assays including soil microbial biomass carbon, basal soil respiration, soil urease activity, earthworm assays, and seed assays were used to evaluate their ecological risks. The results showed that the bioassay data from the treatment variants did not supposedly reflecting the decreased soil Cu concentrations after the electrokinetic treatment, but were highly correlated with some soil physicochemical characteristics. It suggests that bioassays are necessary to assess the ecotoxicity of soil after electrokinetic treatment. - There has been a motivation towards using biological indicators for risk assessment of contaminated soil after electrokinetic remediation

  17. Dermal oncogenicity bioassays of monofunctional and multifunctional acrylates and acrylate-based oligomers.

    Science.gov (United States)

    DePass, L R; Maronpot, R R; Weil, C S

    1985-01-01

    Several important components of photocurable coatings were studied for dermal tumorigenic activity by repeated application to the skin of mice. The substances tested were 2-ethylhexyl acrylate (EHA) and methylcarbamoyloxyethyl acrylate (MCEA) (monomers); neopentyl glycol diacrylate (NPGDA), esterdiol-204-diacrylate (EDDA), and pentaerythritol tri(tetra)acrylate (PETA) (cross-linkers); and three acrylated urethane oligomers. For each bioassay, 40 C3H/HeJ male mice were dosed 3 times weekly on the dorsal skin for their lifetime with the highest dose of the test agent that caused no local irritation or reduction in body weight gain. Two negative control groups received acetone (diluent) only. A positive control group received 0.2% methylcholanthrene (MC). NPGDA and EHA had significant tumorigenic activity with tumor yields of eight and six tumor-bearing mice (three and two malignancies), respectively. The MC group had 34 mice with carcinomas and 1 additional mouse with a papilloma. MCEA had no dermal tumorigenic activity but resulted in early mortality. No skin tumors in the treatment area were observed in the other groups. Additional studies will be necessary to elucidate possible relationships between structure and tumorigenic activity for the acrylates.

  18. Lanthanide-containing polymer microspheres by multiple-stage dispersion polymerization for highly multiplexed bioassays.

    Science.gov (United States)

    Abdelrahman, Ahmed I; Dai, Sheng; Thickett, Stuart C; Ornatsky, Olga; Bandura, Dmitry; Baranov, Vladimir; Winnik, Mitchell A

    2009-10-28

    We describe the synthesis and characterization of metal-encoded polystyrene microspheres by multiple-stage dispersion polymerization with diameters on the order of 2 mum and a very narrow size distribution. Different lanthanides were loaded into these microspheres through the addition of a mixture of lanthanide salts (LnCl(3)) and excess acrylic acid (AA) or acetoacetylethyl methacrylate (AAEM) dissolved in ethanol to the reaction after about 10% conversion of styrene, that is, well after the particle nucleation stage was complete. Individual microspheres contain ca. 10(6)-10(8) chelated lanthanide ions, of either a single element or a mixture of elements. These microspheres were characterized one-by-one utilizing a novel mass cytometer with an inductively coupled plasma (ICP) ionization source and time-of-flight (TOF) mass spectrometry detection. Microspheres containing a range of different metals at different levels of concentration were synthesized to meet the requirements of binary encoding and enumeration encoding protocols. With four different metals at five levels of concentration, we could achieve a variability of 624, and the strategy we report should allow one to obtain much larger variability. To demonstrate the usefulness of element-encoded beads for highly multiplexed immunoassays, we carried out a proof-of-principle model bioassay involving conjugation of mouse IgG to the surface of La and Tm containing particles and its detection by an antimouse IgG bearing a metal-chelating polymer with Pr.

  19. Development of a bioassay system for investigating insulin resistance factors of pregnancy

    International Nuclear Information System (INIS)

    Hausman, D.B.; Singh, R.; Martin, R.J.

    1986-01-01

    To determine if late-term pregnant serum and/or placenta could induce insulin resistance in normal adipose cells, the authors have developed an insulin sensitive bioassay system. Cells isolated from epididymal fat pads of 250-275 g Sprague Dawley rats are preincubated for 3 hours at 37 0 in media 199 and serum or placental extract. The cells are washed free of serum and tested for metabolic activity in a 2 hour incubation which measures the conversion of U- 14 C-glucose to 14 CO 2 and to 14 C-triglyceride fatty acids under basal and insulin stimulated conditions. Maximal insulin responsiveness (350-450% basal for CO 2 and 1400-1700% basal for fatty acids) is achieved using Worthington Type II collagenase and a 45-60 minute digestion period for cell isolations and Krebs-Ringer bicarbonate buffer containing 0.5 mM glucose, 2% Armour bovine serum albumin (CRG-7), 1000 μU/ml insulin and 110,000 to 120,000 cells in the 2 hour incubations. Using this bioasssay system the authors have found that insulin responsiveness, in terms of glucose conversion to fatty acids, is unchanged when cells are preincubated with 5% control pig serum but reduced following preincubation with late pregnant (110 day) pig serum. In future experiments the authors hope to further characterize the factor(s) in pregnant serum responsible for inducing this metabolic effect

  20. Toxicity assessment for petroleum-contaminated soil using terrestrial invertebrates and plant bioassays.

    Science.gov (United States)

    Hentati, Olfa; Lachhab, Radhia; Ayadi, Mariem; Ksibi, Mohamed

    2013-04-01

    The assessment of soil quality after a chemical or oil spill and/or remediation effort may be measured by evaluating the toxicity of soil organisms. To enhance our understanding of the soil quality resulting from laboratory and oil field spill remediation, we assessed toxicity levels by using earthworms and springtails testing and plant growth experiments. Total petroleum hydrocarbons (TPH)-contaminated soil samples were collected from an oilfield in Sfax, Tunisia. Two types of bioassays were performed. The first assessed the toxicity of spiked crude oil (API gravity 32) in Organization for Economic Co-operation and Development artificial soil. The second evaluated the habitat function through the avoidance responses of earthworms and springtails and the ability of Avena sativa to grow in TPH-contaminated soils diluted with farmland soil. The EC50 of petroleum-contaminated soil for earthworms was 644 mg of TPH/kg of soil at 14 days, with 67 % of the earthworms dying after 14 days when the TPH content reached 1,000 mg/kg. The average germination rate, calculated 8 days after sowing, varied between 64 and 74 % in low contaminated soils and less than 50 % in highly contaminated soils.

  1. Submicron polymer particles containing fluorescent semiconductor nanocrystals CdSe/ZnS for bioassays.

    Science.gov (United States)

    Generalova, Alla N; Sizova, Svetlana V; Zdobnova, Tatiana A; Zarifullina, Margarita M; Artemyev, Michail V; Baranov, Alexander V; Oleinikov, Vladimir A; Zubov, Vitaly P; Deyev, Sergey M

    2011-02-01

    This study aimed to design a panel of uniform particulate biochemical reagents and to test them in specific bioassays. These reagents are polymer particles of different sizes doped with semiconductor nanocrystals and conjugated with either full-size antibodies or recombinant mini-antibodies (4D5 scFv fragment) designed by genetic engineering approaches. A panel of highly fluorescent polymer particles (150-800 nm) were formed by embedding CdSe/ZnS nanocrystals (quantum dots) into preformed polyacrolein and poly(acrolein-co-styrene) particles. Morphology, content and fluorescence characteristics of the prepared materials were studied by laser correlation spectroscopy, spectrophotometry, optical and fluorescent microscopy and fluorimetry. The obtained fluorescent particles sensitized by anti-Yersinia pestis antibodies were used for rapid agglutination glass test suitable for screening analysis of Y. pestis antigen and for microtiter particle agglutination, which, owing to its speed and simplicity, is very beneficial for diagnostic detection of Y. pestis antigen. Recombinant 4D5 scFv antibodies designed and conjugated with polymer particles containing quantum dots provide multipoint highly specific binding with cancer marker HER2/neu on the surface of SKOV-3 cell.

  2. Methods to improve routine bioassay monitoring for freshly separated, poorly transported plutonium

    International Nuclear Information System (INIS)

    Bihl, D.E.; Lynch, T.P.; Carbaugh, E.H.; Sula, M.J.

    1988-09-01

    Several human cases involving inhalation of plutonium oxide at Hanford have shown clearance half-times from the lung that are much longer than the 500-day half-time recommended for class Y plutonium in Publication 30 of the International Commission on Radiological Protection(ICRP). The more tenaciously retained material is referred to as super class Y plutonium. The ability to detect super class Y plutonium by current routine bioassay measurements is shown to be poor. Pacific Northwest Laboratory staff involved in the Hanford Internal Dosimetry Program investigated four methods to se if improvements in routine monitoring of workers for fresh super class Y plutonium are feasible. The methods were lung counting, urine sampling, fecal sampling, and use of diethylenetriaminepentaacetate (DTPA) to enhance urinary excretion. Use of DTPA was determined to be not feasible. Routine fecal sampling was found to be feasible but not recommended. Recommendations were made to improve the detection level for routine annual urinalysis and routine annual lung counting. 12 refs., 9 figs., 7 tabs

  3. Effects of Jatropha curcas oil in Lactuca sativa root tip bioassays.

    Science.gov (United States)

    Andrade-Vieira, Larissa F; Botelho, Carolina M; Laviola, Bruno G; Palmieri, Marcel J; Praça-Fontes, Milene M

    2014-03-01

    Jatropha curcas L. (Euphorbiaceae) is important for biofuel production and as a feed ingredient for animal. However, the presence of phorbol esters in the oil and cake renders the seeds toxic. The toxicity of J. curcas oil is currently assessed by testing in animals, leading to their death. The identification of toxic and nontoxic improved varieties is important for the safe use of J. curcas seeds and byproducts to avoid their environmental toxicity. Hence, the aim of this study was to propose a short-term bioassay using a plant as a model to screen the toxicity of J. curcas oil without the need to sacrifice any animals. The toxicity of J. curcas oil was evident in germination, root elongation and chromosomal aberration tests in Lactuca sativa. It was demonstrated that J. curcas seeds contain natural compounds that exert phyto-, cyto- and genotoxic effects on lettuce, and that phorbol esters act as aneugenic agents, leading to the formation of sticky chromosomes and c-metaphase cells. In conclusion, the tests applied have shown reproducibility, which is important to verify the extent of detoxification and to determine toxic doses, thus reducing the numbers of animals that would be used for toxicity tests.

  4. Effects of Jatropha curcas oil in Lactuca sativa root tip bioassays

    Directory of Open Access Journals (Sweden)

    LARISSA F. ANDRADE-VIEIRA

    2014-03-01

    Full Text Available Jatropha curcas L. (Euphorbiaceae is important for biofuel production and as a feed ingredient for animal. However, the presence of phorbol esters in the oil and cake renders the seeds toxic. The toxicity of J. curcas oil is currently assessed by testing in animals, leading to their death. The identification of toxic and nontoxic improved varieties is important for the safe use of J. curcas seeds and byproducts to avoid their environmental toxicity. Hence, the aim of this study was to propose a short-term bioassay using a plant as a model to screen the toxicity of J. curcas oil without the need to sacrifice any animals. The toxicity of J. curcas oil was evident in germination, root elongation and chromosomal aberration tests in Lactuca sativa. It was demonstrated that J. curcas seeds contain natural compounds that exert phyto-, cyto- and genotoxic effects on lettuce, and that phorbol esters act as aneugenic agents, leading to the formation of sticky chromosomes and c-metaphase cells. In conclusion, the tests applied have shown reproducibility, which is important to verify the extent of detoxification and to determine toxic doses, thus reducing the numbers of animals that would be used for toxicity tests.

  5. Bioassay-directed fractionation of a blood coagulation factor Xa inhibitor, betulinic acid from Lycopus lucidus

    Directory of Open Access Journals (Sweden)

    Tan Yin-Feng

    2018-03-01

    Full Text Available Thrombosis is a major cause of morbidity and mortality worldwide and plays a pivotal role in the pathogenesis of several cardiovascular disorders, including acute coronary syndrome, unstable angina, myocardial infarction, sudden cardiac death, peripheral arterial occlusion, ischemic stroke, deep-vein thrombosis, and pulmonary embolism. Anticoagulants, antiplatelet agents, and fibrinolytics can reduce the risks of these clinical events. Especially, the blood coagulation factor Xa (FXa inhibitor is a proven anticoagulant. Promoting blood circulation, using traditional Chinese medicine (TCM, for the treatment of these diseases has been safely used for thousands of years in clinical practice. Therefore, highly safe and effective anticoagulant ingredients, including FXa inhibitors, could be found in TCM for activating the blood circulation. One FXa inhibitor, a pentacyclic triterpene (compound 1, betulinic acid characterized by IR, MS and NMR analyses, was isolated from the ethyl acetate fraction of Lycopus lucidus by bioassay-directed fractionation. Compound 1 exhibited an inhibitory effect on FXa with IC50 25.05 μmol/L and reduced the thrombus weight in an animal model at 25-100 mg/kg. These results indicate that betulinic acid could be the potential for anticoagulant therapy.

  6. Assessment of toxicological profiles of the municipal wastewater effluents using chemical analyses and bioassays.

    Science.gov (United States)

    Smital, Tvrtko; Terzic, Senka; Zaja, Roko; Senta, Ivan; Pivcevic, Branka; Popovic, Marta; Mikac, Iva; Tollefsen, Knut Erik; Thomas, Kevin V; Ahel, Marijan

    2011-05-01

    The hazardous chemical contamination of untreated wastewater and secondary effluent from the wastewater treatment plant (WWTP) of the city of Zagreb, Croatia was comprehensively characterized using large-volume solid-phase extraction (SPE) and silica gel fractionation, followed by a detailed analysis of the resulting extracts by a combination of chemical and bioassay methods. Over 100 individual contaminants or closely related-contaminant groups were identified by high-resolution gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-QTOF). Ecotoxicity profiling of the investigated samples, including cytotoxicity, chronic toxicity and EROD activity; inhibition of the multixenobiotic resistance (MXR), genotoxicity and estrogenic potential, revealed the most significant contribution of toxic compounds to be present in polar fractions. Wastewater treatment using conventional activated sludge process reduced the initial toxicity of raw wastewater to various extents, ranging from 28% for algal toxicity to 73.2% for an estrogenic activity. The most efficient toxicity removal was observed for the polar compounds. Copyright © 2010 Elsevier Inc. All rights reserved.

  7. Seed germination bioassay using maize seeds for phytoxicity evaluation of different composted materials

    International Nuclear Information System (INIS)

    Haq, T.; Begum, R.; Ali, T.A.

    2014-01-01

    In this paper we evaluated the phytotoxicity of different composts obtained by two different composting methods using seed germination bioassay. Seeds of Zea mays were sown in 1:5 extract of composts and these were compared with the control (100% distilled water) for each type of material. Composting of herbal pharmaceutical solid waste (HPSW) was carried out using both conventional bin and pit method. HPSW was mixed separately with poultry manure, cow-manure and goat manure in three different ratios. Uncomposted and composted HPSW were tested to study the Phytotoxicity on Zea mays seed germination, after composting increase in percent germination as well as germination index (GI) values were observed in all combinations regardless, composted by pit or bin method. The results clearly showed that composting reduced Phytotoxicity. The results showed that use of completely composted organic waste reduces the phytotoxicity and is better than the use of uncomposted waste. It was found that pit method was more suitable than bin method. Herbal waste with goat manure in 1:1 ratio was found to be the most effective combination as compared to other combinations here. Germination was 100% and the germination index was 1.4 whereas uncomposted HPSW showed the lowest percent germination i.e., 77% and germination index 52.31 respectively. (author)

  8. Isolation of active constituents from cherry laurel (Laurocerasus officinalis Roem.) leaves through bioassay-guided procedures.

    Science.gov (United States)

    Akkol, Esra Küpeli; Kırmızıbekmez, Hasan; Küçükboyacı, Nurgün; Gören, Ahmet C; Yesilada, Erdem

    2012-01-31

    The fresh leaves of Laurocerasus officinalis Roem. (Rosaceae) are externally used against pain and feverish symptoms in Turkish folk medicine. Effects of the extracts, fractions and isolated compounds from the leaves of L. officinalis were investigated using in vivo models of inflammation and pain in mice. The crude ethanolic extract from the leaves of plant was sequentially fractionated into five subextracts; explicitly, n-hexane, chloroform, ethyl acetate (EtOAc), n-butanol, and remaining water extracts. Further studies were carried out on the most active EtOAc subextract was further subjected to fractionation through column chromatography. For the anti-inflammatory activity, carrageenan-induced hind paw edema and acetic acid-induced increase in capillary permeability models, and for the antinociceptive activity p-benzoquinone-induced writhing test in mice were employed. Ethanolic extract of the leaves was shown to possess significant inhibitory activity in the assay methods without inducing any gastric damage. Through bioassay-guided fractionation and isolation procedures three phenolic compounds, 2-O-β-D-glucopyranosyl-2-hydroxyphenyl-acetic acid (1), kaempferol-3-O-β-D-xylopyranosyl-(1→2)-O-β-D-glucopyranoside (2) and (+)-catechin (3) were isolated from the active fraction and their structures were elucidated by spectral techniques (1D and 2D NMR, ESIMS). The experimental data verified that Laurocerasus officinalis leaves displayed remarkable anti-inflammatory and antinociceptive activity. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  9. Development of a bioassay to assess the toxicity of oil sands sediments to pike (Esox lucius)

    Energy Technology Data Exchange (ETDEWEB)

    Turcotte, D.; Yuan, H.; Tumber, V.; Parrott, J. [Environment Canada, Ottawa, ON (Canada); Raine, J. [Saskatchewan Univ., Saskatoon, SK (Canada)

    2010-07-01

    Pike (Esox lucius) are a commercially sought fish species that inhabit the Athabasca River, which flows through the Athabasca oil sands. The fish are exposed to natural sources of bitumen from the McMurray formation. This study was conducted to design and implement a daily-renewal bioassay to assess the toxicity of oil sands to this fish species and to obtain information regarding the development of pike exposed to bitumen. Eggs were collected and fertilized with milt from spawning wild pike captured from Lake Diefenbaker in Saskatchewan. The fertilized eggs were exposed to different concentrations of sediments or culture water only (negative controls) until complete yolk absorption of control fish, approximately 15 days post-hatch. For the rest of the experiment, brine shrimp were fed to the walleye embryos every day after hatching. The developing fish were examined for morphological deformities, survival, hatching success, and changes in weight and length. The research findings indicated that pike is less sensitive than walleye and fathead minnow to the toxicity of oil sands sediments.

  10. Development of a bioassay using walleye (Sander vitreus) to assess the toxicity of oil sands sediments

    Energy Technology Data Exchange (ETDEWEB)

    Turcotte, D.; Yuan, H.; Tumber, V.; Parrott, J. [Environment Canada, Ottawa, ON (Canada); Raine, J. [Saskatchewan Univ., Saskatoon, SK (Canada)

    2010-07-01

    This study examined the effects of sediments from the Athabasca oil sands area on fish development and survival. Walleye (Sander vitreus) which inhabit the Athabasca River are exposed to natural sources of bitumen eroding from the McMurray formation. This study described the design and implementation of a daily-renewal bioassay to evaluate the potential effects of toxicants on walleye development. Eggs were collected and fertilized with milt from spawning wild walleye captured from Lake Diefenbaker in Saskatchewan. The fertilized eggs were exposed to different concentrations of sediments or culture water only (negative controls) until complete yolk absorption of control fish. The walleye embryos were fed brine shrimp daily after hatching and the developing fish were examined for morphological deformities, survival, hatching success, and changes in weight and length between treatments. Organics concentrations in fish tissues and water were measured when possible. Fathead minnows and northern pikes will also be exposed to the same sediments in order to compare the relative sensitivity of the three species.

  11. Methotrexate intercalated layered double hydroxides with the mediation of surfactants: Mechanism exploration and bioassay study

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Chao-Fan; Tian, De-Ying; Li, Shu-Ping, E-mail: lishuping@njnu.edu.cn; Li, Xiao-Dong

    2015-12-01

    Methotrexatum intercalated layered double hydroxides (MTX/LDHs) hybrids were synthesized by the co-precipitation method and three kinds of nonionic surfactants with different hydrocarbon chain lengths were used. The resulting hybrids were then characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM). XRD and FTIR investigations manifest the successful intercalation of MTX anions into the interlayer of LDHs. TEM graphs indicate that the morphology of the hybrids changes with the variation of the chain length of the surfactants, i.e., the particles synthesized using polyethylene glycol (PEG-7) present regular disc morphology with good monodispersity, while samples with the mediation of alkyl polyglycoside (APG-14) are heavily aggregated and samples with the addition of polyvinylpyrrolidone (PVP-10) exhibit irregular branches. Furthermore, the release and bioassay experiments show that monodisperse MTX/LDHs present good controlled-release and are more efficient in the suppression of the tumor cells. - Highlights: • Surfactants could be used to modify the dispersing state of MTX/LDHs hybrids. • Surfactants have great effect on the morphology of MTX/LDHs hybrids. • MTX/LDHs with good monodisperse degree are more efficient in the suppression of the tumor cells.

  12. Joint use of laboratory bioassays and field-collected plants to evaluate toxicity and contaminant bioaccumulation

    International Nuclear Information System (INIS)

    Long, S.P.; Byron, E.R.; Ohlendorf, H.M.

    1995-01-01

    Soil toxicity tests using lettuce (Latuca saliva) were conducted using soil samples collected as part of ecological risk assessments at two facilities in California. At some sites, terrestrial plants were collected in the field for chemical analysis. Ecological concerns focused on exposures to plants, phytophagous insects, and their secondary consumers, such as birds and small mammals. The toxicity tests were used to assess potential exposures to a variety of site-specific contaminants including organochlorine pesticides, PCBs, PAHs, petroleum hydrocarbons, heavy metals, and other inorganic substances. Site soils were combined with clean control soils to produce toxicity test soil dilutions containing 100%, 75%, 50%, 25%, and 0% site soils. Observations of seed germination and growth were made at day 0, 7, 14, 21 and 28. Toxicity test results were combined with soil chemical analytical results and physical characteristics to establish NOAELs and LOAELs. Bioaccumulation in the lettuce and field-collected plants was evaluated by comparing plant contaminant to soil contaminant concentrations. Allometric equations and sublethal toxicity data were used to predict potential effects on birds and small mammals. Whole-body contaminant concentrations in insects collected on some of the plants in the field were also considered in evaluating the potential for toxicity to insectivorous birds. The study indicated that contaminant uptake was occurring in the field-collected and bioassay plants but not the insects. Site factors in addition to soil contaminant concentration influenced the potential for plant toxicity and bioaccumulation

  13. Bio-assay guided isolation of α-glucosidase inhibitory constituents from Hibiscus mutabilis leaves.

    Science.gov (United States)

    Kumar, Deepak; Kumar, Hemanth; Vedasiromoni, J R; Pal, Bikas C

    2012-01-01

    The increasing demand for natural-product-based medicines and health-care products for the management of diabetes encouraged investigation of this commonly available Indian plant. To establish the anti-diabetic (α-glucosidase inhibitory) activity of H. mutabilis leaf extract, isolate and identify the constituents responsible for the activity, and validate a HPLC method for quantification of the active constituents for standardisation of the extract. The methanolic extract of leaves was partitioned between water, n-butanol and ethyl acetate. Bio-assay guided fractionation, based on inhibition of α-glucosidase, allowed isolation and identification of the active components. The active components were quantified using RP-HPLC-DAD validated for linearity, limit of detection, limit of quantification, precision, accuracy and robustness for this plant extract and the partitioned fractions. Ferulic acid and caffeic acid were identified as the α-glucosidase inhibitors present in H. mutabilis. They were partitioned into an ethyl acetate fraction. The HPLC-DAD calibration curve showed good linearity (r² > 0.99). For the recovery studies the %RSD was less than 2%. The interday and intraday variations were found to be less than 4% RSD for retention time and response. The identification of α-glucosidase inhibition activity in H. mutabilis supports further investigations into the possible use of the plant for the management of diabetes. The HPLC method validated for these extracts will be useful in future research with the plant. Copyright © 2011 John Wiley & Sons, Ltd.

  14. Stability of the intra- and extracellular toxins of Prymnesium parvum using a microalgal bioassay

    DEFF Research Database (Denmark)

    Blossom, Hannah Eva; Andersen, Nikolaj Gedsted; Rasmussen, Silas Anselm

    2014-01-01

    easily maintained. Reducing oxidation by storing the supernatant with no headspace in the vials significantly slowed loss of activity when stored at 4°C. We show that the lytic activity of the intracellular toxins, when released by sonication, is not as high as the extracellular toxins, however...... of P. parvum toxins before attempting to isolate and characterize them. The extracellular toxin in the supernatant is highly unstable, and it loses significant lytic effects after 3 days despite storage at −20°C and after only 24h stored at 4°C. However, when stored at −80°C, lytic activity is more...... the stability of the intracellular toxins when kept as a cell pellet at −20°C is excellent, which proves this is a sufficient storage method for less than 3 months. Our results provide an ecologically appropriate algal bioassay to quantify lytic activity of P. parvum toxins and we have advanced our knowledge...

  15. Assessment of the ecotoxicity of urban estuarine sediment using benthic and pelagic copepod bioassays

    Directory of Open Access Journals (Sweden)

    Maria P. Charry

    2018-05-01

    Full Text Available Urban estuarine sediments are sinks to a range of contaminants of anthropogenic origin, and a key challenge is to characterize the risk of these compounds to receiving environments. In this study, the toxicity of urban estuarine sediments was tested using acute and chronic bioassays in the benthic harpacticoid Quinquelaophonte sp., and in the planktonic calanoid Gladioferens pectinatus, two New Zealand copepod species. The sediment samples from the estuary tributary sites significantly impacted reproduction in Quinquelaophonte sp. However, results from one of the estuary sites were not significantly different to those from the tributaries sites, suggesting that chemicals other than trace metals, polycyclic aromatic hydrocarbons and ammonia may be the causative stressors. Sediment elutriate samples had significant effects on reproductive endpoints in G. pectinatus, and on the induction of DNA damage in cells, as shown by the comet assay. The results indicate that sediment contamination at the Ahuriri Estuary has the potential to impact biological processes of benthic and pelagic organisms. The approach used provides a standardized methodology to assess the toxicity of estuarine sediments.

  16. Detection of genotoxic effects of drinking water disinfection by-products using Vicia faba bioassay.

    Science.gov (United States)

    Hu, Yu; Tan, Li; Zhang, Shao-Hui; Zuo, Yu-Ting; Han, Xue; Liu, Na; Lu, Wen-Qing; Liu, Ai-Lin

    2017-01-01

    Plant-based bioassays have gained wide use among the toxicological and/or ecotoxicological assessment procedures because of their simplicity, sensitivity, low cost, and reliability. The present study describes the use of Vicia faba (V. faba) micronucleus (MN) test and V. faba comet assay in the evaluation of the genotoxic potential of disinfection by-products (DBPs) commonly found in chlorine-disinfected drinking water. Five haloacetic acids and three halogenated acetonitriles were chosen as representatives of DBPs in this study because they are of potentially great public health risk. Results of the MN test indicated that monochloroacetic acid (MCA), monobromoacetic acid (MBA), dichloroacetic acid (DCA), dibromoacetic acid (DBA), trichloroacetic acid (TCA), and trichloroacetonitrile (TCAN) caused a statistically significant increase in MN frequency in V. faba root tip cells. However, no genotoxic response was observed for dichloroacetonitrile (DCAN) and dibromoacetonitrile (DBAN). Results of the comet assay showed that all tested DBPs induced a statistically significant increase in genomic DNA damage to V. faba root tip cells. On considering the capacity to detect genomic damage of a different nature, we suggest that a combination of V. faba MN test and V. faba comet assay is a useful tool for the detection of genotoxic effects of DBPs. It is worthy of assessing the feasibility of using V. faba comet assay combined with V. faba MN test to screen for the genotoxic activity of chlorinated drinking water in future work.

  17. Application of in-situ bioassays with macrophytes in aquatic mesocosm studies.

    Science.gov (United States)

    Coors, Anja; Kuckelkorn, Jochen; Hammers-Wirtz, Monika; Strauss, Tido

    2006-10-01

    Aquatic mesocosm studies assess ecotoxicological effects of chemicals by using small artificial ponds as models of lentic ecosystems. In this study, methods of controlled insertion of macrophytes within an outdoor mesocosm study were explored. Although analytically confirmed concentrations of the model herbicide terbuthylazine were high enough to expect direct effects on phytoplankton, functional parameters and dominant taxa abundance indicated only minor and transient effects. In-situ assays with Lemna minor, Myriophyllum spicatum, Potamogeton lucens and Chara globularis revealed adverse effects at concentrations in accordance with literature data. Complex interactions such as nutrient limitation and competition were possible reasons for the observed growth promotion at the lower concentration of about 5 microg/l terbuthylazine. The approach of macrophyte in-situ bioassays within a mesocosm study proved to be applicable. Presumed advantages are simultaneous acquisition of toxicity data for several species of aquatic plants under more realistic conditions compared to laboratory tests and inclusion of macrophytes as important structural and functional components in mesocosms while limiting their domination of the model ecosystem.

  18. Bioassay data and a retention-excretion model for systemic plutonium

    International Nuclear Information System (INIS)

    Leggett, R.W.

    1984-05-01

    The estimation of systemic burdens from urinalyses has been the most common and useful method of quantifying occupational exposures to plutonium. Problems arise in using this technique, however, because of inadequate modeling of human retention, translocation, and excretion of this element. Present methods for estimating the systemic burden from urinalyses were derived to a large extent from patterns observed in the first few months after exposure, but there is now evidence that these same patterns do not persist over long periods. In this report we collect and discuss data needed for the interpretation of bioassay results for Pu. These data are used to develop a model that describes the movement, retention, and excretion of systemic Pu in the human body in terms of explicitly identified anatomical compartments. This model may be used in conjunction with existing models and/or case-specific information concerning the translocation of Pu from the respiratory or gastrointestinal tract or from wounds to the bloodstream. Attention is restricted to the behavior of Pu after it has gained access to the bloodstream. There remain significant uncertainties concerning some aspects of the movement of Pu, particularly its translocation from the liver. An attempt has been made to construct the model in such a way as to elucidate those areas needing further attention. 98 references, 18 figures, 16 tables

  19. Quantum Dot-Based Luminescent Oxygen Channeling Assay for Potential Application in Homogeneous Bioassays.

    Science.gov (United States)

    Zhuang, Si-Hui; Guo, Xin-Xin; Wu, Ying-Song; Chen, Zhen-Hua; Chen, Yao; Ren, Zhi-Qi; Liu, Tian-Cai

    2016-01-01

    The unique photoproperties of quantum dots are promising for potential application in bioassays. In the present study, quantum dots were applied to a luminescent oxygen channeling assay. The reaction system developed in this study was based on interaction of biotin with streptavidin. Carboxyl-modified polystyrene microspheres doped with quantum dots were biotinylated and used as acceptors. Photosensitizer-doped carboxyl-modified polystyrene microspheres were conjugated with streptavidin and used as donors. The results indicated that the singlet oxygen that was released from the donor beads diffused into the acceptor beads. The acceptor beads were then exited via thioxene, and were subsequently fluoresced. To avoid generating false positives, a high concentration (0.01 mg/mL) of quantum dots is required for application in homogeneous immunoassays. Compared to a conventional luminescent oxygen channeling assay, this quantum dots-based technique requires less time, and would be easier to automate and miniaturize because it requires no washing to remove excess labels.

  20. Towards the development of an embryotoxicity bioassay with terrestrial snails: Screening approach for cadmium and pesticides

    Energy Technology Data Exchange (ETDEWEB)

    Druart, Coline; Scheifler, Renaud [Department of Chrono-Environment, University of Franche-Comte, UMR UFC/CNRS 6249 usc INRA, Place Leclerc, F-25030 Besancon Cedex (France); Vaufleury, Annette de, E-mail: annette.devaufleury@univ-fcomte.fr [Department of Chrono-Environment, University of Franche-Comte, UMR UFC/CNRS 6249 usc INRA, Place Leclerc, F-25030 Besancon Cedex (France)

    2010-12-15

    Currently no bioassays are available to assess the embryotoxicity of chemicals with terrestrial soil invertebrates. We therefore presented a new method for embryotoxicity testing with snail eggs: a relevant biological material that incubates in soil and that can be exposed to contaminants from leachates and soil solution. The effects of aqueous solutions of two herbicide formulations, Reglone (active ingredient (a.i.), diquat) and Roundup or its a.i., glyphosate, of a surfactant (Agral 90, a.i., nonylphenol polyethoxylates) and of cadmium (Cd) were studied. Endpoints were the hatching success and observations of embryo abnormalities after exposure. Roundup was found to be more toxic than its a.i. alone (EC50{sub a.i.} = 18 mg/l and about 1300 mg/l, respectively). Reglone (EC50{sub a.i.} = 0.72 mg/l) and Agral (EC50{sub a.i.} {approx} 50 mg/l) were also tested together, revealing that Reglone accounted for more than 99% of the mixture's toxicity. An antagonistic interaction between the two substances was found. For Cd (EC50 = 3.9 mg/l), a significant transfer from exposure medium to eggs was emphasized, particularly affecting the albumen. Abnormalities of embryogenesis in non-hatched embryos depended on the substance and the concentration considered.

  1. Toxicity assessment of untreated/treated electroplating sludge using human and plant bioassay.

    Science.gov (United States)

    Orescanin, Visnja; Durgo, Ksenija; Mikelic, Ivanka Lovrencic; Halkijevic, Ivan; Kuspilic, Marin

    2018-04-30

    The purpose of this work was to assess the risk to the environment arising from the electroplating sludge from both chemical and toxicological point of view. Both approaches were used for the assessment of the treatment efficiency which consisted of CaO based solidification followed by thermal treatment at 400°C. The elemental composition was determined in the bulk samples and the leachates of untreated sludge. The toxicity of the leachate was determined using two human colorectal adenocarcinoma cell lines (Caco-2 and SW 480) and Hordeum vulgare L. based plant bioassay. The same toxicity tests were employed to the leachate of the treated sludge. Untreated sludge showed extremely high cytotoxic effect to both human and plant bio-system in dose-dependent manner. The percentages higher than 0.5% and 0.05% of the leachate caused significant cytotoxic effect on Caco-2 and SW 480 cells, respectively. The percentages of the leachate higher than 0.05% also showed significant toxic effect to H. vulgare L. bio-system with complete arrest of seed germination following the treatment with 100% to 5% of the leachate. The leachate of the treated sludge showed no toxicity to any of the test systems confirming the efficiency and justification of the employed procedures for the detoxification of electroplating sludge.

  2. Assessment of the ecotoxicity of urban estuarine sediment using benthic and pelagic copepod bioassays.

    Science.gov (United States)

    Charry, Maria P; Keesing, Vaughan; Costello, Mark; Tremblay, Louis A

    2018-01-01

    Urban estuarine sediments are sinks to a range of contaminants of anthropogenic origin, and a key challenge is to characterize the risk of these compounds to receiving environments. In this study, the toxicity of urban estuarine sediments was tested using acute and chronic bioassays in the benthic harpacticoid Quinquelaophonte sp., and in the planktonic calanoid Gladioferens pectinatus , two New Zealand copepod species. The sediment samples from the estuary tributary sites significantly impacted reproduction in Quinquelaophonte sp. However, results from one of the estuary sites were not significantly different to those from the tributaries sites, suggesting that chemicals other than trace metals, polycyclic aromatic hydrocarbons and ammonia may be the causative stressors. Sediment elutriate samples had significant effects on reproductive endpoints in G. pectinatus , and on the induction of DNA damage in cells, as shown by the comet assay. The results indicate that sediment contamination at the Ahuriri Estuary has the potential to impact biological processes of benthic and pelagic organisms. The approach used provides a standardized methodology to assess the toxicity of estuarine sediments.

  3. Eco- and genotoxicity profiling of a rapeseed biodiesel using a battery of bioassays.

    Science.gov (United States)

    Eck-Varanka, Bettina; Kováts, Nora; Horváth, Eszter; Ferincz, Árpád; Kakasi, Balázs; Nagy, Szabolcs Tamás; Imre, Kornélia; Paulovits, Gábor

    2018-04-30

    Biodiesel is considered an important renewable energy source but still there is some controversy about its environmental toxicity, especially to aquatic life. In our study, the toxicity of water soluble fraction of biodiesel was evaluated in relatively low concentrations using a battery of bioassays: Vibrio fischeri bioluminescence inhibition, Sinapis alba root growth inhibition, Daphnia magna immobilization, boar semen live/dead ratio and DNA fragmentation and Unio pictorum micronucleus test. While the S. alba test indicated nutritive (stimulating) effect of the sample, the biodiesel exerted toxic effect in the aquatic tests. D. magna was the most sensitive with EC 50 value of 0.0226%. For genotoxicity assessment, the mussel micronucleus test (MNT) was applied, detecting considerable genotoxic potential of the biodiesel sample: it elucidated micronuclei formation already at low concentration of 3.3%. Although this test has never been employed in biodiesel eco/genotoxicity assessments, it seems a promising tool, based on its appropriate sensitivity, and representativity. Copyright © 2018 Elsevier Inc. All rights reserved.

  4. Influence of cAMP on reporter bioassays for dioxin and dioxin-like compounds

    International Nuclear Information System (INIS)

    Kasai, Ayumi; Yao, Jian; Yamauchi, Kozue; Hiramatsu, Nobuhiko; Hayakawa, Kunihiro; Meng, Yiman; Maeda, Shuichiro; Kitamura, Masanori

    2006-01-01

    In reporter assays for detection of dioxins, the dioxin-responsive element (DRE) is generally used as a sensor sequence. In several systems, the CYP1A1 promoter containing DREs (DRE cyp ) is inserted into a part of the long terminal repeat of mouse mammary tumor virus (LTR MMTV ) to improve sensitivity of assays. We found that DRE cyp -LTR MMTV responds not only to dioxins and dioxin-like compounds but also to forskolin, a cAMP-elevating agent. This effect was dose-dependent and reproduced by other cAMP-elevating agents including 8-bromo-cAMP and 3-isobutyl-methylxanthine. The cAMP response element (CRE) and CRE-like sequences were absent in DRE cyp -LTR MMTV and not involved in this process. In contrast to the effect of dioxin, the activation of DRE cyp -LTR MMTV by cAMP was independent of the aryl hydrocarbon receptor (AhR), a ligand-dependent transcription factor for DRE. Furthermore, neither DRE cyp , LTR MMTV nor the consensus sequence of DRE alone was activated in response to cAMP. These data elucidated for the first time that the combination of DRE cyp with LTR MMTV causes a peculiar response to cAMP and suggested that use of AhR antagonists is essential to exclude false-positive responses of DRE cyp -LTR MMTV -based bioassays for detection and quantification of dioxins and dioxin-like compounds

  5. Applicability of the CALUX bioassay for screening of dioxin levels in human milk samples

    DEFF Research Database (Denmark)

    Laier, P.; Cederberg, Tommy Licht; Larsen, John Christian

    2003-01-01

    The CALUX (chemically activated luciferase expression) bioassay based on rat hepatoma (H4IIE) cells is a sensitive assay for the detection of Ah receptor agonists like 2,3,7,8-substituted chlorinated dibenzo-p-dioxins and dibenzofurans and related PCBs. In this paper, the assay was optimized...... and applied for monitoring levels of dioxins in human milk samples. Combination effects of dioxin-like compounds were evaluated by testing potential mechanisms of interaction between seven of the major dioxin-like compounds in human milk using the isobole method. Results showed that the compounds acted...... lower REP in CALUX. The total dioxin-like activity was determined in 16 Danish human milk samples and was in the range 20.5-55.8 pg TEQ g(-1) fat. These values were compared with TEQs obtained from GC/MS analysis (range 14.8-43.6 pg TEQ-g(-1) fat) that overall were a little lower than CALUX TEQs...

  6. Bioassay-Guided Isolation of Neuroprotective Compounds from Uncaria rhynchophylla against Beta-Amyloid-Induced Neurotoxicity.

    Science.gov (United States)

    Xian, Yan-Fang; Lin, Zhi-Xiu; Mao, Qing-Qiu; Hu, Zhen; Zhao, Ming; Che, Chun-Tao; Ip, Siu-Po

    2012-01-01

    Uncaria rhynchophylla is a component herb of many Chinese herbal formulae for the treatment of neurodegenerative diseases. Previous study in our laboratory has demonstrated that an ethanol extract of Uncaria rhynchophylla ameliorated cognitive deficits in a mouse model of Alzheimer's disease induced by D-galactose. However, the active ingredients of Uncaria rhynchophylla responsible for the anti-Alzheimer's disease activity have not been identified. This study aims to identify the active ingredients of Uncaria rhynchophylla by a bioassay-guided fractionation approach and explore the acting mechanism of these active ingredients by using a well-established cellular model of Alzheimer's disease, beta-amyloid- (Aβ-) induced neurotoxicity in PC12 cells. The results showed that six alkaloids, namely, corynoxine, corynoxine B, corynoxeine, isorhynchophylline, isocorynoxeine, and rhynchophylline were isolated from the extract of Uncaria rhynchophylla. Among them, rhynchophylline and isorhynchophylline significantly decreased Aβ-induced cell death, intracellular calcium overloading, and tau protein hyperphosphorylation in PC12 cells. These results suggest that rhynchophylline and isorhynchophylline are the major active ingredients responsible for the protective action of Uncaria rhynchophylla against Aβ-induced neuronal toxicity, and their neuroprotective effect may be mediated, at least in part, by inhibiting intracellular calcium overloading and tau protein hyperphosphorylation.

  7. Bioassay-Guided Isolation of Neuroprotective Compounds from Uncaria rhynchophylla against Beta-Amyloid-Induced Neurotoxicity

    Directory of Open Access Journals (Sweden)

    Yan-Fang Xian

    2012-01-01

    Full Text Available Uncaria rhynchophylla is a component herb of many Chinese herbal formulae for the treatment of neurodegenerative diseases. Previous study in our laboratory has demonstrated that an ethanol extract of Uncaria rhynchophylla ameliorated cognitive deficits in a mouse model of Alzheimer’s disease induced by D-galactose. However, the active ingredients of Uncaria rhynchophylla responsible for the anti-Alzheimer’s disease activity have not been identified. This study aims to identify the active ingredients of Uncaria rhynchophylla by a bioassay-guided fractionation approach and explore the acting mechanism of these active ingredients by using a well-established cellular model of Alzheimer’s disease, beta-amyloid- (Aβ- induced neurotoxicity in PC12 cells. The results showed that six alkaloids, namely, corynoxine, corynoxine B, corynoxeine, isorhynchophylline, isocorynoxeine, and rhynchophylline were isolated from the extract of Uncaria rhynchophylla. Among them, rhynchophylline and isorhynchophylline significantly decreased Aβ-induced cell death, intracellular calcium overloading, and tau protein hyperphosphorylation in PC12 cells. These results suggest that rhynchophylline and isorhynchophylline are the major active ingredients responsible for the protective action of Uncaria rhynchophylla against Aβ-induced neuronal toxicity, and their neuroprotective effect may be mediated, at least in part, by inhibiting intracellular calcium overloading and tau protein hyperphosphorylation.

  8. Use of Whole-Cell Bioassays for Screening Quorum Signaling, Quorum Interference, and Biofilm Dispersion.

    Science.gov (United States)

    Thornhill, Starla G; McLean, Robert J C

    2018-01-01

    In most bacteria, a global level of regulation, termed quorum sensing (QS), exists involving intercellular communication via the production and response to cell density-dependent signal molecules. QS has been associated with a number of important features in bacteria including virulence regulation and biofilm formation. Consequently, there is considerable interest in understanding, detecting, and inhibiting QS. N-acylated homoserine lactones (AHLs) are used as extracellular QS signals by a variety of Gram-negative bacteria. Chromobacterium violaceum, commonly found in soil and water, produces the characteristic purple pigment violacein, regulated by AHL-mediated QS. Based on this readily observed pigmentation phenotype, C. violaceum strains can be used to detect various aspects of AHL-mediated QS activity. In another commonly used bioassay organism, Agrobacterium tumefaciens, QS can be detected by the use of a reporter gene such as lacZ. Here, we describe several commonly used approaches incorporating C. violaceum and A. tumefaciens that can be used to detect AHL and QS inhibitors. Due to the inherent low susceptibility of biofilm bacteria to antimicrobial agents, biofilm dispersion, whereby bacteria reenter the planktonic community, is another increasingly important area of research. At least one signal, distinct from traditional QS, has been identified and there are a variety of other environmental factors that also trigger dispersion. We describe a microtiter-based experimental strategy whereby potential biofilm dispersion compounds can be screened.

  9. Analysis of 'wet-landscape' surface water fractions using medaka embryo-toxicity bioassay

    Energy Technology Data Exchange (ETDEWEB)

    Peters, L. E.; McConkey, B. J.; Vanden Heuvel, M. R. (Waterloo, Univ., Dept, of Biology, Waterloo, ON (Canada)); MacKinnon, M. D. (Syncrude Canada Ltd., Fort McMurray, AB (Canada)) Munkittricx, K. (Environment Canada, Burlington, ON (Canada))

    1998-01-01

    The self-sustaining biological potential of Syncrude's 'wetland-scape' waste disposal method was evaluated by testing water extracts from experimental pits of different ages and fine tailings/natural water compositions. This waste disposal method involves capping fine tailings with a layer of surface water. Preliminary estimates suggests a higher incidence of mortality and deformity in Japanese Medaka embryos incubated in pit waters containing elevated concentrations of naphthenates. Another study on adult perch stocked in the demonstration pit indicated the presence of PAHs in the fish bile at biologically relevant concentrations. This study was designed to determine the causative agents of the fish embryo toxicity and the level of concentrations at which chronic effects occur. The water extracts were fractionated into acid (containing naphthenates) and base-neutral (containing PAHs) components and tested using the Japanese Medaka bioassay. Endpoints measured were the presence of deformity, hatch success, swim-bladder inflation, length at hatch and time to mortality. HPLC analysis showed that PAHs were present at concentrations in the part/billion and the parts/million range. This is being taken as an indication that PAHs are not directly responsible for the observed toxicity to the embryos.

  10. Influence of the statistical distribution of bioassay measurement errors on the intake estimation

    International Nuclear Information System (INIS)

    Lee, T. Y; Kim, J. K

    2006-01-01

    The purpose of this study is to provide the guidance necessary for making a selection of error distributions by analyzing influence of statistical distribution for a type of bioassay measurement error on the intake estimation. For this purpose, intakes were estimated using maximum likelihood method for cases that error distributions are normal and lognormal, and comparisons between two distributions for the estimated intakes were made. According to the results of this study, in case that measurement results for lung retention are somewhat greater than the limit of detection it appeared that distribution types have negligible influence on the results. Whereas in case of measurement results for the daily excretion rate, the results obtained from assumption of a lognormal distribution were 10% higher than those obtained from assumption of a normal distribution. In view of these facts, in case where uncertainty component is governed by counting statistics it is considered that distribution type have no influence on intake estimation. Whereas in case where the others are predominant, it is concluded that it is clearly desirable to estimate the intake assuming a lognormal distribution

  11. Performance characteristics of a Bioassay method for plutonium using thermal ionisation mass spectrometry (TIMS)

    Energy Technology Data Exchange (ETDEWEB)

    Linauskas, S.H.; Elliot, N.L.; Paterson, L.M.; Totland, M.M

    2003-01-01

    Plutonium-in-urine analysis by radiochemical isolation of Pu followed by thermal ionisation mass spectrometry (TIMS) is capable of high sensitivity and precision measurements {sup 239}Pu and {sup 240}Pu. Bias and precision estimates for the TIMS bioassay program at Chalk River Laboratories easily met the ANSI N13.30 performance criteria standards with {sup 239}Pu results of 1.5% and 3.0%, respectively. Analytical blanks derived from water, artificial urine and true urine samples did not produce any statistically different results. During a four-year period of development and implementation of {sup 239}Pu measurements by TIMS, average sample blank values were reduced from 3.9 fg (9.0 {mu}Bq) to 0.57 fg (1.3 {mu}Bq). This reduction was achieved through rigorous application of clean-room handling techniques throughout sample processing. Blank data were found to follow a Iognormal distribution, and current detection limit parameters for L{sub c} and L{sub d} at the 95% significance levels are 0.85 fg {sup 239}Pu (2.0 {mu}Bq) and 1.3 fg {sup 239}Pu (3.0 {mu}Bq), respectively. Detection limits in this range are expected to be sufficient to identify intakes of Pu/Am mixtures at levels that are around one-twentieth of an ALl or better under routine monitoring situations for ICRP Type S and Type M inhalation solubility classes. (author)

  12. Spinal cord neuronotrophic factors (SCNTFs): I. Bioassay of schwannoma and other conditioned media.

    Science.gov (United States)

    Longo, F M; Manthorpe, M; Varon, S

    1982-02-01

    We present a procedure for the dissociation and growth in serum-free defined culture medium of 4-day chick embryo lumbar spinal cord (LC4) neurons. LC4 neurons will not survive for even 24 h without the addition of trophic supplements (putative spinal cord neuronotrophic factors, SCNTFs). Serum-free medium conditioned over chick embryo heart and skeletal muscle, mouse Schwann and rat RN22 Schwannoma cell cultures were found to contain SCNTF activity which could be quantitated using a convenient neuronal survival bioassay. RN22 conditioned medium also contains polyornithine-binding neurite promoting factors (PNPFs) which can be physically separated from SCNTF. When SCNTF and PNPF were presented to LC4 neurons individually or in combination (i) SCNTF, but not PNPF, supported neuronal survival whereas (ii) PNPF, but not SCNTF, induced neurite production. When LC4 neurons were grown in SCNTF alone, nearly all of them exhibited a flattened, circular, 'fried-egg' morphology. The subsequent addition of PNPF caused these cells to extend long neurites with characteristic terminal growth-cone-like structures.

  13. Comparisons of uniform and discrete source distributions for use in bioassay laboratory performance testing

    International Nuclear Information System (INIS)

    Scherpelz, R.I.; MacLellan, J.A.

    1987-09-01

    The Pacific Northwest Laboratory (PNL) is sending a torso phantom with radioactive material uniformly distributed in the lungs to in vivo bioassay laboratories for analysis. Although the radionuclides ultimately chosen for the studies had relatively long half-lives, future accreditation testing will require repeated tests with short half-life test nuclides. Computer modeling was used to simulate the major components of the phantom. Radiation transport calculations were then performed using the computer models to calculate dose rates either 15 cm from the chest or at its surface. For 144 Ce and 60 Co, three configurations were used for the lung comparison tests. Calculations show that, for most detector positions, a single plug containing 40 K located in the back of the heart provides a good approximation to a uniform distribution of 40 K. The approximation would lead, however, to a positive bias for the detector reading if the detector were located at the chest surface near the center. Loading the 40 K in a uniform layer inside the chest wall is not a good approximation of the uniform distribution in the lungs, because most of the radionuclides would be situated close to the detector location and the only shielding would be the thickness of the chest wall. The calculated dose rates for 60 Co and 144 Ce were similar at all calculated reference points. 3 refs., 5 figs., 10 tabs

  14. Larval development ratio test with the calanoid copepod Acartia tonsa as a new bioassay to assess marine sediment quality.

    Science.gov (United States)

    Buttino, Isabella; Vitiello, Valentina; Macchia, Simona; Scuderi, Alice; Pellegrini, David

    2018-03-01

    The copepod Acartia tonsa was used as a model species to assess marine sediment quality. Acute and chronic bioassays, such as larval development ratio (LDR) and different end-points were evaluated. As a pelagic species, A. tonsa is mainly exposed to water-soluble toxicants and bioassays are commonly performed in seawater. However, an interaction among A. tonsa eggs and the first larval stages with marine sediments might occur in shallow water environments. Here we tested two different LDR protocols by incubating A. tonsa eggs in elutriates and sediments coming from two areas located in Tuscany Region (Central Italy): Livorno harbour and Viareggio coast. The end-points analyzed were larval mortality (LM) and development inhibition (DI) expressed as the percentage of copepods that completed the metamorphosis from nauplius to copepodite. Aims of this study were: i) to verify the suitability of A. tonsa copepod for the bioassay with sediment and ii) to compare the sensitivity of A. tonsa exposed to different matrices, such as water and sediment. A preliminary acute test was also performed. Acute tests showed the highest toxicity of Livorno's samples (two out of three) compared to Viareggio samples, for which no effect was observed. On the contrary, LDR tests with sediments and elutriates revealed some toxic effects also for Viareggio's samples. Results were discussed with regards to the chemical characterization of the samples. Our results indicated that different end-points were affected in A. tonsa, depending on the matrices to which the copepods were exposed and on the test used. Bioassays with elutriates and sediments are suggested and LDR test could help decision-makers to identify a more appropriate management of dredging materials. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Chemical Exacerbation of Light-induced Retinal Degeneration in F344/N Rats in National Toxicology Program Rodent Bioassays

    OpenAIRE

    Yamashita, Haruhiro; Hoenerhoff, Mark J.; Peddada, Shyamal D.; Sills, Robert C.; Pandiri, Arun R.

    2016-01-01

    Retinal degeneration due to chronic ambient light exposure is a common spontaneous age-related finding in albino rats, but it can also be related to exposures associated with environmental chemicals and drugs. Typically, light induced retinal degeneration has a central/hemispherical localization where as chemical induced retinal degeneration has a diffuse localization. This study was conducted to identify National Toxicology Program (NTP) rodent bioassays with treatment-related retinal degene...

  16. Comparison of bioassays using the anostracan crustaceans Artemia salina and Thamnocephalus platyurus for plant extract toxicity screening

    OpenAIRE

    Mayorga,Pablo; Pérez,Karen R.; Cruz,Sully M.; Cáceres,Armando

    2010-01-01

    Three lethality bioassays, using the salt-water crustacean Artemia salina Leach, Artemiidae, (conventional 96 microwell plate test and the Artoxkit M microbiotest) and the freshwater crustacean Thamnocephalus platyurus Packard, Thamnocephalidae, (Thamnotoxkit F microbiotest), were compared using extracts of ten Guatemalan plant species. It was previously observed that five of them have anti-Artemia activity. These were: Solanum americanum Mill., Solanaceae, Gliricidia sepium (Jacq.) Kunth ex ...

  17. Comparative performances of eggs and embryos of sea urchin (Paracentrotus lividus) in toxicity bioassays used for assessment of marine sediment quality.

    Science.gov (United States)

    Khosrovyan, A; Rodríguez-Romero, A; Salamanca, M J; Del Valls, T A; Riba, I; Serrano, F

    2013-05-15

    The potential toxicity of sediments from various ports was assessed by means of two different liquid-phase toxicity bioassays (acute and chronic) with embryos and eggs of sea urchin Paracentrotus lividus. Performances of embryos and eggs of P. lividus in these bioassays were compared for their interchangeable applicability in integrated sediment quality assessment. The obtained endpoints (percentages of normally developed plutei and fertilized eggs) were linked to physical and chemical properties of sediments and demonstrated dependence on sediment contamination. The endpoints in the two bioassays were strongly correlated and generally exhibited similar tendency throughout the samples. Therein, embryos demonstrated higher sensitivity to elutriate exposure, compared to eggs. It was concluded that these tests could be used interchangeably for testing toxicity of marine sediments. Preferential use of any of the bioassays can be determined by the discriminatory capacity of the test or vulnerability consideration of the test subject to the surrounding conditions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Different bioassays for investigating orientation responses of the banana weevil, Cosmopolites sordidus, show additive effects of host plant volatiles and a synthetic male-produced aggregation pheromone

    NARCIS (Netherlands)

    Tinzaara, W.; Dicke, M.; Huis, van A.; Loon, van J.A.; Gold, C.S.

    2003-01-01

    Three different bioassay methods to investigate the orientation behaviour of the banana weevil, Cosmopolites sordidus (Germar) (Coleoptera: Curculionidae), to host plant volatiles and a synthetic pheromone (cosmolure+) were compared. A locomotion compensator was used to separately record walking

  19. Biomarkers and bioassays as alternative screening methods for the presence and effects of PCDD, PCDF and PCB

    International Nuclear Information System (INIS)

    Bosveld, A.T.C.B.; Berg, M.V.

    1994-01-01

    Polyhalogenated aromatic hydrocarbons (PHAH) are wide spread, highly toxic, environmental contaminants. As such they pose risks for both humans and wildlife. For risk assessment purposes, concentrations are generally analyzed by HRGC-HR/LRMS. With the analytical data, mixture toxicity is calculated using the TEF concept. With this method only the defined congeners are taken into account and additivity for all congeners is assumed, whereas synergistic and antagonistic effects for several PCDD/F in combination with PCB have also been reported. To avoid these problems and high analytical costs, bioassays can be used for screening purposes. Cytochrome P 450 1 A 1 induction and vitamin A and thyroid hormone levels are shown to be useful markers for PHAH exposure. When bioassays based on cytochrome P 450 1 A 1 induction, in cultured cells, in multi-well culturing plates, are used, 2,3,7,8-TCDD detection limits <0.2 pg are possible. As such these bioassays are highly sensitive, cost effective and time saving. This application can be used as a pre-screening method to determine total ''dioxin'' content of environmental samples. (orig.)

  20. Linking in situ bioassays and population dynamics of macroinvertebrates to assess agricultural contamination in streams of the Argentine pampa.

    Science.gov (United States)

    Jergentz, S; Pessacq, P; Mugni, H; Bonetto, C; Schulz, R

    2004-10-01

    The two local crustacean species Hyalella curvispina and Macrobrachium borelli were chosen for assessment of agricultural contamination in two streams (Horqueta and Maguire) in the Argentine pampa. In parallel with in situ bioassays of both species, the population dynamics and the organismic drift of H. curvispina were investigated throughout the main period of insecticide application, from December 2001 to March 2002. In Maguire none of the current-use insecticides (chlorpyrifos, alpha-cypermethrin, and endosulfan) in question were detected throughout the sampling period. During 1-week intervals with no contamination by insecticides the survival rate of H. curvispina varied between 77 +/- 6% (+/- SE, n = 4) and 85 +/- 3%. In Horqueta during a week with a peak insecticide contamination of 64 microg/kg chlorpyrifos in the suspended particles, a mortality of 100% was observed in the in situ bioassays for both species, H. curvispina and M. borelli. At the same time, in Maguire H. curvispina showed reduced survival rates of 23 +/- 5% and 25 +/- 18% at the two sites, while the survival rate of M. borelli was 60 +/- 11% upstream and 93 +/- 5% downstream, below a wetland. During the period with 100% mortality of H. curvispina in Horqueta, the population density of this species decreased correspondingly, from 106 +/- 26 to 0 individuals/m(2). We conclude that in situ bioassays can be successfully linked to in-stream population dynamics for the same species and that this link is very useful for interpreting causal exposure-effect relationships.

  1. Assessment of sediment contamination by spermiotoxicity and embryotoxicity bioassays with sea urchins (Paracentrotus lividus) and oysters (Crassostrea gigas).

    Science.gov (United States)

    Geffard, O; Budzinski, H; Augagneur, S; Seaman, M N; His, E

    2001-07-01

    Gametes (sperm) and fertilized eggs (embryos) of the Mediterranean sea urchin, Paracentrotus lividus, and the Japanese oyster, Crassostrea gigas, were used to investigate the toxicity of two marine sediments, one polluted by polycyclic aromatic hydrocarbons (PAH) and the other by heavy metals. The sediment samples were freeze-dried for storage, and three different treatments were used for analysis: whole sediment, unfiltered elutriate, and filtered elutriate. The two sediments were toxic to sea urchin spermatozoa but not to oyster spermatozoa, and embryotoxicity was almost always the more sensitive endpoint for toxicity assessment. As a rule, whole sediment was more toxic than the elutriates by nearly two orders of magnitude. With respect to embryotoxicity, the whole sediments and the elutriates of the PAH-contaminated sediment were more toxic to oyster embryos, whereas the elutriates of the sediment polluted by heavy metals had stronger effects on sea urchin embryos. The results confirm that bioassays with Japanese oyster embryos provide a more sensitive appraisal of toxicity in the marine environment than bioassays with other developmental stages. As a whole, Mediterranean sea urchins and Japanese oysters were similar in overall sensitivity and are therefore both equally suited as bioassay organisms, but tests with oysters are more reproducible because of the better performance of the controls.

  2. Use of plant and earthworm bioassays to evaluate remediation of soil from a site contaminated with polychlorinated biphenyls

    Energy Technology Data Exchange (ETDEWEB)

    Meier, J.R.; Chang, L.W.; Meckes, M.C.; Smith, M.K. [Environmental Protection Agency, Cincinnati, OH (United States); Jacobs, S. [DynCorp, Cincinnati, OH (United States); Torsella, J. [Oak Ridge Inst. of Science and Education, Cincinnati, OH (United States)

    1997-05-01

    Soil from a site heavily contaminated with polychlorinated biphenyls (PCBs) was treated with a pilot-scale, solvent extraction technology. Bioassays in earthworms and plants were used to examine the efficacy of the remediation process for reducing the toxicity of the soil. The earthworm toxicity bioassays were the 14-d survival test and 21-d reproduction test, using Lumbricus terrestris and Eisenia fetida andrei. The plant bioassays included phytotoxicity tests for seed germination and root elongation in lettuce and oats, and a genotoxicity test (anaphase aberrations) in Allium cepa (common onion). Although the PCB content of the soil was reduced by 99% (below the remediation goal), toxicity to earthworm reproduction remained essentially unchanged following remediation. Furthermore, phytotoxicity and genotoxicity were higher for the remediated soil compared to the untreated soil. The toxicity remaining after treatment appeared to be due to residual solvent introduced during the remediation process, and/or to heavy metals or other inorganic contaminants not removed by the treatment. Mixture studies involving isopropanol and known toxicants indicated possible synergistic effects of the extraction solvent and soil contaminants. The toxicity in plants was essentially eliminated by a postremediation, water-rinsing step. These results demonstrate a need for including toxicity measurements in the evaluation of technologies used in hazardous waste site remediations, and illustrate the potential value of such measurements for making modifications to remediation processes.

  3. Comparing the impacts of sediment-bound bifenthrin on aquatic macroinvertebrates in laboratory bioassays and field microcosms.

    Science.gov (United States)

    Boyle, Rhianna L; Hoak, Molly N; Pettigrove, Vincent J; Hoffmann, Ary A; Long, Sara M

    2016-11-01

    We conducted two laboratory bioassays and two field microcosm exposures with bifenthrin (a synthetic pyrethroid) in order to evaluate the capacity of single-species laboratory bioassays to predict lethal and sublethal impacts on aquatic invertebrates in microcosms. For the laboratory species, Chironomus tepperi, larval survival was reduced by 24% at 53.66µg/g OC, while adult emergence was reduced at concentrations of 33.33µg/g OC and higher, with a 61% decrease at 77.78µg/g OC and no emergence at 126.67µg/g OC. The abundance of several other microcosm taxa was reduced in the microcosms at a similar concentration range (33.33µg/g OC and above), however there was no impact on the abundance of the congeneric species, Chironomus oppositus. The differences in impacts between test systems were potentially due to both differing species sensitivity and the interaction of ambient temperature with bifenthrin toxicity. Bifenthrin also was associated with early emergence of Chironomus sp. in both test systems, at concentrations of 10µg/g OC and higher (laboratory) and 43.90µg/g OC (microcosm), and with a significant decrease in the proportion of C. oppositus males in a microcosm. These findings indicate that while laboratory bioassays accurately predict many impacts in the field, there are some limitations to the predictive capacity of these tests. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Monitoring resistance to Bacillus thuringiensis subsp. israelensis in the field by performing bioassays with each Cry toxin separately

    Directory of Open Access Journals (Sweden)

    Guillaume Tetreau

    2013-11-01

    Full Text Available Bacillus thuringiensis subsp. israelensis (Bti is increasingly used worldwide for mosquito control and is the only larvicide used in the French Rhône-Alpes region since decades. The artificial selection of mosquitoes with field-persistent Bti collected in breeding sites from this region led to a moderate level of resistance to Bti, but to relatively high levels of resistance to individual Bti Cry toxins. Based on this observation, we developed a bioassay procedure using each Bti Cry toxin separately to detect cryptic Bti-resistance evolving in field mosquito populations. Although no resistance to Bti was detected in none of the three mosquito species tested (Aedes rusticus, Aedes sticticus and Aedes vexans, an increased tolerance to Cry4Aa (3.5-fold and Cry11Aa toxins (8-fold was found in one Ae. sticticus population compared to other populations of the same species, suggesting that resistance to Bti may be arising in this population. This study confirms previous works showing a lack of Bti resistance in field mosquito populations treated for decades with this bioinsecticide. It also provides a first panorama of their susceptibility status to individual Bti Cry toxins. In combination with bioassays with Bti, bioassays with separate Cry toxins allow a more sensitive monitoring of Bti-resistance in the field.

  5. An aggregated perylene-based broad-spectrum, efficient and label-free quencher for multiplexed fluorescent bioassays.

    Science.gov (United States)

    Liu, Tao; Hu, Rong; Lv, Yi-Fan; Wu, Yuan; Liang, Hao; Huan, Shuang-Yan; Zhang, Xiao-Bing; Tan, Weihong; Yu, Ru-Qin

    2014-08-15

    Fluorescent sensing systems based on the quenching of fluorophores have found wide applications in bioassays. An efficient quencher will endow the sensing system a high sensitivity. The frequently used quenchers are based on organic molecules or nanomaterials, which usually need tedious synthesizing and modifying steps, and exhibit different quenching efficiencies to different fluorophores. In this work, we for the first time report that aggregated perylene derivative can serve as a broad-spectrum and label-free quencher that is able to efficiently quench a variety of fluorophores, such as green, red and far red dyes labeled on DNA. By choosing nucleases as model biomolecules, such a broad-spectrum quencher was then employed to construct a multiplexed bioassay platform through a label-free manner. Due to the high quenching efficiency of the aggregated perylene, the proposed platform could detect nuclease with high sensitivity, with a detection limit of 0.03U/mL for EcoRV, and 0.05U/mL for EcoRI. The perylene quencher does not affect the activity of nuclease, which makes it possible to design post-addition type bioassay platform. Moreover, the proposed platform allows simultaneous and multicolor analysis of nucleases in homogeneous solution, demonstrating its value of potential application in rapid screening of multiple bio-targets. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Detection of estrogen receptor endocrine disruptor potency of commonly used organochlorine pesticides using the LUMI-CELL ER bioassay

    Energy Technology Data Exchange (ETDEWEB)

    Gordon, J D; Chu, A C; Clark, G C [Xenobiotic Detection Systems, Inc., Durham, NC (United States); Chu, M D [Alta Analytical Perspectives, Wilmington, NC (United States); Denison, M S [Dept. of Environmental Toxicology, Univ. of California, Davis, CA (United States)

    2004-09-15

    In order to detect the endocrine disrupting potency of organochlorine pesticides and other compounds, BG-1 (human ovarian carcinoma) cells containing a stably transfected estrogenresponsive luciferase reporter gene plasmid (BG1Luc4E2), was used. This cell line, termed the LUMI-CELL trademark ER estrogenic cell bioassay system, responds in a time-, dose dependent- and chemical-specific manner with the induction of luciferase gene expression in response to exposure to estrogen (but not other steroid hormones) and estrogenic chemicals in a high-throughput screening (HTPS) format6. Here we describe studies in which the LUMI-CELL trademark ER estrogenic cell bioassay system was used for high throughput screening (HTPS) analysis of the estrogenic disrupting potency of several commonly used pesticides and organochlorines: p,p'DDT; p,p'-DDE; DDD; {alpha}a-chlordane; {psi}-chlordane; Kepone; Methoxychlor; Vinclozolin; Fenarimol; 2,4,5-Trichlorophenoxyacetic Acid; and Dieldrin. Our results demonstrate the utility of XDS's LUMI-CELL trademark ER bioassay HTPS system for screening chemicals for estrogenic activity.

  7. Evaluation of genotoxic effects caused by extracts of chlorinated drinking water using a combination of three different bioassays.

    Science.gov (United States)

    Zeng, Qiang; Zhang, Shao-Hui; Liao, Jing; Miao, Dong-Yue; Wang, Xin-Yi; Yang, Pan; Yun, Luo-Jia; Liu, Ai-Lin; Lu, Wen-Qing

    2015-10-15

    Potential genotoxic effects of chlorinated drinking water now are of a great concern. In this study, raw water, finished water, and tap water from a water plant in Wuhan, China were collected in two different sampling times of the year (January and July). Genotoxic effects of water extracts were evaluated using a combination of three different bioassays: SOS/umu test, HGPRT gene mutation assay, and micronucleus assay, which were separately used to detect DNA damage, gene mutation, and chromosome aberration. The results of three different bioassays showed that all water samples in January and July induced at least one types of genotoxic effects, of which the DNA-damage effects were all detectable. The levels of DNA-damage effects and gene-mutation effects of finished water and tap water in January were higher than those in July. Chlorination could increase the DNA-damage effects of drinking water in January and the gene-mutation effects of drinking water in both January and July, but did not increase the chromosome-aberration effects of drinking water in both January and July. Our results highlighted the importance of using a combination of different bioassays to evaluate the genotoxicity of water samples in different seasons. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Laboratory algal bioassays using PAM fluorometry: effects of test conditions on the determination of herbicide and field sample toxicity.

    Science.gov (United States)

    Sjollema, Sascha B; van Beusekom, Sebastiaan A M; van der Geest, Harm G; Booij, Petra; de Zwart, Dick; Vethaak, A Dick; Admiraal, Wim

    2014-05-01

    Pulse Amplitude Modulation (PAM) fluorometry, based on chlorophyll a fluorescence, is a frequently used technique in algal bioassays to assess toxicity of single compounds or complex field samples. Several test conditions can influence the test results, and because a standardized test protocol is currently lacking, linking the results of different studies is difficult. Therefore, the aim of the present study was to gain insight into the effects of test conditions of laboratory algal bioassays using PAM fluorometry on the outcome of toxicity tests. To this purpose, we described the results from several pilot studies on test development in which information is provided on the effects of the main test factors during the pretest phase, the test preparation, the exposure period, and the actual measurement. The experiments were focused on individual herbicides and complex field samples and included the effects of culturing conditions, cell density, solvent concentration, exposure time, and the presence of actinic light. Several of these test conditions were found to influence the outcome of the toxicity test, and the presented information provides important background information for the interpretation of toxicity results and describes which test conditions should be taken into account when using an algal bioassay with PAM fluorometry. Finally, the application of PAM fluorometry in algal toxicity testing is discussed. © 2014 SETAC.

  9. Detection of estrogen receptor endocrine disruptor potency of commonly used organochlorine pesticides using the LUMI-CELL ER bioassay

    Energy Technology Data Exchange (ETDEWEB)

    Gordon, J.D.; Chu, A.C.; Clark, G.C. [Xenobiotic Detection Systems, Inc., Durham, NC (United States); Chu, M.D. [Alta Analytical Perspectives, Wilmington, NC (United States); Denison, M.S. [Dept. of Environmental Toxicology, Univ. of California, Davis, CA (United States)

    2004-09-15

    In order to detect the endocrine disrupting potency of organochlorine pesticides and other compounds, BG-1 (human ovarian carcinoma) cells containing a stably transfected estrogenresponsive luciferase reporter gene plasmid (BG1Luc4E2), was used. This cell line, termed the LUMI-CELL trademark ER estrogenic cell bioassay system, responds in a time-, dose dependent- and chemical-specific manner with the induction of luciferase gene expression in response to exposure to estrogen (but not other steroid hormones) and estrogenic chemicals in a high-throughput screening (HTPS) format6. Here we describe studies in which the LUMI-CELL trademark ER estrogenic cell bioassay system was used for high throughput screening (HTPS) analysis of the estrogenic disrupting potency of several commonly used pesticides and organochlorines: p,p'DDT; p,p'-DDE; DDD; {alpha}a-chlordane; {psi}-chlordane; Kepone; Methoxychlor; Vinclozolin; Fenarimol; 2,4,5-Trichlorophenoxyacetic Acid; and Dieldrin. Our results demonstrate the utility of XDS's LUMI-CELL trademark ER bioassay HTPS system for screening chemicals for estrogenic activity.

  10. Assessing soil ecotoxicity of methyl tert-butyl ether using earthworm bioassay; closed soil microcosm test for volatile organic compounds

    International Nuclear Information System (INIS)

    An, Youn-Joo

    2005-01-01

    An earthworm bioassay was conducted to assess ecotoxicity in methyl tert-butyl ether (MTBE)-amended soils. Ecotoxicity of MTBE to earthworms was evaluated by a paper contact method, natural field soil test, and an OECD artificial soil test. All tests were conducted in closed systems to prevent volatilization of MTBE out of test units. Test earthworm species were Perionyx excavatus and Eisenia andrei. Mortality and abnormal morphology of earthworms exposed to different concentrations of MTBE were examined. MTBE was toxic to both earthworm species and the severity of response increased with increasing MTBE concentrations. Perionyx excavatus was more sensitive to MTBE than Eisenia andrei in filter papers and two different types of soils. MTBE toxicity was more severe in OECD artificial soils than in field soils, possibly due to the burrowing behavior of earthworms into artificial soils. The present study demonstrated that ecotoxicity of volatile organic compounds such as MTBE can be assessed using an earthworm bioassay in closed soil microcosm with short-term exposure duration. - Earthworm bioassay can be a good protocol to assess soil ecotoxicity of volatile organic compounds such as MTBE

  11. The sensitivity of an hydroponic lettuce root elongation bioassay to metals, phenol and wastewaters.

    Science.gov (United States)

    Park, Jihae; Yoon, Jeong-Hyun; Depuydt, Stephen; Oh, Jung-Woo; Jo, Youn-Min; Kim, Kyungtae; Brown, Murray T; Han, Taejun

    2016-04-01

    The root elongation bioassay is one of the most straightforward test methods used for environmental monitoring in terms of simplicity, rapidity and economy since it merely requires filter paper, distilled water and Petri dishes. However, filter paper as a support material is known to be problematic as it can reduce the sensitivity of the test. The newly developed hydroponic method reported here differs from the conventional root elongation method (US EPA filter paper method) in that no support material is used and the exposure time is shorter (48 h in this test versus 120 h in the US EPA test). For metals, the hydroponic test method was 3.3 (for Hg) to 57 (for Cu) times more sensitive than the US EPA method with the rank orders of sensitivity, estimated from EC50 values, being Cu≥Cd>Ni≥Zn≥Hg for the former and Hg≥Cu≥Ni≥Cd≥Zn for the latter methods. For phenol, the results did not differ significantly; EC50 values were 124 mg L(-1) and 108-180 mg L(-1) for the hydroponic and filter paper methods, respectively. Lettuce was less sensitive than daphnids to wastewaters, but the root elongation response appears to be wastewater-specific and is especially sensitive for detecting the presence of fluorine. The new hydroponic test thus provides many practical advantages, especially in terms of cost and time-effectiveness requiring only a well plate, a small volume of distilled water and short exposure period; furthermore, no specialist expertise is required. The method is simpler than the conventional EPA technique in not using filter paper which can influence the sensitivity of the test. Additionally, plant seeds have a long shelf-life and require little or no maintenance. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Using Bioassays and Species Sensitivity Distributions to Assess Herbicide Toxicity towards Benthic Diatoms

    Science.gov (United States)

    Larras, Floriane; Bouchez, Agnès; Rimet, Frédéric; Montuelle, Bernard

    2012-01-01

    Although benthic diatoms are widely used in ecological studies of aquatic systems, there is still a dearth of data concerning species sensitivities towards several contaminants. Within the same community, different species may respond differently depending on their physiological and ecological characteristics. This lack of knowledge makes specific appropriate risk assessment impossible. To find out whether species sensitivity distribution (SSD) could be used to estimate the risk of herbicide toxicity for diatoms, we need to know whether their sensitivity depends on their physiological and ecological characteristics. We carried out single-species bioassays on 11 diatom species exposed to 8 herbicides. Dose-responses relationships were used to extrapolate the Effective Concentration 5 (EC5) and the Effective Concentration 50 (EC50) for each exposure. These data were used to fit a SSD curve for each herbicide, and to determine the Hazardous concentration 5 (HC5) and 50 (HC50). Our results revealed a high level of variability of the sensitivity in the set of species tested. For photosystem-II inhibitor (PSII) herbicides, diatoms species displayed a typical grouping of sensitivity levels consistent with their trophic mode and their ecological guild. N-heterotroph and “motile” guild species were more tolerant of PSII inhibitors, while N-autotroph and “low profile” guild species were more sensitive. Comprehensive SSD curves were obtained for 5 herbicides, but not for sulfonylurea herbicides or for dimetachlor, which had toxicity levels that were below the range of concentration tested. The SSD curves provided the following ranking of toxicity: diuron> terbutryn> isoproturon> atrazine> metolachlor. The HC that affected 5% of the species revealed that, even at the usual environmental concentrations of herbicides, diatom assemblages could be affected, especially by isoproturon, terbutryn, and diuron. PMID:22952981

  13. Ultralow-level measurement of organically-bound tritium in bioassay samples

    International Nuclear Information System (INIS)

    Kotzer, T.; Trivedi, A.; Waito, G.; Workman, W.

    1998-12-01

    An intercomparison study of urine samples having high levels (5 Bq·L -1 ) of organically-bound tritium (OBT) was conducted, in conjunction with the oxygen combustion-liquid scintillation counting (LSC) method, to evaluate the suitability and sensitivity of the 3 He-ingrowth mass spectrometry (MS) technique for OBT in bioassay samples. The study established that 3 He ingrowth-MS has the required sensitivity to measure ultralow levels of OBT-in-urine (∼0.1 Bq·L -1 ). Cumulative 24 h urine samples from a few members of the general population, living in the vicinity of the heavy-water research reactor facility at Chalk River Laboratories (CRL) at Chalk River, were collected and analyzed for tritiated water (HTO) and OBT. The participants were from Ottawa (200 km east), Deep River (10 km west) and an occasionally occupationally HTO-exposed worker at CRL. HTO-in-urine values were 6.5 Bq·L -1 for the Ottawa resident, 15.8 Bq·L -1 for the Deep River resident, and 1260 Bq·L -1 for the exposed worker. OBT-in-urine levels from these same individuals were 0.06 Bq·L -1 (Ottawa), 0.29 Bq·L -1 (Deep River), and 2.2 Bq·L -1 (exposed worker). With a model developed for calculating OBT dose fraction from the measured ratio of HTO to OBT in urine, we estimated that the dose arising from OBT in the body was about 26% of the total tritium dose for the Ottawa resident and 50% for the Deep River resident. The CRL individual had a 5% dose contribution from OBT, but had higher overall tritium dose due to frequent intakes of HTO. The study indicates that the bulk of the tritium dose to the population is the result of HTO intakes and not due to dietary intake of OBT. (author)

  14. Calibration of in vitro bioassay methodology for determination “1”3”1I urine

    International Nuclear Information System (INIS)

    Carvalho, C.B.; Hazin, C.; Lima, A.R.

    2015-01-01

    The use of unsealed radioactive sources in institutions practicing nuclear medicine poses a significant risk of internal exposure of workers. In this context, handling of “1”3”1I plays an important role in relation to other radionuclides due to its wide application, particularly in medical diagnosis and therapy of diseases related to the thyroid gland. Given the increasing number of services using “1”3”1I in their examination protocols, the probability of accidental incorporation of this radionuclide has increased. The present study aimed to implement methodologies for in vitro bioassay at the Centro Regional de Ciências Nucleares do Nordeste (CRCN-NE/CNEN), Recife, Brazil, for internal monitoring of individuals occupationally exposed to “1”3”1I. For in vitro system calibration, a coaxial HPGe detector model GC1018 and a standard “1”3”3Ba source were used. Upon obtaining the calibration factor, it was possible to determine the minimum detectable activities (MDA) for the system by using direct measurements of distilled water simulating urine (in vitro). Then, by using the biokinetic models provided by the International Commission on Radiological Protection, edited with the AIDE software version 6.0, it was possible to estimate the Minimum Detectable Effective Dose (MDED). MDED values obtained were compared to the record level of 1 mSv recommended by the International Atomic Energy Agency in the urine compartment 24 h. The values found were lower than the record level of 1 mSv in all simulated incorporation scenarios: inhalation of vapor and particles with AMAD of 1 μ and 5 μ, type F compound, and ingestion. The results of this work show that the implemented technique is suitable for conducting internal monitoring of workers to “1”3”1I. It is intended to continue the work aiming the monitoring of occupationally exposed individuals from Nuclear Medicine Services in Recife, Brazil. (authors)

  15. Multistep bioassay to predict recolonization potential of emerging parasitoids after a pesticide treatment.

    Science.gov (United States)

    Desneux, Nicolas; Ramirez-Romero, Ricardo; Kaiser, Laure

    2006-10-01

    Neurotoxic pyrethroid insecticides are widely used for crop protection, and lethal and sublethal perturbations can be expected in beneficial insects. Under laboratory conditions, the lethal and sublethal effects of deltamethrin on the aphid parasitoid Diaeretiella rapae M'Intosh (Hymenoptera: Braconidae) were studied at the mummy stage and in emerging adults. Following a multistep bioassay, analyses were aimed at evaluating the effects of deltamethrin at various crucial steps in the recolonization process following a deltamethrin treatment: Parasitoid pupal development (emergence from the mummies), adult survival, and host-searching capacity. A four-armed olfactometer was used to investigate the effect of deltamethrin on host-searching behavior (a range of concentrations causing 0.4-79.4% mortality was tested), and a Potter tower was used to test the deltamethrin effect with a realistic application method (four concentrations were tested: 0.5, 5.0, 6.25, and 50 g active ingredient [a.i.]/ha). Deltamethrin reduced the percentage of emergence from mummies, but only when exposed to the 50 g a.i./ha concentration. However, for all concentrations tested, the insecticide induced a decrease in longevity after emergence from sprayed mummies and significant adult mortality when parasitoids walked on fresh residues on leaves. Indices were defined and predicted a high mortality and, thus, reduction of recolonization capacities. However, deltamethrin had no effect on orientation behavior toward aphid-infested plants for adults that survived a residual exposure to the insecticide. The impact of deltamethrin on recolonization via pupal emergence and interest in the methodology used are discussed.

  16. Application of a canine 238Pu dosimetry model to human bioassay data

    Energy Technology Data Exchange (ETDEWEB)

    Hickman, Jr., A. W. [Florida Univ., Gainesville, FL (United States)

    1991-08-01

    Associated with the use of 2238Pu in thermoelectric power sources for space probes and power supplies for cardiac devices is the potential for human exposure to 238Pu, primarily by inhalation. In the event of human internal exposure, a means is needed for assessing the level of intake and calculating radiation doses. Several bioassay/dosimetry models have been developed for 239Pu. However, results from studies with laboratory animals have indicated that the biokinetics, and therefore the descriptive models, of 238Pu are significantly different from those for 239Pu. A canine model accounting for these differences has been applied in this work to urinary excretion data from seven humans occupationally exposed to low levels of an insoluble 238Pu compound. The modified model provides a good description of the urinary excretion kinetics observed in the exposed humans. The modified model was also used to provide estimates of the initial intakes of 238Pu for the seven individuals; these estimates ranged from 4.5 nCi (170 Bq) to 87 nCi (3200 Bq). Autopsy data on the amount and distribution of 238Pu retained in the organs may be used in the future to validate or refute both these estimates and the assumptions used to formulate the human model. Modification of the human model to simulate an injection exposure to 239Pu gave patterns of retention in the organs and urinary excretion comparable to those seen previously in humans; further modification of the model using fecal data (unavailable for the subjects of this study) is indicated.

  17. Trypanocidal Activity of Smallanthus sonchifolius: Identification of Active Sesquiterpene Lactones by Bioassay-Guided Fractionation

    Directory of Open Access Journals (Sweden)

    F. M. Frank

    2013-01-01

    Full Text Available In order to find novel plant-derived biologically active compounds against Trypanosoma cruzi, we isolated, from the organic extract of Smallanthus sonchifolius, the sesquiterpene lactones enhydrin (1, uvedalin (2, and polymatin B (3 by bioassay-guided fractionation technique. These compounds showed a significant trypanocidal activity against the epimastigote forms of the parasite with IC50 values of 0.84 μM (1, 1.09 μM (2, and 4.90 μM (3. After a 24 h treatment with 10 μg/mL of enhydrin or uvedalin, parasites were not able to recover their replication rate. Compounds 1 and 2 showed IC50 values of 33.4 μM and 25.0 μM against T. cruzi trypomastigotes, while polymatin B was not active. When the three compounds were tested against the intracellular forms of T. cruzi, they were able to inhibit the amastigote replication with IC50 of 5.17 μM, 3.34 μM, and 9.02 μM for 1, 2, and 3, respectively. The cytotoxicity of the compounds was evaluated in Vero cells obtaining CC50 values of 46.5 μM (1, 46.8 μM (2, and 147.3 μM (3 and the selectivity index calculated. According to these results, enhydrin and uvedalin might have potentials as agents against Chagas disease and could serve as lead molecules to develop new drugs.

  18. Microcontact printing with aminosilanes: creating biomolecule micro- and nanoarrays for multiplexed microfluidic bioassays.

    Science.gov (United States)

    Sathish, Shivani; Ricoult, Sébastien G; Toda-Peters, Kazumi; Shen, Amy Q

    2017-05-21

    aminosilane patterning technique enables the creation of functional microfluidic micro- and nano-biomolecule arrays, laying the foundation for high-throughput multiplexed bioassays.

  19. Chip-Scale Bioassays Based on Surface-Enhanced Raman Scattering: Fundamentals and Applications

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hye-Young [Iowa State Univ., Ames, IA (United States)

    2005-01-01

    This work explores the development and application of chip-scale bioassays based on surface-enhanced Raman scattering (SERS) for high throughput and high sensitivity analysis of biomolecules. The size effect of gold nanoparticles on the intensity of SERS is first presented. A sandwich immunoassay was performed using Raman-labeled immunogold nanoparticles with various sizes. The SERS responses were correlated to particle densities, which were obtained by atomic force microscopy (AFM). The response of individual particles was also investigated using Raman-microscope and an array of gold islands on a silicon substrate. The location and the size of individual particles were mapped using AFM. The next study describes a low-level detection of Escherichia coli 0157:H7 and simulants of biological warfare agents in a sandwich immunoassay format using SERS labels, which have been termed Extrinsic Raman labels (ERLs). A new ERL scheme based on a mixed monolayer is also introduced. The mixed monolayer ERLs were created by covering the gold nanoparticles with a mixture of two thiolates, one thiolate for covalently binding antibody to the particle and the other thiolate for producing a strong Raman signal. An assay platform based on mixed self-assembled monolayers (SAMs) on gold is then presented. The mixed SAMs were prepared from dithiobis(succinimidyl undecanoate) (DSU) to covalently bind antibodies on gold substrate and oligo(ethylene glycol)-terminated thiol to prevent nonspecific adsorption of antibodies. After the mixed SAMs surfaces, formed from various mole fraction of DSU were incubated with antibodies, AFM was used to image individual antibodies on the surface. The final study presents a collaborative work on the single molecule adsorption of YOYO-I labeled {lambda}-DNA at compositionally patterned SAMs using total internal reflection fluorescence microscopy. The role of solution pH, {lambda}-DNA concentration, and domain size was investigated. This work also revealed

  20. Chemically induced immunotoxicity in a medium-term multiorgan bioassay for carcinogenesis with Wistar rats

    International Nuclear Information System (INIS)

    Spinardi-Barbisan, Ana Lucia Tozzi; Kaneno, Ramon; Barbisan, Luis Fernando; Viana de Camargo, Joao Lauro; Rodrigues, Maria Aparecida Marchesan

    2004-01-01

    A variety of chemicals can adversely affect the immune system and influence tumor development. The modifying potential of chemical carcinogens on the lymphoid organs and cytokine production of rats submitted to a medium-term initiation-promotion bioassay for carcinogenesis was investigated. Male Wistar rats were sequentially initiated with N-nitrosodiethylamine (DEN), N-methyl-N-nitrosourea (MNU), N-butyl-N-(4hydroxybutyl)nitrosamine (BBN), dihydroxy-di-n-propylnitrosamine (DHPN), and 1,2-dimethylhydrazine (DMH) during 4 weeks. Two initiated groups received phenobarbital (PB) or 2-acetylaminofluorene (2-AAF) for 25 weeks and two noninitiated groups received only PB or 2-AAF. A nontreated group was used as control. Lymphohematopoietic organs, liver, kidneys, lung, intestines, and Zymbal's gland were removed for histological analysis. Interleukin (IL)-2, IL-12, interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), IL-10, and transforming growth factor beta1 (TGF-β1) levels were determined by ELISA in spleen cell culture supernatants. At the fourth week, exposure to the initiating carcinogens resulted in cell depletion of the thymus, spleen and bone marrow, and impairment of IL-2, IL-12, and IFN-γ production. However, at the 30th week, no important alterations were observed both in lymphoid organs and cytokine production in the different groups. The results indicate that the initiating carcinogens used in the present protocol exert toxic effects on the lymphoid organs and affect the production of cytokines at the initiation step of carcinogenesis. This early and reversible depression of the immune surveillance may contribute to the survival of initiated cells facilitating the development of future neoplasia

  1. Screening for unicellular algae as possible bioassay organisms for monitoring marine water samples.

    Science.gov (United States)

    Millán de Kuhn, Rosmary; Streb, Christine; Breiter, Roman; Richter, Peter; Neesse, Thomas; Häder, Donat-Peter

    2006-08-01

    ECOTOX is an automatic early warning system to monitor potential pollution of freshwater, municipal or industrial waste waters or aquatic ecosystems. It is based on a real time image analysis of the motility and orientation parameters of the unicellular, photosynthetic flagellate Euglena gracilis. In order to widen the use of the device to marine habitats and saline waters nine marine flagellates were evaluated as putative bioassay organisms, viz. Dunaliella salina, Dunaliella viridis, Dunaliella bardawil, Prorocentrum minimum Kattegat, P. minimum Lissabon, Tetraselmis suecica, Heterocapsa triquetra, Gyrodinium dorsum and Cryptomonas maculata. Because of their slow growth the last three strains were excluded from further evaluation. Selection criteria were ease of culture, density of cell suspension, stability of motility and gravitactic orientation. The sensitivity toward toxins was tested using copper(II) ions. The instrument allows the user to automatically determine effect-concentration (EC) curves from which the EC(50) values can be calculated. For the interpretation of the EC curves a sigmoid logistic model was proposed which proved to be satisfactory for all tested strains. The inhibition of the motility was considered as the most appropriate movement parameter as an endpoint. The Dunaliella species had the lowest sensitivity to copper with EC(50) values of 220, 198 and 176 mg/L for D. salina, D. bardawil and D. viridis, respectively, followed by T. suecica with an EC(50) value of 40 mg/L. The Prorocentrum species were found to be the most sensitive with an EC(50) value of 13.5 mg/L for P. minimum Lissabon and 7.5 mg/L for P. minimum Kattegat.

  2. Practical issues in discriminating between environmental and occupational sources in a uranium urinalysis bioassay program

    International Nuclear Information System (INIS)

    Long, M.P.; Carbaugh, E.H.; Fairrow, N.L.

    1994-11-01

    Workers at two Department of Energy facilities, the Pantex Plant in Texas and the Hanford Site in Washington, are potentially exposed to class Y depleted or natural uranium. Since trace amounts of uranium are naturally present in urine excretion, site bioassay programs must be able to discern occupational exposure from naturally occurring uranium exposure. In 1985 Hanford established a 0.2-μg/d environmental screening level for elemental uranium in urine; the protocol was based on log-normal probability analysis of unexposed workers. A second study of background uranium levels commenced in 1990, and experiences in the field indicated that there seemed to be an excessive number of urine samples with uranium above the screening level and that the environmental screening level should be reviewed. Due to unforeseen problems, that second study was terminated before the complete data could be obtained. Natural uranium in rock (by weight, 99.27% 288 U, 0.72% 235 U, and 0.006% 234 U) has approximately equal activity concentrations of 238 U and 234 U. Earlier studies, summarized by the U.S. Environmental Protection Agency in 51 FR 32068, have indicated that 234 U (via 234 Th) has a greater environmental mobility than 238 U and may well have a higher concentration in ground water. By assuming that the 238 U-to 234 U ratio in the urine of nonoccupationally exposed persons should reflect the ratio of environmental levels, significant occupational exposure to depleted uranium would shift that ratio in favor of 238 U, allowing use of the ratio as a co-indicator of occupational exposure in addition to the isotope-specific screening levels. This approach has been adopted by Pantex. The Pacific Northwest Laboratory is studying the feasibility of applying this method to the natural and recycled uranium mixtures encountered at Hanford. The Hanford data included in this report represent work-in-progress

  3. Using bioassays and species sensitivity distributions to assess herbicide toxicity towards benthic diatoms.

    Directory of Open Access Journals (Sweden)

    Floriane Larras

    Full Text Available Although benthic diatoms are widely used in ecological studies of aquatic systems, there is still a dearth of data concerning species sensitivities towards several contaminants. Within the same community, different species may respond differently depending on their physiological and ecological characteristics. This lack of knowledge makes specific appropriate risk assessment impossible. To find out whether species sensitivity distribution (SSD could be used to estimate the risk of herbicide toxicity for diatoms, we need to know whether their sensitivity depends on their physiological and ecological characteristics. We carried out single-species bioassays on 11 diatom species exposed to 8 herbicides. Dose-responses relationships were used to extrapolate the Effective Concentration 5 (EC(5 and the Effective Concentration 50 (EC(50 for each exposure. These data were used to fit a SSD curve for each herbicide, and to determine the Hazardous concentration 5 (HC(5 and 50 (HC(50. Our results revealed a high level of variability of the sensitivity in the set of species tested. For photosystem-II inhibitor (PSII herbicides, diatoms species displayed a typical grouping of sensitivity levels consistent with their trophic mode and their ecological guild. N-heterotroph and "motile" guild species were more tolerant of PSII inhibitors, while N-autotroph and "low profile" guild species were more sensitive. Comprehensive SSD curves were obtained for 5 herbicides, but not for sulfonylurea herbicides or for dimetachlor, which had toxicity levels that were below the range of concentration tested. The SSD curves provided the following ranking of toxicity: diuron> terbutryn> isoproturon> atrazine> metolachlor. The HC that affected 5% of the species revealed that, even at the usual environmental concentrations of herbicides, diatom assemblages could be affected, especially by isoproturon, terbutryn, and diuron.

  4. U.S. bioassay Intercomparison Studies Program at Oak Ridge National Lab

    International Nuclear Information System (INIS)

    Payne, G.F.; Bores, N.; Melton, K.K.; Rankin, J.M.

    1998-06-01

    The Intercomparison Studies Program (ISP) for in-vitro bioassay at Oak Ridge National Laboratory (ORNL) has been in place since May 1991. The ISP was originally created to fill a need in the Radiobioassay area at ORNL, specifically in the areas of Quality Control, Quality Assurance, and Performance Testing. In the beginning, this consisted of two or three laboratories working in a pilot intercomparison program. Once it was determined that this could work effectively, the program began to seek additional members to broaden the scope of the effort. The program became formalized with a quarterly report in January 1992. The ISP currently provides cross-check blind/double-blind samples spiked with known amounts of radioactivity to various Department of Energy (DOE) facilities, universities, and private industry organizations throughout the US. These samples can be packaged according to ORNL procedures (ORNL sample bottles, ORNL chain-of-custody forms, tamper seals etc.), for a single blind sample or according to the needs of a particular facility if the double-blind sample mode is to be maintained. In 1998, the customer base was broadened to include European facilities. In January 1993, the whole-body count program was added. This involves each participating facility receiving a block phantom from the ISP and determining a geometry factor using a known standard. At quarterly intervals, each participant receives an unknown sample for analysis. The sample is counted and the data is collected for publication in an annual report. In October 1994, the fecal program was added. This involves spiking an artificial matrix with known amounts of radioactivity. Laboratories receive unknown samples on a quarterly basis. The sample is counted and the data is collected and published in a quarterly report. The ISP maintains archive samples which can be analyzed in the QC laboratory at the request of any participants if a conflict or discrepancy in a sample analysis/result occurs

  5. Rapid bioassay-guided screening of toxic substances in vegetable oils that shorten the life of SHRSP rats

    Directory of Open Access Journals (Sweden)

    Lewandowski Paul

    2010-02-01

    Full Text Available Abstract It has been consistently reported that vegetable oils including canola oil have a life shortening effect in Stroke-Prone Spontaneously Hypertensive Rats (SHRSP and this toxic effect is not due to the fatty acid composition of the oil. Although it is possible that the phytosterol content or type of phytosterol present in vegetable oils may play some role in the life shortening effect observed in SHRSP rats this is still not completely resolved. Furthermore supercritical CO2 fractionation of canola oil with subsequent testing in SHRSP rats identified safe and toxic fractions however, the compounds responsible for life shortening effect were not characterised. The conventional approach to screen toxic substances in oils using rats takes more than six months and involves large number of animals. In this article we describe how rapid bioassay-guided screening could be used to identify toxic substances derived from vegetable oils and/or processed foods fortified with vegetable oils. The technique incorporates sequential fractionation of oils/processed foods and subsequent treatment of human cell lines that can be used in place of animal studies to determine cytotoxicity of the fractions with structural elucidation of compounds of interest determined via HPLC-MS and GC-MS. The rapid bioassay-guided screening proposed would require two weeks to test multiple fractions from oils, compared with six months if animal experiments were used to screen toxic effects. Fractionation of oil before bio-assay enhances the effectiveness of the detection of active compounds as fractionation increases the relative concentration of minor components.

  6. Optimisation of the microplate resazurin assay for screening and bioassay-guided fractionation of phytochemical extracts against Mycobacterium tuberculosis.

    Science.gov (United States)

    O'Neill, Taryn E; Li, Haoxin; Colquhoun, Caitlyn D; Johnson, John A; Webster, Duncan; Gray, Christopher A

    2014-01-01

    Because of increased resistance to current drugs, there is an urgent need to discover new anti-mycobacterial compounds for the development of novel anti-tuberculosis drugs. The microplate resazurin assay (MRA) is commonly used to evaluate natural products and synthetic compounds for anti-mycobacterial activity. However, the assay can be problematic and unreliable when screening methanolic phytochemical extracts. To optimise the MRA for the screening and bioassay-guided fractionation of phytochemical extracts using Mycobacterium tuberculosis H37Ra. The effects of varying assay duration, resazurin solution composition, solvent (dimethyl sulphoxide - DMSO) concentration and type of microtitre plate used on the results and reliability of the MRA were investigated. The optimal bioassay protocol was applied to methanolic extracts of medicinal plants that have been reported to possess anti-mycobacterial activity. The variables investigated were found to have significant effects on the results obtained with the MRA. A standardised procedure that can reliably quantify anti-mycobacterial activity of phytochemical extracts in as little as 48 h was identified. The optimised MRA uses 2% aqueous DMSO, with an indicator solution of 62.5 µg/mL resazurin in 5% aqueous Tween 80 over 96 h incubation. The study has identified an optimal procedure for the MRA when used with M. tuberculosis H37Ra that gives rapid, reliable and consistent results. The assay procedure has been used successfully for the screening and bioassay-guided fractionation of anti-mycobacterial compounds from methanol extracts of Canadian medicinal plants. Copyright © 2014 John Wiley & Sons, Ltd.

  7. Bioassays with caged hyalella azteca to determine in situ toxicity downstream of two Saskatchewan, Canada, uranium operations.

    Science.gov (United States)

    Robertson, Erin L; Liber, Karsten

    2007-11-01

    The main objectives of this in situ study were to evaluate the usefulness of an in situ bioassay to determine if downstream water bodies at the Key Lake and Rabbit Lake uranium operations (Saskatchewan, Canada) were toxic to Hyalella azteca and, if toxicity was observed, to differentiate between the contribution of surface water and sediment contamination to in situ toxicity. These objectives were achieved by performing 4-d in situ bioassays with laboratory-reared H. azteca confined in specially designed, paired, surface water and sediment exposure chambers. Results from the in situ bioassays revealed significant mortality, relative to the respective reference site, at the exposure sites at both Key Lake (p

  8. Comparison and validation of methods to quantify Cry1Ab toxin from Bacillus thuringiensis for standardization of insect bioassays.

    Science.gov (United States)

    Crespo, André L B; Spencer, Terence A; Nekl, Emily; Pusztai-Carey, Marianne; Moar, William J; Siegfried, Blair D

    2008-01-01

    Standardization of toxin preparations derived from Bacillus thuringiensis (Berliner) used in laboratory bioassays is critical for accurately assessing possible changes in the susceptibility of field populations of target pests. Different methods were evaluated to quantify Cry1Ab, the toxin expressed by 80% of the commercially available transgenic maize that targets the European corn borer, Ostrinia nubilalis (Hübner). We compared three methods of quantification on three different toxin preparations from independent sources: enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis and densitometry (SDS-PAGE/densitometry), and the Bradford assay for total protein. The results were compared to those obtained by immunoblot analysis and with the results of toxin bioassays against susceptible laboratory colonies of O. nubilalis. The Bradford method resulted in statistically higher estimates than either ELISA or SDS-PAGE/densitometry but also provided the lowest coefficients of variation (CVs) for estimates of the Cry1Ab concentration (from 2.4 to 5.4%). The CV of estimates obtained by ELISA ranged from 12.8 to 26.5%, whereas the CV of estimates obtained by SDS-PAGE/densitometry ranged from 0.2 to 15.4%. We standardized toxin concentration by using SDS-PAGE/densitometry, which is the only method specific for the 65-kDa Cry1Ab protein and is not confounded by impurities detected by ELISA and Bradford assay for total protein. Bioassays with standardized Cry1Ab preparations based on SDS-PAGE/densitometry showed no significant differences in LC(50) values, although there were significant differences in growth inhibition for two of the three Cry1Ab preparations. However, the variation in larval weight caused by toxin source was only 4% of the total variation, and we conclude that standardization of Cry1Ab production and quantification by SDS-PAGE/densitometry may improve data consistency in monitoring efforts to identify changes in

  9. A versatile and low-cost open source pipetting robot for automation of toxicological and ecotoxicological bioassays.

    Directory of Open Access Journals (Sweden)

    Sebastian Steffens

    Full Text Available In the past decades, bioassays and whole-organism bioassay have become important tools not only in compliance testing of industrial chemicals and plant protection products, but also in the monitoring of environmental quality. With few exceptions, such test systems are discontinuous. They require exposure of the biological test material in small units, such as multiwell plates, during prolonged incubation periods, and do not allow online read-outs. It is mostly due to these shortcomings that applications in continuous monitoring of, e.g., drinking or surface water quality are largely missing. We propose the use of pipetting robots that can be used to automatically exchange samples in multiwell plates with fresh samples in a semi-static manner, as a potential solution to overcome these limitations. In this study, we developed a simple and low-cost, versatile pipetting robot constructed partly using open-source hardware that has a small footprint and can be used for online monitoring of water quality by means of an automated whole-organism bioassay. We tested its precision in automated 2-fold dilution series and used it for exposure of zebrafish embryos (Danio rerio-a common model species in ecotoxicology-to cadmium chloride and permethrin. We found that, compared to conventional static or semi-static exposure scenarios, effects of the two chemicals in zebrafish embryos generally occurred at lower concentrations, and analytically verified that the increased frequency of media exchange resulted in a greater availability of the chemical. In combination with advanced detection systems this custom-made pipetting robot has the potential to become a valuable tool in future monitoring strategies for drinking and surface water.

  10. A versatile and low-cost open source pipetting robot for automation of toxicological and ecotoxicological bioassays.

    Science.gov (United States)

    Steffens, Sebastian; Nüßer, Leonie; Seiler, Thomas-Benjamin; Ruchter, Nadine; Schumann, Mark; Döring, Ricarda; Cofalla, Catrina; Ostfeld, Avi; Salomons, Elad; Schüttrumpf, Holger; Hollert, Henner; Brinkmann, Markus

    2017-01-01

    In the past decades, bioassays and whole-organism bioassay have become important tools not only in compliance testing of industrial chemicals and plant protection products, but also in the monitoring of environmental quality. With few exceptions, such test systems are discontinuous. They require exposure of the biological test material in small units, such as multiwell plates, during prolonged incubation periods, and do not allow online read-outs. It is mostly due to these shortcomings that applications in continuous monitoring of, e.g., drinking or surface water quality are largely missing. We propose the use of pipetting robots that can be used to automatically exchange samples in multiwell plates with fresh samples in a semi-static manner, as a potential solution to overcome these limitations. In this study, we developed a simple and low-cost, versatile pipetting robot constructed partly using open-source hardware that has a small footprint and can be used for online monitoring of water quality by means of an automated whole-organism bioassay. We tested its precision in automated 2-fold dilution series and used it for exposure of zebrafish embryos (Danio rerio)-a common model species in ecotoxicology-to cadmium chloride and permethrin. We found that, compared to conventional static or semi-static exposure scenarios, effects of the two chemicals in zebrafish embryos generally occurred at lower concentrations, and analytically verified that the increased frequency of media exchange resulted in a greater availability of the chemical. In combination with advanced detection systems this custom-made pipetting robot has the potential to become a valuable tool in future monitoring strategies for drinking and surface water.

  11. Development and characterization of a green fluorescent protein-based rat cell bioassay system for detection of AH receptor ligands

    Energy Technology Data Exchange (ETDEWEB)

    Zhao Bin; Denison, M. [California Univ., Davis, CA (United States). Dept. of Environmental Toxicology

    2004-09-15

    Proper epidemiological, risk assessment and exposure analysis of TCDD and related HAHs requires accurate measurements of these chemicals both in the species of interest and in various exposure matrices (i.e. biological, environmental, food and feed). While high-resolution instrumental analysis techniques are established for these chemicals, these procedures are very costly, time-consuming and are impractical for large scale sampling studies. Accordingly, numerous bioanalytical methods have been developed for the detection of these chemicals in extracts from a variety of matrices, the majority of which take the advantage of the ability of these chemicals to activate one or more aspects of the AhR-dependent mechanism of action. One of the most sensitive bioassay systems developed to date is the so-called CALUX (Chemically Activated Luciferase Expression) assay, which is based on novel recombinant cell lines that contain a stably transfected dioxin (AhR)-responsive firefly luciferase gene. Treatment of these cells with TCDD and related HAHs and polycyclic aromatic hydrocarbons (PAHs), as well as other AhR ligands, results in induction of reporter gene expression in a time-, dose-, AhR-, and chemical-specific manner. The level of reporter gene expression correlates with the total concentration of the TCDD-like AhR inducers (agonists) present in the sample. Although the firefly luciferase reporter gene contributes to the high degree of sensitivity of the assay, it also has limitations with respect to our need for a rapid and inexpensive bioassay for high-throughput screening analysis. Accordingly, we previously developed a stably transfected murine cell line containing an AhRresponsive enhanced green fluorescent protein (EGFP) reporter gene. This cell line provided us with a high-throughput cell bioassay system for identification and characterization of AhR agonists and antagonists. Here we have extended these studies and describe the development, optimization, and

  12. In situ bioassays with Chironomus riparius larvae to biomonitor metal pollution in rivers and to evaluate the efficiency of restoration measures in mine areas

    Energy Technology Data Exchange (ETDEWEB)

    Faria, Mafalda S. [CESAM and Departamento de Biologia da Universidade de Aveiro, Campus de Santiago, 3810-193 Aveiro (Portugal)], E-mail: mafaldafaria@sapo.pt; Lopes, Ricardo J. [CIBIO, Centro de Investigacao em Biodiversidade e Recursos Geneticos, Campus Agrario de Vairao, 4485-661 Vairao (Portugal); Malcato, Joao; Nogueira, Antonio J.A.; Soares, Amadeu M.V.M. [CESAM and Departamento de Biologia da Universidade de Aveiro, Campus de Santiago, 3810-193 Aveiro (Portugal)

    2008-01-15

    In this study we evaluate the ability of an in situ bioassay with Chironomus riparius larvae, using larval development and growth as endpoints, to biomonitor water quality and to assess the biological recovery of metal contaminated freshwater ecosystems of mine areas that are subject of restoration measures. The bioassay was carried out in streams located near an abandoned goldmine in North Portugal, throughout an environmental rehabilitation of the mine (2002-2004). During this period, a decrease in the inhibition of larval growth in the metal contaminated stream was observed. The bioassay was also performed in streams located near an active tungsten mine in Central Portugal. Larval growth and development were highly inhibited in the stream that receives acid drainage from the tungsten mine and treated water from the AMD treatment station. The results indicate that the bioassay can be used to evaluate the efficiency of environmental restoration measures in mining areas. - In situ bioassays with Chironomus riparius larvae can be a suitable tool to monitor restoration efficiency after a long time of metallic sediment contamination.

  13. In situ and laboratory bioassays with Chironomus riparius larvae to assess toxicity of metal contamination in rivers: the relative toxic effect of sediment versus water contamination.

    Science.gov (United States)

    Faria, Mafalda S; Lopes, Ricardo J; Nogueira, António J A; Soares, Amadeu M V M

    2007-09-01

    We used bioassays employing head capsule width and body length increase of Chironomus riparius larvae as end points to evaluate metal contamination in streams. Bioassays were performed in situ near an abandoned Portuguese goldmine in the spring of 2003 and 2004. Bioassays also were performed under laboratory conditions with water and sediment collected from each stream to verify if laboratory bioassays could detect in situ toxicity and to evaluate the relative contribution of sediment and water to overall toxicity. We used field sediments with control water and control sediments with field water to discriminate between metal contamination in water and sediment. Field water with dry and sieved, organic matter-free, and nontreated sediments was used to determine the toxicity of heavy metals that enter the organism through ingested material. In both in situ and laboratory bioassays, body length increase was significantly inhibited by metal contamination, whereas head capsule width was not affected. Body length increase was more affected by contaminated sediment compared to contaminated water. The lowest-effect level of heavy metals was observed in the dry and sieved sediment that prevented ingestion of sediment particles by larvae. These results suggest that body length increase of C. riparius larvae can be used to indicate the impact of metal contamination in rivers. Chironomus riparius larvae are more affected by heavy metals that enter the organism through ingested sediment than by heavy metals dissolved in the water column. Nevertheless, several factors, such as the particle size and organic matter of sediment, must be taken into account.

  14. In situ bioassays with Chironomus riparius larvae to biomonitor metal pollution in rivers and to evaluate the efficiency of restoration measures in mine areas

    International Nuclear Information System (INIS)

    Faria, Mafalda S.; Lopes, Ricardo J.; Malcato, Joao; Nogueira, Antonio J.A.; Soares, Amadeu M.V.M.

    2008-01-01

    In this study we evaluate the ability of an in situ bioassay with Chironomus riparius larvae, using larval development and growth as endpoints, to biomonitor water quality and to assess the biological recovery of metal contaminated freshwater ecosystems of mine areas that are subject of restoration measures. The bioassay was carried out in streams located near an abandoned goldmine in North Portugal, throughout an environmental rehabilitation of the mine (2002-2004). During this period, a decrease in the inhibition of larval growth in the metal contaminated stream was observed. The bioassay was also performed in streams located near an active tungsten mine in Central Portugal. Larval growth and development were highly inhibited in the stream that receives acid drainage from the tungsten mine and treated water from the AMD treatment station. The results indicate that the bioassay can be used to evaluate the efficiency of environmental restoration measures in mining areas. - In situ bioassays with Chironomus riparius larvae can be a suitable tool to monitor restoration efficiency after a long time of metallic sediment contamination

  15. Design and Proof-of-Concept Use of a Circular PMMA Platform with 16-Well Sample Capacity for Microwave-Accelerated Bioassays.

    Science.gov (United States)

    Mohammed, Muzaffer; Aslan, Kadir

    2013-01-01

    We demonstrate the design and the proof-of-concept use of a new, circular poly(methyl methacrylate)-based bioassay platform (PMMA platform), which affords for the rapid processing of 16 samples at once. The circular PMMA platform (5 cm in diameter) was coated with a silver nanoparticle film to accelerate the bioassay steps by microwave heating. A model colorimetric bioassay for biotinylated albumin (using streptavidin-labeled horse radish peroxidase) was performed on the PMMA platform coated with and without silver nanoparticles (a control experiment), and at room temperature and using microwave heating. It was shown that the simulated temperature profile of the PMMA platform during microwave heating were comparable to the real-time temperature profile during actual microwave heating of the constructed PMMA platform in a commercial microwave oven. The model colorimetric bioassay for biotinylated albumin was successfully completed in ~2 min (total assay time) using microwave heating, as compared to 90 min at room temperature (total assay time), which indicates a ~45-fold decrease in assay time. Our PMMA platform design afforded for significant reduction in non-specific interactions and low background signal as compared to non-silvered PMMA surfaces when employed in a microwave-accelerated bioassay carried out in a conventional microwave cavity.

  16. Ecotoxicological bioassays of sediment leachates in a river bed flanked by decommissioned pesticide plants in Nantong City, East China.

    Science.gov (United States)

    Zhou, Yan; Wang, Fenghe; Wan, Jinzhong; He, Jian; Li, Qun; Qiang Chen; Gao, Jay; Lin, Yusuo; Zhang, Shengtian

    2017-03-01

    Traditionally, the toxicity of river contaminants is analyzed chemically or physically through river bed sediments. The biotoxicity of polluted sediment leachates has not caught our attention. This study aims to overcome this deficiency through a battery of biotests which were conducted to monitor comprehensive toxicity of sediment leachates for the Yaogang River in East Jiangsu Province of China, which is in close proximity to former pesticide plants. The general physical and chemical parameters of major pollutants were analyzed from river bed sediments collected at five strategic locations. The ecotoxicity analyses undertaken include overall fish (adult zebrafish) acute toxicity, luminescent bacteria (Vibrio fischeri) bioassay, and zebrafish embryo toxicity assay. Compared with the control group, sediment leachates increased the lethality, inhibited the embryos hatching and induced development abnormalities of zebrafish embryos, and inhibited the luminescence of V. fischeri. The results show that sediment leachates may assume various toxic effects, depending on the test organism. This diverse toxicity to aquatic organisms reflects their different sensitivity to sediment leachates. It is found clearly that V. fischeri was the organism which was characterized by the highest sensitivity to the sediment leachates. The complicated toxicity of leachates was not caused by one single factor but by multiple pollutants together. This indicates the need of estimations of sediment leachate not only taking into account chemical detection but also of applying the biotests to the problem. Thus, multigroup bioassays are necessary to realistically evaluate river ecological risks imposed by leachates.

  17. Selection of a bioassay battery to assess toxicity in the affluents and effluents of three water-treatment plants

    Directory of Open Access Journals (Sweden)

    Paola Bohórquez-Echeverry

    2012-08-01

    Full Text Available The assessment of water quality includes the analysis of both physical-chemical and microbiological parameters. However,none of these evaluates the biological effect that can be generated in ecosystems or humans. In order to define the most suitable organismsto evaluate the toxicity in the affluent and effluent of three drinking-water treatment plants, five acute toxicity bioassays were used,incorporating three taxonomic groups of the food chain. Materials and methods. The bioassays used were Daphnia magna and Hydraattenuata as animal models, Lactuca sativa and Pseudokirchneriella subcapitata as plant models, and Photobacterium leioghnathi asbacterial model. To meet this objective, selection criteria of the organisms evaluated and cluster analysis were used to identify the mostsensitive in the affluent and effluent of each plant. Results. All organisms are potentially useful in the assessment of water quality bymeeting four essential requirements and 17 desirable requirements equivalent to 100% acceptability, except P. leioghnathi which doesnot meet two essential requirements that are the IC50 for the toxic reference and the confidence interval. The animal, plant and bacterialmodels showed different levels of sensitivity at the entrance and exit of the water treatment systems. Conclusions. H. attenuata, P.subcapitata and P. leioghnathi were the most effective organisms in detecting toxicity levels in the affluents and D. magna, P. subcapitataand P. leioghnathi in the effluents.

  18. Stomatal development in barley as a bioassay for cell differentation: its use with X-rays and gibberellic acid

    Energy Technology Data Exchange (ETDEWEB)

    Zeiger, E; Rafalowsky, J [Chile Univ., Santiago. Departamento de Biologia y Genetica

    1976-01-01

    A bioassay for cell differentiation during stomatal development in barley (Hordeum vulgare L.) has been defined. It uses cell kinetics analysis to follow the temporal course of cell divisions in the developmental sequence. The rate of displacement of the divisions along the stomatal rows provides a measure of differentiation. Physical factors affecting differentiation may be tested with intact seedlings. The bioassay showed that X-ray irradiation inhibited the divisions leading to stomatal formation. The inhibition kinetics was similar to the one observed in root meristems. Chemical substances are tested by culturing excised shoots in a synthetic medium. Detached leaves responded to sucrose and light with increasing rates of stomatal divisions. Gibberellic acid (GA/sub 3/) was assayed for its effects on the growth of the leaf and the differentiation of stomata. GA/sub 3/ increased the overall length of the leaves without affecting the rates of cell division. The treated cells responded with increased elongation rates and a precocious initiation and completion of cell enlargement. GA/sub 3/ had no specific effect on stomatal differentiation.

  19. Characterization of quality of sediments from Paranaguá Bay (Brazil) by combined in vitro bioassays and chemical analyses.

    Science.gov (United States)

    Rizzi, Juliane; Pérez-Albaladejo, Elisabet; Fernandes, Denise; Contreras, Javier; Froehner, Sandro; Porte, Cinta

    2017-07-01

    The present study characterizes the quality of sediments from the Paranaguá Estuarine Complex (South Brazil). Polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and organochlorine pesticides (OCPs) were determined in sediment samples together with a series of different in vitro bioassays. The fish hepatoma cell line (PLHC-1) was used to determine the presence of cytotoxic compounds and CYP1A- and oxidative stress-inducing agents in sediment extracts. Ovarian microsomal fractions from sea bass (Dicentrarchus labrax) were used to detect the presence of endocrine disrupters that interfered with the synthesis of estrogens (ovarian CYP19). Despite the relatively low levels of pollutants and no evidence of negative effects based on guideline levels, sediments collected close to harbors were enriched with CYP1A-inducing agents and they showed higher cytotoxicity. In contrast, sediments from internal areas inhibited CYP19 activity, which suggests the presence of endocrine disrupters at these sites. Overall, the selected bioassays and the chemistry data led to the identification of potentially impacted areas along the Paranaguá Estuarine Complex that would require further action to improve their environmental quality. Environ Toxicol Chem 2017;36:1811-1819. © 2016 SETAC. © 2016 SETAC.

  20. Fundulus heteroclitus gonadotropins.5: Small scale chromatographic fractionation of pituitary extracts into components with different steroidogenic activities using homologous bioassays

    Directory of Open Access Journals (Sweden)

    Petrino Teresa R

    2004-03-01

    Full Text Available Abstract Fractionation and characterization of gonadotropins (GtH from Fundulus heteroclitus pituitary extracts were carried out using a biocompatible liquid chromatographic procedure (Pharmacia FPLC system. Chromatographic fractions were monitored for gonadotropic activities (induction of oocyte maturation and steroid production using homologous follicle bioassays in vitro. Size-exclusion chromatography eluted gonadotropic activity in one major protein peak (Mr ~ 30,000. Anion-exchange and hydrophobic-interaction chromatography (HIC yielded two distinct peaks of 17beta-estradiol (E2- and 17alpha-hydroxy,20beta-dihydroprogesterone (DHP-promoting activity with associated oocyte maturation. Two-dimensional chromatography (chromatofocusing followed by HIC resolved pituitary extracts into two active fractions; both induced E2 synthesis, but one was relatively poor in eliciting DHP and testosterone production. Thus, using homologous bioassays, at least two quantitatively different gonadotropic (steroidogenic activities: an E2-promoting gonadotropin (GtH I-like and a DHP-promoting gonadotropin (GtH II-like, which has a lower isoelectric point but greater hydrophobicity than the former, can be distinguished from F. heteroclitus pituitaries by a variety of chromatographic procedures. This study complements previous biochemical and molecular data in F. heteroclitus and substantiates the duality of GtH function in a multiple-spawning teleost.

  1. Evaluation of genotoxicity of liquid effluents from gas washing systems by means of bioassay Trad-MCN

    International Nuclear Information System (INIS)

    Machado, Alessandra Carla Fattori Ergesse; Alves, Edenise Segala

    2007-01-01

    In the gas washing systems the gaseous emissions from a facility are forced through an absorbing liquid preventing pollutants to be dispersed into the atmosphere. In the Centro Tecnologico da Marinha em Sao Paulo/Centro Experimental Aramar (CEA), the gas washing are used to control the emissions from the uranium enrichment facilities. Uranium. fluoride, ammonia and hydrogen fluoride are the main contaminants, all heavily toxic. Biological assays, using plants or other living organisms, have been used to assess genotoxic agents in the environment. Among the bioassays using plants, the Trad-MCN has been used extensively, as it allows the evaluation of liquid or gaseous contaminants. The species Tradescantia pallida (Rose) was exposed in a dynamic system to liquid effluents from CEA. A positive control was the exposure to formaldehyde 10% in water, known as a very toxic solution, and the negative control was the exposure to filtered air. The protocol established by Ma (1983) for hybrid clones and validated for the T. pallida by Guimaraes (2003) was used to perform the Trad-MCN assays. Only preparations containing early tetrads were scored. In that context, the present study objectifies to evaluate, by the Trad-MCN bioassay, the genotoxicity of the solution from the gas washing and, also, evaluate the efficiency of that system. The results obtained show that the T. pallida is a sensitive bioindicator for the pollutants tested and can be useful for in vitro environmental monitoring under controlled conditions. (author)

  2. Determination of the viability of Toxoplasma gondii in cured ham using bioassay: influence of technological processing and food safety implications.

    Science.gov (United States)

    Bayarri, Susana; Gracia, María J; Lázaro, Regina; Pe Rez-Arquillué, Consuelo; Barberán, Montserrat; Herrera, Antonio

    2010-12-01

    Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii and distributed worldwide. Ingestion of viable cysts from infected raw or undercooked meat is an important route of horizontal transmission of the parasite to humans. Little information is available concerning the effect of commercial curing on cysts of T. gondii. This study is the first in which the influence of processing of cured ham on the viability of T. gondii has been evaluated, using bioassay to assess the risk of infection from eating this meat product. Naturally infected pigs were selected for the study, and a mouse concentration bioassay technique was used to demonstrate viable bradyzoites of T. gondii in porcine tissues and hams. No viable parasites were found in the final product (14 months of curing) based on results of the indirect immunofluorescence assay and histological and PCR analyses. Our results indicate that the consumption of hams cured as described here poses an insignificant risk of acquiring toxoplasmosis. However, additional studies are required to evaluate the safety of ham products cured under different conditions of curing time, salt, and nitrite concentration.

  3. Genotoxic and mutagenic effects of polluted surface water in the midwestern region of Brazil using animal and plant bioassays

    Directory of Open Access Journals (Sweden)

    Priscila Leocádia Rosa Dourado

    Full Text Available Abstract This study aimed to evaluate DNA damage in animal and plant cells exposed to water from the Água Boa stream (Dourados, Mato Grosso do Sul, Brazil by using bioassays, and to identify the chemical compounds in the water to determine the water quality in the area. Through the cytotoxicity bioassay with Allium cepa, using micronucleus test, and comet assay, using Astyanax altiparanae fish, the results indicated that biological samples were genetically altered. Micronuclei were observed in erythrocytes of A. altiparanae after exposure to water from locations close to industrial waste discharge. The highest DNA damage observed with the comet assay in fish occurred with the exposure to water from locations where the presence of metals (Cu, Pb, Cd, Ni was high, indicating the possibility of genotoxic effects of these compounds. Thus, these results reinforce the importance of conducting genotoxicity tests for developing management plans to improve water quality, and indicate the need for waste management before domestic and industrial effluents are released into the rivers and streams.

  4. Spawning and multiple end points of the embryo-larval bioassay of the blue mussel Mytilus galloprovincialis (Lmk).

    Science.gov (United States)

    Resgalla, Charrid

    2016-10-01

    Since the 1960s, little has been done to improve and simulate the use of short-duration chronic bioassays of bivalve embryos, particularly in mussels. However, these test organisms offer great advantages in relation to other groups, due to the ease of obtaining breeders in cultivation systems, in the environment and any time, and due to their high sensitivity to chemicals or contaminants. To contribute some methodological aspects, this study uses techniques to stimulate spawning or improve the obtaining of gametes for use in bioassays with the mussel Mytilus galloprovincialis. It also evaluates different criteria for determining the effect on the larvae, for estimation of EC 50 and NOEC values, based on morphological analysis of developmental delay and the biometrics of the larvae. KCl proved to be a reliable inducer of spawning, with positive responses in 10 of the 12 months of the year tested. Moreover, this chemical, in association with NH 4 Cl, demonstrated the capacity to activate immature oocytes obtained from extirpated gonads, enabling an improvement in fertilization rates. The different criteria adopted to determine the effects on the larvae in the assays with reference toxicants (SDS and K 2 Cr 2 O 7 ) resulted in EC 50 and NOEC values without significant differences, indicating reliability in the results and freedom in the choice of criteria of effect to be adopted in the trials.

  5. PASSPORT-seq: A Novel High-Throughput Bioassay to Functionally Test Polymorphisms in Micro-RNA Target Sites

    Directory of Open Access Journals (Sweden)

    Joseph Ipe

    2018-06-01

    Full Text Available Next-generation sequencing (NGS studies have identified large numbers of genetic variants that are predicted to alter miRNA–mRNA interactions. We developed a novel high-throughput bioassay, PASSPORT-seq, that can functionally test in parallel 100s of these variants in miRNA binding sites (mirSNPs. The results are highly reproducible across both technical and biological replicates. The utility of the bioassay was demonstrated by testing 100 mirSNPs in HEK293, HepG2, and HeLa cells. The results of several of the variants were validated in all three cell lines using traditional individual luciferase assays. Fifty-five mirSNPs were functional in at least one of three cell lines (FDR ≤ 0.05; 11, 36, and 27 of them were functional in HEK293, HepG2, and HeLa cells, respectively. Only four of the variants were functional in all three cell lines, which demonstrates the cell-type specific effects of mirSNPs and the importance of testing the mirSNPs in multiple cell lines. Using PASSPORT-seq, we functionally tested 111 variants in the 3′ UTR of 17 pharmacogenes that are predicted to alter miRNA regulation. Thirty-three of the variants tested were functional in at least one cell line.

  6. Evaluation of a combined macrophyte–epiphyte bioassay for assessing nutrient enrichment in the Portneuf River, Idaho, USA

    Science.gov (United States)

    Ray, Andrew M.; Mebane, Christopher A.; Raben, Flint; Irvine, Kathryn M.; Marcarelli, Amy M.

    2014-01-01

    We describe and evaluate a laboratory bioassay that uses Lemna minor L. and attached epiphytes to characterize the status of ambient and nutrient-enriched water from the Portneuf River, Idaho. Specifically, we measured morphological (number of fronds, longest surface axis, and root length) and population-level (number of plants and dry mass) responses of L. minor and community-level (ash-free dry mass [AFDM] and chlorophyll a [Chl a]) responses of epiphytes to nutrient enrichment. Overall, measures of macrophyte biomass and abundance increased with increasing concentrations of dissolved phosphorus (P) and responded more predictably to nutrient enrichment than morphological measures. Epiphyte AFDM and Chl a were also greatest in P-enriched water; enrichments of N alone produced no measurable epiphytic response. The epiphyte biomass response did not directly mirror macrophyte biomass responses, illustrating the value of a combined macrophyte–epiphyte assay to more fully evaluate nutrient management strategies. Finally, the most P-enriched waters not only supported greater standing stocks of macrophyte and epiphytes but also had significantly higher water column dissolved oxygen and dissolved organic carbon concentrations and a lower pH. Advantages of this macrophyte–epiphyte bioassay over more traditional single-species assays include the use of a more realistic level of biological organization, a relatively short assay schedule (~10 days), and the inclusion of multiple biological response and water-quality measures.

  7. Evaluation of genotoxicity of liquid effluents from gas washing systems by means of bioassay Trad-MCN

    Energy Technology Data Exchange (ETDEWEB)

    Machado, Alessandra Carla Fattori Ergesse [Centro Tecnologico da Marinha em Sao Paulo (CTMSP), SP (Brazil). Div. de Monitoracao Ambiental], E-mail: alessandra@ctmsp.mar.mil.br; Alves, Edenise Segala [Instituto de Botanica de Sao Paulo, SP (Brazil). Secao de Anatomia], E-mail: ealves@ibot.sp.gov.br

    2007-07-01

    In the gas washing systems the gaseous emissions from a facility are forced through an absorbing liquid preventing pollutants to be dispersed into the atmosphere. In the Centro Tecnologico da Marinha em Sao Paulo/Centro Experimental Aramar (CEA), the gas washing are used to control the emissions from the uranium enrichment facilities. Uranium. fluoride, ammonia and hydrogen fluoride are the main contaminants, all heavily toxic. Biological assays, using plants or other living organisms, have been used to assess genotoxic agents in the environment. Among the bioassays using plants, the Trad-MCN has been used extensively, as it allows the evaluation of liquid or gaseous contaminants. The species Tradescantia pallida (Rose) was exposed in a dynamic system to liquid effluents from CEA. A positive control was the exposure to formaldehyde 10% in water, known as a very toxic solution, and the negative control was the exposure to filtered air. The protocol established by Ma (1983) for hybrid clones and validated for the T. pallida by Guimaraes (2003) was used to perform the Trad-MCN assays. Only preparations containing early tetrads were scored. In that context, the present study objectifies to evaluate, by the Trad-MCN bioassay, the genotoxicity of the solution from the gas washing and, also, evaluate the efficiency of that system. The results obtained show that the T. pallida is a sensitive bioindicator for the pollutants tested and can be useful for in vitro environmental monitoring under controlled conditions. (author)

  8. Bioassay-guided fractionation of extracts from Easter lily (Lilium longiflorum) flowers reveals unprecedented structural variability of steroidal glycoalkaloids.

    Science.gov (United States)

    Uhlig, Silvio; Hussain, Fozia; Wisløff, Helene

    2014-12-15

    Several Lilium species are nephrotoxic in cats (Felis silvestris catus), among them Easter lilies (Lilium longiflorum). Although clinical trials have been carried out, the causative toxic phytochemicals have not yet been identified. We thus aimed to determine the toxic constituents of Easter lily flowers applying a bioassay-guided approach based on a feline kidney cell line model. The bioassay-guided fractionation traced the observed cytotoxicity to a complex mixture of compounds that were tentatively identified as steroidal glycoalkaloids of the solasodine-type, based on multiple-fragmentation ion trap and high-resolution mass spectrometry. The glycoalkaloids in the active fraction possessed trisaccharide chains, and at least 16 different congeners could be separated using liquid chromatography-mass spectrometry. The two principal compounds were solasodine trisaccharides containing two hexose and one deoxy-hexose unit. In the remaining 14 analogues, one or two of the hydroxyl groups of the second hexose from the aglycone were acetylated. In addition, some of the analogues appeared to be carbonate esters. Esterification of steroidal glycoalkaloids in plants has only been reported once and was in accordance with higher antifungal activity of the acetylated versus the parent congener. Our pilot study shows that esterification of steroidal glycoalkaloids in Lilium species might be common resulting in an array of different analogues with largely unexplored structural variability and bioactivity. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Novel bioassay demonstrates attraction of the white potato cyst nematode Globodera pallida (Stone) to non-volatile and volatile host plant cues.

    Science.gov (United States)

    Farnier, Kevin; Bengtsson, Marie; Becher, Paul G; Witzell, Johanna; Witzgall, Peter; Manduríc, Sanja

    2012-06-01

    Potato cyst nematodes (PCNs) are a major pest of solanaceous crops such as potatoes, tomatoes, and eggplants and have been widely studied over the last 30 years, with the majority of earlier studies focusing on the identification of natural hatching factors. As a novel approach, we focused instead on chemicals involved in nematode orientation towards its host plant. A new dual choice sand bioassay was designed to study nematode responses to potato root exudates (PRE). This bioassay, conducted together with a traditional hatching bioassay, showed that biologically active compounds that induce both hatching and attraction of PCNs can be collected by water extraction of incised potato roots. Furthermore, our results demonstrated that PCN also were attracted by potato root volatiles. Further work is needed to fully understand how PCNs use host plant chemical cues to orientate towards hosts. Nevertheless, the simple attraction assay used in this study provides an important tool for the identification of host-emitted attractants.

  10. Fluconazole Pharmacokinetics in Galleria mellonella Larvae and Performance Evaluation of a Bioassay Compared to Liquid Chromatography-Tandem Mass Spectrometry for Hemolymph Specimens

    DEFF Research Database (Denmark)

    Astvad, Karen Marie Thyssen; Meletiadis, Joseph; Whalley, Sarah

    2017-01-01

    The invertebrate model organism Galleria mellonella can be used to assess the efficacy of treatment of fungal infection. The fluconazole dose best mimicking human exposure during licensed dosing is unknown. We validated a bioassay for fluconazole detection in hemolymph and determined...... the fluconazole pharmacokinetics and pharmacodynamics in larval hemolymph in order to estimate a humanized dose for future experiments. A bioassay using 4-mm agar wells, 20 μl hemolymph, and the hypersusceptible Candida albicans DSY2621 was established and compared to a validated liquid chromatography-tandem mass...... spectrometry (LC-MS-MS) method. G. mellonella larvae were injected with fluconazole (5, 10, and 20 mg/kg of larval weight), and hemolymph was harvested for 24 h for pharmacokinetics calculations. The exposure was compared to the human exposure during standard licensed dosing. The bioassay had a linear standard...

  11. Bioassays for TSH Receptor Autoantibodies, from FRTL-5 Cells to TSH Receptor-LH/CG Receptor Chimeras: The Contribution of Leonard D. Kohn.

    Science.gov (United States)

    Giuliani, Cesidio; Saji, Motoyasu; Bucci, Ines; Napolitano, Giorgio

    2016-01-01

    Since the discovery 60 years ago of the "long-acting thyroid stimulator" by Adams and Purves, great progress has been made in the detection of thyroid-stimulating hormone (TSH) receptor (TSHR) autoantibodies (TRAbs) in Graves' disease. Today, commercial assays are available that can detect TRAbs with high accuracy and provide diagnostic and prognostic evaluation of patients with Graves' disease. The present review focuses on the development of TRAbs bioassays, and particularly on the role that Leonard D. Kohn had in this. Indeed, 30 years ago, the Kohn group developed a bioassay based on the use of FRTL-5 cells that was characterized by high reproducibility, feasibility, and diagnostic accuracy. Using this FRTL-5 bioassay, Kohn and his colleagues were the first to develop monoclonal antibodies (moAbs) against the TSHR. Furthermore, they demonstrated the multifaceted functional nature of TRAbs in patients with Graves' disease, with the identification of stimulating and blocking TRAbs, and even antibodies that activated pathways other than cAMP. After the cloning of the TSHR, the Kohn laboratory constructed human TSHR-rat luteinizing hormone/chorionic gonadotropin receptor chimeras. This paved the way to a new bioassay based on the use of non-thyroid cells transfected with the Mc4 chimera. The new Mc4 bioassay is characterized by high diagnostic and prognostic accuracy, greater than for other assays. The availability of a commercial kit based on the Mc4 chimera is spreading the use of this assay worldwide, indicating its benefits for these patients with Graves' disease. This review also describes the main contributions made by other researchers in TSHR molecular biology and TRAbs assay, especially with the development of highly potent moAbs. A comparison of the diagnostic accuracies of the main TRAbs assays, as both immunoassays and bioassays, is also provided.

  12. Diagnostic doses and times for Phlebotomus papatasi and Lutzomyia longipalpis sand flies (Diptera: Psychodidae: Phlebotominae) using the CDC bottle bioassay to assess insecticide resistance.

    Science.gov (United States)

    Denlinger, David S; Creswell, Joseph A; Anderson, J Laine; Reese, Conor K; Bernhardt, Scott A

    2016-04-15

    Insecticide resistance to synthetic chemical insecticides is a worldwide concern in phlebotomine sand flies (Diptera: Psychodidae), the vectors of Leishmania spp. parasites. The CDC bottle bioassay assesses resistance by testing populations against verified diagnostic doses and diagnostic times for an insecticide, but the assay has been used limitedly with sand flies. The objective of this study was to determine diagnostic doses and diagnostic times for laboratory Lutzomyia longipalpis (Lutz & Nieva) and Phlebotomus papatasi (Scopoli) to ten insecticides, including pyrethroids, organophosphates, carbamates, and DDT, that are used worldwide to control vectors. Bioassays were conducted in 1,000-ml glass bottles each containing 10-25 sand flies from laboratory colonies of L. longipalpis or P. papatasi. Four pyrethroids, three organophosphates, two carbamates and one organochlorine, were evaluated. A series of concentrations were tested for each insecticide, and four replicates were conducted for each concentration. Diagnostic doses were determined only during the exposure bioassay for the organophosphates and carbamates. For the pyrethroids and DDT, diagnostic doses were determined for both the exposure bioassay and after a 24-hour recovery period. Both species are highly susceptible to the carbamates as their diagnostic doses are under 7.0 μg/ml. Both species are also highly susceptible to DDT during the exposure assay as their diagnostic doses are 7.5 μg/ml, yet their diagnostic doses for the 24-h recovery period are 650.0 μg/ml for Lu. longipalpis and 470.0 μg/ml for P. papatasi. Diagnostic doses and diagnostic times can now be incorporated into vector management programs that use the CDC bottle bioassay to assess insecticide resistance in field populations of Lu. longipalpis and P. papatasi. These findings provide initial starting points for determining diagnostic doses and diagnostic times for other sand fly vector species and wild populations using the CDC

  13. Development of bioassay for pathogenecity testing of Ureaplasma urealyticum as part of host-pathogen communication

    Directory of Open Access Journals (Sweden)

    Purnomo Soeharso

    2005-12-01

    Full Text Available Bioassay of Ureaplasma urealyticum is necessary for detection as well as determination of pathogenic factors in order to understand the pathogenesis of diseases associate with ureaplasma infection. Cultivation and verification of ureaplasma is the first step of this study in the purpose of discovering sensitive method for ureaplasma detection. Cultivation of ureaplasma either in liquid or in solid media are able to detect the existence of ureaplasma in samples analyzed. However, application of PCR using specific primers to be compatible with urease gene (ure would confirm the presence of ureaplasma. The pathogenicity of ureaplasma is potentially monitored using reporter gene as a marker for gene expression. IceC was chosen as reporter gene for ureaplasma pathogenic determination as the gene has great sensitivity, easily detectable and quantitated in simple method of ice nucleation assay. Transposon 916 (Tn916 was selected as a vector for iceC gene to transform ureaplasma. The application of recombinant Tn916-iceC which is considered as pUI, allow detection of ureaplasma activities when transform ureaplasma is tested by ice nucleation assay. It was expected that ureaplasma transformation is the manifestation of mutagenesis which interfere genes responsible for bacterial pathogenicity, in order pathogenesis of bacterial infection to be analyzed accurately. IgA1 protease is considered to be an important factor for ureaplasma pathogenicity as the enzyme is required for successful colonization. Identification of iga gene and  determination of IgA1 protease activity are important for understanding the pathogenesis of ureaplasma infection. Putative iga gene of Mycoplasma genitalium was used as a reference to identify the presence of iga nucleotide sequence in U. urealyticum. Convincing evidence were obtained after PCR amplification of ureaplasma DNA using primers designed to be compatible with putative iga gene of M. genitalium followed by the

  14. Developmental status of bioassays in genetic toxicology: a report of Phase II of the US Environmental Protection Agency Gene-Tox program

    Energy Technology Data Exchange (ETDEWEB)

    Brusick, D; Auletta, A

    1985-01-01

    The Gene-Tox Program was structured around two phases of genetic test data evaluation. The first phase consisted of 36 Work Group reports, each evaluating the results and performance of a specific bioassay. The second phase consisted of a plan to summarize the information provided by the Work Groups. The Gene-Tox Coordinating Committee was to be responsible for Phase II, and several subgroups were assigned specific goals in implementing this analysis. This report deals with Goal I which is to identify the developmental status of the individual bioassays reviewed by the Gene-Tox Work Groups in the first phase of the Program. 5 references, 6 tables.

  15. Effect-based trigger values for in vitro and in vivo bioassays performed on surface water extracts supporting the environmental quality standards (EQS) of the European Water Framework Directive

    NARCIS (Netherlands)

    Escher, Beate I.; Aїt-Aїssa, Selim; Behnisch, Peter A.; Brack, Werner; Brion, François; Brouwer, Abraham; Buchinger, Sebastian; Crawford, Sarah E.; Du Pasquier, David; Hamers, Timo; Hettwer, Karina; Hilscherová, Klára; Hollert, Henner; Kase, Robert; Kienle, Cornelia; Tindall, Andrew J.; Tuerk, Jochen; van der Oost, Ron; Vermeirssen, Etienne; Neale, Peta A.

    Effect-based methods including cell-based bioassays, reporter gene assays and whole-organism assays have been applied for decades in water quality monitoring and testing of enriched solid-phase extracts. There is no common EU-wide agreement on what level of bioassay response in water extracts is

  16. Evaluation of genotoxic effects of surface waters using a battery of bioassays indicating different mode of action.

    Science.gov (United States)

    Han, Yingnan; Li, Na; Oda, Yoshimitsu; Ma, Mei; Rao, Kaifeng; Wang, Zijian; Jin, Wei; Hong, Gang; Li, Zhiguo; Luo, Yi

    2016-11-01

    With the burgeoning contamination of surface waters threatening human health, the genotoxic effects of surface waters have received much attention. Because mutagenic and carcinogenic compounds in water cause tumors by different mechanisms, a battery of bioassays that each indicate a different mode of action (MOA) is required to evaluate the genotoxic effects of contaminants in water samples. In this study, 15 water samples from two source water reservoirs and surrounding rivers in Shijiazhuang city of China were evaluated for genotoxic effects. Target chemical analyses of 14 genotoxic pollutants were performed according to the Environmental quality standards for surface water of China. Then, the in vitro cytokinesis-block micronucleus (CBMN) assay, based on a high-content screening technique, was used to detect the effect of chromosome damage. The SOS/umu test using strain TA1535/pSK1002 was used to detect effects on SOS repair of gene expression. Additionally, two other strains, NM2009 and NM3009, which are highly sensitive to aromatic amines and nitroarenes, respectively, were used in the SOS/umu test to avoid false negative results. In the water samples, only two of the genotoxic chemicals listed in the water standards were detected in a few samples, with concentrations that were below water quality standards. However, positive results for the CBMN assay were observed in two river samples, and positive results for the induction of umuC gene expression in TA1535/pSK1002 were observed in seven river samples. Moreover, positive results were observed for NM2009 with S9 and NM3009 without S9 in some samples that had negative results using the strain TA1535/pSK1002. Based on the results with NM2009 and NM3009, some unknown or undetected aromatic amines and nitroarenes were likely in the source water reservoirs and the surrounding rivers. Furthermore, these compounds were most likely the causative pollutants for the genotoxic effect of these water samples. Therefore

  17. Assessment of sediment toxicity in the Lagoon of Venice (Italy) using a multi-species set of bioassays.

    Science.gov (United States)

    Picone, Marco; Bergamin, Martina; Losso, Chiara; Delaney, Eugenia; Arizzi Novelli, Alessandra; Ghirardini, Annamaria Volpi

    2016-01-01

    Within the framework of a Weight of Evidence (WoE) approach, a set of four toxicity bioassays involving the amphipod Corophium volutator (10 d lethality test on whole sediment), the sea urchin Paracentrotus lividus (fertilization and embryo toxicity tests on elutriate) and the pacific oyster Crassostrea gigas (embryo toxicity test on elutriate) was applied to sediments from 10 sampling sites of the Venice Lagoon (Italy). Sediments were collected during three campaigns carried out in May 2004 (spring campaign), October 2004 (autumn campaign) and February 2005 (winter campaign). Toxicity tests were performed on all sediment samples. Sediment grain-size and chemistry were measured during spring and autumn campaigns. This research investigated (i) the ability of toxicity tests in discriminating among sites with different contamination level, (ii) the occurrence of a gradient of effect among sampling sites, (iii) the possible correlation among toxicity tests, sediment chemistry, grain size and organic carbon, and (iv) the possible occurrence of toxicity seasonal variability. Sediment contamination levels were from low to moderate. No acute toxicity toward amphipods was observed, while sea urchin fertilization was affected only in few sites in just a single campaign. Short-term effects on larval development of sea urchin and oyster evidenced a clear spatial trend among sites, with increasing effects along the axis connecting the sea-inlets with the industrial area. The set of bioassays allowed the identification of a spatial gradient of effect, with decreasing toxicity from the industrial area toward the sea-inlets. Multivariate data analysis showed that the malformations of oyster embryos were significantly correlated to the industrial contamination (metals, polynuclear aromatic hydrocarbons, hexachlorobenzene and polychlorinated biphenyls), while sea urchin development to sediment concentrations of As, Cr and organic carbon. Both embryo toxicity tests were

  18. The potential of a fluorescent-based approach for bioassay of antifungal agents against chili anthracnose disease in Thailand.

    Science.gov (United States)

    Chutrakul, Chanikul; Khaokhajorn, Pratoomporn; Auncharoen, Patchanee; Boonruengprapa, Tanapong; Mongkolporn, Orarat

    2013-01-01

    Severe chili anthracnose disease in Thailand is caused by Colletotrichum gloeosporioides and C. capsici. To discover anti-anthracnose substances we developed an efficient dual-fluorescent labeling bioassay based on a microdilution approach. Indicator strains used in the assay were constructed by integrating synthetic green fluorescent protein (sGFP) and Discosoma sp. red fluorescent protein (DsRedExp) genes into the genomes of C. gloeosporioides or C. capsici respectively. Survival of co-spore cultures in the presence of inhibitors was determined by the expression levels of these fluorescent proteins. This developed assay has high potential for utilization in the investigation of selective inhibition activity to either one of the pathogens as well as the broad-range inhibitory effect against both pathogens. The value of using the dual-fluorescent assay is rapid, reliable, and consistent identification of anti-anthracnose agents. Most of all, the assay enables the identification of specific inhibitors under the co-cultivation condition.

  19. Chemical investigation of Cyperus distans L. and inhibitory activity of scabequinone in seed germination and seedling growth bioassays.

    Science.gov (United States)

    Vilhena, Karyme S S; Guilhon, Giselle Maria Skelding Pinheiro; Zoghbi, Maria das Graças B; Santos, Lourivaldo Silva; Souza Filho, Antonio Pedro Silva

    2014-01-01

    Chemical investigation of the rhizomes of Cyperus distans (Cyperaceae) led to the identification of α-ciperone, cyperotundone and scabequinone, besides other common constituents. Complete assignment of the (13)C NMR data of scabequinone is being published for the first time. The inhibitory effects of C. distans extracts and scabequinone on the seed germination and seedling growth of Mimosa pudica, Senna obtusifolia and Pueraria phaseoloides were evaluated. Seed germination inhibition bioassay revealed that S. obtusifolia (52-53%) was more sensitive to the hexane and the methanol extracts at 1% than M. pudica (0-10%). Scabequinone at 250 mg L⁻¹ displayed seed germination inhibitions more than 50% and radicle growth reduction of more than 35% of the test species S. obtusifolia and P. phaseoloides, while the hypocotyl growth of M. pudica was significantly affected (>50%) by the quinone at the same concentration. These results demonstrate that scabequinone contributes to the overall inhibitory activities of C. distans.

  20. Phytotoxicity and Cytotoxicity of Essential Oil from Leaves of Plectranthus amboinicus, Carvacrol, and Thymol in Plant Bioassays.

    Science.gov (United States)

    Pinheiro, Patrícia Fontes; Costa, Adilson Vidal; Alves, Thammyres de Assis; Galter, Iasmini Nicoli; Pinheiro, Carlos Alexandre; Pereira, Alexandre Fontes; Oliveira, Carlos Magno Ramos; Fontes, Milene Miranda Praça

    2015-10-21

    The essential oil of Plectranthus amboinicus and its chemotypes, carvacrol and thymol, were evaluated on the germination and root and aerial growth of Lactuca sativa and Sorghum bicolor and in acting on the cell cycle of meristematic root cells of L. sativa. The main component found in the oil by analysis in gas chromatography-mass spectrometry and gas chromatography flame ionization detection was carvacrol (88.61% in area). At a concentration of 0.120% (w v(-1)), the oil and its chemotypes retarded or inhibited the germination and decreased root and aerial growth in monocot and dicot species used in the bioassays. In addition, all substances caused changes in the cell cycle of the meristematic cells of L. sativa, with chromosomal alterations occurring from the 0.015% (w v(-1)) concentration. The essential oil of P. amboinicus, carvacrol, and thymol have potential for use as bioherbicides.

  1. Docking, synthesis and bioassay studies of imine derivatives as potential inhibitors for dengue NS2B/ NS3 serine protease

    Directory of Open Access Journals (Sweden)

    Neni Frimayanti

    2017-11-01

    Full Text Available Objective: To search imine derivatives as new active agents against dengue type 2 NS2B/NS3 using molecular docking, since there is no effective vaccine against flaviviral infections. Methods: In this research, molecular docking was performed for a series of imine derivatives and the information obtained from the docking studies was used to explore the binding modes of these imine derivatives with dengue type 2 NS2B/NS3 serine protease. A set of imine were synthesized and bioassay study of the inhibitory activities of these compounds was then performed. Results: The results indicated that MY8 and MY4 have the ability to inhibit DEN2 NS2B/NS3 proteolytic activity. Conclusions: These two compounds were chosen as the reference for the next stage in drug design as new inhibitor agents against NS2B/NS3.

  2. Allelopathic potential of Artemisia arborescens: isolation, identification and quantification of phytotoxic compounds through fractionation-guided bioassays.

    Science.gov (United States)

    Araniti, Fabrizio; Lupini, Antonio; Sorgonà, Agostino; Conforti, Filomena; Marrelli, Mariangela; Statti, Giancarlo Antonio; Menichini, Francesco; Abenavoli, Maria Rosa

    2013-01-01

    The aerial part of Artemisia arborescens L. (Asteraceae) was extracted with water and methanol, and both extracts were fractionated using n-hexane, chloroform, ethyl acetate and n-butanol. The potential phytotoxicity of both crude extracts and their fractions were assayed in vitro on seed germination and root growth of lettuce (Lactuca sativa L.), a sensitive species largely employed in the allelopathy studies. The inhibitory activities were analysed by dose-response curves and the ED 50 were estimated. Crude extracts strongly inhibited both germination and root growth processes. The fraction-bioassay indicated the following hierarchy of phytotoxicity for both physiological processes: ethyl acetate ≥ n-hexane > chloroform ≥ n-butanol. On the n-hexane fraction, GC-MS analyses were carried out to characterise and quantify some of the potential allelochemicals. Twenty-one compounds were identified and three of them, camphor, trans-caryophyllene and pulegone were quantified.

  3. The relationship of radioimmunoassay to bioassay: In vitro studies with synthetic lysine vasopressin in aqueous solution inactivated by heat

    International Nuclear Information System (INIS)

    Loeve Lemboel, H.

    1978-01-01

    The relationship of radioimmunoassay to pressor assay and antidiuretic assay was investigated in a simple in vitro system of synthetic lysine vasopressin in aqueous solution inactivated by heating at 100 deg C for 9, 18, 27, 36, 54 and 72 h. An apparent dissociation between radioimmunoassay and bioassay was demonstrated, with biological activity being lost more rapidly than immunological activity. The half-times were 32 h for radioimmunoassay, 23 h for antidiuretic assay and 22 h for pressor assay. However, ion-exchange chromatography showed immunological heterogeneity but biological homogeneity of the lysine vasopressin used, and indicated that the presence of impurities in the vasopressin might to some extent explain the discrepancy between assay results. Synthetic arginine vasopressin and arginine vasopressin of pituitary origin showed a similar immunological heterogeneity by ion-exchange chromatography. (author)

  4. Soil ciliate bioassay for the pore water habitat. A missing link between microflora and earthworm testing in soil toxicity assessment

    Energy Technology Data Exchange (ETDEWEB)

    Berthold, A. [Lab. for Ecotoxicology, Univ. of Veterinary Medicine, Vienna (Austria); Jakl, T. [Chemicals Policy Unit, Federal Ministry of Agriculture, Forestry, Environment and Water Management, Vienna (Austria)

    2002-07-01

    Background, Scope and Goal. The chemical, pesticide and biocide legislation of the European Union assembles a variety of bioassays. Among the ecotoxicological tests involved, the testing strategy for the aquatic compartment builds up on three tests reflecting the main trophic levels (algae, Daphnia, fish). For the soil compartment at least one trophic level for a basic food chain is missing, namely between microflora and earthworms. Protozoa are an ideal missing link as they were shown to be the most prominent faunal contributors to nutrient cycling in soil ecosystems and as they represent the lacking first level consumers as well as the highly diverse microfauna. As protozoa inhabit the soil pore water, they can serve as direct indicators for the solved and thus bioavailable portion of xenobiotics. In order to widen the spectrum of available toxicity tests for a meaningful effect assessment for the soil compartment, a test with the soil ciliate Colpoda inflata (Ciliophora, Protozoa), introduced by Pratt et al. (1997), was improved. Methods. The novel improvements comprise a substantially refined inoculation and counting procedure, as well as the adaptation to yeast as a nutritional source. The test was designed to be rapid and easy to perform, in order to have both a higher degree of standardisation and reproducibility, as well as to be in compliance with international test guidelines. Results and Discussion. Five test substances, cadmium chloride, potassium dichromate, acetone, atrazine, and metolachlor, were used in single-compound, static, short-term exposure (24 h, 48 h) tests to examine the effect on the population growth of C. inflata. The median effective concentrations (EC50) were 0.17 to 0.26 mg/l for Cd, 34 to 63 mg/l for Cr, >3000 mg/l for acetone, 91-112 mg/l for atrazine and 83-119 mg/l for metolachlor. The equilibrium partitioning approach was used to extrapolate the results to total soil exposure and thus enable a sensitivity comparison to

  5. Assessment of Sediment Heavy Metals Pollution Using Screening Methods (XRF, TGA/MS, XRPD and Earthworms Bioassay)

    Science.gov (United States)

    Findoráková, Lenka; Šestinová, Ol'ga; Hančul'ák, Jozef; Fedorová, Erika; Zorkovská, Anna

    2016-10-01

    The aim of this study is focused on the use of screening methods (TG/DTA coupled with MS, XRF, AAS, XRPD and earthworm bioassay) for sediments pollution assessing by heavy metals (Cu, Zn, Pb, Hg) coming from the former mining workloads in the central Spis, Eastern Slovakia. The screening methods (XRF, AAS) indicated pollution of studied sediments by Cu, Zn, Pb, Hg. The earthworms Dendrobaena veneta caused in some studied samples decrease of heavy metals concentration after their 7 days’ exposure in sediments. The other screening methods such as thermal analysis and XRPD analysis, does not confirm the specifically changes in physicochemical properties comparing the properties before and after 7 days’ earthworm's exposure.

  6. Bioassay battery interlaboratory investigation of emerging contaminants in spiked water extracts e Towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring

    NARCIS (Netherlands)

    Di Paolo, C.; Ottermanns, R.; Keiter, S.; Ait-Aissa, S.; Bluhm, K.; Brack, W.; Breitholz, M.; Buchinger, S.; Carere, M.; Chalon, C.; Cousin, X.; Dulio, V.; Escher, B.I.; Hamers, T.; Jarque, S.; Jonas, A.; Maillot-Marechal, E.; Marneffe, Y.; Nguyen, M.T.; Pandard, P.; Schifferli, A.; Schulze, T.; Seidensticker, S.; Seiler, T.B.; Tang, J.; van der Oost, R.; Vermeirssen, E.; Zounková, R.; Zwart, N.; Hollert, H.

    2016-01-01

    Bioassays are particularly useful tools to link the chemical and ecological assessments in water quality monitoring. Different methods cover a broad range of toxicity mechanisms in diverse organisms, and account for risks posed by non-target compounds and mixtures. Many tests are already applied in

  7. The use of the DR CALUX bioassay and indicator polychlorinated biphenyls for screening of elevated levels of dioxins and dioxin-like polychlorinated biphenyls in eel.

    NARCIS (Netherlands)

    Hoogenboom, Ron; Bovee, Toine; Traag, Win A; Hoogerbrugge, Ronald; Baumann, Bert; Portier, Liza; Weg, Guido van de; Vries, Jaap de

    2006-01-01

    The DR CALUX bioassay is a very suitable screening method for dioxins and dioxin-like-PCBs in feed and food. This was, e. g. demonstrated in a survey in the Netherlands to control the dioxin levels in eel. The DR CALUX assay, but also indicator polychlorinated biphenyls (PCB) were evaluated as a

  8. A RIKILT yeast estrogen bioassay (REA) for estrogen residue detection in urine of calves experimentally treated with 17ß-estradiol

    NARCIS (Netherlands)

    Divari, S.; Maria, De R.; Cannizzo, F.T.; Spada, F.; Mulasso, C.; Bovee, T.F.H.; Capra, P.; Leporati, M.; Biolatti, B.

    2010-01-01

    17ß-Estradiol is one of the most powerful sex steroids illegally used in bovine production. The objective of this study was to evaluate the application and the specificity of the RIKILT yeast estrogen bioassay (REA) for the detection of molecules with estrogenic activities in the urine of calves

  9. Cancer risk assessment of polycyclic aromatic hydrocarbon contaminated soils determined using bioassay-derived levels of benzo[a]pyrene equivalents.

    Science.gov (United States)

    Lemieux, Christine L; Long, Alexandra S; Lambert, Iain B; Lundstedt, Staffan; Tysklind, Mats; White, Paul A

    2015-02-03

    Here we evaluate the excess lifetime cancer risk (ELCR) posed by 10 PAH-contaminated soils using (i) the currently advocated, targeted chemical-specific approach that assumes dose additivity for carcinogenic PAHs and (ii) a bioassay-based approach that employs the in vitro mutagenic activity of the soil fractions to determine levels of benzo[a]pyrene equivalents and, by extension, ELCR. Mutagenic activity results are presented in our companion paper.1 The results show that ELCR values for the PAH-containing fractions, determined using the chemical-specific approach, are generally (i.e., 8 out of 10) greater than those calculated using the bioassay-based approach; most are less than 5-fold greater. Only two chemical-specific ELCR estimates are less than their corresponding bioassay-derived values; differences are less than 10%. The bioassay-based approach, which permits estimation of ELCR without a priori knowledge of mixture composition, proved to be a useful tool to evaluate the chemical-specific approach. The results suggest that ELCR estimates for complex PAH mixtures determined using a targeted, chemical-specific approach are reasonable, albeit conservative. Calculated risk estimates still depend on contentious PEFs and cancer slope factors. Follow-up in vivo mutagenicity assessments will be required to validate the results and their relevance for human health risk assessment of PAH-contaminated soils.

  10. Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays.

    Science.gov (United States)

    Minouflet, Marion; Ayrault, Sophie; Badot, Pierre-Marie; Cotelle, Sylvie; Ferard, Jean-François

    2005-01-01

    Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137Cs pollution.

  11. A novel bioassay to evaluate Beauveria bassiana strain NI8 and the insect growth regulator, novaluron, against Lygus lineolaris on a non-autoclaved solid artificial diet

    Science.gov (United States)

    Detailed life-tables studies or more quantitative estimates of the impact of control agents on TPB life history require a bioassay option to study the impact of prolonged exposure for weeks following contact with the control agent. This is difficult with plant tissue that must be routinely replaced ...

  12. BROMOETHANE, CHLOROETHANE AND ETHYLENE OXIDE INDUCED UTERINE NEOPLASMS IN B6C3F1 MICE FROM 2-YEAR NTP INHALATION BIOASSAYS: PATHOLOGY AND INCIDENCE DATA REVISITED

    Science.gov (United States)

    SUMMARY: Chloroethane, bromoethane and etjulene oxide represent a unique set of three chemicals that induce endometrial neoplasms in the uterus of B6C3F1 mice following an inhalation route of exposure. The results of the NTP's chronic bioassays with these three compounds resu...

  13. A preliminary evaluation of the toxic equivalence factor (TEF) for 2,3,4,7,8-PCDF (4-PCDF) using data from the recent NTP dioxin bioassays

    Energy Technology Data Exchange (ETDEWEB)

    Budinsky, R.; Landenberger, B.; Fontaine, D. [The Dow Chemical Company (United States); Starr, T.B. [TBS Associates, Raleigh, NC (United States); Paustenbach, D. [ChemRisk (United States)

    2004-09-15

    PCDD/PCDF toxic equivalency factors (TEFs) are being routinely applied now in human cancer risk assessments for dioxin-like compounds despite the fact that until very recently only two congeners (1,2,3,6,7,8- and 1,2,3,7,8,9-hexachlorodibenzo-p-dioxin) other than 2,3,7,8-TCDD had been evaluated for carcinogenicity in a standard cancer bioassay. However, in December 2003, draft reports from the National Toxicology Program (NTP) cancer bioassays were made available for 2,3,7,8-TCDD, 4-PCDF, PCB 126, and a (presumably) equipotent mixture of these three compounds. These data provide important new information for evaluating the accuracy of TEFs in predicting the potential human cancer hazard for 4-PCDF and PCB126 alone and in combination with TCDD. We present herein results from a series of simple statistical tests demonstrating that the current TEF of 0.5 for 4-PCDF, when combined appropriately with the new TCDD bioassay data, fails to predict accurately the results from the new 4-PCDF bioassay.

  14. A Choline Oxidase Amperometric Bioassay for the Detection of Mustard Agents Based on Screen-Printed Electrodes Modified with Prussian Blue Nanoparticles

    Directory of Open Access Journals (Sweden)

    Fabiana Arduini

    2015-02-01

    Full Text Available In this work a novel bioassay for mustard agent detection was proposed. The bioassay is based on the capability of these compounds to inhibit the enzyme choline oxidase. The enzymatic activity, which is correlated to the mustard agents, was electrochemically monitored measuring the enzymatic product, hydrogen peroxide, by means of a screen-printed electrode modified with Prussian Blue nanoparticles. Prussian Blue nanoparticles are able to electrocatalyse the hydrogen peroxide concentration reduction at low applied potential (−50 mV vs. Ag/AgCl, thus allowing the detection of the mustard agents with no electrochemical interferences. The suitability of this novel bioassay was tested with the nitrogen mustard simulant bis(2-chloroethylamine and the sulfur mustard simulants 2-chloroethyl ethyl sulfide and 2-chloroethyl phenyl sulfide. The bioassay proposed in this work allowed the detection of mustard agent simulants with good sensitivity and fast response, which are excellent premises for the development of a miniaturised sensor well suited for an alarm system in case of terrorist attacks.

  15. A small-volume bioassay for quantification of the esterase inhibiting potency of mixtures of organophosphate and carbamate insecticides in rainwater : development and optimization

    NARCIS (Netherlands)

    Hamers, T.; Molin, K.R.J.; Koeman, J.H.; Murk, A.J.

    2000-01-01

    The goal of this study was to develop a sensitive in vitro bioassay for quantification of the total esterase inhibiting potency of low concentrations of organophosphate and carbamate insecticides in relatively small rainwater samples. Purified acetylcholinesterase (AChE) from electric eel

  16. Screening for estrogen residues in calf urine: Comparison of a validated yeast estrogen bioassay and gas chromatography-tandem mass spectrometry

    NARCIS (Netherlands)

    Nielen, M.W.F.; Bovee, T.F.H.; Heskamp, H.H.; Lasaroms, J.J.P.; Sanders, M.B.; Rhijn, van J.A.; Groot, M.J.; Hoogenboom, L.A.P.

    2006-01-01

    Within the European Union, the control for residues of illegal hormones in food-producing animals is based on urine analysis for a few target analytes using gas chromatography/mass spectrometry and/or liquid chromatography¿tandem mass spectrometry. Recently, we developed a robust yeast bioassay

  17. Third-generation Ah receptor-responsive luciferase reporter plasmids: amplification of dioxin-responsive elements dramatically increases CALUX bioassay sensitivity and responsiveness.

    Science.gov (United States)

    He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S; Zhao, Jing; Heath-Pagliuso, Sharon; Denison, Michael S

    2011-10-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related dioxin-like chemicals are widespread and persistent environmental contaminants that produce diverse toxic and biological effects through their ability to bind to and activate the Ah receptor (AhR) and AhR-dependent gene expression. The chemically activated luciferase expression (CALUX) system is an AhR-responsive recombinant luciferase reporter gene-based cell bioassay that has been used in combination with chemical extraction and cleanup methods for the relatively rapid and inexpensive detection and relative quantitation of dioxin and dioxin-like chemicals in a wide variety of sample matrices. Although the CALUX bioassay has been validated and used extensively for screening purposes, it has some limitations when screening samples with very low levels of dioxin-like chemicals or when there is only a small amount of sample matrix for analysis. Here, we describe the development of third-generation (G3) CALUX plasmids with increased numbers of dioxin-responsive elements, and stable transfection of these new plasmids into mouse hepatoma (Hepa1c1c7) cells has produced novel amplified G3 CALUX cell bioassays that respond to TCDD with a dramatically increased magnitude of luciferase induction and significantly lower minimal detection limit than existing CALUX-type cell lines. The new G3 CALUX cell lines provide a highly responsive and sensitive bioassay system for the detection and relative quantitation of very low levels of dioxin-like chemicals in sample extracts.

  18. Using the dioxin receptor-calux in vitro bioassay to screen marine harbor sediments for compounds with a dioxin-like mode of action

    NARCIS (Netherlands)

    Stronkhorst, J.; Leonards, P.E.G.; Murk, A.J.

    2002-01-01

    The presence of dioxin-like compounds in sediments from harbors and reference sites along the Dutch coast was investigated using the dioxin receptor–chemically activated luciferase gene expression (DR-CALUX) bioassay. The DR-CALUX response varied between 0.2 and 136 ng/kg dry weight expressed in

  19. Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays

    International Nuclear Information System (INIS)

    Minouflet, Marion; Ayrault, Sophie; Badot, Pierre-Marie; Cotelle, Sylvie; Ferard, Jean-Francois

    2005-01-01

    Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137 Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137 Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137 Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137 Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137 Cs pollution

  20. Fluconazole Pharmacokinetics in Galleria mellonella Larvae and Performance Evaluation of a Bioassay Compared to Liquid Chromatography-Tandem Mass Spectrometry for Hemolymph Specimens

    Science.gov (United States)

    Astvad, Karen Marie Thyssen; Meletiadis, Joseph; Whalley, Sarah

    2017-01-01

    ABSTRACT The invertebrate model organism Galleria mellonella can be used to assess the efficacy of treatment of fungal infection. The fluconazole dose best mimicking human exposure during licensed dosing is unknown. We validated a bioassay for fluconazole detection in hemolymph and determined the fluconazole pharmacokinetics and pharmacodynamics in larval hemolymph in order to estimate a humanized dose for future experiments. A bioassay using 4-mm agar wells, 20 μl hemolymph, and the hypersusceptible Candida albicans DSY2621 was established and compared to a validated liquid chromatography-tandem mass spectrometry (LC–MS-MS) method. G. mellonella larvae were injected with fluconazole (5, 10, and 20 mg/kg of larval weight), and hemolymph was harvested for 24 h for pharmacokinetics calculations. The exposure was compared to the human exposure during standard licensed dosing. The bioassay had a linear standard curve between 1 and 20 mg/liter. Accuracy and coefficients of variation (percent) values were below 10%. The Spearman coefficient between assays was 0.94. Fluconazole larval pharmacokinetics followed one-compartment linear kinetics, with the 24-h area under the hemolymph concentration-time curve (AUC24 h) being 93, 173, and 406 mg · h/liter for the three doses compared to 400 mg · h/liter in humans under licensed treatment. In conclusion, a bioassay was validated for fluconazole determination in hemolymph. The pharmacokinetics was linear. An exposure comparable to the human exposure during standard licensed dosing was obtained with 20 mg/kg. PMID:28760893

  1. A “walker” tool to place Diaphorina citri (Hemiptera: Liviidae) adults at predetermined sites for bioassays of behavior in citrus (Sapindales: Rutacease) trees

    Science.gov (United States)

    A walker tool was developed to assist placement of D. citri on citrus host trees in behavioral bioassays. The walker performs better than a commonly used paintbrush tool in the proportion of successful placements and in the reduction of jumps away from the citrus leaf, although it takes about two mi...

  2. Screening for the presence of lipophilic marine biotoxins in shellfish samples using the neuro-2a bioassay.

    Science.gov (United States)

    Bodero, Marcia; Bovee, Toine F H; Wang, Si; Hoogenboom, Ron L A P; Klijnstra, Mirjam D; Portier, Liza; Hendriksen, Peter J M; Gerssen, Arjen

    2018-02-01

    The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10%. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.

  3. Enhancement of preneoplastic lesion yield by Chios Mastic Gum in a rat liver medium-term carcinogenesis bioassay

    International Nuclear Information System (INIS)

    Doi, Kenichiro; Wei, Min; Kitano, Mitsuaki; Uematsu, Naomi; Inoue, Masayo; Wanibuchi, Hideki

    2009-01-01

    The mastic (Pistacia lentiscus var. chia) tree is native throughout the Mediterranean region and has long proved a source of food additives and medical treatments. To investigate the modifying effects of Chios Mastic Gum on rat liver carcinogenesis, 6-week-old male F344 rats were subjected to the established rat liver medium-term carcinogenesis bioassay (Ito-test). At the commencement, rats (groups 1-4) were intraperitoneally injected with 200 mg/kg body weight of diethylnitrosamine (DEN). After two weeks, mastic was added to CRF (Charles River Formula)-1 powdered basal diet at doses of 0, 0.01, 0.1 and 1% in groups 1-4, respectively. At week 3, all rats were underwent two-thirds partial hepatectomy. The experiment was terminated at week 8. As results show, liver weights were significantly increased in a mastic dose-dependent manner among groups 1-4. The numbers (/cm 2 ) and the areas (mm 2 /cm 2 ) of glutathione S-transferase placental form (GST-P)-positive cell foci (≥ 0.2 mm in diameter) were significantly increased in the DEN-1% group compared to the DEN-alone group, along with the average areas per foci and larger-sized foci (≥ 0.4 mm). 5-Bromo-2'-deoxyuridine (BrdU) + GST-P double-immunohistochemistry showed the highest BrdU-labeling indices within GST-P foci in the DEN-1% group. 8-hydroxydeoxyguanosine (8-OHdG) levels in liver DNA did not vary, while real-time quantitative polymerase chain reaction (PCR) analysis of livers revealed many up- or down-regulated genes in the DEN-1% group. In conclusion, this is the first report to display a promotion potential of Chios Mastic Gum on the formation of preneoplastic lesions in the established rat liver medium-term carcinogenesis bioassay

  4. Results of long-term carcinogenicity bioassays on Coca-Cola administered to Sprague-Dawley rats.

    Science.gov (United States)

    Belpoggi, Fiorella; Soffritti, Morando; Tibaldi, Eva; Falcioni, Laura; Bua, Luciano; Trabucco, Francesca

    2006-09-01

    Coca-Cola was invented in May 1886 in Atlanta, Georgia by a pharmacist who, by accident or design, mixed carbonated water with the syrup of sugar, phosphoric acid, caffeine, and other natural flavors to create what is known as "the world's favorite soft drink." Coca-Cola is currently sold in more than 200 countries and in early 2000, the company sold its 10 billionth unit case of Coca-Cola branded products. Given the worldwide consumption of Coca-Cola, a project of experimental bioassays to study its long-term effects when administered as substitute for drinking water on male and female Sprague-Dawley rats was planned and executed. The objective of the project was to study whether and how long-term consumption of Coca-Cola affects the basic tumorigram of test animals. The bioassays were performed on rats beginning at different ages, namely: (a) on males and females exposed since embryonic life or from 7 weeks of age; and (b) on males and females exposed from 30, 39, or 55 weeks of age. Overall, the project included 1999 rats. During the biophase, data were collected on fluid and feed consumption, body weight, and survival. Animals were kept under observation until spontaneous death and underwent complete necropsy. The results indicate: (a) an increase in body weight in all treated animals; (b) a statistically significant increase of the incidence in females, both breeders and offspring, bearing malignant mammary tumors; (c) a statistically significant increase in the incidence of exocrine ademonas of the pancreas in both male and female breeders and offspring; and (d) an increased incidence, albeit not statistically significant, of pancreatic islet cell carcinomas in females, a malignant tumor which occurs very rarely in our historical controls. On the basis of the results of this study, excessive consumption of regular soft-drinks should be generally discouraged, in particular for children and adolescents.

  5. Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability.

    Directory of Open Access Journals (Sweden)

    Emily Peak

    2010-07-01

    Full Text Available Schistosomiasis, caused by infection with the blood fluke Schistosoma, is responsible for greater than 200,000 human deaths per annum. Objective high-throughput screens for detecting novel anti-schistosomal targets will drive 'genome to drug' lead translational science at an unprecedented rate. Current methods for detecting schistosome viability rely on qualitative microscopic criteria, which require an understanding of parasite morphology, and most importantly, must be subjectively interpreted. These limitations, in the current state of the art, have significantly impeded progress into whole schistosome screening for next generation chemotherapies.We present here a microtiter plate-based method for reproducibly detecting schistosomula viability that takes advantage of the differential uptake of fluorophores (propidium iodide and fluorescein diacetate by living organisms. We validate this high-throughput system in detecting schistosomula viability using auranofin (a known inhibitor of thioredoxin glutathione reductase, praziquantel and a range of small compounds with previously-described (gambogic acid, sodium salinomycin, ethinyl estradiol, fluoxetidine hydrochloride, miconazole nitrate, chlorpromazine hydrochloride, amphotericin b, niclosamide or suggested (bepridil, ciclopirox, rescinnamine, flucytosine, vinblastine and carbidopa anti-schistosomal activities. This developed method is sensitive (200 schistosomula/well can be assayed, relevant to industrial (384-well microtiter plate compatibility and academic (96-well microtiter plate compatibility settings, translatable to functional genomics screens and drug assays, does not require a priori knowledge of schistosome biology and is quantitative.The wide-scale application of this fluorescence-based bioassay will greatly accelerate the objective identification of novel therapeutic lead targets/compounds to combat schistosomiasis. Adapting this bioassay for use with other parasitic worm species

  6. Endocrine effects of contaminated sediments on the freshwater snail Potamopyrgus antipodarum in vivo and in the cell bioassays in vitro.

    Science.gov (United States)

    Mazurová, E; Hilscherová, K; Jálová, V; Köhler, H-R; Triebskorn, R; Giesy, J P; Bláha, L

    2008-09-17

    Lake Pilnok located in the black coal-mining region Ostrava-Karvina, Czech Republic, contains sediments highly contaminated with powdered waste coal. Moreover, population of the endangered species of narrow-clawed crayfish Pontastacus leptodactylus with high proportion of intersex individuals (18%) was observed at this site. These findings motivated our work that aimed to evaluate contamination, endocrine disruptive potency using in vitro assays and in vivo effects of contaminated sediments on reproduction of sediment-dwelling invertebrates. Chemical analyses revealed low concentrations of persistent chlorinated compounds and heavy metals but concentrations of polycyclic aromatic hydrocarbons (PAH) were high (sum of 16 PAHs 10 microg/g dw). Organic extracts from sediments caused significant in vitro AhR-mediated activity in the bioassay with H4IIE-luc cells, estrogenicity in MVLN cells and anti-androgenicity in recombinant yeast assay, and these effects could be attributed to non-persistent compounds derived from the waste coal. We have also observed significant in vivo effects of the sediments in laboratory experiments with the Prosobranchian euryhaline mud snail Potamopyrgus antipodarum. Sediments from Lake Pilnok as well as organic extracts of the sediments (externally added to the control sediment) significantly affected fecundity during 8 weeks of exposure. The effects were stimulations of fecundity at lower concentrations at the beginning of the experiment followed by inhibitions of fecundity and general toxicity. Our study indicates presence of chemicals that affected endocrine balance in invertebrates, and emphasizes the need for integrated approaches combining in vitro and in vivo bioassays with identification of chemicals to elucidate ecotoxicogical impacts of contaminated sediment samples.

  7. Relationships among exceedences of metals criteria, the results of ambient bioassays, and community metrics in mining-impacted streams.

    Science.gov (United States)

    Griffith, Michael B; Lazorchak, James M; Herlihy, Alan T

    2004-07-01

    If bioassessments are to help diagnose the specific environmental stressors affecting streams, a better understanding is needed of the relationships between community metrics and ambient criteria or ambient bioassays. However, this relationship is not simple, because metrics assess responses at the community level of biological organization, while ambient criteria and ambient bioassays assess or are based on responses at the individual level. For metals, the relationship is further complicated by the influence of other chemical variables, such as hardness, on their bioavailability and toxicity. In 1993 and 1994, U.S. Environmental Protection Agency (U.S. EPA) conducted a Regional Environmental Monitoring and Assessment Program (REMAP) survey on wadeable streams in Colorado's (USA) Southern Rockies Ecoregion. In this ecoregion, mining over the past century has resulted in metals contamination of streams. The surveys collected data on fish and macroinvertebrate assemblages, physical habitat, and sediment and water chemistry and toxicity. These data provide a framework for assessing diagnostic community metrics for specific environmental stressors. We characterized streams as metals-affected based on exceedence of hardness-adjusted criteria for cadmium, copper, lead, and zinc in water; on water toxicity tests (48-h Pimephales promelas and Ceriodaphnia dubia survival); on exceedence of sediment threshold effect levels (TELs); or on sediment toxicity tests (7-d Hyalella azteca survival and growth). Macroinvertebrate and fish metrics were compared among affected and unaffected sites to identify metrics sensitive to metals. Several macroinvertebrate metrics, particularly richness metrics, were less in affected streams, while other metrics were not. This is a function of the sensitivity of the individual metrics to metals effects. Fish metrics were less sensitive to metals because of the low diversity of fish in these streams.

  8. A new bioassay for the ecotoxicological testing of VOCs on groundwater invertebrates and the effects of toluene on Niphargus inopinatus

    Energy Technology Data Exchange (ETDEWEB)

    Avramov, Maria; Schmidt, Susanne I. [Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Groundwater Ecology, Ingolstädter Landstrasse 1, D-85764 Neuherberg (Germany); Griebler, Christian, E-mail: christian.griebler@helmholtz-muenchen.de [Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Groundwater Ecology, Ingolstädter Landstrasse 1, D-85764 Neuherberg (Germany)

    2013-04-15

    Highlights: ► A new bioassay for testing the toxicity of VOCs on groundwater fauna is presented. ► Results on the toxicity of toluene to Niphargus inopinatus are now available. ► Henry equilibrium needs to be considered when bioassays with VOCs are designed. ► Methodological aspects related to “difficult-to-test substances” are discussed. -- Abstract: A protocol was developed for testing the ecotoxicological effects of volatile organic compounds (VOCs) on groundwater invertebrates. Test substance volatility was addressed in a “closed from start to analysis”-design. Since manifestation of toxic effects may be delayed in ‘slower metabolizing’ organisms such as groundwater fauna, a time-independent (TI-) approach was adopted. Toluene was used as a model substance and its toxicity to the groundwater amphipod Niphargus inopinatus was assessed as an example. The method evaluation process considered various methodological issues such as partitioning of the toxicant between the water and the gas phase (Henry equilibrium), the possible depletion of oxygen in closed test vials, as well as microbial biodegradation of the test substance. For N. inopinatus, an LC{sub 50},{sub 14} {sub days} of 46.6 mg L{sup −1} toluene was obtained. The ultimate LC{sub 50} value was estimated at 23.3 mg L{sup −1} toluene. No oxygen depletion occurred in the test vials and Henry equilibrium was found to be established after 6 h. The new test system proposed now awaits broad practical application.

  9. The activity of the pyrrole insecticide chlorfenapyr in mosquito bioassay: towards a more rational testing and screening of non-neurotoxic insecticides for malaria vector control.

    Science.gov (United States)

    Oxborough, Richard M; N'Guessan, Raphael; Jones, Rebecca; Kitau, Jovin; Ngufor, Corine; Malone, David; Mosha, Franklin W; Rowland, Mark W

    2015-03-24

    The rapid selection of pyrethroid resistance throughout sub-Saharan Africa is a serious threat to malaria vector control. Chlorfenapyr is a pyrrole insecticide which shows no cross resistance to insecticide classes normally used for vector control and is effective on mosquito nets under experimental hut conditions. Unlike neurotoxic insecticides, chlorfenapyr owes its toxicity to disruption of metabolic pathways in mitochondria that enable cellular respiration. A series of experiments explored whether standard World Health Organization (WHO) guidelines for evaluation of long-lasting insecticidal nets, developed through testing of pyrethroid insecticides, are suitable for evaluation of non-neurotoxic insecticides. The efficacy of WHO recommended cone, cylinder and tunnel tests was compared for pyrethroids and chlorfenapyr. To establish bioassay exposure times predictive of insecticide-treated net (ITN) efficacy in experimental hut trials, standard three-minute bioassays of pyrethroid and chlorfenapyr ITNs were compared with longer exposures. Mosquito behaviour and response to chlorfenapyr ITN in bioassays conducted at night were compared to day and across a range of temperatures representative of highland and lowland transmission. Standard three-minute bioassay of chlorfenapyr produced extremely low levels of mortality compared to pyrethroids. Thirty-minute day-time bioassay produced mortality closer to hut efficacy of chlorfenapyr ITN but still fell short of the WHO threshold. Overnight tunnel test with chlorfenapyr produced 100% mortality and exceeded the WHO threshold of 80%. The endogenous circadian activity rhythm of anophelines results in inactivity by day and raised metabolism and flight activity by night. A model which explains improved toxicity of chlorfenapyr ITN when tested at night, and during the day at higher ambient temperature, is that activation of chlorfenapyr and disruption of respiratory pathways is enhanced when the insect is more metabolically

  10. Development of a bioassay to screen for chemicals mimicking the anti-aging effects of calorie restriction

    Energy Technology Data Exchange (ETDEWEB)

    Chiba, Takuya, E-mail: takuya@nagasaki-u.ac.jp [Department of Investigative Pathology, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523 (Japan); Tsuchiya, Tomoshi [Division of Surgical Oncology, Graduate School of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki 852-8501 (Japan); Komatsu, Toshimitsu; Mori, Ryoichi; Hayashi, Hiroko [Department of Investigative Pathology, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523 (Japan); Shimano, Hitoshi [Department of Internal Medicine (Endocrinology and Metabolism), Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba Ibaraki 305-8575 (Japan); Spindler, Stephen R. [Department of Biochemistry, Room 5478, Boyce Hall, University of California - Riverside, Riverside, CA 92521 (United States); Shimokawa, Isao [Department of Investigative Pathology, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523 (Japan)

    2010-10-15

    Research highlights: {yields} We identified four sequence motifs lying upstream of putative pro-longevity genes. {yields} One of these motifs binds to HNF-4{alpha}. {yields} HNF-4{alpha}/PGC-1{alpha} could up-regulate the transcription of a reporter gene linked to this motif. {yields} The reporter system described here could be used to screen candidate anti-aging molecules. -- Abstract: Suppression of the growth hormone/insulin-like growth factor-I pathway in Ames dwarf (DF) mice, and caloric restriction (CR) in normal mice extends lifespan and delays the onset of age-related disorders. In combination, these interventions have an additive effect on lifespan in Ames DF mice. Therefore, common signaling pathways regulated by DF and CR could have additive effects on longevity. In this study, we tried to identity the signaling mechanism and develop a system to assess pro-longevity status in cells and mice. We previously identified genes up-regulated in the liver of DF and CR mice by DNA microarray analysis. Motif analysis of the upstream sequences of those genes revealed four major consensus sequence motifs, which have been named dwarfism and calorie restriction-responsive elements (DFCR-REs). One of the synthesized sequences bound to hepatocyte nuclear factor-4{alpha} (HNF-4{alpha}), an important transcription factor involved in liver metabolism. Furthermore, using this sequence information, we developed a highly sensitive bioassay to identify chemicals mimicking the anti-aging effects of CR. When the reporter construct, containing an element upstream of a secreted alkaline phosphatase (SEAP) gene, was co-transfected with HNF-4{alpha} and its regulator peroxisome proliferator-activated receptor (PPAR) {gamma} coactivator-1{alpha} (PGC-1{alpha}), SEAP activity was increased compared with untransfected controls. Moreover, transient transgenic mice established using this construct showed increased SEAP activity in CR mice compared with ad libitum-fed mice. These data

  11. Rapid screening of aquatic toxicity of several metal-based nanoparticles using the MetPLATE™ bioassay

    International Nuclear Information System (INIS)

    Pokhrel, Lok R.; Silva, Thilini; Dubey, Brajesh; El Badawy, Amro M.; Tolaymat, Thabet M.; Scheuerman, Phillip R.

    2012-01-01

    Current understanding of potential toxicity of engineered nanomaterials to aquatic microorganisms is limited for risk assessment and management. Here we evaluate if the MetPLATE™ test can be used as an effective and rapid screening tool to test for potential aquatic toxicity of various metal-based nanoparticles (NPs). The MetPLATE bioassay is a heavy metal sensitive test based on β-galactosidase activity in Escherichia coli. Five different types of metal-based NPs were screened for toxicity: (1) citrate coated nAg (Citrate-nanosilver), (2) polyvinylpyrrolidone coated nAg (PVP-nAg), (3) uncoated nZnO, (4) uncoated nTiO 2 and (5) 1-Octadecylamine coated CdSe Quantum Dots (CdSe QDs); and compared with their corresponding ionic salt toxicity. Citrate-nAg was further fractionated into clean Citrate-nAg, unclean Citrate-nAg and permeate using a tangential flow filtration (TFF) system to eliminate residual ions and impurities from the stock Citrate-nAg suspension and also to differentiate between ionic- versus nano-specific toxicity. Our results showed that nAg, nZnO and CdSe QDs were less toxic than their corresponding ionic salts tested, while nano- or ionic form of TiO 2 was not toxic as high as 2.5 g L −1 to the MetPLATE™ bacteria. Although coating-dependent toxicity was noticeable between two types of Ag NPs evaluated, particle size and surface charge were not adequate to explain the observed toxicity; hence, the toxicity appeared to be material-specific. Overall, the toxicity followed the trend: CdCl 2 > AgNO 3 > PVP-nAg > unclean Citrate-nAg > clean Citrate-nAg > ZnSO 4 > nZnO > CdSe QDs > nTiO 2 /TiO 2 . These results indicate that an evaluation of β-galactosidase inhibition in MetPLATE™ E. coli can be an important consideration for rapid screening of metal-based NP toxicity, and should facilitate ecological risk assessment of these emerging contaminants. - Highlights: ► MetPLATE bioassay was evaluated as a rapid screening tool for nanotoxicity.

  12. Determination of bacteriocin activity with bioassays carried out on solid and liquid substrates: assessing the factor "indicator microorganism"

    Directory of Open Access Journals (Sweden)

    Ambrosiadis Ioannis

    2006-10-01

    Full Text Available Abstract Background Successful application of growth inhibition techniques for quantitative determination of bacteriocins relies on the sensitivity of the applied indicator microorganism to the bacteriocin to which is exposed. However, information on indicator microorganisms' performance and comparisons in bacteriocin determination with bioassays is almost non-existing in the literature. The aim of the present work was to evaluate the parameter "indicator microorganism" in bioassays carried out on solid -agar diffusion assay- and liquid -turbidometric assay- substrates, applied in the quantification of the most studied bacteriocin nisin. Results The performance of characterized microorganisms of known sources, belonging to the genera of Lactobacillus, Pediococcus, Micrococcus and Leuconostoc, has been assessed in this work in the assays of plate agar diffusion and turbidometry. Dose responses and sensitivities were examined and compared over a range of assay variables in standard bacteriocin solutions, fermentation broth filtrates and processed food samples. Measurements on inhibition zones produced on agar plates were made by means of digital image analysis. The data produced were analyzed statistically using the ANOVA technique and pairwise comparisons tests. Sensitivity limits and linearity of responses to bacteriocin varied significantly among different test-microorganisms in both applied methods, the lower sensitivity limits depending on both the test-microorganism and the applied method. In both methods, however, only two of the nine tested microorganisms (Lactobacillus curvatus ATCC 51436 and Pediococcus acidilactici ATCC 25740 were sensitive to very low concentrations of the bacteriocin and produced a linear-type of response in all kinds of samples used in this work. In all cases, very low bacteriocin concentrations, e.g. 1 IU/ml nisin, were more accurately determined in the turbidometric assay. Conclusion The present work shows that in

  13. Seasonal changes in food quantity and quality of the common North Sea copepods Temora longicornis and Pseudocalanus elongatus: a bioassay approach

    DEFF Research Database (Denmark)

    Koski, Marja; Dutz, Jörg; Klein Breteler, W.

    2010-01-01

    We evaluated the food quantity and quality over a seasonal cycle for the development and egg production of the common North Sea copepods Temora longicornis and Pseudocalanus elongatus, using a bioassay approach. Seston was sampled from December to October from a well-mixed water column of the Mar......We evaluated the food quantity and quality over a seasonal cycle for the development and egg production of the common North Sea copepods Temora longicornis and Pseudocalanus elongatus, using a bioassay approach. Seston was sampled from December to October from a well-mixed water column...... with the seston from the spring bloom in March-April. The juveniles of both species were able to complete their development only in spring experiments. A multiple regression analyses and comparison to a good-quality standard food of the same concentration suggested that, in an annual scale, the egg production...

  14. A novel bioassay using the barnacle Amphibalanus amphitrite to evaluate chronic effects of aluminium, gallium and molybdenum in tropical marine receiving environments.

    Science.gov (United States)

    van Dam, Joost W; Trenfield, Melanie A; Harries, Simon J; Streten, Claire; Harford, Andrew J; Parry, David; van Dam, Rick A

    2016-11-15

    A need exists for appropriate tools to evaluate risk and monitor potential effects of contaminants in tropical marine environments, as currently impact assessments are conducted by non-representative approaches. Here, a novel bioassay is presented that allows for the estimation of the chronic toxicity of contaminants in receiving tropical marine environments. The bioassay is conducted using planktonic larvae of the barnacle Amphibalanus amphitrite and is targeted at generating environmentally relevant, chronic toxicity data for water quality guideline derivation or compliance testing. The developmental endpoint demonstrated a consistently high control performance, validated through the use of copper as a reference toxicant. In addition, the biological effects of aluminium, gallium and molybdenum were assessed. The endpoint expressed high sensitivity to copper and moderate sensitivity to aluminium, whereas gallium and molybdenum exhibited no discernible effects, even at high concentrations, providing valuable information on the toxicity of these elements in tropical marine waters. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

  15. Comparative toxicity of eugenol and its quinone methide metabolite in cultured liver cells using kinetic fluorescence bioassays.

    Science.gov (United States)

    Thompson, D C; Barhoumi, R; Burghardt, R C

    1998-03-01

    Comparative kinetic analyses of the mechanisms of toxicity of the alkylphenol eugenol and its putative toxic metabolite (quinone methide, EQM) were carried out in cultured rat liver cells (Clone 9, ATCC) using a variety of vital fluorescence bioassays with a Meridian Ultima laser cytometer. Parameters monitored included intracellular GSH and calcium levels ([Ca2+]i), mitochondrial and plasma membrane potentials (MMP and PMP), intracellular pH, reactive oxygen species (ROS) generation, and gap junction-mediated intercellular communication (GJIC). Cells were exposed to various concentrations of test compounds (1 to 1000 microM) and all parameters monitored directly after addition at 15 s intervals for at least 10 min. Eugenol depleted intracellular GSH, inhibited GJIC and generation of ROS, and had a modest effect on MMP at concentrations of 10 to 100 microM. At high concentrations (1000 microM), eugenol also affected [Ca2+]i, PMP, and pH. Effects of EQM were seen at lower concentrations (1 to 10 microM). The earliest and most potent effects of either eugenol or EQM were seen on GSH levels and GJIC. Coadministration of glutathione ethyl ester enhanced intracellular GSH levels by almost 100% and completely protected cells from cell death caused by eugenol and EQM. These results suggest that eugenol mediates its hepatotoxic effects primarily through depletion of cytoprotective thiols and interference in thiol-dependent processes such as GJIC. Furthermore, our results support the hypothesis that the toxic effects of eugenol are mediated through its quinone methide metabolite.

  16. Inhibition of Oxidative Stress and Lipid Peroxidation by Anthocyanins from Defatted Canarium odontophyllum Pericarp and Peel Using In Vitro Bioassays

    Science.gov (United States)

    Khoo, Hock Eng; Azlan, Azrina; Ismail, Amin; Abas, Faridah; Hamid, Muhajir

    2014-01-01

    Canarium odontophyllum, also known as CO, is a highly nutritious fruit. Defatted parts of CO fruit are potent sources of nutraceutical. This study aimed to determine oxidative stress and lipid peroxidation effects of defatted CO pericarp and peel extracts using in vitro bioassays. Cell cytotoxic effect of the CO pericarp and peel extracts were also evaluated using HUVEC and Chang liver cell lines. The crude extracts of defatted CO peel and pericarp showed cytoprotective effects in t-BHP and 40% methanol-induced cell death. The crude extracts also showed no toxic effect to Chang liver cell line. Using CD36 ELISA, NAD+ and LDL inhibition assays, inhibition of oxidative stress were found higher in the crude extract of defatted CO peel compared to the pericarp extract. Hemoglobin and LDL oxidation assays revealed both crude extracts had significantly reduced lipid peroxidation as compared to control. TBARS values among defatted CO pericarp, peel, and cyanidin-3-glucoside showed no significant differences for hemoglobin and LDL oxidation assays. The protective effects of defatted CO parts, especially its peel is related to the presence of high anthocyanin that potentially offers as a pharmaceutical ingredient for cardioprotection. PMID:24416130

  17. Comparison of bioassays using the anostracan crustaceans Artemia salina and Thamnocephalus platyurus for plant extract toxicity screening

    Directory of Open Access Journals (Sweden)

    Pablo Mayorga

    2010-10-01

    Full Text Available Three lethality bioassays, using the salt-water crustacean Artemia salina Leach, Artemiidae, (conventional 96 microwell plate test and the Artoxkit M microbiotest and the freshwater crustacean Thamnocephalus platyurus Packard, Thamnocephalidae, (Thamnotoxkit F microbiotest, were compared using extracts of ten Guatemalan plant species. It was previously observed that five of them have anti-Artemia activity. These were: Solanum americanum Mill., Solanaceae, Gliricidia sepium (Jacq. Kunth ex Walp., Fabaceae, Neurolaena lobata (L. Cass., Asteraceae, Petiveria alliacea L., Phytolaccaceae, and Ocimum campechianum Mill., Lamiaceae. The five others: Curatella americana L., Dilleniaceae, Prunus barbata Koehne, Rosaceae, Quercus crispifolia Trel., Fagaceae, Rhizophora mangle L., Rhizophoraceae, and Smilax domingensis Willd., Smilacaceae, do not. All plants without anti-Artemia activity had no lethal effects in both assays with A. salina. For the plants with anti-Artemia activity the Artoxkit M was not sensitive to G. sepium and the conventional Artemia test was not sensitive to S. americanum, G. sepium and N. lobata. All the plant extracts, except for that of C. americana, had lethal effects on T. platyurus and the lethal median concentration (LC50 levels for this organism were in all cases substantially lower than those of the salt-water test species. This study revealed that T. platyurus is a promising test species worth further in depth investigation for toxicity screening of plant extracts with potential medicinal properties.

  18. Three-dimensional spheroid culture targeting versatile tissue bioassays using a PDMS-based hanging drop array.

    Science.gov (United States)

    Kuo, Ching-Te; Wang, Jong-Yueh; Lin, Yu-Fen; Wo, Andrew M; Chen, Benjamin P C; Lee, Hsinyu

    2017-06-29

    Biomaterial-based tissue culture platforms have emerged as useful tools to mimic in vivo physiological microenvironments in experimental cell biology and clinical studies. We describe herein a three-dimensional (3D) tissue culture platform using a polydimethylsiloxane (PDMS)-based hanging drop array (PDMS-HDA) methodology. Multicellular spheroids can be achieved within 24 h and further boosted by incorporating collagen fibrils in PDMS-HDA. In addition, the spheroids generated from different human tumor cells exhibited distinct sensitivities toward drug chemotherapeutic agents and radiation as compared with two-dimensional (2D) cultures that often lack in vivo-like biological insights. We also demonstrated that multicellular spheroids may enable key hallmarks of tissue-based bioassays, including drug screening, tumor dissemination, cell co-culture, and tumor invasion. Taken together, these results offer new opportunities not only to achieve the active control of 3D multicellular spheroids on demand, but also to establish a rapid and cost-effective platform to study anti-cancer therapeutics and tumor microenvironments.

  19. Bioassay-guided investigation of two Monarda essential oils as repellents of yellow fever mosquito Aedes aegypti.

    Science.gov (United States)

    Tabanca, Nurhayat; Bernier, Ulrich R; Ali, Abbas; Wang, Mei; Demirci, Betul; Blythe, Eugene K; Khan, Shabana I; Baser, K Husnu Can; Khan, Ikhlas A

    2013-09-11

    As part of an ongoing research program to identify active mosquito repellents, Monarda bradburiana Beck and Monarda fistulosa L. essential oils showed good repellent activity with minimum effective dosages (MED) of 0.055 ± 0.036 and 0.078 ± 0.027 mg/cm(2), respectively, compared to reference standard N,N-diethyl-3-methylbenzamide (DEET) (0.039 ± 0.014 mg/cm(2)). Systematic bioassay-guided fractionation of essential oils of both Monarda species was performed to identify the active repellent compounds, and isolated pure compounds were individually tested for repellency. Of the isolated compounds, carvacrol, thymol, eugenol, and carvacrol methyl ether were found to be the repellent compounds with MEDs in the range of 0.013-0.063 mg/cm(2). Active repellent compounds were also tested for larvicidal activity against 1-day-old Aedes aegypti larvae. Thymol was the best larvicide among the tested individual compounds (LD50 of 13.9 ppm). None of the individual compounds showed cytotoxicity against mammalian cells; however, the essential oils were toxic to all cell lines.

  20. Implementation of the NCRP wound model for interpretation of bioassay data for intake of radionuclides through contaminated wounds

    International Nuclear Information System (INIS)

    Ishigure, Nobuhito

    2009-01-01

    Emergency response preparedness for radiological accidents involving wound contamination has become more important, considering the current extending tendency in the nuclear industry related to the nuclear fuel cycle. The US National Council on Radiation Protection and Measurements (NCRP) proposed a biokinetic and dosimetric model for the intake of radionuclides through contaminated wounds in 2007. The present paper describes the implementation of this NCRP wound model for the prediction of systemic behaviour of some important radioactive elements encountered in workplaces related to the nuclear industry. The NCRP wound model was linked to the current ICRP systemic model at each blood compartment and simultaneous differential equations for the content of radioactivity in each compartment and excreta were solved with the Runge-Kutta method. The results of the calculation of wound, whole-body or specific organ retention and daily urinary or faecal excretion rate of some selected elements will be useful for the interpretation of bioassay data and dose assessment for cases of wound contamination. (author)

  1. Evaluation of the antiproliferative activity of the leaves from Arctium lappa by a bioassay-guided fractionation.

    Science.gov (United States)

    Machado, Fabio Bahls; Yamamoto, Rafael Eidi; Zanoli, Karine; Nocchi, Samara Requena; Novello, Cláudio Roberto; Schuquel, Ivânia Teresinha Albrecht; Sakuragui, Cássia Mônica; Luftmann, Heinrich; Ueda-Nakamura, Tânia; Nakamura, Celso Vataru; de Mello, João Carlos Palazzo

    2012-02-14

    Arctium lappa L. (Asteraceae) is used in folk medicine around the World, and shows several kinds of biological activity, particularly in vitro antitumor activity in different cell lines. This study evaluated the antiproliferative activity of the crude extract, semipurified fractions, and isolated compounds from the leaves of A. lappa, through bioassay-guided testing in Caco-2 cells. The crude extract was obtained with a 50% hydroethanolic extract and then partitioned with hexane, ethyl acetate, and n-butanol. The ethyl-acetate fraction (EAF) showed antiproliferative activity. This fraction was subjected to sequential column chromatography over silica gel to afford onopordopicrin (1), mixtures of 1 with dehydromelitensin-8-(4'-hydroxymethacrylate) (2), a mixture of 2 with dehydromelitensin (3), mixture of 1 with melitensin (4), dehydrovomifoliol (5), and loliolide (6). The compounds were identified by spectroscopic methods (NMR, MS) and comparison with literature data. This is the first description of compounds 2-5 from this species. The compounds tested in Caco-2 cells showed the following CC(50) (µg/mL) values: 1: 19.7 ± 3.4, 1 with 2: 24.6 ± 1.5, 2 with 3: 27 ± 11.7, 1 with 4: 42 ± 13.1, 6 30 ± 6.2; compound 5 showed no activity.

  2. Evaluation of the Antiproliferative Activity of the Leaves from Arctium lappa by a Bioassay-Guided Fractionation

    Directory of Open Access Journals (Sweden)

    Celso Vataru Nakamura

    2012-02-01

    Full Text Available Arctium lappa L. (Asteraceae is used in folk medicine around the World, and shows several kinds of biological activity, particularly in vitro antitumor activity in different cell lines. This study evaluated the antiproliferative activity of the crude extract, semipurified fractions, and isolated compounds from the leaves of A. lappa, through bioassay-guided testing in Caco-2 cells. The crude extract was obtained with a 50% hydroethanolic extract and then partitioned with hexane, ethyl acetate, and n-butanol. The ethyl-acetate fraction (EAF showed antiproliferative activity. This fraction was subjected to sequential column chromatography over silica gel to afford onopordopicrin (1, mixtures of 1 with dehydromelitensin-8-(4'-hydroxymethacrylate (2, a mixture of 2 with dehydromelitensin (3, mixture of 1 with melitensin (4, dehydrovomifoliol (5, and loliolide (6. The compounds were identified by spectroscopic methods (NMR, MS and comparison with literature data. This is the first description of compounds 2–5 from this species. The compounds tested in Caco-2 cells showed the following CC50 (µg/mL values: 1: 19.7 ± 3.4, 1 with 2: 24.6 ± 1.5, 2 with 3: 27 ± 11.7, 1 with 4: 42 ± 13.1, 6 30 ± 6.2; compound 5 showed no activity.

  3. Genotoxicity biomonitoring of sewage in two municipal wastewater treatment plants using the Tradescantia pallida var. purpurea bioassay.

    Science.gov (United States)

    Thewes, Márcia Regina; Junior, Delio Endres; Droste, Annette

    2011-10-01

    The genotoxicity of untreated and treated sewage from two municipal wastewater treatment plants (WTP BN and WTP SJN) in the municipality of Porto Alegre, in the southern Brazilian state of Rio Grande do Sul, was evaluated over a one-year period using the Tradescantia pallida var. purpurea (Trad-MCN) bioassay. Inflorescences of T. pallida var. purpurea were exposed to sewage samples in February (summer), April (autumn), July (winter) and October (spring) 2009, and the micronuclei (MCN) frequencies were estimated in each period. The high genotoxicity of untreated sewage from WTP BN in February and April was not observed in treated sewage, indicating the efficiency of treatment at this WTP. However, untreated and treated sewage samples from WTP SJN had high MCN frequencies, except in October, when rainfall may have been responsible for reducing these frequencies at both WTPs. Physicochemical analyses of sewage from both WTPs indicated elevated concentrations of organic matter that were higher at WTP SJN than at WTP BN. Chromium was detected in untreated and treated sewage from WTP SJN, but not in treated sewage from WTP BN. Lead was found in all untreated sewage samples from WTP SJN, but only in the summer and autumn at WTP BN. These results indicate that the short-term Trad-MCN genotoxicity assay may be useful for regular monitoring of municipal WTPs.

  4. Bioassay-guided fractionation of Melastoma malabathricum Linn. leaf solid phase extraction fraction and its anticoagulant activity.

    Science.gov (United States)

    Khoo, Li Teng; Abdullah, Janna Ong; Abas, Faridah; Tohit, Eusni Rahayu Mohd; Hamid, Muhajir

    2015-02-24

    The aims of this study were to examine the bioactive component(s) responsible for the anticoagulant activity of M. malabathricum Linn. leaf hot water crude extract via bioassay-guided fractionation and to evaluate the effect of bioactive component(s) on the intrinsic blood coagulation pathway. The active anticoagulant fraction of F3 was subjected to a series of chromatographic separation and spectroscopic analyses. Furthermore, the effect of the bioactive component(s) on the intrinsic blood coagulation pathway was studied through immediate and time incubation mixing studies. Through Activated Partial Thromboplastin Time (APTT) assay-guided fractionation, Subfraction B was considered the most potent anticoagulant fraction. Characterisation of Subfraction B indicated that anticoagulant activity could partly be due to the presence of cinnamic acid and a cinnamic acid derivative. APTT assays for both the immediate and time incubation mixing were corrected back into normal clotting time range (35.4-56.3 s). In conclusion, cinnamic acid and cinnamic acid derivative from Subfraction B were the first such compounds to be discovered from M. malabathricum Linn. leaf hot water crude extract that possess anticoagulant activity. This active anticoagulant Subfraction B prolonged blood clotting time by causing factor(s) deficiency in the intrinsic blood coagulation pathway.

  5. Development of a Simple and Effective Bioassay Method to Evaluate Resistance of Watermelon Plants to Fusarium oxysporum f. sp. niveum

    Directory of Open Access Journals (Sweden)

    Eun Ju Jo

    2017-06-01

    Full Text Available Root-dipping inoculation method has been widely used to determine the resistance of watermelon to Fusarium oxysporum f. sp. niveum causing Fusarium wilt. Although this method leads to the precise results of plant disease responses, more rapid and efficient assay methods have been still required because the root-dipping inoculation method is labor-intensive and time-consuming. In this study, we established a simple and effective bioassay method based on the comparison of various inoculation methods and growth conditions. To develop the system, the occurrence of Fusarium wilt on four resistant and susceptible cultivars was investigated by four different inoculation methods, root-dipping, scalpel, tip and soil-drenching methods. Of these inoculation methods, scalpel method resulted in clear plant disease resistance responses with the simplicity. With the use of scalpel method, we also explored the disease development of the cultivars depending on inoculum concentration, growth stage of seedlings, and incubation temperature after inoculation. Furthermore, we found that the resistance degrees of 23 cultivars derived by scalpel inoculation method were similar to the results by root-dipping method established previously.

  6. Bioassay responses and effects on benthos after pilot remediations in the delta of the rivers Rhine and Meuse

    International Nuclear Information System (INIS)

    Besten, Pieter J. den; Brink, Paul J. van den

    2005-01-01

    Chemical and biological monitoring was carried out for 5 years following pilot remediations at two locations in the Rhine-Meuse delta. The remediations consisted of partial excavation of the contaminated sediments, followed by applying a clean layer of sandy material on top. After the remediation, a new silty sediment top layer was formed exhibiting a lower toxicity in five sediment/sediment pore water bioassays. Compared to the unremediated sites, lower metal and PAH concentrations were found at the remediated sites, but in one location at the same time elevated HCH, PCB and HCB levels were recorded. One year after the remediation, the differences became smaller, although effects-based classification showed that the remediated site showed a higher quality up to the last year. In both remediated sites a rapid recolonization of nematodes, oligochaetes and chironomids was observed, while the recolonization by bivalves was slower. A few years after the remediation the differences decrease. - Capping contaminated sediments can be an effective remediation measure in two large river deltas

  7. Bioassay responses and effects on benthos after pilot remediations in the delta of the rivers Rhine and Meuse

    Energy Technology Data Exchange (ETDEWEB)

    Besten, Pieter J. den [Institute for Inland Water Management and Waste Water Treatment (RIZA), Ministry of Transport, Public Works and Water Management, PO Box 17, 8200 AA Lelystad (Netherlands)]. E-mail: p.dbesten@riza.rws.minvenw.nl; Brink, Paul J. van den [Alterra Green World Research, Wageningen University and Research Centre, PO Box 47, 6700 AA Wageningen (Netherlands)

    2005-07-15

    Chemical and biological monitoring was carried out for 5 years following pilot remediations at two locations in the Rhine-Meuse delta. The remediations consisted of partial excavation of the contaminated sediments, followed by applying a clean layer of sandy material on top. After the remediation, a new silty sediment top layer was formed exhibiting a lower toxicity in five sediment/sediment pore water bioassays. Compared to the unremediated sites, lower metal and PAH concentrations were found at the remediated sites, but in one location at the same time elevated HCH, PCB and HCB levels were recorded. One year after the remediation, the differences became smaller, although effects-based classification showed that the remediated site showed a higher quality up to the last year. In both remediated sites a rapid recolonization of nematodes, oligochaetes and chironomids was observed, while the recolonization by bivalves was slower. A few years after the remediation the differences decrease. - Capping contaminated sediments can be an effective remediation measure in two large river deltas.

  8. Detection of endocrine active substances in the aquatic environment in southern Taiwan using bioassays and LC-MS/MS.

    Science.gov (United States)

    Chen, Kuang-Yu; Chou, Pei-Hsin

    2016-06-01

    Endocrine active substances, including naturally occurring hormones and various synthetic chemicals have received much concern owing to their endocrine disrupting potencies. It is essential to monitor their environmental occurrence since these compounds may pose potential threats to biota and human health. In this study, yeast-based reporter assays were carried out to investigate the presence of (anti-)androgenic, (anti-)estrogenic, and (anti-)thyroid compounds in the aquatic environment in southern Taiwan. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was also used to measure the environmental concentrations of selected endocrine active substances for assessing potential ecological risks and characterizing contributions to the endocrine disrupting activities. Bioassay results showed that anti-androgenic (ND-7489 μg L(-1) flutamide equivalent), estrogenic (ND-347 ng L(-1) 17β-estradiol equivalent), and anti-thyroid activities were detected in the dissolved and particulate phases of river water samples, while anti-estrogenic activities (ND-10 μg L(-1) 4-hydroxytamoxifen equivalent) were less often found. LC-MS/MS analysis revealed that anti-androgenic and estrogenic contaminants, such as bisphenol A, triclosan, and estrone were frequently detected in Taiwanese rivers. In addition, their risk quotient values were often higher than 1, suggesting that they may pose an ecological risk to the aquatic biota. Further identification of unknown anti-androgenic and estrogenic contaminants in Taiwanese rivers may be necessary to protect Taiwan's aquatic environment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Use of cyanopropyl-bonded hplc column for bioassay-directed fractionation of organic extracts from incinerator emissions

    International Nuclear Information System (INIS)

    DeMarini, D.M.; Williams, R.W.; Brooks, L.R.; Taylor, M.S.

    1992-01-01

    The present study has shown that cyanopropyl-(CN) bonded silica HPLC columns are applicable for the fractionation of mass and mutagenic activity of organic extracts from some incinerator emissions. Dichloromethane-extractable organics from particles emitted by two different municipal waste incinerators and by a pilot-scale rotary kiln incinerator that was combusting polyethylene were fractionated by HPLC, and the mutagenicity of the fractions was determined by means of a microsuspension mutagenicity assay with Salmonella TA98. The CN-bonded silica columns provided high (80-100 percent) mass and mutagenicity recoveries for most emission extracts, and it fractionated the mutagenic activity. The results suggest that the emissions from municipal waste incinerators contain a high amount of direct-acting (-S9) mutagenic activity that is resolvable by HPLC using CN-bonded silica. Sub-fractionation of selected mutagenic HPLC fractions and subsequent analysis by gas chromatography/mass spectroscopy can be used to identify mutagenic species within complex incinerator emissions. The coupling of microsuspension bioassays to HPLC fractionation should be a useful tool for this type of analysis

  10. Genetic mouse embryo assay: improving performance and quality testing for assisted reproductive technology (ART) with a functional bioassay.

    Science.gov (United States)

    Gilbert, Rebecca S; Nunez, Brandy; Sakurai, Kumi; Fielder, Thomas; Ni, Hsiao-Tzu

    2016-03-24

    Growing concerns about safety of ART on human gametes, embryos, clinical outcomes and long-term health of offspring require improved methods of risk assessment to provide functionally relevant assays for quality control testing and pre-clinical studies prior to clinical implementation. The one-cell mouse embryo assay (MEA) is the most widely used for development and quality testing of human ART products; however, concerns exist due to the insensitivity/variability of this bioassay which lacks standardization and involves subjective analysis by morphology alone rather than functional analysis of the developing embryos. We hypothesized that improvements to MEA by the use of functional molecular biomarkers could enhance sensitivity and improve detection of suboptimal materials/conditions. Fresh one-cell transgenic mouse embryos with green fluorescent protein (GFP) expression driven by Pou6f1 or Cdx2 control elements were harvested and cultured to blastocysts in varied test and control conditions to compare assessment by standard morphology alone versus the added dynamic expression of GFP for screening and selection of critical raw materials and detection of suboptimal culture conditions. Transgenic mouse embryos expressing functionally relevant biomarkers of normal early embryo development can be used to monitor the developmental impact of culture conditions. This novel approach provides a superior MEA that is more meaningful and sensitive for detection of embryotoxicity than morphological assessment alone.

  11. Detection scheme for bioassays based on 2,6-pyridinedicarboxylic acid derivatives and enzyme-amplified lanthanide luminescence

    Energy Technology Data Exchange (ETDEWEB)

    Steinkamp, Tanja [Department of Chemical Analysis, MESA Institute for Nanotechnology, University of Twente, P.O. Box 217, 7500 AE Enschede (Netherlands); Karst, Uwe [Department of Chemical Analysis, MESA Institute for Nanotechnology, University of Twente, P.O. Box 217, 7500 AE Enschede (Netherlands)]. E-mail: u.karst@utwente.nl

    2004-11-15

    2,6-Pyridinedicarboxylic acid (PDC) and its derivatives are introduced as a new sensitizer system for enzyme-amplified lanthanide luminescence (EALL), a detection scheme for bioassays, which combines enzymatic amplification with time-resolved luminescence measurements of lanthanide chelates. Various PDC esters have been synthesized as esterase substrates that are cleaved to PDC in the presence of the enzyme. PDC forms luminescent complexes with Tb(III) or Eu(III), and the evaluation of the reaction is used for the selective and sensitive detection of esterases. For an esterase from hog liver a limit of detection of 10{sup -3} u/mL (equivalent to 10{sup -9} mol/L) and a limit of quantification of 3 x 10{sup -3} u/mL (equivalent to 3 x 10{sup -9} mol/L) could be achieved. As a second model reaction, xanthine oxidase (XOD) catalyzes the oxidation of 2,6-pyridinedicarboxaldehyde to PDC. Here, the limit of detection was 3 x 10{sup -3} u/mL and the limit of quantification 10{sup -2} u/mL for XOD from microorganisms. Major advantage of the tridentate PDC ligand is the possibility to perform all steps of the assay within or close to the physiological pH range, while the established EALL schemes based on bidentate salicylates or bisphenols have to be carried out at strongly alkaline pH to ensure sufficient complexation with the lanthanides.

  12. A Simple and Sensitive Plant-Based Western Corn Rootworm Bioassay Method for Resistance Determination and Event Selection.

    Science.gov (United States)

    Wen, Zhimou; Chen, Jeng Shong

    2018-05-26

    We report here a simple and sensitive plant-based western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), bioassay method that allows for examination of multiple parameters for both plants and insects in a single experimental setup within a short duration. For plants, injury to roots can be visually examined, fresh root weight can be measured, and expression of trait protein in plant roots can be analyzed. For insects, in addition to survival, larval growth and development can be evaluated in several aspects including body weight gain, body length, and head capsule width. We demonstrated using the method that eCry3.1Ab-expressing 5307 corn was very effective against western corn rootworm by eliciting high mortality and significantly inhibiting larval growth and development. We also validated that the method allowed determination of resistance in an eCry3.1Ab-resistant western corn rootworm strain. While data presented in this paper demonstrate the usefulness of the method for selection of events of protein traits and for determination of resistance in laboratory populations, we envision that the method can be applied in much broader applications.

  13. Radiomutagenesis and an in vitro conductimetric bioassay: it's application to avocado improvement for Phytophthora spp. resistance

    International Nuclear Information System (INIS)

    Álvarez, A.; Santiago, L.; Coto, O.; Machado, M.; Ramos, M.

    2009-01-01

    Avocado root-rot, caused by Phytophtora cinnamomi is among the main biotic stresses causing important economic losses to the crop. Therefore, development and identification of new genotypes resistant to the pathogen is important for an integrated disease management. However, limitations to avocado breeding and production are also related to the long juvenile period, large cultivation areas and extensive natural cross-pollination. In this sense, radiomutagenesis has proven to be a valuable tool to improve disease resistance in fruit trees. For this, the determination of mutagenic doses, combined with in vitro selection and conductimetric bioassays are necessary to accelerate the mutation breeding schemes. In avocado, the measurement of electrical conductivity, based on the release of microelectrolytes to the medium due to cell permeability damage, has been performed on inoculated roots and leaves. In the present work is showed a report of mutagenic doses against gamma rays (LD50 and LD20) determined for in vitro propagated zygotic embryos of three avocado cultivars. Additionally, electric conductivity was measured in leaf discs and root segments from zygotic embryos of Catalina cv. challenged with culture filtrates of isolates from different regions of the country. The results indicated a differential response of the genotype depending on the strain used. Also, differential response was observed, depending on tissue type

  14. Inhibition of oxidative stress and lipid peroxidation by anthocyanins from defatted Canarium odontophyllum pericarp and peel using in vitro bioassays.

    Directory of Open Access Journals (Sweden)

    Hock Eng Khoo

    Full Text Available Canarium odontophyllum, also known as CO, is a highly nutritious fruit. Defatted parts of CO fruit are potent sources of nutraceutical. This study aimed to determine oxidative stress and lipid peroxidation effects of defatted CO pericarp and peel extracts using in vitro bioassays. Cell cytotoxic effect of the CO pericarp and peel extracts were also evaluated using HUVEC and Chang liver cell lines. The crude extracts of defatted CO peel and pericarp showed cytoprotective effects in t-BHP and 40% methanol-induced cell death. The crude extracts also showed no toxic effect to Chang liver cell line. Using CD36 ELISA, NAD(+ and LDL inhibition assays, inhibition of oxidative stress were found higher in the crude extract of defatted CO peel compared to the pericarp extract. Hemoglobin and LDL oxidation assays revealed both crude extracts had significantly reduced lipid peroxidation as compared to control. TBARS values among defatted CO pericarp, peel, and cyanidin-3-glucoside showed no significant differences for hemoglobin and LDL oxidation assays. The protective effects of defatted CO parts, especially its peel is related to the presence of high anthocyanin that potentially offers as a pharmaceutical ingredient for cardioprotection.

  15. Gamma-interferon bioassay for detection of bovine tuberculosis in cattle: kinetics of production and dose response in whole blood culture

    International Nuclear Information System (INIS)

    Bhatia, Sandeep; Das, S.K.

    1999-01-01

    Stimulation with mycobacterium bovis PPD sensitised lymphocytes (whole blood or peripheral blood lymphocytes) results in release of gamma-interferon that can be detected by simple bioassay. The optimum concentration of bovine PPD was 20 μg ml and the optimum incubation period was 24 hr for maximum production of gamma-interferon in whole blood culture (128 units/ml) and peripheral blood culture (64 units/ml). (author)

  16. Summary report of bioassays for the city of Hollywood water plant membrane reject water as it mixed with WWTP effluent in an ocean outfall environment

    Energy Technology Data Exchange (ETDEWEB)

    Fergen, R.E.; Vinci, P.; Bloetscher, F.

    1999-07-01

    A special bioassay study was conducted to review the impact of the City of Hollywood's Membrane Softening Water Treatment Plant (WRP) reject water as it mixes with the City's Wastewater Treatment Plant (WWTP) effluent. Three sampling periods occurred during 1997. The purpose of this study was to determine potential toxicity of the WTP reject water, pre-chlorinated effluent, and combined effluent, and to demonstrate if the combined effluent was acceptable for ocean discharge on the basis of no potential toxicity. Effluent was acceptable for ocean discharge on the basis of no potential toxicity. Effluent samples were collected at six sampling points; three were in the plant, while the other three were along the outfall pipeline. Definitive, static renewal bioassay tests were performed using Mysidopsis bahia and Menidia beryllina as indicators of potential toxicity. The bioassay tests at 30% effluent concentration indicate that there is not potential toxicity for the pre-chlorinated WTP effluent, WTP reject water, dechlorinate combined effluent at the plant, and chlorinated combined effluent at Holland Park, the riser, and the terminus. The results indicate that the WTP reject water (100%) is not toxic to Menidia beryllina but was toxic to Mysidopsis bahia. When combined with the WWRP effluent, the reject water's impact on the potential toxicity of the commingled effluent was insignificant. All of the tests indicate the combined effluents are not toxic to the species tested at the 30% effluent level. Therefore, potential toxicity concerns were not demonstrated for this outfall discharge and did not prevent FDEP from issuing a permit to the City of Hollywood for the disposal of the combined effluent. Furthermore, these results, in combination with the previous results, indicated that individual bioassay testing for the reject water for regulatory compliance is not required.

  17. Potency determination of follitropin, lutropin And thyrotropin: a comparison between the quantification by reversed-phase high-performance liquid chromatography and in vivo bioassay

    International Nuclear Information System (INIS)

    Almeida, Beatriz Elane de

    2013-01-01

    With the intention of setting up physico-chemical methods as an alternative to in vivo bioassay for determining biological activity, the hFSH, hTSH and hLH content of native and recombinant preparations was determined by reversed-phase high-performance liquid chromatography (RP-HPLC) and compared with the data obtained by the classical mouse or rat in vivo bioassays (BA). A linear relationship between the two methods was found for these hormones: hFSH BA U= 0.9925 RP-HPLC U– 1.3165, r = 0.9371, p IU = 0.8771 RP-HPLC IU + 12.41, r = 0.9786, p < 0.01, n = 5. For nine other hFSH and eleven hTSH preparations, the mean difference ( ) between the bioactivity predicted from RP-HPLC data via these equations and the mean of the bioactivities obtained with the two methods was as follows. For hLH this difference could not be estimated due to lack of different samples. In the case of hFSH, ± SD = -2.11 ± 3.49% with a precision of 1.16% and in the case of hTSH, ± SD = -2.01 ± 5.56 %, with precision of 1.68%. Partly-degraded hFSH, hTSH and hLH samples presented different activity degrees that could be predicted by RP-HPLC, with an acceptable agreement with the in vivo bioassays. These results demonstrate that the employment of a non-animal physico-chemical assay, such as RP-HPLC, is a viable alternative to the use of an in vivo bioassay for hFSH and hTSH potency determination, thus reducing the number of animals currently used for assuring quality and efficacy of a pharmaceutical product. (author)

  18. Third-Generation Ah Receptor–Responsive Luciferase Reporter Plasmids: Amplification of Dioxin-Responsive Elements Dramatically Increases CALUX Bioassay Sensitivity and Responsiveness

    Science.gov (United States)

    He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S.; Zhao, Jing; Heath-Pagliuso, Sharon; Denison, Michael S.

    2011-01-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related dioxin-like chemicals are widespread and persistent environmental contaminants that produce diverse toxic and biological effects through their ability to bind to and activate the Ah receptor (AhR) and AhR-dependent gene expression. The chemically activated luciferase expression (CALUX) system is an AhR-responsive recombinant luciferase reporter gene–based cell bioassay that has been used in combination with chemical extraction and cleanup methods for the relatively rapid and inexpensive detection and relative quantitation of dioxin and dioxin-like chemicals in a wide variety of sample matrices. Although the CALUX bioassay has been validated and used extensively for screening purposes, it has some limitations when screening samples with very low levels of dioxin-like chemicals or when there is only a small amount of sample matrix for analysis. Here, we describe the development of third-generation (G3) CALUX plasmids with increased numbers of dioxin-responsive elements, and stable transfection of these new plasmids into mouse hepatoma (Hepa1c1c7) cells has produced novel amplified G3 CALUX cell bioassays that respond to TCDD with a dramatically increased magnitude of luciferase induction and significantly lower minimal detection limit than existing CALUX-type cell lines. The new G3 CALUX cell lines provide a highly responsive and sensitive bioassay system for the detection and relative quantitation of very low levels of dioxin-like chemicals in sample extracts. PMID:21775728

  19. Flower inhibition in Kalanchoe blossfeldiana. Bioassay of an endogenous long-day inhibitor and inhibition by (±) abscisic acid and xanthoxin.

    Science.gov (United States)

    Schwabe, W W

    1972-03-01

    The inhibition of flowering in Kalanchoe by crude sap expressed from leaves held in non-inductive long-day conditions is described, using a bioassay technique of leaf injection, which confirms the existence of a transferable inhibitor.This technique has also revealed that ± abscisic acid and Xanthoxin are inhibitory to flowering at 50 and 100 ppm respectively. The previously known inhibitory effects of gibberellic acid on flowering have also been confirmed.

  20. Microbial Detoxification of Deoxynivalenol (DON), Assessed via a Lemna minor L. Bioassay, through Biotransformation to 3-epi-DON and 3-epi-DOM-1.

    Science.gov (United States)

    Vanhoutte, Ilse; De Mets, Laura; De Boevre, Marthe; Uka, Valdet; Di Mavungu, José Diana; De Saeger, Sarah; De Gelder, Leen; Audenaert, Kris

    2017-02-13

    Mycotoxins are toxic metabolites produced by fungi. To mitigate mycotoxins in food or feed, biotransformation is an emerging technology in which microorganisms degrade toxins into non-toxic metabolites. To monitor deoxynivalenol (DON) biotransformation, analytical tools such as ELISA and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) are typically used. However, these techniques do not give a decisive answer about the remaining toxicity of possible biotransformation products. Hence, a bioassay using Lemna minor L. was developed. A dose-response analysis revealed significant inhibition in the growth of L. minor exposed to DON concentrations of 0.25 mg/L and higher. Concentrations above 1 mg/L were lethal for the plant. This bioassay is far more sensitive than previously described systems. The bioassay was implemented to screen microbial enrichment cultures, originating from rumen fluid, soil, digestate and activated sludge, on their biotransformation and detoxification capability of DON. The enrichment cultures originating from soil and activated sludge were capable of detoxifying and degrading 5 and 50 mg/L DON. In addition, the metabolites 3-epi-DON and the epimer of de-epoxy-DON (3-epi-DOM-1) were found as biotransformation products of both consortia. Our work provides a new valuable tool to screen microbial cultures for their detoxification capacity.

  1. Microbial Detoxification of Deoxynivalenol (DON, Assessed via a Lemna minor L. Bioassay, through Biotransformation to 3-epi-DON and 3-epi-DOM-1

    Directory of Open Access Journals (Sweden)

    Ilse Vanhoutte

    2017-02-01

    Full Text Available Mycotoxins are toxic metabolites produced by fungi. To mitigate mycotoxins in food or feed, biotransformation is an emerging technology in which microorganisms degrade toxins into non-toxic metabolites. To monitor deoxynivalenol (DON biotransformation, analytical tools such as ELISA and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS are typically used. However, these techniques do not give a decisive answer about the remaining toxicity of possible biotransformation products. Hence, a bioassay using Lemna minor L. was developed. A dose–response analysis revealed significant inhibition in the growth of L. minor exposed to DON concentrations of 0.25 mg/L and higher. Concentrations above 1 mg/L were lethal for the plant. This bioassay is far more sensitive than previously described systems. The bioassay was implemented to screen microbial enrichment cultures, originating from rumen fluid, soil, digestate and activated sludge, on their biotransformation and detoxification capability of DON. The enrichment cultures originating from soil and activated sludge were capable of detoxifying and degrading 5 and 50 mg/L DON. In addition, the metabolites 3-epi-DON and the epimer of de-epoxy-DON (3-epi-DOM-1 were found as biotransformation products of both consortia. Our work provides a new valuable tool to screen microbial cultures for their detoxification capacity.

  2. The nanotoxicology of a newly developed zero-valent iron nanomaterial for groundwater remediation and its remediation efficiency assessment combined with in vitro bioassays for detection of dioxin-like environmental pollutants

    OpenAIRE

    Schiwy, Andreas Herbert

    2016-01-01

    The assessment of chemicals and new compounds is an important task of ecotoxicology. In this thesis a newly developed zero-valent iron material for nanoremediation of groundwater contaminations was investigated and in vitro bioassays for high throughput screening were developed. These two elements of the thesis were combined to assess the remediation efficiency of the nanomaterial on the groundwater contaminant acridine. The developed in vitro bioassays were evaluated for quantification of th...

  3. Bioassay battery interlaboratory investigation of emerging contaminants in spiked water extracts - Towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring.

    Science.gov (United States)

    Di Paolo, Carolina; Ottermanns, Richard; Keiter, Steffen; Ait-Aissa, Selim; Bluhm, Kerstin; Brack, Werner; Breitholtz, Magnus; Buchinger, Sebastian; Carere, Mario; Chalon, Carole; Cousin, Xavier; Dulio, Valeria; Escher, Beate I; Hamers, Timo; Hilscherová, Klára; Jarque, Sergio; Jonas, Adam; Maillot-Marechal, Emmanuelle; Marneffe, Yves; Nguyen, Mai Thao; Pandard, Pascal; Schifferli, Andrea; Schulze, Tobias; Seidensticker, Sven; Seiler, Thomas-Benjamin; Tang, Janet; van der Oost, Ron; Vermeirssen, Etienne; Zounková, Radka; Zwart, Nick; Hollert, Henner

    2016-11-01

    Bioassays are particularly useful tools to link the chemical and ecological assessments in water quality monitoring. Different methods cover a broad range of toxicity mechanisms in diverse organisms, and account for risks posed by non-target compounds and mixtures. Many tests are already applied in chemical and waste assessments, and stakeholders from the science-police interface have recommended their integration in regulatory water quality monitoring. Still, there is a need to address bioassay suitability to evaluate water samples containing emerging pollutants, which are a current priority in water quality monitoring. The presented interlaboratory study (ILS) verified whether a battery of miniaturized bioassays, conducted in 11 different laboratories following their own protocols, would produce comparable results when applied to evaluate blinded samples consisting of a pristine water extract spiked with four emerging pollutants as single chemicals or mixtures, i.e. triclosan, acridine, 17α-ethinylestradiol (EE2) and 3-nitrobenzanthrone (3-NBA). Assays evaluated effects on aquatic organisms from three different trophic levels (algae, daphnids, zebrafish embryos) and mechanism-specific effects using in vitro estrogenicity (ER-Luc, YES) and mutagenicity (Ames fluctuation) assays. The test battery presented complementary sensitivity and specificity to evaluate the different blinded water extract spikes. Aquatic organisms differed in terms of sensitivity to triclosan (algae > daphnids > fish) and acridine (fish > daphnids > algae) spikes, confirming the complementary role of the three taxa for water quality assessment. Estrogenicity and mutagenicity assays identified with high precision the respective mechanism-specific effects of spikes even when non-specific toxicity occurred in mixture. For estrogenicity, although differences were observed between assays and models, EE2 spike relative induction EC 50 values were comparable to the literature, and E2/EE2

  4. Transcriptomic analyses in a benthic fish exposed to contaminated estuarine sediments through laboratory and in situ bioassays.

    Science.gov (United States)

    Costa, Pedro M; Miguel, Célia; Caeiro, Sandra; Lobo, Jorge; Martins, Marta; Ferreira, Ana M; Caetano, Miguel; Vale, Carlos; DelValls, T A; Costa, Maria H

    2011-11-01

    The transcription of contaminant response-related genes was investigated in juvenile Senegalese soles exposed to sediments from three distinct sites (a reference plus two contaminated) of a Portuguese estuary (the Sado, W Portugal) through simultaneous 28-day laboratory and in situ bioassays. Transcription of cytochrome P450 1A (CYP1A), metallothionein 1 (MT1), glutathione peroxidase (GPx), catalase (CAT), caspase 3 (CASP3) and 90 kDa heat-shock protein alpha (HSP90AA) was surveyed in the liver by real-time PCR. CASP3 transcription analysis was complemented by surveying apoptosis through the TUNEL reaction. After 14 days of exposure, relative transcription was either reduced or decreased in fish exposed to the contaminated sediments, revealing a disturbance stress phase during which animals failed to respond to insult. After 28 days of exposure all genes' transcription responded to contamination but laboratory and in situ assays depicted distinct patterns of regulation. Although sediments revealed a combination of organic and inorganic toxicants, transcription of the CYP1A gene was consistently correlated to organic contaminants. Metallothionein regulation was found correlated to metallic and organic xenobiotic contamination in the laboratory and in situ, respectively. The transcription of oxidative stress-related genes can be a good indicator of general stress but caution is mandatory when interpreting the results since regulation may be influenced by multiple factors. As for MT1, HSP90 up-regulation has potential to be a good indicator for total contamination, as well as the CASP3 gene, even though hepatocyte apoptosis depicted values inconsistent with sediment contamination, showing that programmed cell death did not directly depend on caspase transcription alone.

  5. Screening of hormone-like activities in bottled waters available in Southern Spain using receptor-specific bioassays.

    Science.gov (United States)

    Real, Macarena; Molina-Molina, José-Manuel; Jiménez-Díaz, Inmaculada; Arrebola, Juan Pedro; Sáenz, José-María; Fernández, Mariana F; Olea, Nicolás

    2015-01-01

    Bottled water consumption is a putative source of human exposure to endocrine-disrupting chemicals (EDCs). Research has been conducted on the presence of chemicals with estrogen-like activity in bottled waters and on their estrogenicity, but few data are available on the presence of hormonal activities associated with other nuclear receptors (NRs). The aim of this study was to determine the presence of endocrine activities dependent on the activation of human estrogen receptor alpha (hERa) and/or androgen receptor (hAR) in water in glass or plastic bottles sold to consumers in Southern Spain. Hormone-like activities were evaluated in 29 bottled waters using receptor-specific bioassays based on reporter gene expression in PALM cells [(anti-)androgenicity] and cell proliferation assessment in MCF-7 cells [(anti-)estrogenicity] after optimized solid phase extraction (SPE). All of the water samples analyzed showed hormonal activity. This was estrogenic in 79.3% and anti-estrogenic in 37.9% of samples and was androgenic in 27.5% and anti-androgenic in 41.3%, with mean concentrations per liter of 0.113pM 17β-estradiol (E2) equivalent units (E2Eq), 11.01pM anti-estrogen (ICI 182780) equivalent units (ICI 182780Eq), 0.33pM methyltrienolone (R1881) equivalent units (R1881Eq), and 0.18nM procymidone equivalent units (ProcEq). Bottled water consumption contributes to EDC exposure. Hormone-like activities observed in waters from both plastic and glass bottles suggest that plastic packaging is not the sole source of contamination and that the source of the water and bottling process may play a role, among other factors. Further research is warranted on the cumulative effects of long-term exposure to low doses of EDCs. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Estimating Prion Adsorption Capacity of Soil by BioAssay of Subtracted Infectivity from Complex Solutions (BASICS)

    Science.gov (United States)

    Wyckoff, A. Christy; Lockwood, Krista L.; Meyerett-Reid, Crystal; Michel, Brady A.; Bender, Heather; VerCauteren, Kurt C.; Zabel, Mark D.

    2013-01-01

    Prions, the infectious agent of scrapie, chronic wasting disease and other transmissible spongiform encephalopathies, are misfolded proteins that are highly stable and resistant to degradation. Prions are known to associate with clay and other soil components, enhancing their persistence and surprisingly, transmissibility. Currently, few detection and quantification methods exist for prions in soil, hindering an understanding of prion persistence and infectivity in the environment. Variability in apparent infectious titers of prions when bound to soil has complicated attempts to quantify the binding capacity of soil for prion infectivity. Here, we quantify the prion adsorption capacity of whole, sandy loam soil (SLS) typically found in CWD endemic areas in Colorado; and purified montmorillonite clay (Mte), previously shown to bind prions, by BioAssay of Subtracted Infectivity in Complex Solutions (BASICS). We incubated prion positive 10% brain homogenate from terminally sick mice infected with the Rocky Mountain Lab strain of mouse-adapted prions (RML) with 10% SLS or Mte. After 24 hours samples were centrifuged five minutes at 200×g and soil-free supernatant was intracerebrally inoculated into prion susceptible indicator mice. We used the number of days post inoculation to clinical disease to calculate the infectious titer remaining in the supernatant, which we subtracted from the starting titer to determine the infectious prion binding capacity of SLS and Mte. BASICS indicated SLS bound and removed ≥ 95% of infectivity. Mte bound and removed lethal doses (99.98%) of prions from inocula, effectively preventing disease in the mice. Our data reveal significant prion-binding capacity of soil and the utility of BASICS to estimate prion loads and investigate persistence and decomposition in the environment. Additionally, since Mte successfully rescued the mice from prion disease, Mte might be used for remediation and decontamination protocols. PMID:23484043

  7. Expression Patterns of Cancer Stem Cell Markers During Specific Celecoxib Therapy in Multistep Rat Colon Carcinogenesis Bioassays.

    Science.gov (United States)

    Salim, Elsayed I; Hegazi, Mona M; Kang, Jin Seok; Helmy, Hager M

    2016-01-01

    The purpose of this study was to investigate the role of colon cancer stem cells (CSCs) during chemicallyinduced rat multi-step colon carcinogenesis with or without the treatment with a specific cyclooxygenase-2 inhibitor drug (celecoxib). Two experiments were performed, the first, a short term 12 week colon carcinogenesis bioassay in which only surrogate markers for colon cancer, aberrant crypt foci (ACF) lesions, were formed. The other experiment was a medium term colon cancer rat assay in which tumors had developed after 32 weeks. Treatment with celecoxib lowered the numbers of ACF, as well as the tumor volumes and multiplicities after 32 weeks. Immunohistochemical proliferating cell nuclear antigen (PCNA) labeling indexes LI (%) were downregulated after treatment by celecoxib. Also different cell surface antigens known to associate with CSCs such as the epithelial cell adhesion molecule (EpCAM), CD44 and CD133 were compared between the two experiments and showed differential expression patterns depending on the stage of carcinogenesis and treatment with celecoxib. Flow cytometric analysis demonstrated that the numbers of CD133 cells were increased in the colonic epithelium after 12 weeks while those of CD44 but not CD133 cells were increased after 32 weeks. Moreover, aldehyde dehydrogenase-1 activity levels in the colonic epithelium (a known CSC marker) detected by ELISA assay were found down-regulated after 12 weeks, but were up-regulated after 32 weeks. The data have also shown that the protective effect of celecoxib on these specific markers and populations of CSCs and on other molecular processes such as apoptosis targeted by this drug may vary depending on the genetic and phenotypic stages of carcinogenesis. Therefore, uncovering these distinction roles of CSCs during different phases of carcinogenesis and during specific treatment could be useful for targeted therapy.

  8. Determination of beta-ray emitter concentrations in bioassay samples of the victims in JCO criticality accident

    International Nuclear Information System (INIS)

    Yukawa, Masae; Nishimura, Yoshikazu; Watanabe, Yoshito

    2001-01-01

    Concentrations of neutron-induced β-ray emitters in the hair, blood, urine and bone of three workers severely exposed to neutrons in JCO criticality accident were measured with a low background β-ray spectrometer (Pico β) and liquid scintillation counter for the purpose of neutron dose estimation. 32 P is generated by the fast neutron of 2.5 MeV and over in sulfur with (n, p) reaction. Since content of sulfur in hair is high as compared with the other human tissues, we tried to estimate fast neutron fluence to the body surface of the victims using concentrations of 32 P and stable sulfur in their hair. The result shows that two workers, who were exposed to the higher neutron radiation than another worker, received higher doses of irradiation to the frontal side of their trunks than to the heads. For a more detailed mapping of neutron fluence in the body, the measurements of 32 P and 45 Ca induced by (n, γ) reaction in bone were carried out. The results show that one worker (worker A) received a higher dose of neutrons at the frontal right side of the trunk, and that the dose decreased with the distance from the central part of the body. The other (worker B) seems to have gotten a higher dose of irradiation in the face, hands and waist. High amount of 32 P was detected in urine of the workers, and the concentration gradient among three workers showed a similar tendency to the estimated neutron dose from 24 Na in blood. Therefore, radioactivity of 32 P in urine could be used for estimating the neutron exposure dose. Moreover, the activity can be easily determined by scintillation counting, and urine is less invading bioassay sample that can be collected by non-medical stuffs. (author)

  9. Calibrating Self-Reported Measures of Maternal Smoking in Pregnancy via Bioassays Using a Monte Carlo Approach

    Directory of Open Access Journals (Sweden)

    Lauren S. Wakschlag

    2009-06-01

    Full Text Available Maternal smoking during pregnancy is a major public health problem that has been associated with numerous short- and long-term adverse health outcomes in offspring. However, characterizing smoking exposure during pregnancy precisely has been rather difficult: self-reported measures of smoking often suffer from recall bias, deliberate misreporting, and selective non-disclosure, while single bioassay measures of nicotine metabolites only reflect recent smoking history and cannot capture the fluctuating and complex patterns of varying exposure of the fetus. Recently, Dukic et al. [1] have proposed a statistical method for combining information from both sources in order to increase the precision of the exposure measurement and power to detect more subtle effects of smoking. In this paper, we extend the Dukic et al. [1] method to incorporate individual variation of the metabolic parameters (such as clearance rates into the calibration model of smoking exposure during pregnancy. We apply the new method to the Family Health and Development Project (FHDP, a small convenience sample of 96 predominantly working-class white pregnant women oversampled for smoking. We find that, on average, misreporters smoke 7.5 cigarettes more than what they report to smoke, with about one third underreporting by 1.5, one third under-reporting by about 6.5, and one third underreporting by 8.5 cigarettes. Partly due to the limited demographic heterogeneity in the FHDP sample, the results are similar to those obtained by the deterministic calibration model, whose adjustments were slightly lower (by 0.5 cigarettes on average. The new results are also, as expected, less sensitive to assumed values of cotinine half-life.

  10. Albino mutation rates in red mangroves (Rhizophora mangle L.) as a bioassay of contamination history in Tampa Bay, Florida, USA

    Science.gov (United States)

    Proffitt, C.E.; Travis, S.E.

    2005-01-01

    We assessed the sensitivity of a viviparous estuarine tree species, Rhizophora mangle, to historic sublethal mutagenic stress across a fine spatial scale by comparing the frequency of trees producing albino propagules in historically contaminated (n=4) and uncontaminated (n=11) forests in Tampa Bay, Florida, USA. Data from uncontaminated forests were used to provide estimates of background mutation rates. We also determined whether other fitness parameters were negatively correlated with mutagenic stress (e.g., degree of outcrossing and numbers of reproducing trees km-1). Contaminated sites in Tampa Bay had significantly higher frequencies of trees that were heterozygous for albinism per 1000 total reproducing trees (FHT) than uncontaminated forests (mean ?? SE: 11.4 ?? 4.3 vs 4.3 ?? 0.73, P 25 yrs of subsequent recruitment and tree replacement may have allowed an initial elevation in the FHT to decay. Patterns of FHT were not explained by distance from the bay mouth or the degree of urbanization. However, there was a significant positive relationship between tree size and FHT (r=0.83, P<0.018), which suggests that forests with older or larger trees provide a more lasting record of cumulative mutagenic stress. No other fitness parameters correlated with FHT. There was a difference in FHT between two latitudes, as determined by comparing Tampa Bay with literature values for Puerto Rico. The sensitivity of this bioassay for the effects of mutagens will facilitate future monitoring of contamination events and comparisons of bay-wide recovery in future decades. Development of a database of FHT values for a range of subtropical and tropical estuaries is underway that will provide a baseline against which to compare mutational consequences of global change. ?? 2005, The Society of Wetland Scientists.

  11. Solid-phase extraction clean-up of ciguatoxin-contaminated coral fish extracts for use in the mouse bioassay.

    Science.gov (United States)

    Wong, Chun Kwan; Hung, Patricia; Lee, Kellie L H; Kam, Kai Man

    2009-02-01

    Florisil solid-phase extraction (SPE) cartridges were used for purifying ciguatoxin (CTX)-contaminated coral fish extracts, with the aim of removing extracted lipid but retaining optimal level of CTXs in the purified fractions. The CTX-containing fraction (target fraction) in fish ether extract was isolated and purified by eluting through a commercially available Florisil cartridge with hexane-acetone-methanol solvent mixtures of increasing polarity (hexane-acetone (4:1, v/v) < acetone-methanol (7:3, v/v) < 100% methanol). Application of Florisil SPE using acetone-methanol (7:3, v/v) condition facilitated the separation of 4.2 +/- 0.4 mg (mean +/- standard error of the mean (SEM)) of purified target fraction from 20 mg ether extract with good retention of CTXs. The mouse bioassay was used to demonstrate that the average CTX recovery of the target fraction from CTX-spiked samples was 75.8% +/- 3.3%, which was significantly increased by 96.7% +/- 15% when compared with CTX recovery from ether extracts (44.8% +/- 5.2%) without performing SPE purification. Over 70% of non-target lipids were removed in which no CTX toxicity was found. Moreover, the target fractions of both CTX-spiked and naturally CTX-contaminated samples gave more prominent toxic responses of hypothermia and/or induced more rapid death of the mice. The use of acetone-methanol (7:3, v/v) condition in the elution could significantly improve overall recovery of CTXs, while minimizing the possible interferences of lipid matrix from co-extractants on mice.

  12. Primate polonium metabolic models and their use in estimation of systemic radiation doses from bioassay data. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Cohen, N. [New York Univ. Medical Center, Tuxedo, NY (United States). Dept. of Environmental Medicine

    1989-03-15

    A Polonium metabolic model was derived and incorporated into a Fortran algorithm which estimates the systemic radiation dose from {sup 210}Po when applied to occupational urine bioassay data. The significance of the doses estimated are examined by defining the degree of uncertainty attached to them through comprehensive statistical testing procedures. Many parameters necessary for dosimetry calculations (such as organ partition coefficients and excretion fractions), were evaluated from metabolic studies of {sup 210}Po in non-human primates. Two tamarins and six baboons were injected intravenously with {sup 210}Po citrate. Excreta and blood samples were collected. Five of the baboons were sacrificed at times ranging from 1 day to 3 months post exposure. Complete necropsies were performed and all excreta and the majority of all skeletal and tissue samples were analyzed radiochemically for their {sup 210}Po content. The {sup 210}Po excretion rate in the baboon was more rapid than in the tamarin. The biological half-time of {sup 210}Po excretion in the baboon was approximately 15 days while in the tamarin, the {sup 210}Po excretion rate was in close agreement with the 50 day biological half-time predicted by ICRP 30. Excretion fractions of {sup 210}Po in the non-human primates were found to be markedly different from data reported elsewhere in other species, including man. A thorough review of the Po urinalysis procedure showed that significant recovery losses resulted when metabolized {sup 210}Po was deposited out of raw urine. Polonium-210 was found throughout the soft tissues of the baboon but not with the partition coefficients for liver, kidneys, and spleen that are predicted by the ICRP 30 metabolic model. A fractional distribution of 0.29 for liver, 0.07 for kidneys, and 0.006 for spleen was determined. Retention times for {sup 210}Po in tissues are described by single exponential functions with biological half-times ranging from 15 to 50 days.

  13. Laboratory bioassays to estimate the lethal and sublethal effects of various insecticides and fungicides on Deraeocoris brevis (Hemiptera: Miridae).

    Science.gov (United States)

    Amarasekare, K G; Shearer, P W

    2013-04-01

    This laboratory bioassay focused on lethal and sublethal effects of five insecticides (chlorantraniliprole, cyantraniliprole, spinetoram, novaluron, and lambda-cyhalothrin) and two fungicide treatments (sulfur and a mixture of copper hydroxide and mancozeb) on the predatory mired bug, Deraeocoris brevis (Uhler) (Hemiptera: Miridae) using second instars and adult males and females. Formulated pesticides were tested using concentrations that were equivalent to the high label rate (1x) (high rate) and 1/10th of that amount (0.1x) (low rate) dissolved in 378.5 liters of water. Lambda-cyhalothrin was highly toxic to D. brevis nymphs and adults at both rates, whereas both rates of novaluron were highly toxic to nymphs. Cyantraniliprole, chlorantraniliprole, and novaluron were less toxic to adults, and chlorantraniliprole and spinetoram were less toxic to nymphs. Both rates of spinetoram caused significant mortality to adults. Fecundity of adult females was affected negatively by the high rates of either novaluron or spinetoram, whereas the fertility was affected only by the high rate of novaluron. The high rate of spinetoram reduced survival of nymphs. Adults treated with spinetoram had reduced longevity. Cyantraniliprole caused some mortality to nymphs and affected their survival. Both rates of sulfur were toxic to nymphs and affected emergence to adults. The mixture of copper hydroxide and mancozeb was less toxic to D. brevis. Neither adult longevity nor sex ratio was affected by the fungicides. The r values for D. brecis treated with lambda-cyhalothrin, novaluron, spinetoram, and sulfur were low, indicating that these products may have negative impact on population growth.

  14. Evaluation of the removal of antiestrogens and antiandrogens via ozone and granular activated carbon using bioassay and fluorescent spectroscopy.

    Science.gov (United States)

    Ma, Dehua; Chen, Lujun; Wu, Yuchao; Liu, Rui

    2016-06-01

    Antiestrogens and antiandrogens are relatively rarely studied endocrine disrupting chemicals which can be found in un/treated wastewaters. Antiestrogens and antiandrogens in the wastewater treatment effluents could contribute to sexual disruption of organisms. In this study, to assess the removal of non-specific antiestrogens and antiandrogens by advanced treatment processes, ozonation and adsorption to granular activated carbon (GAC), the biological activities and excitation emission matrix fluorescence spectroscopy of wastewater were evaluated. As the applied ozone dose increased to 12 mg/L, the antiestrogenic activity dramatically decreased to 3.2 μg 4-hydroxytamoxifen equivalent (4HEQ)/L, with a removal efficiency of 84.8%, while the antiandrogenic activity was 23.1 μg flutamide equivalent (FEQ)/L, with a removal efficiency of 75.5%. The removal of antiestrogenic/antiandrogenic activity has high correlation with the removal of fulvic acid-like materials and humic acid-like organics, suggesting that they can be used as surrogates for antiestrogenic/antiandrogenic activity during ozonation. The adsorption kinetics of antiestrogenic activity and antiandrogenic activity were well described by pseudo-second-order kinetics models. The estimated equilibrium concentration of antiestrogenic activity is 7.9 μg 4HEQ/L with an effective removal efficiency of 70.5%, while the equilibrium concentration of antiandrogenic activity is 33.7 μg FEQ/L with a removal efficiency of 67.0%. Biological activity evaluation of wastewater effluents is an attractive way to assess the removal of endocrine disrupting chemicals by different treatment processes. Fluorescence spectroscopy can be used as a surrogate measure of bioassays during ozonation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. BIOASSAY-GUIDED FRACTINATION OF ANTIMITOTIC COMPOUND FROM ONGKEA CORTEX (MEZZETTIA PARVIFLORA BECC TOWARDS SEA URCHIN EGGS

    Directory of Open Access Journals (Sweden)

    Muh. Akbar Bahar

    2015-06-01

    Full Text Available Ongkea cortex, the wood bark of Mezzettia Parviflora Becc, is a traditional medicine originated from Southeastern Sulawesi (Indonesia. It has been empirically known to have antitumor property. In this study, we examined the antiproliferative activity and obtained the antimitotic compound of ongkea cortex. Antimitotic activity was ultimately determined by the inhibition of cleavage-stage of newly fertilized sea urchin (Lytechinus variegatus eggs. A bioassay-guided fractination was performed in order to find the bioactive substance of ongkea cortex. The IC50 values of methanolic extract, ethyl acetate-soluble part of metanolic extract and ethil acetat insoluble part of metanolic extract were      1221.68 µg/mL,  2.69 µg/mL, and 15.15 µg/mL, respectively. Ethyl acetate-soluble part of metanolic extract was further investigated. It was partitionated using vacuum liquid column chromatoghraphy with different solvent system by increasing their polarities. There were three different fractions obtained. Fraction III exerted the highest inhibition activity with IC50 value of 1.33 µg/mL.  It was separated subsequently to result four groups of compounds. III-C group presented the most potent inhibition activity with IC50 value of  0.7147 µg/mL. It was then subjected to preparative TLC and yieldedsix groups of subfractions. III-C-3 subfraction was indicated as the most potent compound with IC50 value of 0.3378 µg/mL. It was ten times weaker compared with antimitotic activity of Vincristine with IC50 of 0.0351 µg/mL. As a conclusian, ongkea cortex might have antimitotic property with the highest rate inhibition activity exhibited by III-C-3 compound. Keywords: ongkea cortex, Mezzettia Parviflora Becc, sea urchin eggs, antimitotic compound, antiproliverative activity

  16. Temperature and food quantity effects on the harpacticoid copepod Nitocra spinipes: Combining in vivo bioassays with population modeling

    Science.gov (United States)

    Bui, Thuy T.; Lundström Belleza, Elin; Brinkmann, Markus; Hollert, Henner; Breitholtz, Magnus

    2017-01-01

    The harpacticoid copepod Nitocra spinipes has become a popular model species for toxicity testing over the past few decades. However, the combined influence of temperature and food shortage, two climate change-related stressors, has never be