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Sample records for bacteria lysis processes

  1. Process for the lysis of a culture of lactic acid bacteria by means of a lysin, and uses of the resulting lysed culture

    NARCIS (Netherlands)

    Buist, Girbe; Venema, Gerard; Kok, Jan; Ledeboer, Aat M.

    1995-01-01

    The invention provides a process for the lysis of a culture of lactic acid bacteria, or a product containing such culture e.g. cheese, by means of a lysin through the in situ production of a homologous autolysin, or a heterologous autolysin obtainable from Gram-positive bacteria esp. from lactic aci

  2. Study of the phage production efficiency in the bacteria lysis processes; Estudio del rendimiento en fagos para los procesos de lisis bacteriana

    Energy Technology Data Exchange (ETDEWEB)

    Vidania Munoz, R. de; Garces, F.; Davila, C. A.

    1979-07-01

    In this work we present a search for the best production conditions of {lambda}vir and{lambda} clear phages In E coli K12 and E coli C{sub 6}00 infected cells respectively. By keeping fixed some parameters of the process as the bacterial and phage generation times and the bacterial burst side, we have found that the lysis yield is strongly dependent on the multiplicity and in a lesser degree on the infection time. It appears from the experimental results that other variables are important, as infection efficiency and approach time from phages to bacteria. We will try to describe the lysis phenomenon by a numerical model on the bases of the se experimental results. (Author) 11 refs.

  3. Plasma nanotextured polymeric lab-on-a-chip for highly efficient bacteria capture and lysis.

    Science.gov (United States)

    Tsougeni, K; Papadakis, G; Gianneli, M; Grammoustianou, A; Constantoudis, V; Dupuy, B; Petrou, P S; Kakabakos, S E; Tserepi, A; Gizeli, E; Gogolides, E

    2016-01-07

    We describe the design, fabrication, and successful demonstration of a sample preparation module comprising bacteria cell capture and thermal lysis on-chip with potential applications in food sample pathogen analysis. Plasma nanotexturing of the polymeric substrate allows increase of the surface area of the chip and the antibody binding capacity. Three different anti-Salmonella antibodies were directly and covalently linked to plasma treated chips without any additional linker chemistry or other treatment. Then, the Ab-modified chips were tested for their capacity to bind bacteria in the concentration range of 10(2)-10(8) cells per mL; the module exhibited 100% efficiency in Salmonella enterica serovar Typhimurium bacteria capture for cell suspensions below 10(5) cells per mL (10(4) cells injected with a 100 μL sample volume) and efficiency higher than 50% for 10(7) cells per mL. Moreover, thermal lysis achieved on-chip from as low as 10 captured cells was demonstrated and shown to compare well with off-chip lysis. Excellent selectivity (over 1 : 300) was obtained in a sample containing, in addition to S. Typhimurium and E. coli bacteria.

  4. Critical cell wall hole size for lysis in Gram-positive bacteria

    Science.gov (United States)

    Mitchell, Gabriel; Wiesenfeld, Kurt; Nelson, Daniel; Weitz, Joshua

    2013-03-01

    Gram-positive bacteria transport molecules necessary for their survival through holes in their cell wall. The holes in cell walls need to be large enough to let critical nutrients pass through. However, the cell wall must also function to prevent the bacteria's membrane from protruding through a large hole into the environment and lysing the cell. As such, we hypothesize that there exists a range of cell wall hole sizes that allow for molecule transport but prevent membrane protrusion. Here we develop and analyze a biophysical theory of the response of a Gram-positive cell's membrane to the formation of a hole in the cell wall. We predict a critical hole size in the range 15-24nm beyond which lysis occurs. To test our theory, we measured hole sizes in Streptococcus pyogenes cells undergoing enzymatic lysis via transmission electron microscopy. The measured hole sizes are in strong agreement with our theoretical prediction. Together, the theory and experiments provide a means to quantify the mechanisms of death of Gram-positive cells via enzymatically mediated lysis and provides insight into the range of cell wall hole sizes compatible with bacterial homeostasis.

  5. Influence of a bacteriophage on the population dynamics of toxic dinoflagellates by lysis of algicidal bacteria.

    Science.gov (United States)

    Cai, Wenwei; Wang, Hui; Tian, Yun; Chen, Feng; Zheng, Tianling

    2011-11-01

    A lytic phage (øZCW1) was isolated from an algicidal bacterium Pseudoalteromonas sp. strain SP48 that specifically kills the toxic dinoflagellate Alexandrium tamarense. We demonstrated that øZCW1 could trigger the growth of A. tamarense by inhibiting the growth of algicidal bacterium SP48. In contrast, the growth of A. tamarense was suppressed when cocultured with either SP48 or the øZCW1-resistant mutant of SP48. This study provides the first evidence of the indirect impact of bacteriophage on bloom-forming microalgae via phage lysis of alga-killing bacteria.

  6. One-step cell lysis suitable for quantitative bacteria detection in inhibitor-laden sands

    Science.gov (United States)

    Lim, Hyun Jeong; Choi, Jung-Hyun; Son, Ahjeong

    2015-04-01

    Complexity and heterogeneity of soils often hinder effective DNA extraction from the soil matrix. In particular, conventional DNA extraction techniques require extensive purification which makes DNA extraction time-consuming and labor-intensive. Other drawbacks include lower recovery yield, degradation, and damage of DNA, which are also caused by intensive purifications during DNA extraction. Therefore a rapid and simple and yet effective DNA pretreatment method is preferred for environmental monitoring and screening. This study has evaluated the feasibility of simple physical pretreatment for effective cell lysis of bacteria in sands. Bead beating method was selected as an effective physical cell lysis method in this study. We examined the capability of this physical lysis for Pseudomonas putida seeded sands without additional chemical purification steps. The lysate from the method was analysed by the quantitative polymerase chain reaction (qPCR) assay and subsequently compared to that by commercial DNA extraction kit. The best lysis condition (treatment with 0.1 mm glass beads at 3000 rpm for 3 minutes) was selected. The qPCR results of bead beating treated samples showed the better performance than that of conventional DNA extraction kit. Moreover, the qPCR assay was performed to the sands laden with qPCR inhibitors (humic acids, clay, and magnesium), which generally present in environmental samples. Further experiments with the sands containing less than 10 μg/g of humic acids and 70% of clay showed successful quantification results of qPCR assay. In conclusion, the bead beating method is useful for simplified DNA extraction prior to qPCR analysis for sand samples of particular composition. It is expected that this approach will be beneficial for environmental in-situ analysis or immediate pre-screening. It also provides the groundwork for future studies with real soil samples that have various physico-chemical properties.

  7. Immobilized lysozyme for the continuous lysis of lactic bacteria in wine: Bench-scale fluidized-bed reactor study.

    Science.gov (United States)

    Cappannella, Elena; Benucci, Ilaria; Lombardelli, Claudio; Liburdi, Katia; Bavaro, Teodora; Esti, Marco

    2016-11-01

    Lysozyme from hen egg white (HEWL) was covalently immobilized on spherical supports based on microbial chitosan in order to develop a system for the continuous, efficient and food-grade enzymatic lysis of lactic bacteria (Oenococcus oeni) in white and red wine. The objective is to limit the sulfur dioxide dosage required to control malolactic fermentation, via a cell concentration typical during this process. The immobilization procedure was optimized in batch mode, evaluating the enzyme loading, the specific activity, and the kinetic parameters in model wine. Subsequently, a bench-scale fluidized-bed reactor was developed, applying the optimized process conditions. HEWL appeared more effective in the immobilized form than in the free one, when the reactor was applied in real white and red wine. This preliminary study suggests that covalent immobilization renders the enzyme less sensitive to the inhibitory effect of wine flavans.

  8. Comparison of point-of-care-compatible lysis methods for bacteria and viruses.

    Science.gov (United States)

    Heiniger, Erin K; Buser, Joshua R; Mireles, Lillian; Zhang, Xiaohong; Ladd, Paula D; Lutz, Barry R; Yager, Paul

    2016-09-01

    Nucleic acid sample preparation has been an especially challenging barrier to point-of-care nucleic acid amplification tests in low-resource settings. Here we provide a head-to-head comparison of methods for lysis of, and nucleic acid release from, several pathogenic bacteria and viruses-methods that are adaptable to point-of-care usage in low-resource settings. Digestion with achromopeptidase, a mixture of proteases and peptidoglycan-specific hydrolases, followed by thermal deactivation in a boiling water bath, effectively released amplifiable nucleic acid from Staphylococcus aureus, Bordetella pertussis, respiratory syncytial virus, and influenza virus. Achromopeptidase was functional after dehydration and reconstitution, even after eleven months of dry storage without refrigeration. Mechanical lysis methods proved to be effective against a hard-to-lyse Mycobacterium species, and a miniature bead-mill, the AudioLyse, is shown to be capable of releasing amplifiable DNA and RNA from this species. We conclude that point-of-care-compatible sample preparation methods for nucleic acid tests need not introduce amplification inhibitors, and can provide amplification-ready lysates from a wide range of bacterial and viral pathogens.

  9. STUDIES ON THE BACTERIOPHAGE OF D'HERELLE : VIII. THE MECHANISM OF LYSIS OF DEAD BACTERIA IN THE PRESENCE OF BACTERIOPHAGE.

    Science.gov (United States)

    Bronfenbrenner, J; Muckenfuss, R

    1927-04-30

    We have been able to confirm the observations of Twort as well as of Gratia, that dead staphylococcus may undergo lysis if, in addition to a suitable bacteriophage, there is also present live staphylococcus. Moreover, we have endeavored to ascertain the mechanism of this phenomenon and have found that in order to elicit it it is necessary to control the numbers of live and dead bacteria in the mixture. An excess of dead bacteria interferes with lysis by adsorbing the bacteriophage before it has the opportunity to initiate necessary changes in the live bacteria, so that all lysis is prevented. The phenomenon is specific, that is, the lysis of live bacteria is accompanied by lysis of dead bacteria of the same species only. Lysis of dead bacteria occurs best with staphylococcus, an organism which easily undergoes spontaneous autolysis under appropriate conditions. In the case of B. coli or B. dysenteriae the lysis of the dead bacteria is uncertain. Dead bacteria need not be present in the mixture at the beginning of the experiment; they will be dissolved if added any time before, during, or after the completion of lysis of live bacteria. If the test is performed so that a suitable semipermeable membrane is interposed between the dead and live bacteria, the dead bacteria are not dissolved, in spite of the lysis of live bacteria on the other side of the membrane. The agent determining the lysis of dead bacteria is not diffusible, while the principle initiating the lysis of live bacteria diffuses freely and is demonstrably present on both sides of the membrane. The complete independence of the agent causing dissolution of dead bacteria from bacteriophage can also be shown by separating the two agents by means of filtration, or by adsorption on bacteria. The ferment-like substance responsible for the lysis of dead bacteria is different from the bacteriophage. It is not diffusible through collodion, it is easily adsorbed on clay filters, it is heat-labile, and is

  10. A super-family of transcriptional activators regulates bacteriophage packaging and lysis in Gram-positive bacteria

    Science.gov (United States)

    Quiles-Puchalt, Nuria; Tormo-Más, María Ángeles; Campoy, Susana; Toledo-Arana, Alejandro; Monedero, Vicente; Lasa, Íñigo; Novick, Richard P.; Christie, Gail E.; Penadés, José R.

    2013-01-01

    The propagation of bacteriophages and other mobile genetic elements requires exploitation of the phage mechanisms involved in virion assembly and DNA packaging. Here, we identified and characterized four different families of phage-encoded proteins that function as activators required for transcription of the late operons (morphogenetic and lysis genes) in a large group of phages infecting Gram-positive bacteria. These regulators constitute a super-family of proteins, here named late transcriptional regulators (Ltr), which share common structural, biochemical and functional characteristics and are unique to this group of phages. They are all small basic proteins, encoded by genes present at the end of the early gene cluster in their respective phage genomes and expressed under cI repressor control. To control expression of the late operon, the Ltr proteins bind to a DNA repeat region situated upstream of the terS gene, activating its transcription. This involves the C-terminal part of the Ltr proteins, which control specificity for the DNA repeat region. Finally, we show that the Ltr proteins are the only phage-encoded proteins required for the activation of the packaging and lysis modules. In summary, we provide evidence that phage packaging and lysis is a conserved mechanism in Siphoviridae infecting a wide variety of Gram-positive bacteria. PMID:23771138

  11. Recyclable Photo-Thermal Nano-Aggregates of Magnetic Nanoparticle Conjugated Gold Nanorods for Effective Pathogenic Bacteria Lysis.

    Science.gov (United States)

    Ramasamy, Mohankandhasamy; Kim, Sanghyo; Lee, Su Seong; Yi, Dong Kee

    2016-01-01

    We describe the nucleophilic hybridization technique for fabricating magnetic nanoparticle (MNP) around gold nanorod (AuNR) for desired photo-thermal lysis on pathogenic bacteria. From the electromagnetic energy conversion into heat to the surrounding medium, a significant and quicker temperature rise was noted after light absorption on nanohybrids, at a controlled laser light output and optimum nanoparticle concentration. We observed a similar photo-thermal pattern for more than three times for the same material up on repeated magnetic separation. Regardless of the cell wall nature, superior pathogenic cell lysis has been observed for the bacteria suspensions of individual and mixed samples of Salmonella typhi (S.typhi) and Bacillus subtilis (B.subtilis) by the photo-heated nanoparticles. The synthesis of short gold nanorod, conjugation with magnetic nanoparticle and its subsequent laser exposure provides a rapid and reiterated photo-thermal effect with enhanced magnetic separation for efficient bactericidal application in water samples. Resultant novel properties of the nano-aggregates makes them a candidate to be used for a rapid, effective, and re-iterated photo-thermal agent against a wide variety of pathogens to attain microbe free water.

  12. Concerning the role of cell lysis-cryptic growth in anaerobic side-stream reactors: the single-cell analysis of viable, dead and lysed bacteria.

    Science.gov (United States)

    Foladori, P; Velho, V F; Costa, R H R; Bruni, L; Quaranta, A; Andreottola, G

    2015-05-01

    In the Anaerobic Side-Stream Reactor (ASSR), part of the return sludge undergoes alternating aerobic and anaerobic conditions with the aim of reducing sludge production. In this paper, viability, enzymatic activity, death and lysis of bacterial cells exposed to aerobic and anaerobic conditions for 16 d were investigated at single-cell level by flow cytometry, with the objective of contributing to the understanding of the mechanisms of sludge reduction in the ASSR systems. Results indicated that total and viable bacteria did not decrease during the anaerobic phase, indicating that anaerobiosis at ambient temperature does not produce a significant cell lysis. Bacteria decay and lysis occurred principally under aerobic conditions. The aerobic decay rate of total bacteria (bTB) was considered as the rate of generation of lysed bacteria. Values of bTB of 0.07-0.11 d(-1) were measured in anaerobic + aerobic sequence. The enzymatic activity was not particularly affected by the transition from anaerobiosis to aerobiosis. Large solubilisation of COD and NH4(+) was observed only under anaerobic conditions, as a consequence of hydrolysis of organic matter, but not due to cell lysis. The observations supported the proposal of two independent mechanisms contributing equally to sludge reduction: (1) under anaerobic conditions: sludge hydrolysis of non-bacterial material, (2) under aerobic conditions: bacterial cell lysis and oxidation of released biodegradable compounds.

  13. Citrullination in Rheumatoid Arthritis—A Process Promoted by Neutrophil Lysis?

    Science.gov (United States)

    Gazitt, Tal; Lood, Christian; Elkon, Keith B.

    2016-01-01

    Anti-citrullinated protein antibodies (ACPAs) are highly specific serologic markers for rheumatoid arthritis (RA) and can pre-date clinical disease onset by up to 10 years, also predicting erosive disease. The process of citrullination, the post-translational conversion of arginine to citrulline residues, is mediated by peptidylarginine deiminase (PAD) enzymes present in polymorphonuclear cells (PMNs). Calcium ions (Ca2+) are required for PAD activation, but the intracellular Ca2+ concentration in normal cells is much lower than the optimal Ca2+ concentration needed for PAD activation. For this reason, it has been proposed that PAD activation, and thus citrullination, occurs only during PMN cell death when PAD enzymes leak out of the cells into the extracellular matrix, or extracellular Ca2+ enters the cells, with the high Ca2+ concentration activating PAD. Recently, using artificial in vitro systems to corroborate their hypothesis, Romero et al. demonstrated that “hypercitrullination,” citrullination of multiple intracellular proteins, occurs within synovial fluid (SF) cells of RA patients, and that only modes of death leading to membranolysis such as perforin-granzyme pathway or complement membrane attack complex activation cause hypercitrullination. In order for Romero’s hypothesis to hold, it is reasonable to surmise that PMN-directed lysis should occur in the rheumatoid joint or the circulation of RA patients. Research conducted thus far has shown that immunoglobulin G (IgG) targeting PMNs are present in RA SF and mediate PMN activation. However, the role of anti-PMN IgG in mediating complement activation and subsequent PMN lysis and hypercitrullination has not been fully evaluated. PMID:27824546

  14. Citrullination in Rheumatoid Arthritis—A Process Promoted by Neutrophil Lysis?

    Directory of Open Access Journals (Sweden)

    Tal Gazitt

    2016-10-01

    Full Text Available Anti-citrullinated protein antibodies (ACPAs are highly specific serologic markers for rheumatoid arthritis (RA and can pre-date clinical disease onset by up to 10 years, also predicting erosive disease. The process of citrullination, the post-translational conversion of arginine to citrulline residues, is mediated by peptidylarginine deiminase (PAD enzymes present in polymorphonuclear cells (PMNs. Calcium ions (Ca2+ are required for PAD activation, but the intracellular Ca2+ concentration in normal cells is much lower than the optimal Ca2+ concentration needed for PAD activation. For this reason, it has been proposed that PAD activation, and thus citrullination, occurs only during PMN cell death when PAD enzymes leak out of the cells into the extracellular matrix, or extracellular Ca2+ enters the cells, with the high Ca2+ concentration activating PAD. Recently, using artificial in vitro systems to corroborate their hypothesis, Romero et al. demonstrated that “hypercitrullination,” citrullination of multiple intracellular proteins, occurs within synovial fluid (SF cells of RA patients, and that only modes of death leading to membranolysis such as perforin-granzyme pathway or complement membrane attack complex activation cause hypercitrullination. In order for Romero’s hypothesis to hold, it is reasonable to surmise that PMN-directed lysis should occur in the rheumatoid joint or the circulation of RA patients. Research conducted thus far has shown that immunoglobulin G (IgG targeting PMNs are present in RA SF and mediate PMN activation. However, the role of anti-PMN IgG in mediating complement activation and subsequent PMN lysis and hypercitrullination has not been fully evaluated.

  15. Artificial Intelligence vs. Statistical Modeling and Optimization of Continuous Bead Milling Process for Bacterial Cell Lysis.

    Science.gov (United States)

    Haque, Shafiul; Khan, Saif; Wahid, Mohd; Dar, Sajad A; Soni, Nipunjot; Mandal, Raju K; Singh, Vineeta; Tiwari, Dileep; Lohani, Mohtashim; Areeshi, Mohammed Y; Govender, Thavendran; Kruger, Hendrik G; Jawed, Arshad

    2016-01-01

    For a commercially viable recombinant intracellular protein production process, efficient cell lysis and protein release is a major bottleneck. The recovery of recombinant protein, cholesterol oxidase (COD) was studied in a continuous bead milling process. A full factorial response surface methodology (RSM) design was employed and compared to artificial neural networks coupled with genetic algorithm (ANN-GA). Significant process variables, cell slurry feed rate (A), bead load (B), cell load (C), and run time (D), were investigated and optimized for maximizing COD recovery. RSM predicted an optimum of feed rate of 310.73 mL/h, bead loading of 79.9% (v/v), cell loading OD600nm of 74, and run time of 29.9 min with a recovery of ~3.2 g/L. ANN-GA predicted a maximum COD recovery of ~3.5 g/L at an optimum feed rate (mL/h): 258.08, bead loading (%, v/v): 80%, cell loading (OD600nm): 73.99, and run time of 32 min. An overall 3.7-fold increase in productivity is obtained when compared to a batch process. Optimization and comparison of statistical vs. artificial intelligence techniques in continuous bead milling process has been attempted for the very first time in our study. We were able to successfully represent the complex non-linear multivariable dependence of enzyme recovery on bead milling parameters. The quadratic second order response functions are not flexible enough to represent such complex non-linear dependence. ANN being a summation function of multiple layers are capable to represent complex non-linear dependence of variables in this case; enzyme recovery as a function of bead milling parameters. Since GA can even optimize discontinuous functions present study cites a perfect example of using machine learning (ANN) in combination with evolutionary optimization (GA) for representing undefined biological functions which is the case for common industrial processes involving biological moieties.

  16. Palytoxin induces cell lysis by priming a two-step process in mcf-7 cells.

    Science.gov (United States)

    Prandi, Simone; Sala, Gian Luca; Bellocci, Mirella; Alessandrini, Andrea; Facci, Paolo; Bigiani, Albertino; Rossini, Gian Paolo

    2011-08-15

    The cytolytic action of palytoxin (PlTX) was recognized long ago, but its features have remained largely undetermined. We used biochemical, morphological, physiological, and physical tools, to study the cytolytic response in MCF-7 cells, as our model system. Cytolysis represented a stereotyped response induced by the addition of isotonic phosphate buffer (PBS) to cells that had been exposed to PlTX, after toxin removal and under optimal and suboptimal experimental conditions. Cytolysis was sensitive to osmolytes present during cell exposure to PlTX but not in the course of the lytic phase. Fluorescence microscopy showed that PlTX caused cell rounding and rearrangement of the actin cytoskeleton. Atomic force microscopy (AFM) was used to monitor PlTX effects in real time, and we found that morphological and mechanical properties of MCF-7 cells did not change during toxin exposure, but increased cell height and decreased stiffness at its surface were observed when PBS was added to PlTX-treated cells. The presence of an osmolyte during PlTX treatment prevented the detection of changes in morphological and mechanical properties caused by PBS addition to toxin-treated cells, as detected by AFM. By patch-clamp technique, we confirmed that PlTX action involved the transformation of the Na(+),K(+)-ATPase into a channel and found that cell membrane capacitance was not changed by PlTX, indicating that the membrane surface area was not greatly affected in our model system. Overall, our findings show that the cytolytic response triggered by PlTX in MCF-7 cells includes a first phase, which is toxin-dependent and osmolyte-sensitive, priming cells to lytic events taking place in a separate phase, which does not require the presence of the toxin and is osmolyte-insensitive but is accompanied by marked reorganization of actin-based cytoskeleton and altered mechanical properties at the cell's surface. A model of the two-step process of PlTX-induced cytolysis is presented.

  17. Learning from bacteria about natural information processing.

    Science.gov (United States)

    Ben-Jacob, Eshel

    2009-10-01

    Under natural growth conditions, bacteria live in complex hierarchical communities. To conduct complex cooperative behaviors, bacteria utilize sophisticated communication to the extent that their chemical language includes semantic and even pragmatic aspects. I describe how complex colony forms (patterns) emerge through the communication-based interplay between individual bacteria and the colony. Individual cells assume newly co-generated traits and abilities that are not prestored in the genetic information of the cells, that is, not all the information required for efficient responses to all environmental conditions is stored. To solve newly encountered problems, they assess the problem via collective sensing, recall stored information of past experience, and then execute distributed information processing of the 10(9)-10(12) bacteria in the colony--transforming the colony into a "super-brain." I show illuminating examples of swarming intelligence of live bacteria in which they solve optimization problems that are beyond what human beings can solve. This will lead to a discussion about the special nature of bacterial computational principles compared to Turing algorithm computational principles, in particular about the role of distributed information processing.

  18. Same day identification and full panel antimicrobial susceptibility testing of bacteria from positive blood culture bottles made possible by a combined lysis-filtration method with MALDI-TOF VITEK mass spectrometry and the VITEK2 system.

    Science.gov (United States)

    Machen, Alexandra; Drake, Tim; Wang, Yun F Wayne

    2014-01-01

    Rapid identification and antimicrobial susceptibility testing of microorganisms causing bloodstream infections or sepsis have the potential to improve patient care. This proof-of-principle study evaluates the Lysis-Filtration Method for identification as well as antimicrobial susceptibility testing of bacteria directly from positive blood culture bottles in a clinical setting. A total of 100 non-duplicated positive blood cultures were tested and 1012 microorganism-antimicrobial combinations were assessed. An aliquot of non-charcoal blood culture broth was incubated with lysis buffer briefly before being filtered and washed. Microorganisms recovered from the filter membrane were first identified by using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight VITEK® Mass Spectrometry (VITEK MS). After quick identification from VITEK MS, filtered microorganisms were inoculated to VITEK®2 system for full panel antimicrobial susceptibility testing analysis. Of 100 bottles tested, the VITEK MS resulted in 94.0% correct organism identification to the species level. Compared to the conventional antimicrobial susceptibility testing methods, direct antimicrobial susceptibility testing from VITEK®2 resulted in 93.5% (946/1012) category agreement of antimicrobials tested, with 3.6% (36/1012) minor error, 1.7% (7/1012) major error, and 1.3% (13/1012) very major error of antimicrobials. The average time to identification and antimicrobial susceptibility testing was 11.4 hours by using the Lysis-Filtration method for both VITEK MS and VITEK®2 compared to 56.3 hours by using conventional methods (pblood culture can be achieved and can be used for appropriate antibiotic therapy and antibiotic stewardship.

  19. Same day identification and full panel antimicrobial susceptibility testing of bacteria from positive blood culture bottles made possible by a combined lysis-filtration method with MALDI-TOF VITEK mass spectrometry and the VITEK2 system.

    Directory of Open Access Journals (Sweden)

    Alexandra Machen

    Full Text Available Rapid identification and antimicrobial susceptibility testing of microorganisms causing bloodstream infections or sepsis have the potential to improve patient care. This proof-of-principle study evaluates the Lysis-Filtration Method for identification as well as antimicrobial susceptibility testing of bacteria directly from positive blood culture bottles in a clinical setting. A total of 100 non-duplicated positive blood cultures were tested and 1012 microorganism-antimicrobial combinations were assessed. An aliquot of non-charcoal blood culture broth was incubated with lysis buffer briefly before being filtered and washed. Microorganisms recovered from the filter membrane were first identified by using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight VITEK® Mass Spectrometry (VITEK MS. After quick identification from VITEK MS, filtered microorganisms were inoculated to VITEK®2 system for full panel antimicrobial susceptibility testing analysis. Of 100 bottles tested, the VITEK MS resulted in 94.0% correct organism identification to the species level. Compared to the conventional antimicrobial susceptibility testing methods, direct antimicrobial susceptibility testing from VITEK®2 resulted in 93.5% (946/1012 category agreement of antimicrobials tested, with 3.6% (36/1012 minor error, 1.7% (7/1012 major error, and 1.3% (13/1012 very major error of antimicrobials. The average time to identification and antimicrobial susceptibility testing was 11.4 hours by using the Lysis-Filtration method for both VITEK MS and VITEK®2 compared to 56.3 hours by using conventional methods (p<0.00001. Thus, the same-day results of microorganism identification and antimicrobial susceptibility testing directly from positive blood culture can be achieved and can be used for appropriate antibiotic therapy and antibiotic stewardship.

  20. Maintenance, endogeneous, respiration, lysis, decay and predation

    DEFF Research Database (Denmark)

    loosdrecht, Marc C. M. Van; Henze, Mogens

    1999-01-01

    In activated sludge processes an increased sludge age is associated with a decreased sludge production. This phenomenon is generally interpreted as a result of endogenous respiration processes. In the activated sludge models cell lysis (or decay) is incorporated. The lysis is modelled such that i......In activated sludge processes an increased sludge age is associated with a decreased sludge production. This phenomenon is generally interpreted as a result of endogenous respiration processes. In the activated sludge models cell lysis (or decay) is incorporated. The lysis is modelled...... mechanism is microbiologically correct. The lysis/decay model mechanism is a strongly simplified representation of reality. This paper tries to review the processes grouped under endogenous respiration in activated sludge models. Mechanisms and processes such as maintenance, lysis, internal and external...... and maintenance processes. This conversion will in general be denoted as endogenous respiration. Based on the literature review the phenomena are discussed and organised, in order to create a working platform for discussing more detailed activated sludge models, one of which is being sketched. (C) 1999 IAWQ...

  1. Factors influencing lysis time stochasticity in bacteriophage λ

    Directory of Open Access Journals (Sweden)

    Dennehy John J

    2011-08-01

    Full Text Available Abstract Background Despite identical genotypes and seemingly uniform environments, stochastic gene expression and other dynamic intracellular processes can produce considerable phenotypic diversity within clonal microbes. One trait that provides a good model to explore the molecular basis of stochastic variation is the timing of host lysis by bacteriophage (phage. Results Individual lysis events of thermally-inducible λ lysogens were observed using a temperature-controlled perfusion chamber mounted on an inverted microscope. Both mean lysis time (MLT and its associated standard deviation (SD were estimated. Using the SD as a measure of lysis time stochasticity, we showed that lysogenic cells in controlled environments varied widely in lysis times, and that the level of lysis time stochasticity depended on allelic variation in the holin sequence, late promoter (pR' activity, and host growth rate. In general, the MLT was positively correlated with the SD. Both lower pR' activities and lower host growth rates resulted in larger SDs. Results from premature lysis, induced by adding KCN at different time points after lysogen induction, showed a negative correlation between the timing of KCN addition and lysis time stochasticity. Conclusions Taken together with results published by others, we conclude that a large fraction of λ lysis time stochasticity is the result of random events following the expression and diffusion of the holin protein. Consequently, factors influencing the timing of reaching critical holin concentrations in the cell membrane, such as holin production rate, strongly influence the mean lysis time and the lysis time stochasticity.

  2. Rapid identification of bacteria and yeasts from positive-blood-culture bottles by using a lysis-filtration method and matrix-assisted laser desorption ionization-time of flight mass spectrum analysis with the SARAMIS database.

    Science.gov (United States)

    Fothergill, Amy; Kasinathan, Vyjayanti; Hyman, Jay; Walsh, John; Drake, Tim; Wang, Yun F Wayne

    2013-03-01

    Rapid identification of microorganisms causing bloodstream infections directly from a positive blood culture would decrease the time to directed antimicrobial therapy and greatly improve patient care. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a fast and reliable method for identifying microorganisms from positive culture. This study evaluates the performance of a novel filtration-based method for processing positive-blood-culture broth for immediate identification of microorganisms by MALDI-TOF with a Vitek MS research-use-only system (VMS). BacT/Alert non-charcoal-based blood culture bottles that were flagged positive by the BacT/Alert 3D system were included. An aliquot of positive-blood-culture broth was incubated with lysis buffer for 2 to 4 min at room temperature, the resulting lysate was filtered through a membrane, and harvested microorganisms were identified by VMS. Of the 259 bottles included in the study, VMS identified the organisms in 189 (73%) cultures to the species level and 51 (19.7%) gave no identification (ID), while 6 (2.3%) gave identifications that were considered incorrect. Among 131 monomicrobic isolates from positive-blood-culture bottles with one spot having a score of 99.9%, the IDs for 131 (100%) were correct to the species level. In 202 bottles where VMS was able to generate an ID, the IDs for 189 (93.6%) were correct to the species level, whereas the IDs provided for 7 isolates (3.5%) were incorrect. In conclusion, this method does not require centrifugation and produces a clean spectrum for VMS analysis in less than 15 min. This study demonstrates the effectiveness of the new lysis-filtration method for identifying microorganisms directly from positive-blood-culture bottles in a clinical setting.

  3. Phytoplankton, Bacteria and Heterotrophic Nanoflagellate studies using ship and OCM-2 data along a coastal transect in the Arabian Sea

    Digital Repository Service at National Institute of Oceanography (India)

    Matondkar, S.G.P.; Basu, S.; Parab, S.G.; Pednekar, S.; Niyati Hede, N.; Dwivedi, R.M.; Raman, M.; Babu, K.N.; Shukla, A.K.; Shirdhankar, M.M.

    ). This relationship develops primarily when bacteria assimilate dissolved organic matter (DOM) generated from phytoplankton via processes extracellular exudation like rupture and degradation of phytoplankton cells during grazing, viral lysis and/or direct...

  4. Optimization of cell lysis conditions of soybean dregs in the process of DNA extraction by response surface methodology%响应面法优化豆渣DNA提取中的细胞裂解工艺

    Institute of Scientific and Technical Information of China (English)

    柴菜建; 李娜; 江秀霞; 樊星; 邵美丽

    2012-01-01

    采用响应面法对豆渣DNA提取中的细胞裂解工艺进行优化。在单因素实验基础上.选取SDS(十二烷基硫酸钠)浓度、裂解液pH、裂解温度为影响因子,利用Box-Behnken中心组合进行三因素三水平的实验设计,以DNA提取率为响应值,进行响应面分析。结果表明,裂解时间70min时,豆渣细胞裂解的最优条件为:SDS浓度1.84%、裂解液pH5.95、裂解温度81℃。在此条件下,DNA提取率的理论值为0.63%,实测值为0.625%。%Response surface methodology (RSM) was used for optimizing the cell lysis conditions of soybean dregs in the process of DNA extraction. Based on single-factor experiment,three independent variables, namely the concentration of Sodium dodecy sulfate(SDS),pH and lysis temperature,were selected as affecting factors during lysis. The experiment was arranged according to Box-Behnken central composite design. The results showed that the optimum lysis conditions for soybean dregs were as follows.concentration of SDS was 1.84% ,pH was 5.95,1ysis temperature was 81℃ when lysis time was 70min. The predicted DNA extraction yield was 0.63% ,while the actual extraction yield was 0.625%.

  5. Miniaturized Cell Lysis Device Using Spherically Focused Ultrasound

    Institute of Scientific and Technical Information of China (English)

    李刚; 肖宏; 郭旻; 程京

    2003-01-01

    A prototype of a miniaturized cell lysis device developed using a concave spherical transducer is capable of lysing bacteria without added chemical denaturants, enzymes or microparticles and is capable of efficiently lysing yeast without any mechanical or enzymatic pretreatment.The device is designed for miniature bio-analysis systems where cell lysing is needed to obtain intracellular materials for further analysis such as DNA identification.The device lysis efficiency was evaluated using viable cell counts and microscopy.Additionally, the device efficiency was compared with that of traditional chemical cell lysis methods using standard molecular biological techniques such as agarose gels and ultraviolet (UV) spectroscopy.The results indicate that efficient bacteria and cell disruption can be achieved through a low-voltage-driven and spherically focused high-frequency ultrasonic device.

  6. Fluorescent method for monitoring cheese starter permeabilization and lysis

    NARCIS (Netherlands)

    Bunthof, C.J.; Schalkwijk, van S.; Meijer, W.; Abee, T.; Hugenholtz, J.

    2001-01-01

    A fluorescence method to monitor lysis of cheese starter bacteria using dual staining with the LIVE/DEAD BacLight bacterial viability kit is described. This kit combines membrane-permeant green fluorescent nucleic acid dye SYTO 9 and membrane-impermeant red fluorescent nucleic acid dye propidium iod

  7. An integratable microfluidic cartridge for forensic swab samples lysis.

    Science.gov (United States)

    Yang, Jianing; Brooks, Carla; Estes, Matthew D; Hurth, Cedric M; Zenhausern, Frederic

    2014-01-01

    Fully automated rapid forensic DNA analysis requires integrating several multistep processes onto a single microfluidic platform, including substrate lysis, extraction of DNA from the released lysate solution, multiplexed PCR amplification of STR loci, separation of PCR products by capillary electrophoresis, and analysis for allelic peak calling. Over the past several years, most of the rapid DNA analysis systems developed started with the reference swab sample lysate and involved an off-chip lysis of collected substrates. As a result of advancement in technology and chemistry, addition of a microfluidic module for swab sample lysis has been achieved in a few of the rapid DNA analysis systems. However, recent reports on integrated rapid DNA analysis systems with swab-in and answer-out capability lack any quantitative and qualitative characterization of the swab-in sample lysis module, which is important for downstream forensic sample processing. Maximal collection and subsequent recovery of the biological material from the crime scene is one of the first and critical steps in forensic DNA technology. Herein we present the design, fabrication and characterization of an integratable swab lysis cartridge module and the test results obtained from different types of commonly used forensic swab samples, including buccal, saliva, and blood swab samples, demonstrating the compatibility with different downstream DNA extraction chemistries. This swab lysis cartridge module is easy to operate, compatible with both forensic and microfluidic requirements, and ready to be integrated with our existing automated rapid forensic DNA analysis system. Following the characterization of the swab lysis module, an integrated run from buccal swab sample-in to the microchip CE electropherogram-out was demonstrated on the integrated prototype instrument. Therefore, in this study, we demonstrate that this swab lysis cartridge module is: (1) functionally, comparable with routine benchtop lysis

  8. Use of surface enhanced blocking (SEB electrodes for microbial cell lysis in flow-through devices.

    Directory of Open Access Journals (Sweden)

    Abdossamad Talebpour

    Full Text Available By simultaneously subjecting microbial cells to high amplitude pulsed electric fields and flash heating of the cell suspension fluid, effective release of intracellular contents was achieved. The synergistic effect of the applied electric field and elevated temperature on cell lysis in a flow-through device was demonstrated for Gram-negative and Gram-positive bacteria, and Mycobacterium species. The resulting lysate is suitable for downstream nucleic acid amplification and detection without requiring further preparation. The lysis chamber employs surface enhanced blocking electrodes which possess an etched micro-structured surface and a thin layer of dielectric metal oxide which provides a large effective area and blocks transmission of electrical current. The surface enhanced blocking electrodes enable simultaneous suppression of the rapid onset of electric field screening in the bulk of the cell suspension medium and avoidance of undesired electrochemical processes at the electrode-electrolyte interface. In addition the blocking layer ensures the robustness of the cell lysis device in applications involving prolonged flow-through processing of the microbial cells.

  9. INTRAPERITONEAL LYSIS OF TUBERCLE BACILLI.

    Science.gov (United States)

    Manwaring, W H; Bronfenbrenner, J

    1913-12-01

    1. Tubercle bacilli injected into the peritoneal cavities of tuberculous guinea pigs, rats, rabbits, dogs, and monkeys, rapidly disappear from the peritoneal fluids, while persisting in the peritoneal fluids of normal control animals. 2. This disappearance is in part due to an adhesion of the injected bacilli to the peritoneal leucocytes and a fixation of the leucocytes on the omentum. 3. The injected tubercle bacilli can be recovered quantitatively from the peritoneal cavities of normal guinea pigs from one and one half to two hours after the injection, while from tuberculous guinea pigs only 65 per cent. of the bacilli can be recovered at this time. 4. Isolated peritoneal tissues from tuberculous guinea pigs have the power of destroying tubercle bacilli in vitro. 5. A second factor reducing the number of tubercle bacilli free in the peritoneal fluid is therefore an actual lysis of the bacilli. 6. The intraperitoneal lysis is not due solely to substances present in the circulating fluids, since the phenomenon cannot be produced by these fluids in vitro, and since a lytic power cannot be passively conferred even by a direct transfusion of blood from tuberculous to normal animals. 7. The intraperitoneal lysis is apparently due to specific changes in the fixed peritoneal cells of the tuberculous animals.

  10. LACTIC ACID BACTERIA AND THEIR ROLE IN THE MEAT PROCESSING

    Directory of Open Access Journals (Sweden)

    Josef Kameník

    2016-01-01

    Full Text Available Abstract Lactic acid bacteria (LAB play in meat processing a positive but also a negative role. The principal advantage of LAB in the production of dry fermented sausages lies in the fermentation of saccharides, i.e. the creation of lactic acid and the subsequent fall in pH. The role of LAB in fresh meat spoilage is still controversial. Several species are able to dominate the meat system in VP and MAP storage conditions and can release odor-impact molecules. On the other hand they can provide favorable antagonistic activity against other undesired microorganisms. LAB are important spoilage agents in cooked meat products.

  11. Carbon nanotubes for voltage reduction and throughput enhancement of electrical cell lysis on a lab-on-a-chip

    Energy Technology Data Exchange (ETDEWEB)

    Shahini, Mehdi; Yeow, John T W, E-mail: jyeow@uwaterloo.ca [University of Waterloo, 200 University Avenue West, Waterloo, ON (Canada)

    2011-08-12

    We report on the enhancement of electrical cell lysis using carbon nanotubes (CNTs). Electrical cell lysis systems are widely utilized in microchips as they are well suited to integration into lab-on-a-chip devices. However, cell lysis based on electrical mechanisms has high voltage requirements. Here, we demonstrate that by incorporating CNTs into microfluidic electrolysis systems, the required voltage for lysis is reduced by half and the lysis throughput at low voltages is improved by ten times, compared to non-CNT microchips. In our experiment, E. coli cells are lysed while passing through an electric field in a microchannel. Based on the lightning rod effect, the electric field strengthened at the tip of the CNTs enhances cell lysis at lower voltage and higher throughput. This approach enables easy integration of cell lysis with other on-chip high-throughput sample-preparation processes.

  12. Improved lysis efficiency and immunogenicity of Salmonella ghosts mediated by co-expression of λ phage holin-endolysin and ɸX174 gene E

    Science.gov (United States)

    Won, Gayeon; Hajam, Irshad Ahmed; Lee, John Hwa

    2017-01-01

    Bacterial ghosts (BGs) are empty cell envelopes derived from Gram-negative bacteria by bacteriophage ɸX174 gene E mediated lysis. They represent a novel inactivated vaccine platform; however, the practical application of BGs for human vaccines seems to be limited due to the safety concerns on the presence of viable cells in BGs. Therefore, to improve the lysis efficiency of the gene E, we exploited the peptidoglycan hydrolyzing ability of the λ phage holin-endolysins to expedite the process of current BG production system. In this report, we constructed a novel ghost plasmid encoding protein E and holin-endolysins in tandem. We observed that sequential expressions of the gene E and the holin-endolysins elicited rapid and highly efficient Salmonella lysis compared to the lysis mediated by gene E only. These lysed BGs displayed improved immunogenicity in mice compared to the gene E mediated BGs. Consequently, seventy percent of the mice immunized with these novel ghosts survived against a lethal challenge while all the mice vaccinated with gene E mediated ghosts died by day 9 post-infection. We conclude that this novel strategy has the potential to generate highly efficient inactivated candidate vaccines that could replace the currently available bacterial vaccines. PMID:28332591

  13. Streptococcus pneumoniae GAPDH Is Released by Cell Lysis and Interacts with Peptidoglycan.

    Science.gov (United States)

    Terrasse, Rémi; Amoroso, Ana; Vernet, Thierry; Di Guilmi, Anne Marie

    2015-01-01

    Release of conserved cytoplasmic proteins is widely spread among Gram-positive and Gram-negative bacteria. Because these proteins display additional functions when located at the bacterial surface, they have been qualified as moonlighting proteins. The GAPDH is a glycolytic enzyme which plays an important role in the virulence processes of pathogenic microorganisms like bacterial invasion and host immune system modulation. However, GAPDH, like other moonlighting proteins, cannot be secreted through active secretion systems since they do not contain an N-terminal predicted signal peptide. In this work, we investigated the mechanism of GAPDH export and surface retention in Streptococcus pneumoniae, a major human pathogen. We addressed the role of the major autolysin LytA in the delivery process of GAPDH to the cell surface. Pneumococcal lysis is abolished in the ΔlytA mutant strain or when 1% choline chloride is added in the culture media. We showed that these conditions induce a marked reduction in the amount of surface-associated GAPDH. These data suggest that the presence of GAPDH at the surface of pneumococcal cells depends on the LytA-mediated lysis of a fraction of the cell population. Moreover, we demonstrated that pneumococcal GAPDH binds to the bacterial cell wall independently of the presence of the teichoic acids component, supporting peptidoglycan as a ligand to surface GAPDH. Finally, we showed that peptidoglycan-associated GAPDH recruits C1q from human serum but does not activate the complement pathway.

  14. Streptococcus pneumoniae GAPDH Is Released by Cell Lysis and Interacts with Peptidoglycan.

    Directory of Open Access Journals (Sweden)

    Rémi Terrasse

    Full Text Available Release of conserved cytoplasmic proteins is widely spread among Gram-positive and Gram-negative bacteria. Because these proteins display additional functions when located at the bacterial surface, they have been qualified as moonlighting proteins. The GAPDH is a glycolytic enzyme which plays an important role in the virulence processes of pathogenic microorganisms like bacterial invasion and host immune system modulation. However, GAPDH, like other moonlighting proteins, cannot be secreted through active secretion systems since they do not contain an N-terminal predicted signal peptide. In this work, we investigated the mechanism of GAPDH export and surface retention in Streptococcus pneumoniae, a major human pathogen. We addressed the role of the major autolysin LytA in the delivery process of GAPDH to the cell surface. Pneumococcal lysis is abolished in the ΔlytA mutant strain or when 1% choline chloride is added in the culture media. We showed that these conditions induce a marked reduction in the amount of surface-associated GAPDH. These data suggest that the presence of GAPDH at the surface of pneumococcal cells depends on the LytA-mediated lysis of a fraction of the cell population. Moreover, we demonstrated that pneumococcal GAPDH binds to the bacterial cell wall independently of the presence of the teichoic acids component, supporting peptidoglycan as a ligand to surface GAPDH. Finally, we showed that peptidoglycan-associated GAPDH recruits C1q from human serum but does not activate the complement pathway.

  15. Silicon Nitride Bioceramics Induce Chemically Driven Lysis in Porphyromonas gingivalis.

    Science.gov (United States)

    Pezzotti, Giuseppe; Bock, Ryan M; McEntire, Bryan J; Jones, Erin; Boffelli, Marco; Zhu, Wenliang; Baggio, Greta; Boschetto, Francesco; Puppulin, Leonardo; Adachi, Tetsuya; Yamamoto, Toshiro; Kanamura, Narisato; Marunaka, Yoshinori; Bal, B Sonny

    2016-03-29

    Organisms of Gram-negative phylum bacteroidetes, Porphyromonas gingivalis, underwent lysis on polished surfaces of silicon nitride (Si3N4) bioceramics. The antibacterial activity of Si3N4 was mainly the result of chemically driven principles. The lytic activity, although not osmotic in nature, was related to the peculiar pH-dependent surface chemistry of Si3N4. A buffering effect via the formation of ammonium ions (NH4(+)) (and their modifications) was experimentally observed by pH microscopy. Lysis was confirmed by conventional fluorescence spectroscopy, and the bacteria's metabolism was traced with the aid of in situ Raman microprobe spectroscopy. This latter technique revealed the formation of peroxynitrite within the bacterium itself. Degradation of the bacteria's nucleic acid, drastic reduction in phenilalanine, and reduction of lipid concentration were observed due to short-term exposure (6 days) to Si3N4. Altering the surface chemistry of Si3N4 by either chemical etching or thermal oxidation influenced peroxynitrite formation and affected bacteria metabolism in different ways. Exploiting the peculiar surface chemistry of Si3N4 bioceramics could be helpful in counteracting Porphyromonas gingivalis in an alkaline pH environment.

  16. Charge injection through nanocomposite electrode in microfluidic channel for electrical lysis of biological cells

    Science.gov (United States)

    Mishra, Madhusmita; Krishna, Anil; Chandra, Aman; Shenoy, B. M.; Hegde, G. M.; Mahapatra, D. Roy

    2013-03-01

    Several concepts have been developed in the recent years for nanomaterial based integrated MEMS platform in order to accelerate the process of biological sample preparation followed by selective screening and identification of target molecules. In this context, there exist several challenges which need to be addressed in the process of electrical lysis of biological cells. These are due to (i) low resource settings while achieving maximal lysis (ii) high throughput of target molecules to be detected (iii) automated extraction and purification of relevant molecules such as DNA and protein from extremely small volume of sample (iv) requirement of fast, accurate and yet scalable methods (v) multifunctionality toward process monitoring and (vi) downward compatibility with already existing diagnostic protocols. This paper reports on the optimization of electrical lysis process based on various different nanocomposite coated electrodes placed in a microfluidic channel. The nanocomposites are synthesized using different nanomaterials like Zinc nanorod dispersion in polymer. The efficiency of electrical lysis with various different electrode coatings has been experimentally verified in terms of DNA concentration, amplification and protein yield. The influence of the coating thickness on the injection current densities has been analyzed. We further correlate experimentally the current density vs. voltage relationship with the extent of bacterial cell lysis. A coupled multiphysics based simulation model is used to predict the cell trajectories and lysis efficiencies under various electrode boundary conditions as estimated from experimental results. Detailed in-situ fluorescence imaging and spectroscopy studies are performed to validate various hypotheses.

  17. Detergent induced lysis of erythrocytes in kwashiorkor.

    Science.gov (United States)

    Rao, A; Onuora, C U; Cherian, A

    1987-09-15

    The effect of the non-ionic detergent Nonidet P40 on lysis of erythrocytes in children suffering from kwashiorkor was studied. The concentration of the detergent causing 50% haemolysis was significantly reduced in these patients. Detergent haemolysis was more sensitive than osmotic fragility (which was reduced). The abnormality was only slight in marasmic children.

  18. Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control

    Directory of Open Access Journals (Sweden)

    Andrea Meitz

    2016-03-01

    Full Text Available The Bacterial Ghost (BG platform technology evolved from a microbiological expression system incorporating the ϕX174 lysis gene E. E-lysis generates empty but structurally intact cell envelopes (BGs from Gram-negative bacteria which have been suggested as candidate vaccines, immunotherapeutic agents or drug delivery vehicles. E-lysis is a highly dynamic and complex biological process that puts exceptional demands towards process understanding and control. The development of a both economic and robust fed-batch production process for BGs required a toolset capable of dealing with rapidly changing concentrations of viable biomass during the E-lysis phase. This challenge was addressed using a transfer function combining dielectric spectroscopy and soft-sensor based biomass estimation for monitoring the rapid decline of viable biomass during the E-lysis phase. The transfer function was implemented to a feed-controller, which followed the permittivity signal closely and was capable of maintaining a constant specific substrate uptake rate during lysis phase. With the described toolset, we were able to increase the yield of BG production processes by a factor of 8–10 when compared to currently used batch procedures reaching lysis efficiencies >98%. This provides elevated potentials for commercial application of the Bacterial Ghost platform technology.

  19. Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control.

    Science.gov (United States)

    Meitz, Andrea; Sagmeister, Patrick; Lubitz, Werner; Herwig, Christoph; Langemann, Timo

    2016-03-24

    The Bacterial Ghost (BG) platform technology evolved from a microbiological expression system incorporating the ϕX174 lysis gene E. E-lysis generates empty but structurally intact cell envelopes (BGs) from Gram-negative bacteria which have been suggested as candidate vaccines, immunotherapeutic agents or drug delivery vehicles. E-lysis is a highly dynamic and complex biological process that puts exceptional demands towards process understanding and control. The development of a both economic and robust fed-batch production process for BGs required a toolset capable of dealing with rapidly changing concentrations of viable biomass during the E-lysis phase. This challenge was addressed using a transfer function combining dielectric spectroscopy and soft-sensor based biomass estimation for monitoring the rapid decline of viable biomass during the E-lysis phase. The transfer function was implemented to a feed-controller, which followed the permittivity signal closely and was capable of maintaining a constant specific substrate uptake rate during lysis phase. With the described toolset, we were able to increase the yield of BG production processes by a factor of 8-10 when compared to currently used batch procedures reaching lysis efficiencies >98%. This provides elevated potentials for commercial application of the Bacterial Ghost platform technology.

  20. Wireless induction heating in a microfluidic device for cell lysis.

    Science.gov (United States)

    Baek, Seung-ki; Min, Junghong; Park, Jung-Hwan

    2010-04-07

    A wireless induction heating system in a microfluidic device was devised for cell lysis to extract DNA and RNA from Escherichia coli. The thermal responses of nickel, iron and copper heating units were studied by applying an alternating magnetic field as a function of geometry of unit, strength of magnetic field, and kind of metal. Heating units were prepared by cutting metal film using a fiber laser, and the units were integrated into a microchannel system using a soft lithographic process. Variation and distribution of temperature on the surface of the heating units was observed using a thermographic camera and temperature labels. The amount of protein released from E. coli by thermal lysis was determined by protein concentration measurement. Hemoglobin released from red blood cells was observed using colorimetric intensity measurement. Extracted DNA was quantified by real-time polymerase chain reaction, and the profile was compared with that of a positive control of ultrasonically disrupted E. coli. The stability of RNA extracted by induction heating was quantified by the measurement of 23S/16S rRNA ratio and comparison with that by normal RNA extraction kit as a gold standard. A solid-shaped nickel structure was selected as the induction heating element in the microfluidic device because of the relatively small influence of geometries and faster thermal response.The amount of protein extracted from E. coli and hemoglobin released from red blood cells by induction heating of the nickel unit in the microfluidic device was proportional to the strength of the applied magnetic field. The lysis of E. coli by induction heating was as effective as lysis of DNA by the ultrasonication method because the threshold cycle values of the sample were compatible with those of the positive control as measured by ultrasonication. Thermal lysis of E. coli by induction heating represents a reasonable alternative to a commercial RNA extraction method as shown by the comparative

  1. Caspase-11 activation requires lysis of pathogen-containing vacuoles by IFN-induced GTPases.

    Science.gov (United States)

    Meunier, Etienne; Dick, Mathias S; Dreier, Roland F; Schürmann, Nura; Kenzelmann Broz, Daniela; Warming, Søren; Roose-Girma, Merone; Bumann, Dirk; Kayagaki, Nobuhiko; Takeda, Kiyoshi; Yamamoto, Masahiro; Broz, Petr

    2014-05-15

    Lipopolysaccharide from Gram-negative bacteria is sensed in the host cell cytoplasm by a non-canonical inflammasome pathway that ultimately results in caspase-11 activation and cell death. In mouse macrophages, activation of this pathway requires the production of type-I interferons, indicating that interferon-induced genes have a critical role in initiating this pathway. Here we report that a cluster of small interferon-inducible GTPases, the so-called guanylate-binding proteins, is required for the full activity of the non-canonical caspase-11 inflammasome during infections with vacuolar Gram-negative bacteria. We show that guanylate-binding proteins are recruited to intracellular bacterial pathogens and are necessary to induce the lysis of the pathogen-containing vacuole. Lysis of the vacuole releases bacteria into the cytosol, thus allowing the detection of their lipopolysaccharide by a yet unknown lipopolysaccharide sensor. Moreover, recognition of the lysed vacuole by the danger sensor galectin-8 initiates the uptake of bacteria into autophagosomes, which results in a reduction of caspase-11 activation. These results indicate that host-mediated lysis of pathogen-containing vacuoles is an essential immune function and is necessary for efficient recognition of pathogens by inflammasome complexes in the cytosol.

  2. Miniature acoustic wave lysis system and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Branch, Darren W.; Vreeland, Erika Cooley; Smith, Gennifer Tanabe

    2016-12-06

    The present invention relates to an acoustic lysis system including a disposable cartridge that can be reversibly coupled to a platform having a small, high-frequency piezoelectric transducer array. In particular, the system releases viable DNA, RNA, and proteins from human or bacterial cells, without chemicals or additional processing, to enable high-speed sample preparation for clinical point-of-care medical diagnostics and use with nano/microfluidic cartridges. Also described herein are methods of making and using the system of the invention.

  3. Hsp90 inhibition accelerates cell lysis. Anti-Hsp90 ribozyme reveals a complex mechanism of Hsp90 inhibitors involving both superoxide- and Hsp90-dependent events.

    Science.gov (United States)

    Sreedhar, Amere Subbarao; Mihály, Katalin; Pató, Bálint; Schnaider, Tamás; Steták, Attila; Kis-Petik, Katalin; Fidy, Judit; Simonics, Tibor; Maraz, Anna; Csermely, Péter

    2003-09-12

    The 90 kDa heat shock protein, Hsp90, is an abundant molecular chaperone participating in the cytoprotection of eukaryotic cells. Here we analyzed the involvement of Hsp90 in the maintenance of cellular integrity using partial cell lysis as a measure. Inhibition of Hsp90 by geldanamycin, radicicol, cisplatin, and novobiocin induced a significant acceleration of detergent- and hypotonic shock-induced cell lysis. The concentration and time dependence of cell lysis acceleration was in agreement with the Hsp90 inhibition characteristics of the N-terminal inhibitors, geldanamycin and radicicol. Glutathione and other reducing agents partially blocked geldanamycin-induced acceleration of cell lysis but were largely ineffective with other inhibitors. Indeed, geldanamycin treatment led to superoxide production and a change in membrane fluidity. When Hsp90 content was diminished using anti-Hsp90 hammerhead ribozymes, an accelerated cell lysis was also observed. Hsp90 inhibition-induced cell lysis was more pronounced in eukaryotic (yeast, mouse red blood, and human T-lymphoma) cells than in bacteria. Our results indicate that besides the geldanamycin-induced superoxide production, and a consequent increase in cell lysis, inhibition or lack of Hsp90 alone can also compromise cellular integrity. Moreover, cell lysis after hypoxia and complement attack was also enhanced by any type of Hsp90 inhibition used, which shows that the maintenance of cellular integrity by Hsp90 is important in physiologically relevant lytic conditions of tumor cells.

  4. STUDIES ON THE BACTERIOPHAGE OF D'HERELLE : IX. EVIDENCE OF HYDROLYSIS OF BACTERIAL PROTEIN DURING LYSIS.

    Science.gov (United States)

    Hetler, D M; Bronfenbrenner, J

    1928-07-31

    1. During the process of lysis by bacteriophage, there is an appreciable increase in the amount of free amino acid present in the culture. 2. The increase of free amino acid is due to hydrolysis of bacterial protein.

  5. A new device for measurement of fibrin clot lysis: application to the Euglobulin Clot Lysis Time

    Directory of Open Access Journals (Sweden)

    Brohée D

    2002-05-01

    Full Text Available Abstract Background Determination of clot lysis times on whole blood, diluted whole blood, plasma or plasma fraction has been used for many years to assess the overall activity of the fibrinolytic system. We designed a completely computerised semi-automatic 8-channel device for measurement and determination of fibrin clot lysis. The lysis time is evaluated by a mathematical analysis of the lysis curve and the results are expressed in minute (range: 5 to 9999. We have used this new device for Euglobulin Clot Lysis Time (ECLT determination, which is the most common test used in laboratories to estimate plasma fibrinolytic capacity. Results The correlation between ECLT and manual method is very tight : R = 0,99; p -6. The efficiency scores of the method are Conclusions The routine use of this new device could be useful in various situations such as assessment in atherosclerosis and arteriosclerosis associated diseases, coagulation survey of liver transplantations, cardiovascular surgery or pharmacological research. It has already provided highly promising results in preliminary studies on the relation between fibrinolysis and cardiovascular risk factors.

  6. A new device for measurement of fibrin clot lysis: application to the Euglobulin Clot Lysis Time

    Science.gov (United States)

    Boudjeltia, K Zouaoui; Cauchie, Ph; Remacle, Cl; Guillaume, M; Brohée, D; Hubert, JL; Vanhaeverbeek, M

    2002-01-01

    Background Determination of clot lysis times on whole blood, diluted whole blood, plasma or plasma fraction has been used for many years to assess the overall activity of the fibrinolytic system. We designed a completely computerised semi-automatic 8-channel device for measurement and determination of fibrin clot lysis. The lysis time is evaluated by a mathematical analysis of the lysis curve and the results are expressed in minute (range: 5 to 9999). We have used this new device for Euglobulin Clot Lysis Time (ECLT) determination, which is the most common test used in laboratories to estimate plasma fibrinolytic capacity. Results The correlation between ECLT and manual method is very tight : R = 0,99; p < 10-6. The efficiency scores of the method are <4% in intra-assay and <7% in inter-assay. It allows to achieve the tests on hyperlipaemic samples. This new device has been easily integrated in laboratory routine and allows to achieve several ECLT every day without disturbance of laboratory workflow. Conclusions The routine use of this new device could be useful in various situations such as assessment in atherosclerosis and arteriosclerosis associated diseases, coagulation survey of liver transplantations, cardiovascular surgery or pharmacological research. It has already provided highly promising results in preliminary studies on the relation between fibrinolysis and cardiovascular risk factors. PMID:11985782

  7. Processing of heme and heme-containing proteins by bacteria.

    Science.gov (United States)

    Stojiljkovic, Igor; Perkins-Balding, Donna

    2002-04-01

    An extensive amount of new knowledge on bacterial systems involved in heme processing has been accumulated in the last 10 years. We discuss common themes in heme transport across bacterial outer and inner membranes, emphasizing proteins and mechanisms involved. The processing of heme in the bacterial cytoplasm is extensively covered, and a new hypothesis about the fate of heme in the bacterial cell is presented. Auxiliary genes involved in heme utilization, i.e., TonB, proteases, proteins involved in heme storage and pigmentation, as well as genes involved in regulation of heme assimilation are reviewed.

  8. Amount of colicin release in Escherichia coli is regulated by lysis gene expression of the colicin E2 operon.

    Directory of Open Access Journals (Sweden)

    Andreas Mader

    Full Text Available The production of bacteriocins in response to worsening environmental conditions is one means of bacteria to outcompete other microorganisms. Colicins, one class of bacteriocins in Escherichia coli, are effective against closely related Enterobacteriaceae. Current research focuses on production, release and uptake of these toxins by bacteria. However, little is known about the quantitative aspects of these dynamic processes. Here, we quantitatively study expression dynamics of the Colicin E2 operon in E. coli on a single cell level using fluorescence time-lapse microscopy. DNA damage, triggering SOS response leads to the heterogeneous expression of this operon including the cea gene encoding the toxin, Colicin E2, and the cel gene coding for the induction of cell lysis and subsequent colicin release. Advancing previous whole population investigations, our time-lapse experiments reveal that at low exogenous stress levels all cells eventually respond after a given time (heterogeneous timing. This heterogeneous timing is lost at high stress levels, at which a synchronized stress response of all cells 60 min after induction via stress can be observed. We further demonstrate, that the amount of colicin released is dependent on cel (lysis gene expression, independent of the applied exogenous stress level. A heterogeneous response in combination with heterogeneous timing can be biologically significant. It might enable a bacterial population to endure low stress levels, while at high stress levels an immediate and synchronized population wide response can give single surviving cells of the own species the chance to take over the bacterial community after the stress has ceased.

  9. Role of Heterotrophic Bacteria in Marine Ecological Processes

    Digital Repository Service at National Institute of Oceanography (India)

    Ramaiah, N.

    that ubiquitous and self-regulating microbial communities provided the foundation to our understanding on the processes in marine food-web (Landry, 2001). Heterotrophy in the Marine Environment Marine heterotrophy involving all animals and many microbes is a..., distribution, production and, their involvement in nutrient cycling and how they are at the base of microbial food web is essential. Following phytoplankton blooms, the aggregation and sinking flux of organic matter (Takahashi, 1986; Bodungen et al., 1986...

  10. Stochastic holin expression can account for lysis time variation in the bacteriophage λ.

    Science.gov (United States)

    Singh, Abhyudai; Dennehy, John J

    2014-06-06

    The inherent stochastic nature of biochemical processes can drive differences in gene expression between otherwise identical cells. While cell-to-cell variability in gene expression has received much attention, randomness in timing of events has been less studied. We investigate event timing at the single-cell level in a simple system, the lytic pathway of the bacterial virus phage λ. In individual cells, lysis occurs on average at 65 min, with an s.d. of 3.5 min. Interestingly, mutations in the lysis protein, holin, alter both the lysis time (LT) mean and variance. In our analysis, LT is formulated as the first-passage time (FPT) for cellular holin levels to cross a critical threshold. Exact analytical formulae for the FPT moments are derived for stochastic gene expression models. These formulae reveal how holin transcription and translation efficiencies independently modulate the LT mean and variation. Analytical expressions for the LT moments are used to evaluate previously published single-cell LT data for λ phages with mutations in the holin sequence or its promoter. Our results show that stochastic holin expression is sufficient to account for the intercellular LT differences in both wild-type phages, and phage variants where holin transcription and the threshold for lysis have been experimentally altered. Finally, our analysis reveals regulatory motifs that enhance the robustness of lysis timing to cellular noise.

  11. Sub classification and targeted characterization of prophage-encoded two-component cell lysis cassette

    Indian Academy of Sciences (India)

    K V Srividhya; S Krishnaswamy

    2007-08-01

    Bacteriophage induced lysis of host bacterial cell is mediated by a two component cell lysis cassette comprised of holin and lysozyme. Prophages are integrated forms of bacteriophages in bacterial genomes providing a repertoire for bacterial evolution. Analysis using the prophage database (http://bicmku.in:8082) constructed by us showed 47 prophages were associated with putative two component cell lysis genes. These proteins cluster into four different subgroups. In this process, a putative holin (essd) and endolysin (ybcS), encoded by the defective lambdoid prophage DLP12 was found to be similar to two component cell lysis genes in functional bacteriophages like p21 and P1. The holin essd was found to have a characteristic dual start motif with two transmembrane regions and C-terminal charged residues as in class II holins. Expression of a fusion construct of essd in Escherichia coli showed slow growth. However, under appropriate conditions, this protein could be over expressed and purified for structure function studies. The second component of the cell lysis cassette, ybcS, was found to have an N-terminal SAR (Signal Arrest Release) transmembrane domain. The construct of ybcS has been over expressed in E. coli and the purified protein was functional, exhibiting lytic activity against E. coli and Salmonella typhi cell wall substrate. Such targeted sequence-structure-function characterization of proteins encoded by cryptic prophages will help understand the contribution of prophage proteins to bacterial evolution.

  12. The euglobulin clot lysis time to assess the impact of nanoparticles on fibrinolysis

    Energy Technology Data Exchange (ETDEWEB)

    Minet, Valentine, E-mail: valentine.minet@unamur.be; Alpan, Lutfiye; Mullier, François [University of Namur – UNamur, Department of Pharmacy, Namur Thrombosis and Hemostasis Center (NTHC), Namur Nanosafety Center (NNC), NAmur Research Institute for Life Sciences NARILIS (Belgium); Toussaint, Olivier [Laboratory of Cellular Biochemistry and Biology (URBC) (Belgium); Lucas, Stéphane [University of Namur (UNamur), Research Centre for the Physics of Matter and Radiation (PMR-LARN), Namur Nanosafety Center NNC, NAmur Research Institute for Life Sciences NARILIS (Belgium); Dogné, Jean-Michel; Laloy, Julie, E-mail: julie.laloy@unamur.be [University of Namur – UNamur, Department of Pharmacy, Namur Thrombosis and Hemostasis Center (NTHC), Namur Nanosafety Center (NNC), NAmur Research Institute for Life Sciences NARILIS (Belgium)

    2015-07-15

    Nanoparticles (NPs) are developed for many applications in various fields, including nanomedicine. The NPs used in nanomedicine may disturb homeostasis in blood. Secondary hemostasis (blood coagulation) and fibrinolysis are complex physiological processes regulated by activators and inhibitors. An imbalance of this system can either lead to the development of hemorrhages or thrombosis. No data are currently available on the impact of NPs on fibrinolysis. The objectives of this study are (1) to select a screening test to study ex vivo the impact of NPs on fibrinolysis and (2) to test NPs with different physicochemical properties. Euglobulin clot lysis time test was selected to screen the impact of some NPs on fibrinolysis using normal pooled plasma. A dose-dependent decrease in the lysis time was observed with silicon dioxide and silver NPs without disturbing the fibrin network. Carbon black, silicon carbide, and copper oxide did not affect the lysis time at the tested concentrations.

  13. The euglobulin clot lysis time to assess the impact of nanoparticles on fibrinolysis

    Science.gov (United States)

    Minet, Valentine; Alpan, Lutfiye; Mullier, François; Toussaint, Olivier; Lucas, Stéphane; Dogné, Jean-Michel; Laloy, Julie

    2015-07-01

    Nanoparticles (NPs) are developed for many applications in various fields, including nanomedicine. The NPs used in nanomedicine may disturb homeostasis in blood. Secondary hemostasis (blood coagulation) and fibrinolysis are complex physiological processes regulated by activators and inhibitors. An imbalance of this system can either lead to the development of hemorrhages or thrombosis. No data are currently available on the impact of NPs on fibrinolysis. The objectives of this study are (1) to select a screening test to study ex vivo the impact of NPs on fibrinolysis and (2) to test NPs with different physicochemical properties. Euglobulin clot lysis time test was selected to screen the impact of some NPs on fibrinolysis using normal pooled plasma. A dose-dependent decrease in the lysis time was observed with silicon dioxide and silver NPs without disturbing the fibrin network. Carbon black, silicon carbide, and copper oxide did not affect the lysis time at the tested concentrations.

  14. Utilization of graphene electrode in transparent microwell arrays for high throughput cell trapping and lysis.

    Science.gov (United States)

    Ameri, S Kabiri; Singh, P K; Sonkusale, S

    2014-11-15

    Here we present a high-throughput, transparent microfluidic device with embedded microwell arrays sandwiched between transparent electrodes made from graphene (at the bottom) and indium tin oxide (at the top) for dielectrophoretic cell trapping and electrical lysis. Graphene suppresses unwanted faradaic reaction effects on the cells and the medium that is typically observed in ITO based electrodes from application of DC field for electrical lysis. This is because graphene is more electrochemically inert than indium tin oxide (ITO) where ITO undergoes reduction-oxidation (redox) reaction in the presence of electrolyte in most standard cell media. This redox process also compromises ITO's electrical properties and optical transparency over multiple use. The presented microfluidic device shows high efficiency for cell trapping and lysis and an electrochemically stable behavior for long operational life.

  15. Solubilization of proteins: the importance of lysis buffer choice.

    Science.gov (United States)

    Peach, Mandy; Marsh, Noelle; Miskiewicz, Ewa I; MacPhee, Daniel J

    2015-01-01

    The efficient extraction of proteins of interest from cells and tissues is not always straightforward. Here we demonstrate the differences in extraction of the focal adhesion protein Kindlin-2 from choriocarcinoma cells using NP-40 and RIPA lysis buffer. Furthermore, we demonstrate the use of a more denaturing urea/thiourea lysis buffer for solubilization, by comparing its effectiveness for solubilization of small heat-shock proteins from smooth muscle with the often utilized RIPA lysis buffer. Overall, the results demonstrate the importance of establishing the optimal lysis buffer for specific protein solubilization within the experimental workflow.

  16. Culture-independent detection of "TM7" bacteria in a streptomycin-resistant acidophilic nitrifying process

    Science.gov (United States)

    Kurogi, T.; Linh, N. T. T.; Kuroki, T.; Yamada, T.; Hiraishi, A.

    2014-02-01

    Nitrification in biological wastewater treatment processes has been believed for long time to take place under neutral conditions and is inhibited under acidic conditions. However, we previously constructed acidophilic nitrifying sequencing-batch reactors (ANSBRs) being capable of nitrification at level, this study was undertaken to construct streptomycin-resistant acidophilic nitrifying (SRAN) reactors and to demonstrate whether TM7 bacteria are abundant in these reactors. The SRAN reactors were constructed by seeding with nitrifying sludge from an ANSBR and cultivating with ammonium-containing mineral medium (pH 4.0), to which streptomycin at a concentration of 10, 30 and 50 mg L-1 was added. In all reactors, the pH varied between 2.7 and 4.0, and ammonium was completely converted to nitrate in every batch cycle. PCR-aided denaturing gradient gel electrophoresis (DGGE) targeting 16S rRNA genes revealed that some major clones assigned to TM7 bacteria and Gammaproteobacteria were constantly present during the overall period of operation. Fluorescence in situ hybridization (FISH) with specific oligonucleotide probes also showed that TM7 bacteria predominated in all SRAN reactors, accounting for 58% of the total bacterial population on average. Although the biological significance of the TM7 bacteria in the SRAN reactors are unknown, our results suggest that these bacteria are possibly streptomycin-resistant and play some important roles in the acidophilic nitrifying process.

  17. Managing the potential risks of using bacteria-laden water in mineral processing to protect freshwater.

    Science.gov (United States)

    Liu, Wenying; Moran, Chris J; Vink, Sue

    2013-06-18

    The minerals industry is being driven to access multiple water sources and increase water reuse to minimize freshwater withdrawal. Bacteria-laden water, such as treated effluent, has been increasingly used as an alternative to freshwater for mineral processing, in particular flotation, where conditions are favorable for bacterial growth. However, the risk posed by bacteria to flotation efficiency is poorly understood. This could be a barrier to the ongoing use of this water source. This study tested the potential of a previously published risk-based approach as a management tool to both assist mine sites in quantifying the risk from bacteria, and finding system-wide cost-effective solutions for risk mitigation. The result shows that the solution of adjusting the flotation chemical regime could only partly control the risk. The second solution of using tailings as an absorbent was shown to be effective in the laboratory in reducing bacterial concentration and thus removing the threat to flotation recovery. The best solution is likely to combine internal and external approaches, that is, inside and outside processing plants. Findings in this study contribute possible methods applicable to managing the risk from water-borne bacteria to plant operations that choose to use bacteria-containing water, when attempting to minimize freshwater use, and avoiding the undesirable consequences of increasing its use.

  18. Role of water hardness in rinsing bacteria from the skin of processed broiler chickens

    Science.gov (United States)

    The effect of water hardness on the ability of water to rinse bacteria from the skin of processed broiler chickens was examined. Artificial hard water with a total hardness of 200 ppm (very hard water) was prepared by dissolving calcium chloride (CaCl2) and magnesium chloride hexahydrate (MgCl2 •6H2...

  19. Inactivation of Bacteria in Oil Field Injected Water by a Pulsed Plasma Discharge Process

    Science.gov (United States)

    Xin, Qing; Li, Zhongjian; Lei, Lecheng; Yang, Bin

    2016-09-01

    Pulsed plasma discharge was employed to inactivate bacteria in the injection water for an oil field. The effects of water conductivity and initial concentration of bacteria on elimination efficiency were investigated in the batch and continuous flow modes. It was demonstrated that Fe2+ contained in injection water could enhance the elimination efficiency greatly. The addition of reducing agent glutathione (GSH) indicated that active radicals generated by pulsed plasma discharges played an important role in the inactivation of bacteria. Moreover, it was found that the microbial inactivation process for both batch and continuous flow mode well fitted the model based on the Weibull's survival function. supported by Zhejiang Province Welfare Technology Applied Research Project of China (No. 2014C31137), National Natural Science Foundation of China (Nos. 21436007 and U1462201), and the Fundamental Research Funds for the Central Universities of China (No. 2015QNA4032)

  20. Inactivation of bacteria and helminth in wastewater treatment plant effluent using oxidation processes.

    Science.gov (United States)

    Guadagnini, Regiane Aparecida; dos Santos, Luciana Urbano; Franco, Regina Maura Bueno; Guimarães, José Roberto

    2013-01-01

    The contamination of bodies of water by raw and even treated sewage is worrying because pathogens that affect public health and the environment are not fully eliminated in wastewater treatment systems. The disinfection step is an important barrier to adopt to reduce this contamination. However, widely used disinfectants such as chlorine do not guarantee the inactivation of resistant organisms such as spore-forming bacteria and helminth eggs. This study evaluated the effectiveness of processes of peroxidation (H2O2), ultraviolet radiation (UV) and peroxidation assisted by ultraviolet radiation (H2O2/UV) in terms of reduction and inactivation of total coliform bacteria, Escherichia coli, helminth eggs and larvae present in a treated sewage. Doses of UV radiation of 70 mJ cm(-2) and hydrogen peroxide concentration of 30 mg L(-1) were used. The number of bacteria reduced after UV and H2O2/UV processes was 3 and 4 log, respectively. An average reduction of 59% in the number of eggs was verified when using H2O2, UV, and H2O2/UV processes. Helminth larvae were reduced by 24% after H2O2 and UV; the process H2O2/UV did not reduce the number of larvae. Statistically significant differences between the processes for both organisms were not observed.

  1. Rapid thermal lysis of cells using silicon-diamond microcantilever heaters.

    Science.gov (United States)

    Privorotskaya, Natalya; Liu, Yi-Shao; Lee, Jungchul; Zeng, Hongjun; Carlisle, John A; Radadia, Adarsh; Millet, Larry; Bashir, Rashid; King, William P

    2010-05-07

    This paper presents the design and application of microcantilever heaters for biochemical applications. Thermal lysis of biological cells was demonstrated as a specific example. The microcantilever heaters, fabricated from selectively doped single crystal silicon, provide local resistive heating with highly uniform temperature distribution across the cantilevers. Very importantly, the microcantilever heaters were coated with a layer of 100 nm thick electrically insulating ultrananocrystalline diamond (UNCD) layer used for cell immobilization on the cantilever surface. Fibroblast cells or bacterial cells were immobilized on the UNCD/cantilever surfaces and thermal lysis was demonstrated via optical fluorescence microscopy. Upon electrical heating of the cantilever structures to 93 degrees C for 30 seconds, fibroblast cell and nuclear membrane were compromised and the cells were lysed. Over 90% of viable bacteria were also lysed after 15 seconds of heating at 93 degrees C. This work demonstrates the utility of silicon-UNCD heated microcantilevers for rapid cell lysis and forms the basis for other rapid and localized temperature-regulated microbiological experiments in cantilever-based lab on chip applications.

  2. Drinking water biotic safety of particles and bacteria attached to fines in activated carbon process

    Institute of Scientific and Technical Information of China (English)

    CHEN Wei; LIN Tao; WANG Leilei

    2007-01-01

    In this paper,the drinking water biotic safety of particles and bacteria attached to fines in activated carbon process was investigated by actual treatment process and advanced treatment pilot trial with granular activated carbon.In the experiment,the particles were detected by IBR particle calculating instrument,the activated carbon fines were counted on the basis of the most probable number (MPN) with a microscope,the total number of bacteria was analyzed between the conventional agar culture medium and the one with R2A,and the bacteria attached to activated carbon fines was resolved by the homogenization technique.The experimental results showed that the average total number of particles was 205 CNT/mL in the activated carbon effluent during a filter cycle,of which the number of particles with sizes>2μm was 77 CNT/mL more than the present particle control criterion of the American drinking water product standard (50 CNT/mL).The backwash of low density and long duration lowered particle number in the effluent.The MPN of activated carbon frees in the effluent was between 400 and 600 CNT/L,which accounted for less than 5‰ of the total particles from activated carbon filtration for a poor relative level (R2= 0.34).The microorganisms in activated carbon effluent consisted mostly of heterotrophic bacillus and the total bacteria number was five times as high as that of the inflow,i.e.the effluent from sand filter.The actual bacteria number may be truly indicated by the detection technique with R2A culture medium compared with the traditional agar cultivation.The inactivation efficiency of bacteria attached to activated carbon fines was less than 40% under 1.1 mg/L of chlorine contacting for 40 min.Results showed that the particles and bacteria attached to activated carbon fines may influence drinking water biotic safety,and that the effective control measures need to be further investigated.

  3. The influence of fat and monoacylglycerols on growth of spore-forming bacteria in processed cheese.

    Science.gov (United States)

    Hauerlandová, Iva; Lorencová, Eva; Buňka, František; Navrátil, Jan; Janečková, Kristýna; Buňková, Leona

    2014-07-16

    Highly undesirable microbial contaminants of processed cheese are endospore-forming bacteria of the genera Bacillus and Clostridium. Survival of Bacillus subtilis, B. cereus, Clostridium butyricum and C. sporogenes was examined in model processed cheese samples supplemented with monoacylglycerols. In processed cheese samples, monoacylglycerols of undecanoic, undecenoic, lauric and adamantane-1-carboxylic acid at concentration of 0.15% w/w prevented the growth and multiplication of both Bacillus species throughout the storage period. The two species of Clostridium were less affected by monoacylglycerols in processed cheese samples and only partial inhibition was observed. The effect of milk fat content on microbial survival in processed cheese was also evaluated. The growth of Bacillus sp. was affected by the fat level of processed cheese while population levels of Clostridium sp. did not differ in processed cheese samples with 30, 40 and 50% fat in dry matter.

  4. Disposable on-chip microfluidic system for buccal cell lysis, DNA purification, and polymerase chain reaction.

    Science.gov (United States)

    Cho, Woong; Maeng, Joon-Ho; Ahn, Yoomin; Hwang, Seung Yong

    2013-09-01

    This paper reports the development of a disposable, integrated biochip for DNA sample preparation and PCR. The hybrid biochip (25 × 45 mm) is composed of a disposable PDMS layer with a microchannel chamber and reusable glass substrate integrated with a microheater and thermal microsensor. Lysis, purification, and PCR can be performed sequentially on this microfluidic device. Cell lysis is achieved by heat and purification is performed by mechanical filtration. Passive check valves are integrated to enable sample preparation and PCR in a fixed sequence. Reactor temperature is needed to lysis and PCR reaction is controlled within ±1°C by PID controller of LabVIEW software. Buccal epithelial cell lysis, DNA purification, and SY158 gene PCR amplification were successfully performed on this novel chip. Our experiments confirm that the entire process, except the off-chip gel electrophoresis, requires only approximately 1 h for completion. This disposable microfluidic chip for sample preparation and PCR can be easily united with other technologies to realize a fully integrated DNA chip.

  5. A Cell Lysis and Protein Purification - Single Molecule Assay Devices for Evaluation of Genetically Engineered Proteins

    Science.gov (United States)

    Nakyama, Tetsuya; Tabata, Kazuhito; Noji, Hiroyuki; Yokokawa, Ryuji

    We have developed two devices applicable to evaluate genetically engineered proteins in single molecule assay: on-chip cell lysis device, and protein purification - assay device. A motor protein, F1-ATPase expressed in E.coli, was focused in this report as a target protein. Cell lysis was simply performed by applying pulse voltage between Au electrodes patterned by photolithography, and its efficiency was determined by absorptiometry. The subsequent processes, purification and assay of extracted proteins, were demonstrated in order to detect F1-ATPase and to evaluate its activity. The specific bonding between his-tag in F1-ATPase and Ni-NTA coated on a glass surface was utilized for the purification process. After immobilization of F1-ATPase, avidin-coated microspheres and adenosine tri-phosphate (ATP) solution were infused sequentially to assay the protein. Microsphere rotation was realized by activity of F1-ATPase corresponding to ATP hydrolysis. Results show that the cell lysis device, at the optimum condition, extracts enough amount of protein for single molecule assay. Once cell lysate was injected to the purification - assay device, proteins were diffused in the lateral direction in a Y-shape microchannel. The gradient of protein concentratioin provides an optimal concentration for the assay i.e. the highest density of rotating beads. Density of rotating beads is also affected by the initial concentration of protein injected to the device. The optimum concentration was achieved by our cell lysis device not by the conventional method by ultrasonic wave. Rotation speed was analyzed for several microspheres assayed in the purification - assay device, and the results were compatible to that of conventional assay in which F1-ATPase was purified in bulk scale. In conclusion, we have demonstrated on-chip cell lysis and assay appropriate for the sequential analysis without any pretreatment. On-chip devices replacing conventional bioanalytical methods will be

  6. Hygiene Aspects of the Biogas Process with Emphasis on Spore-Forming Bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Bagge, Elisabeth

    2009-07-01

    Biogas is a renewable source of energy which can be obtained from processing of biowaste. The digested residues can be used as fertiliser. Biowaste intended for biogas production contains pathogenic micro-organisms. A pre-pasteurisation step at 70 deg C for 60 min before anaerobic digestion reduces non spore-forming bacteria such as Salmonella spp. To maintain the standard of the digested residues it must be handled in a strictly hygienic manner to avoid recontamination and re-growth of bacteria. The risk of contamination is particularly high when digested residues are transported in the same vehicles as the raw material. However, heat treatment at 70 deg C for 60 min will not reduce spore-forming bacteria such as Bacillus spp. and Clostridium spp. Spore-forming bacteria, including those that cause serious diseases, can be present in substrate intended for biogas production. The number of species and the quantity of Bacillus spp. and Clostridium spp. in manure, slaughterhouse waste and in samples from different stages during the biogas process were investigated. The number of species of clostridia seemed to decrease following digestion, likewise the quantity. However, Bacillus spp. seemed to pass unaffected through the biogas process. In laboratory-scale experiments the effects on clostridia during pasteurisation and digestion were investigated. Pathogenic clostridia were inoculated in substrates from homogenisation tanks and digester tanks. The inoculated clostridia remained after pasteurisation, but the impacts of digestion differ between different species. Culture followed by identification of C. chauvoei by PCR in samples from cattle died from blackleg, is faster and safer than culture followed by biochemical identification of C. chauvoei. However, for environmental samples the PCR method is not practically applicable for detection of C. chauvoei. To avoid spreading of diseases via biogas plants when digested residues are spread on arable land, a pasteurisation

  7. Testing optimality with experimental evolution: lysis time in a bacteriophage.

    Science.gov (United States)

    Heineman, Richard H; Bull, James J

    2007-07-01

    Optimality models collapse the vagaries of genetics into simple trade-offs to calculate phenotypes expected to evolve by natural selection. Optimality approaches are commonly criticized for this neglect of genetic details, but resolution of this disagreement has been difficult. The importance of genetic details may be tested by experimental evolution of a trait for which an optimality model exists and in which genetic details can be studied. Here we evolved lysis time in bacteriophage T7, a virus of Escherichia coli. Lysis time is equivalent to the age of reproduction in an organism that reproduces once and then dies. Delaying lysis increases the number of offspring but slows generation time, and this trade-off renders the optimum sensitive to environmental conditions: earlier lysis is favored when bacterial hosts are dense, later lysis is favored when hosts are sparse. In experimental adaptations, T7 evolved close to the optimum in conditions favoring early lysis but not in conditions favoring late lysis. One of the late lysis adaptations exhibited no detectable phenotypic evolution despite genetic evolution; the other evolved only partly toward the expected optimum. Overall, the lysis time of the adapted phages remained closer to their starting values than predicted by the model. From the perspective of the optimality model, the experimental conditions were expected to select changes only along the postulated trade-off, but a trait outside the trade-off evolved as well. Evidence suggests that the model's failure ultimately stems from a violation of the trade-off, rather than a paucity of mutations.

  8. Halotolerant bacteria in the São Paulo Zoo composting process and their hydrolases and bioproducts

    Directory of Open Access Journals (Sweden)

    Lilian C.G. Oliveira

    2015-06-01

    Full Text Available Halophilic microorganisms are able to grow in the presence of salt and are also excellent source of enzymes and biotechnological products, such as exopolysaccharides (EPSs and polyhydroxyalkanoates (PHAs. Salt-tolerant bacteria were screened in the Organic Composting Production Unit (OCPU of São Paulo Zoological Park Foundation, which processes 4 ton/day of organic residues including plant matter from the Atlantic Rain Forest, animal manure and carcasses and mud from water treatment. Among the screened microorganisms, eight halotolerant bacteria grew at NaCl concentrations up to 4 M. These cultures were classified based on phylogenetic characteristics and comparative partial 16S rRNA gene sequence analysis as belonging to the genera Staphylococcus, Bacillus and Brevibacterium. The results of this study describe the ability of these halotolerant bacteria to produce some classes of hydrolases, namely, lipases, proteases, amylases and cellulases, and biopolymers. The strain characterized as of Brevibacterium avium presented cellulase and amylase activities up to 4 M NaCl and also produced EPSs and PHAs. These results indicate the biotechnological potential of certain microorganisms recovered from the composting process, including halotolerant species, which have the ability to produce enzymes and biopolymers, offering new perspectives for environmental and industrial applications.

  9. THE POPULATION OF BACTERIA AND CO2 RELEASE ON PROCESS OF COMPOSTING MANURE AND SWAMP GRASS

    Directory of Open Access Journals (Sweden)

    Diana Utama

    2017-03-01

    Full Text Available This study aimed to analyze the bacteria population, the release of CO2, pH and organic-C and total-N in the process of compost made from manure and swamp grass. Treatment level consist of 100% manure (K100, 50% of manure + 50% swamp grass (K50R50, 25% of manure + 75% swamp grass (K25R75, and 10% of manure + 90% swamp grass (K10R90. The result of this study indicated the dynamic of different bacteria population on different composting materials by increasing of the composting time. The release of CO2 decrease on all treatment levels by increasing of the composting time. The pH value increased at all levels of treatment, except the composition of 100% manure. The best composition obtained by mixing of 10% manure and 90% swamp grass.

  10. Effect of Inoculating Lactic Acid Bacteria Starter in Low-Salt Pickle Process of Zhacai

    Directory of Open Access Journals (Sweden)

    Shiyang Gao

    2012-10-01

    Full Text Available The traditional pickle process of Zhacai was thought unsafely and unstable, because of the high salt and nitrite contents. The study has shown a method to solve these problems. One Lactic Acid Bacteria (LAB strain, Lactobacillus plantarum was used as starter in the low salt pickle process of Chinese Zhacai. The LAB inoculation amounts were 2, 5, 8%, respectively and the group without LAB inoculation was set as the control. The inoculation groups were added with 8% salt, while the control group 10%. The dynamic changes of physical and chemical parameters of two groups were detected during the pickle process. The results showed that: pH value of the inoculated groups declined rapidly, the lowest pH value was 3.61; the LAB became the predominant bacteria strains during the pickle and the colony amounts of the inoculated groups could be up to 6.32 log10 cfu/mL; the nitrite content of each group kept rising and the nitrite peaks of the inoculated groups appeared 3 days in advance, but the peak values were nearly half of the control group; amino nitrogen contents were lower but finally higher than control group and the highest value could reach 0.46 g/100 mL finally. Therefore, the inoculated groups with the lower salt could be more safety and better flavor. This study provided important reference to optimize the pickle fermentation process of Chinese Zhacai.

  11. The Spheroplast Lysis Assay for Yeast in Microtiter Plate Format

    Science.gov (United States)

    Ovalle, Rafael; Spencer, Moyah; Thiwanont, Monthiwa; Lipke, Peter N.

    1999-01-01

    A yeast lysis assay in the microtiter plate format improved precision and throughput and led to an improved algorithm for estimating lag time. The assay reproducibly revealed differences of 10% or greater in the maximal lysis rate and 50% or greater in the lag time. Clonal differences were determined to be the major source of variation. Microtiter-based assays should be useful for screening for drug susceptibility and for analyzing mutant phenotypes. PMID:10427014

  12. Coexistence of Lactic Acid Bacteria and Potential Spoilage Microbiota in a Dairy Processing Environment.

    Science.gov (United States)

    Stellato, Giuseppina; De Filippis, Francesca; La Storia, Antonietta; Ercolini, Danilo

    2015-11-01

    Microbial contamination in food processing plants can play a fundamental role in food quality and safety. In this study, the microbiota in a dairy plant was studied by both 16S rRNA- and 26S rRNA-based culture-independent high-throughput amplicon sequencing. Environmental samples from surfaces and tools were studied along with the different types of cheese produced in the same plant. The microbiota of environmental swabs was very complex, including more than 200 operational taxonomic units with extremely variable relative abundances (0.01 to 99%) depending on the species and sample. A core microbiota shared by 70% of the samples indicated a coexistence of lactic acid bacteria with a remarkable level of Streptococcus thermophilus and possible spoilage-associated bacteria, including Pseudomonas, Acinetobacter, and Psychrobacter, with a relative abundance above 50%. The most abundant yeasts were Kluyveromyces marxianus, Yamadazyma triangularis, Trichosporon faecale, and Debaryomyces hansenii. Beta-diversity analyses showed a clear separation of environmental and cheese samples based on both yeast and bacterial community structure. In addition, predicted metagenomes also indicated differential distribution of metabolic pathways between the two categories of samples. Cooccurrence and coexclusion pattern analyses indicated that the occurrence of potential spoilers was excluded by lactic acid bacteria. In addition, their persistence in the environment can be helpful to counter the development of potential spoilers that may contaminate the cheeses, with possible negative effects on their microbiological quality.

  13. Streptomyces lunalinharesii Strain 235 Shows the Potential to Inhibit Bacteria Involved in Biocorrosion Processes

    Directory of Open Access Journals (Sweden)

    Juliana Pacheco da Rosa

    2013-01-01

    Full Text Available Four actinomycete strains previously isolated from Brazilian soils were tested for their antimicrobial activity against Bacillus pumilus LF-4 and Desulfovibrio alaskensis NCIMB 13491, bacteria that are well known to be involved in biofilm formation and biocorrosion. Strain 235, belonging to the species Streptomyces lunalinharesii, inhibited the growth of both bacteria. The antimicrobial activity was seen over a wide range of pH, and after treatment with several chemicals and heat but not with proteinase K and trypsin. The antimicrobial substances present in the concentrated supernatant from growth media were partially characterized by SDS-PAGE and extracellular polypeptides were seen. Bands in the size range of 12 to 14.4 kDa caused antimicrobial activity. Transmission electron microscopy of D. alaskensis cells treated with the concentrated supernatant containing the antimicrobial substances revealed the formation of prominent bubbles, the spherical double-layered structures on the cell membrane, and the periplasmic space completely filled with electron-dense material. This is the first report on the production of antimicrobial substances by actinomycetes against bacteria involved in biocorrosion processes, and these findings may be of great relevance as an alternative source of biocides to those currently employed in the petroleum industry.

  14. Streptomyces lunalinharesii strain 235 shows the potential to inhibit bacteria involved in biocorrosion processes.

    Science.gov (United States)

    Pacheco da Rosa, Juliana; Korenblum, Elisa; Franco-Cirigliano, Marcella Novaes; Abreu, Fernanda; Lins, Ulysses; Soares, Rosângela M A; Macrae, Andrew; Seldin, Lucy; Coelho, Rosalie R R

    2013-01-01

    Four actinomycete strains previously isolated from Brazilian soils were tested for their antimicrobial activity against Bacillus pumilus LF-4 and Desulfovibrio alaskensis NCIMB 13491, bacteria that are well known to be involved in biofilm formation and biocorrosion. Strain 235, belonging to the species Streptomyces lunalinharesii, inhibited the growth of both bacteria. The antimicrobial activity was seen over a wide range of pH, and after treatment with several chemicals and heat but not with proteinase K and trypsin. The antimicrobial substances present in the concentrated supernatant from growth media were partially characterized by SDS-PAGE and extracellular polypeptides were seen. Bands in the size range of 12 to 14.4 kDa caused antimicrobial activity. Transmission electron microscopy of D. alaskensis cells treated with the concentrated supernatant containing the antimicrobial substances revealed the formation of prominent bubbles, the spherical double-layered structures on the cell membrane, and the periplasmic space completely filled with electron-dense material. This is the first report on the production of antimicrobial substances by actinomycetes against bacteria involved in biocorrosion processes, and these findings may be of great relevance as an alternative source of biocides to those currently employed in the petroleum industry.

  15. Bacterial Lysis through Interference with Peptidoglycan Synthesis Increases Biofilm Formation by Nontypeable Haemophilus influenzae

    Science.gov (United States)

    Puig, Carmen; Merlos, Alexandra; Viñas, Miguel; de Jonge, Marien I.; Liñares, Josefina; Ardanuy, Carmen

    2017-01-01

    ABSTRACT Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen that mainly causes otitis media in children and community-acquired pneumonia or exacerbations of chronic obstructive pulmonary disease in adults. A large variety of studies suggest that biofilm formation by NTHi may be an important step in the pathogenesis of this bacterium. However, the underlying mechanisms involved in this process are poorly elucidated. In this study, we used a transposon mutant library to identify bacterial genes involved in biofilm formation. The growth and biofilm formation of 4,172 transposon mutants were determined, and the involvement of the identified genes in biofilm formation was validated in in vitro experiments. Here, we present experimental data showing that increased bacterial lysis, through interference with peptidoglycan synthesis, results in elevated levels of extracellular DNA, which increased biofilm formation. Interestingly, similar results were obtained with subinhibitory concentrations of β-lactam antibiotics, known to interfere with peptidoglycan synthesis, but such an effect does not appear with other classes of antibiotics. These results indicate that treatment with β-lactam antibiotics, especially for β-lactam-resistant NTHi isolates, might increase resistance to antibiotics by increasing biofilm formation. IMPORTANCE Most, if not all, bacteria form a biofilm, a multicellular structure that protects them from antimicrobial actions of the host immune system and affords resistance to antibiotics. The latter is especially disturbing with the increase in multiresistant bacterial clones worldwide. Bacterial biofilm formation is a multistep process that starts with surface adhesion, after which attached bacteria divide and give rise to biomass. The actual steps required for Haemophilus influenzae biofilm formation are largely not known. We show that interference with peptidoglycan biosynthesis increases biofilm formation because of the release

  16. Electrical lysis: dynamics revisited and advances in On-chip operation.

    Science.gov (United States)

    Morshed, Bashir; Shams, Maitham; Mussivand, Tofy

    2013-01-01

    Electrical lysis (EL) is the process of breaking the cell membrane to expose the internal contents under an applied high electric field. Lysis is an important phenomenon for cellular analysis, medical treatment, and biofouling control. This paper aims to review, summarize, and analyze recent advancements on EL. Major databases including PubMed, Ei Engineering Village, IEEE Xplore, and Scholars Portal were searched using relevant keywords. More than 50 articles published in English since 1997 are cited in this article. EL has several key advantages compared to other lysis techniques such as chemical, mechanical, sonication, or laser, including rapid speed of operation, ability to control, miniaturization, low cost, and low power requirement. A variety of cell types have been investigated for including protoplasts, E. coli, yeasts, blood cells, and cancer cells. EL has been developed and applied for decontamination, cytology, genetics, single-cell analysis, cancer treatment, and other applications. On-chip EL is a promising technology for multiplexed automated implementation of cell-sample preparation and processing with micro- or nanoliter reagents.

  17. Direct cell lysis for single-cell gene expression profiling

    Directory of Open Access Journals (Sweden)

    David eSvec

    2013-11-01

    Full Text Available The interest to analyze single and few cell samples is rapidly increasing. Numerous extraction protocols to purify nucleic acids are available, but most of them compromise severely on yield to remove contaminants and are therefore not suitable for the analysis of samples containing small numbers of transcripts only. Here, we evaluate 17 direct cell lysis protocols for transcript yield and compatibility with downstream reverse transcription quantitative real-time PCR. Four endogenously expressed genes are assayed together with RNA and DNA spikes in the samples. We found bovine serum albumin (BSA to be the best lysis agent, resulting in efficient cell lysis, high RNA stability and enhanced reverse transcription efficiency. Furthermore, we found direct cell lysis with BSA superior to standard column based extraction methods, when analyzing from 1 up to 512 mammalian cells. In conclusion, direct cell lysis protocols based on BSA can be applied with most cell collection methods and are compatible with most analytical workflows to analyze single cells as well as samples composed of small numbers of cells.

  18. Biomineralization processes of calcite induced by bacteria isolated from marine sediments.

    Science.gov (United States)

    Wei, Shiping; Cui, Hongpeng; Jiang, Zhenglong; Liu, Hao; He, Hao; Fang, Nianqiao

    2015-06-01

    Biomineralization is a known natural phenomenon associated with a wide range of bacterial species. Bacterial-induced calcium carbonate precipitation by marine isolates was investigated in this study. Three genera of ureolytic bacteria, Sporosarcina sp., Bacillus sp. and Brevundimonas sp. were observed to precipitate calcium carbonate minerals. Of these species, Sporosarcina sp. dominated the cultured isolates. B. lentus CP28 generated higher urease activity and facilitated more efficient precipitation of calcium carbonate at 3.24 ± 0.25 × 10(-4) mg/cell. X-ray diffraction indicated that the dominant calcium carbonate phase was calcite. Scanning electron microscopy showed that morphologies of the minerals were dominated by cubic, rhombic and polygonal plate-like crystals. The dynamic process of microbial calcium carbonate precipitation revealed that B. lentus CP28 precipitated calcite crystals through the enzymatic hydrolysis of urea, and that when ammonium ion concentrations reached 746 mM and the pH reached 9.6, that favored calcite precipitation at a higher level of 96 mg/L. The results of this research provide evidence that a variety of marine bacteria can induce calcium carbonate precipitation, and may influence the marine carbonate cycle in natural environments.

  19. Biomineralization processes of calcite induced by bacteria isolated from marine sediments

    Directory of Open Access Journals (Sweden)

    Shiping Wei

    2015-06-01

    Full Text Available Biomineralization is a known natural phenomenon associated with a wide range of bacterial species. Bacterial-induced calcium carbonate precipitation by marine isolates was investigated in this study. Three genera of ureolytic bacteria, Sporosarcina sp., Bacillus sp. and Brevundimonas sp. were observed to precipitate calcium carbonate minerals. Of these species, Sporosarcina sp. dominated the cultured isolates. B. lentus CP28 generated higher urease activity and facilitated more efficient precipitation of calcium carbonate at 3.24 ± 0.25 × 10−4 mg/cell. X-ray diffraction indicated that the dominant calcium carbonate phase was calcite. Scanning electron microscopy showed that morphologies of the minerals were dominated by cubic, rhombic and polygonal plate-like crystals. The dynamic process of microbial calcium carbonate precipitation revealed that B. lentus CP28 precipitated calcite crystals through the enzymatic hydrolysis of urea, and that when ammonium ion concentrations reached 746 mM and the pH reached 9.6, that favored calcite precipitation at a higher level of 96 mg/L. The results of this research provide evidence that a variety of marine bacteria can induce calcium carbonate precipitation, and may influence the marine carbonate cycle in natural environments.

  20. The Increasing Interest of ANAMMOX Research in China: Bacteria, Process Development, and Application

    Directory of Open Access Journals (Sweden)

    Mohammad Ali

    2013-01-01

    Full Text Available Nitrogen pollution created severe environmental problems and increasingly has become an important issue in China. Since the first discovery of ANAMMOX in the early 1990s, this related technology has become a promising as well as sustainable bioprocess for treating strong nitrogenous wastewater. Many Chinese research groups have concentrated their efforts on the ANAMMOX research including bacteria, process development, and application during the past 20 years. A series of new and outstanding outcomes including the discovery of new ANAMMOX bacterial species (Brocadia sinica, sulfate-dependent ANAMMOX bacteria (Anammoxoglobus sulfate and Bacillus benzoevorans, and the highest nitrogen removal performance (74.3–76.7 kg-N/m3/d in lab scale granule-based UASB reactors around the world were achieved. The characteristics, structure, packing pattern and floatation mechanism of the high-rate ANAMMOX granules in ANAMMOX reactors were also carefully illustrated by native researchers. Nowadays, some pilot and full-scale ANAMMOX reactors were constructed to treat different types of ammonium-rich wastewater including monosodium glutamate wastewater, pharmaceutical wastewater, and leachate. The prime objective of the present review is to elucidate the ongoing ANAMMOX research in China from lab scale to full scale applications, comparative analysis, and evaluation of significant findings and to set a design to usher ANAMMOX research in culmination.

  1. Erythrocyte lysis in isotonic solution of ammonium chloride: theoretical modeling and experimental verification.

    Science.gov (United States)

    Chernyshev, Andrey V; Tarasov, Peter A; Semianov, Konstantin A; Nekrasov, Vyacheslav M; Hoekstra, Alfons G; Maltsev, Valeri P

    2008-03-07

    A mathematical model of erythrocyte lysis in isotonic solution of ammonium chloride is presented in frames of a statistical approach. The model is used to evaluate several parameters of mature erythrocytes (volume, surface area, hemoglobin concentration, number of anionic exchangers on membrane, elasticity and critical tension of membrane) through their sphering and lysis measured by a scanning flow cytometer (SFC). SFC allows measuring the light-scattering pattern (indicatrix) of an individual cell over the angular range from 10 degrees to 60 degrees . Comparison of the experimentally measured and theoretically calculated light scattering patterns allows discrimination of spherical from non-spherical erythrocytes and evaluation of volume and hemoglobin concentration for individual spherical cells. Three different processes were applied for erythrocytes sphering: (1) colloid osmotic lysis in isotonic solution of ammonium chloride, (2) isovolumetric sphering in the presence of sodium dodecyl sulphate and albumin in neutrally buffered isotonic saline, and (3) osmotic fragility test in hypotonic media. For the hemolysis in ammonium chloride, the evolution of distributions of sphered erythrocytes on volume and hemoglobin content was monitored in real-time experiments. The analysis of experimental data was performed in the context of a statistical approach, taking into account that parameters of erythrocytes vary from cell to cell.

  2. Detecting cell lysis using viscosity monitoring in E. coli fermentation to prevent product loss.

    Science.gov (United States)

    Newton, Joseph M; Schofield, Desmond; Vlahopoulou, Joanna; Zhou, Yuhong

    2016-07-01

    Monitoring the physical or chemical properties of cell broths to infer cell status is often challenging due to the complex nature of the broth. Key factors indicative of cell status include cell density, cell viability, product leakage, and DNA release to the fermentation broth. The rapid and accurate prediction of cell status for hosts with intracellular protein products can minimise product loss due to leakage at the onset of cell lysis in fermentation. This article reports the rheological examination of an industrially relevant E. coli fermentation producing antibody fragments (Fab'). Viscosity monitoring showed an increase in viscosity during the exponential phase in relation to the cell density increase, a relatively flat profile in the stationary phase, followed by a rapid increase which correlated well with product loss, DNA release and loss of cell viability. This phenomenon was observed over several fermentations that a 25% increase in broth viscosity (using induction-point viscosity as a reference) indicated 10% product loss. Our results suggest that viscosity can accurately detect cell lysis and product leakage in postinduction cell cultures, and can identify cell lysis earlier than several other common fermentation monitoring techniques. This work demonstrates the utility of rapidly monitoring the physical properties of fermentation broths, and that viscosity monitoring has the potential to be a tool for process development to determine the optimal harvest time and minimise product loss. © 2016 The Authors. Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers, 32:1069-1076, 2016.

  3. Comparison of different disinfection processes in the effective removal of antibiotic-resistant bacteria and genes.

    Science.gov (United States)

    Oh, Junsik; Salcedo, Dennis Espineli; Medriano, Carl Angelo; Kim, Sungpyo

    2014-06-01

    This study compared three different disinfection processes (chlorination, E-beam, and ozone) and the efficacy of three oxidants (H2O2, S2O(-)8, and peroxymonosulfate (MPS)) in removing antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in a synthetic wastewater. More than 30 mg/L of chlorine was needed to remove over 90% of ARB and ARG. For the E-beam method, only 1 dose (kGy) was needed to remove ARB and ARG, and ozone could reduce ARB and ARG by more than 90% even at 3 mg/L ozone concentration. In the ozone process, CT values (concentration × time) were compared for ozone alone and combined with different catalysts based on the 2-log removal of ARB and ARG. Ozone treatment yielded a value of 31 and 33 (mg·min)/L for ARB and ARGs respectively. On the other hand, ozone with persulfate yielded 15.9 and 18.5 (mg·min)/L while ozone with monopersulfate yielded a value of 12 and 14.5 (mg·min)/L. This implies that the addition of these catalysts significantly reduces the contact time to achieve a 2-log removal, thus enhancing the process in terms of its kinetics.

  4. Performance and dye-degrading bacteria isolation of a hybrid membrane process

    Energy Technology Data Exchange (ETDEWEB)

    You, Sheng-Jie, E-mail: sjyou@cycu.edu.tw [Department of Bioenvironmental Engineering and R and D Center for Membrane Technology, Chung Yuan Christian University, No. 200, Rd. Chung-Pei, Chungli 320, Taiwan (China); Teng, Jun-Yu, E-mail: nickprometheus@yahoo.com.tw [Department of Civil Engineering, Chung Yuan Christian University, Chungli 320, Taiwan (China)

    2009-12-15

    Textile dyeing wastewater contains harmful compounds, which are toxic to both marine organisms and human beings if it discharged into an aquatic environmental without suitable treatment. In this study, the wastewater containing the azo dye, Reactive Black 5 (RB5), was partially treated in an anaerobic sequencing batch reactor which was further treated either in an aerobic membrane bioreactors (AOMBR) or in combined aerobic membrane bioreactor/reverse osmosis (AOMBR/RO) process. The results showed that in the anaerobic sequencing batch reactor the RB5 dye was degraded to form aromatic amine intermediate metabolites, which were further mineralized in the AOMBR. It was also observed that although all effluents from the AOMBR and AOMBR/RO processes met the Taiwan EPA's effluent criteria, irrespective of which membranes were used in the aerobic tank, the effluent from the AOMBR/RO process met the criteria for reuse for toilet flushing, landscaping, irrigation, and cooling water purposes, where as the AOMBR effluent only met the criteria for cooling water due to incomplete color removal. Five anaerobic high dye-degrading bacteria were isolated, which were identified to be the same species of Lactococcus lactis by 16S rRNA sequencing. The L. lactis showed complete degradation of RB5 and further studies showed that it can also able to degrade Reactive Red 120 and Reactive Yellow 84 efficiently within 6 h.

  5. Excretion of cytoplasmic proteins in Staphylococcus is most likely not due to cell lysis.

    Science.gov (United States)

    Ebner, Patrick; Rinker, Janina; Götz, Friedrich

    2016-02-01

    The excretion of cytoplasmic proteins (ECP) is a long-known phenomenon in bacteria and eukaryotes. So far, it was not possible to associate either a signal peptide-dependent or a signal peptide-independent pathway to ECP. Nevertheless 25% of the proteins found in Staphylococcus aureus supernatants were cytoplasmic proteins. Because the excreted proteins do not possess a common motive, the most widespread opinion is that ECP is due to cell lysis. This explanation seems to be too easy since several indications imply that there exists a yet unknown mechanism for ECP. Certainly, the up-regulation of autolysins as well as decreased peptidoglycan cross-linking increased ECP. However, in recent years, several evidences arose that cell lysis is not the only reason for ECP. It seems that ECP is a part of the normal cell cycle of S. aureus as it turned out that ECP with several model proteins occurs mainly during cell growth. It has common features as proteins secreted via the Sec translocon and finally the excretion site is the cross wall of dividing cells.

  6. Intermittent filtration of bacteria and colloids in porous media

    Science.gov (United States)

    Auset, Maria; Keller, Arturo A.; Brissaud, François; Lazarova, Valentina

    2005-09-01

    Intermittent filtration through porous media used for water and wastewater treatment can achieve high pathogen and colloid removal efficiencies. To predict the removal of bacteria, the effects of cyclic infiltration and draining events (transient unsaturated flow) were investigated. Using physical micromodels, we visualized the intermittent transport of bacteria and other colloids in unsaturated porous media. Column experiments provided quantitative measurements of the phenomena observed at the pore scale. Tagged Escherichia coli and a conservative tracer (NaI) were introduced in an initial pulse into a 1.5 m sand column. Subsequent hydraulic flushes without tagged bacteria or tracer were repeated every 4 hours for the next 4 days, during which outflow concentrations were monitored. Breakthrough behavior between colloids and dissolved tracer differed significantly, reflecting the differences in transport processes. Advancement of the wetting front remobilized bacteria which were held in thin water films, attached to the air-water interface (AWI), or entrapped in stagnant pore water between gas bubbles. In contrast, the tracer was only remobilized by diffusion from immobile to mobile water. Remobilization led to successive concentration peaks of bacteria and tracer in the effluent but with significant temporal differences. Observations at the pore-scale indicated that the colloids were essentially irreversibly attached to the solid-water interface, which explained to some extent the high removal efficiency of microbes in the porous media. Straining, cluster filtration, cell lysis, protozoa grazing, and bacteriophage parasitism could also contribute to the removal efficiency of bacteria.

  7. Process for Generation of Hydrogen Gas from Various Feedstocks Using Thermophilic Bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Ooteghem Van, Suellen

    2005-09-13

    A method for producing hydrogen gas is provided comprising selecting a bacteria from the Order Thermotogales, subjecting the bacteria to a feedstock and to a suitable growth environment having an oxygen concentration below the oxygen concentration of water in equilibrium with air; and maintaining the environment at a predetermined pH and at a temperature of at least approximately 45 degrees C. for a time sufficient to allow the bacteria to metabolize the feedstock.

  8. Specific biofilter process design using bacteria capable of removing hydrogen sulphide from air emissions

    Energy Technology Data Exchange (ETDEWEB)

    Coleman, R.N. [Alberta Environmental Centre, Vegreville, AB (Canada)

    1996-05-01

    The design and development of a biofilter unit which can remove hydrogen sulphide (H{sub 2}S) from air emissions, such as those found in the oil and gas industry and the Kraft wood pulping process, was discussed. Biofilters which contain species of bacteria which are able to oxidize H{sub 2}S have been very successful in removing high concentrations of H{sub 2}S from contaminated air streams. The biofilters could control noxious or offensive odours through aerobic metabolism of contaminants by microorganisms attached to such surfaces as peat or wood bark. Microbial oxidation of low molecular weight inorganic and volatile organic compounds were shown to result in their degradation to end products such as carbon dioxide, sulphate, water and new cell biomass. Research was directed toward the treatment of sulphur containing emissions generated during the sulphide recovery cycle in the Kraft wood pulping process, but since emissions are similar to those present in oil and gas industry, this technology was said to be equally applicable in the petroleum industry. 14 refs., 5 tabs., 8 figs.

  9. Tracking spore-forming bacteria in food: from natural biodiversity to selection by processes.

    Science.gov (United States)

    Postollec, Florence; Mathot, Anne-Gabrielle; Bernard, Muriel; Divanac'h, Marie-Laure; Pavan, Sonia; Sohier, Danièle

    2012-08-01

    Sporeforming bacteria are ubiquitous in the environment and exhibit a wide range of diversity leading to their natural prevalence in foodstuff. The state of the art of sporeformer prevalence in ingredients and food was investigated using a multiparametric PCR-based tool that enables simultaneous detection and identification of various genera and species mostly encountered in food, i.e., Alicyclobacillus, Anoxybacillus flavithermus, Bacillus, B. cereus group, B. licheniformis, B. pumilus, B. sporothermodurans, B. subtilis, Brevibacillus laterosporus, Clostridium, Geobacillus stearothermophilus, Moorella and Paenibacillus species. In addition, 16S rDNA sequencing was used to extend identification to other possibly present contaminants. A total of 90 food products, with or without visible trace of spoilage were analysed, i.e., 30 egg-based products, 30 milk and dairy products and 30 canned food and ingredients. Results indicated that most samples contained one or several of the targeted genera and species. For all three tested food categories, 30 to 40% of products were contaminated with both Bacillus and Clostridium. The percentage of contaminations associated with Clostridium or Bacillus represented 100% in raw materials, 72% in dehydrated ingredients and 80% in processed foods. In the last two product types, additional thermophilic contaminants were identified (A. flavithermus, Geobacillus spp., Thermoanaerobacterium spp. and Moorella spp.). These results suggest that selection, and therefore the observed (re)-emergence of unexpected sporeforming contaminants in food might be favoured by the use of given food ingredients and food processing technologies.

  10. A mathematical model for expected time to extinction of pathogenic bacteria through antibiotic

    Science.gov (United States)

    Ghosh, M. K.; Nandi, S.; Roy, P. K.

    2016-04-01

    Application of antibiotics in human system to prevent bacterial diseases like Gastritis, Ulcers, Meningitis, Pneumonia and Gonorrhea are indispensable. Antibiotics saved innumerable lives and continue to be a strong support for therapeutic application against pathogenic bacteria. In human system, bacterial diseases occur when pathogenic bacteria gets into the body and begin to reproduce and crowd out healthy bacteria. In this process, immature bacteria releases enzyme which is essential for bacterial cell-wall biosynthesis. After complete formation of cell wall, immature bacteria are converted to mature or virulent bacteria which are harmful to us during bacterial infections. Use of antibiotics as drug inhibits the bacterial cell wall formation. After application of antibiotics within body, the released bacterial enzyme binds with antibiotic molecule instead of its functional site during the cell wall synthesis in a competitive inhibition approach. As a consequence, the bacterial cell-wall formation as well as maturation process of pathogenic bacteria is halted and the disease is cured with lysis of bacterial cells. With this idea, a mathematical model has been developed in the present research investigation to review the inhibition of biosynthesis of bacterial cell wall by the application of antibiotics as drug in the light of enzyme kinetics. This approach helps to estimate the expected time to extinction of the pathogenic bacteria. Our mathematical approach based on the enzyme kinetic model for finding out expected time to extinction contributes favorable results for understanding of disease dynamics. Analytical and numerical results based on simulated findings validate our mathematical model.

  11. Sulfate reducing bacteria and their activities in oil sands process-affected water biofilm

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Hong; Yu, Tong, E-mail: tong.yu@ualberta.ca; Liu, Yang, E-mail: yang.liu@ualberta.ca

    2015-12-01

    Biofilm reactors were constructed to grow stratified multispecies biofilm in oil sands process-affected water (OSPW) supplemented with growth medium. The development of sulfate reducing bacteria (SRB) within the biofilm and the biofilm treatment of OSPW were evaluated. The community structure and potential activity of SRB in the biofilm were investigated with H{sub 2}S microsensor measurements, dsrB gene-based denaturing gradient gel electrophoresis (DGGE), and the real time quantitative polymerase chain reaction (qPCR). Multispecies biofilm with a thickness of 1000 μm was successfully developed on engineered biocarriers. H{sub 2}S production was observed in the deeper anoxic zone of the biofilm from around 750 μm to 1000 μm below the bulk water-biofilm interface, revealing sulfate reduction in the deeper zone of the stratified biofilm. The biofilm removed chemical oxygen demand (COD), sulfate, and nitrogen. The study expands current knowledge of biofilm treatment of OSPW and the function of anaerobic SRB in OSPW biofilm, and thus provides information for future bioreactor development in the reclamation of OSPW. - Graphical abstract: The development of sulfate reducing bacteria (SRB) within Oil Sands Process-affected Water (OSPW) biofilm and the biofilm treatment of OSPW were evaluated by Liu and coworkers. Combined microsensor and molecular biology techniques were utilized in this study. Their results demonstrated that multispecies biofilm with a thickness of 1000 μm was successfully developed on engineered biocarriers. H{sub 2}S production was observed in the deeper anoxic zone of the biofilm from around 750 μm to 1000 μm below the bulk water-biofilm interface, revealing sulfate reduction in the deeper zone of the biofilm. The biofilm removed chemical oxygen demand (COD), sulfate, and nitrogen. - Highlights: • Biofilm in oil sands wastewater was developed on engineered biocarriers. • Bacterial community and in situ activity of SRB were studied in the

  12. Sulfate reducing bacteria and their activities in oil sands process-affected water biofilm.

    Science.gov (United States)

    Liu, Hong; Yu, Tong; Liu, Yang

    2015-12-01

    Biofilm reactors were constructed to grow stratified multispecies biofilm in oil sands process-affected water (OSPW) supplemented with growth medium. The development of sulfate reducing bacteria (SRB) within the biofilm and the biofilm treatment of OSPW were evaluated. The community structure and potential activity of SRB in the biofilm were investigated with H2S microsensor measurements, dsrB gene-based denaturing gradient gel electrophoresis (DGGE), and the real time quantitative polymerase chain reaction (qPCR). Multispecies biofilm with a thickness of 1000 μm was successfully developed on engineered biocarriers. H2S production was observed in the deeper anoxic zone of the biofilm from around 750 μm to 1000 μm below the bulk water-biofilm interface, revealing sulfate reduction in the deeper zone of the stratified biofilm. The biofilm removed chemical oxygen demand (COD), sulfate, and nitrogen. The study expands current knowledge of biofilm treatment of OSPW and the function of anaerobic SRB in OSPW biofilm, and thus provides information for future bioreactor development in the reclamation of OSPW.

  13. Influence of water hardness on the ability of water to rinse bacteria from the skin of processed broilers

    Science.gov (United States)

    Experiments were conducted to examine the effect of water hardness on the ability of water to rinse bacteria from the skin of processed broiler chickens. Very hard water (200 ppm total hardness) was prepared by dissolving 0.38 g calcium chloride (CaCl2) and 0.175 g magnesium chloride hexahydrate (Mg...

  14. Effect of water hardness on the ability of water to rinse bacteria from the skin of processed broilers

    Science.gov (United States)

    The effect of water hardness on the ability of water to rinse bacteria from the skin of processed broiler chickens was examined. Artificial hard water with a total hardness of 200 ppm (very hard water) was prepared by dissolving calcium chloride (CaCl2) and magnesium chloride hexahydrate (MgCl2 •6H2...

  15. Metagenomics as a tool to obtain full genomes of process-critical bacteria in engineered systems

    DEFF Research Database (Denmark)

    Albertsen, Mads; Hugenholtz, Philip; Tyson, Gene W.;

    Bacteria play a pivotal role in engineered systems such as wastewater treatment plants. Obtaining genomes of the bacteria provides the genetic potential of the system and also allows studies of in situ functions through transcriptomics and proteomics. Hence, it enables correlations of operational...... parameters with functions of specific bacteria within the ecosystems in order to decipher principles that might be used to control and predict ecosystem performance. The main bottleneck in obtaining genomes from the environment is that the vast majority of bacteria are not readily cultured. Metagenomics......, the sequencing of bulk genomic DNA from environmental samples, has the potential to provide genomes of this uncultured majority. However, so far only few bacterial genomes have been obtained from metagenomic data. In this study we present a new approach to obtain individual genomes from metagenomes. We deeply...

  16. Characterization of aerobic spore-forming bacteria associated with industrial dairy processing environments and product spoilage.

    Science.gov (United States)

    Lücking, Genia; Stoeckel, Marina; Atamer, Zeynep; Hinrichs, Jörg; Ehling-Schulz, Monika

    2013-09-02

    Due to changes in the design of industrial food processing and increasing international trade, highly thermoresistant spore-forming bacteria are an emerging problem in food production. Minimally processed foods and products with extended shelf life, such as milk products, are at special risk for contamination and subsequent product damages, but information about origin and food quality related properties of highly heat-resistant spore-formers is still limited. Therefore, the aim of this study was to determine the biodiversity, heat resistance, and food quality and safety affecting characteristics of aerobic spore-formers in the dairy sector. Thus, a comprehensive panel of strains (n=467), which originated from dairy processing environments, raw materials and processed foods, was compiled. The set included isolates associated with recent food spoilage cases and product damages as well as isolates not linked to product spoilage. Identification of the isolates by means of Fourier-transform infrared spectroscopy and molecular methods revealed a large biodiversity of spore-formers, especially among the spoilage associated isolates. These could be assigned to 43 species, representing 11 genera, with Bacillus cereus s.l. and Bacillus licheniformis being predominant. A screening for isolates forming thermoresistant spores (TRS, surviving 100°C, 20 min) showed that about one third of the tested spore-formers was heat-resistant, with Bacillus subtilis and Geobacillus stearothermophilus being the prevalent species. Strains producing highly thermoresistant spores (HTRS, surviving 125°C, 30 min) were found among mesophilic as well as among thermophilic species. B. subtilis and Bacillus amyloliquefaciens were dominating the group of mesophilic HTRS, while Bacillus smithii and Geobacillus pallidus were dominating the group of thermophilic HTRS. Analysis of spoilage-related enzymes of the TRS isolates showed that mesophilic strains, belonging to the B. subtilis and B. cereus

  17. Role of Protein Phosphorylation in the Regulation of Cell Cycle and DNA-Related Processes in Bacteria

    DEFF Research Database (Denmark)

    Garcia-Garcia, Transito; Poncet, Sandrine; Derouiche, Abderahmane;

    2016-01-01

    replication during the cell cycle, as well as in the mechanisms that cope with stress-induced replication blocks. Similar to eukaryotes, bacteria use Hanks-type kinases and phosphatases for signal transduction, and protein phosphorylation is involved in numerous cellular processes. However, it remains unclear...... the bacterial cell cycle. Recent phosphoproteomics and interactomics studies identified numerous phosphoproteins involved in various aspect of DNA metabolism strongly supporting the existence of such level of regulation in bacteria. Similar to eukaryotes, bacterial scaffolding-like proteins emerged as platforms...... for kinase activation and signaling. This review reports the current knowledge on the phosphorylation of proteins involved in the maintenance of genome integrity and the regulation of cell cycle in bacteria that reveals surprising similarities to eukaryotes....

  18. Decreasing ammonium generation using hydrogenotrophic bacteria in the process of nitrate reduction by nanoscale zero-valent iron

    Energy Technology Data Exchange (ETDEWEB)

    An, Yi; Li, Tielong [College of Environmental Science and Engineering/Tianjin Key Laboratory of Environmental Remediation and Pollution Control/Ministry of Education Key Laboratory of Pollution Processed and Environmental Criteria, Nankai University, Tianjin 300071 (China); Jin, Zhaohui, E-mail: jinzh@nankai.edu.cn [College of Environmental Science and Engineering/Tianjin Key Laboratory of Environmental Remediation and Pollution Control/Ministry of Education Key Laboratory of Pollution Processed and Environmental Criteria, Nankai University, Tianjin 300071 (China); Dong, Meiying; Li, Qianqian; Wang, Shuaima [College of Environmental Science and Engineering/Tianjin Key Laboratory of Environmental Remediation and Pollution Control/Ministry of Education Key Laboratory of Pollution Processed and Environmental Criteria, Nankai University, Tianjin 300071 (China)

    2009-10-15

    An integrated nitrate treatment using nanoscale zero-valent iron (NZVI) and Alcaligenes eutrophus, which is a kind of hydrogenotrophic denitrifying bacteria, was conducted to remove nitrate and decrease ammonium generation. Within 8 days, nitrate was removed completely in the reactors containing NZVI particles plus bacteria while the proportion of ammonium generated was only 33%. That is a lower reduction rate but a smaller proportion of ammonium relative to that in abiotic reactors. It was also found that ammonium generation experienced a biphasic process, involving an increasing period and a stable period. After domestication of the bacteria, the combined NZVI-cell system could remove all nitrate without ammonium released when the refreshed nitrate was introduced. Nitrate reduction and the final product distribution were also studied in batch reactors amended with different initial NZVI contents and biomass concentrations, respectively. Both the nitrate removal rate and the ammonium yield decreased when the initial content of NZVI reduced and the initial biomass concentration increased. However, about 27% of the nitrate was converted to ammonium when excess bacteria (OD{sub 422} = 0.026) were used, which was higher than that with appropriate amount of bacteria.

  19. Fibrin clot formation and lysis: basic mechanisms

    DEFF Research Database (Denmark)

    Sidelmann, JJ; Gram, J; Jespersen, J;

    2000-01-01

    consequently is an important substrate in the physiology of hemostasis. This review describes the components and processes involved in fibrin formation and fibrin degradation. Particular emphasis is put on the reactions involved in the conversion of fibrinogen to fibrin, the polymerization of fibrin molecules...... induced by coagulation factor XIII (FXIII), and the degradation of fibrinogen and fibrin mediated by plasmin and elastase. Furthermore, factors influencing fibrin structure and fibrin breakdown are addressed; in particular polymorphisms in the genes coding for fibrinogen and FXIII, but also the physical...

  20. Direct Cell Lysis for Single-Cell Gene Expression Profiling

    OpenAIRE

    David eSvec; Daniel eAndersson; Milos ePekny; Robert eSjöback; Mikael eKubista; Anders eStåhlberg

    2013-01-01

    The interest to analyze single and few cell samples is rapidly increasing. Numerous extraction protocols to purify nucleic acids are available, but most of them compromise severely on yield to remove contaminants and are therefore not suitable for the analysis of samples containing small numbers of transcripts only. Here, we evaluate 17 direct cell lysis protocols for transcript yield and compatibility with downstream reverse transcription quantitative real-time PCR. Four endogenously express...

  1. Dark membrane lysis and photosensitization by 3-carbethoxypsoralen

    Energy Technology Data Exchange (ETDEWEB)

    Muller-Runkel, R.; Grossweiner, L.I. (Illinois Inst. of Tech., Chicago (USA). Dept. of Physics)

    1981-03-01

    Aqueous solutions of 3-carbethoxypsoralen (3-CPs) induced lysis of egg lecithin liposomes and whole human erythrocytes in the dark. Near-UV irradiation of 3-CPs sensitized the inactivation of lysozyme attributed to the production of reactive radical intermediates. The implications of these findings for the use of 3-CPs as a sensitizer in psoralen + UV-A (PUVA) therapy of psoriasis are discussed.

  2. Effect of boiling water carcass immersion on aerobic bacteria counts of poultry skin and processed ground poultry meat.

    Science.gov (United States)

    Tompkins, N M; Avens, J S; Kendall, P A; Salman, M D

    2008-06-01

    This study was conducted to determine the relationship between bacteria destruction on poultry carcass skin and bacteria in raw ground poultry meat from the same carcasses. Immersion time in boiling water of broiler chicken whole carcasses required for maximum reduction of naturally occurring aerobic bacterial count on skin was measured. Treatments for chicken carcasses consisted of immersion in boiling water (approximately 95 degrees C) for 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4 min. Four skin samples taken following treatment and three taken from subsequently ground carcass meat were analyzed for total aerobic plate counts (APC). Analysis of the data indicated a linear increase in bacterial destruction on skin with increased boiling water immersion time from 0 to 4 min. Reduction of skin bacteria to less than 1 log10 occurred at 3 min carcass immersion or longer. The analysis also indicated that treatment with boiling water and removal of skin was effective in reducing bacterial counts in ground meat to similar levels at all treatment times from 0.5 to 4.0 min. Findings from this study indicated that a boiling water immersion intervention and removal of skin could reduce subsequent bacteria contamination of ground meat. This intervention could minimize the risk of pathogen-contaminated primary processed poultry carcasses used in further processing.

  3. [Community Characteristics of ANAMMOX Bacteria in Subsurface Flow Constructed Wetland (SSFCW) for Processing of Aquaculture Waster Water].

    Science.gov (United States)

    Zeng, Xian-lei; Liu, Xing-guo; Wu, Zong-fan; Shi, Xu; Lu, Shi-min

    2016-02-15

    Anaerobic ammonium oxidation (ANAMMOX) is one of the important functions in waste water treatment by subsurface flow constructed wetland (SSFCW), however, there are few studies on ANAMMOX in SSFCW environment at present. The community characteristics of ANAMMOX in the SSFCW of processing aquaculture waste water were explored in this study. In order to analyze the structure, diversity and abundance of ANAMMOX bacteria, several 16S rRNA clone libraries were constructed and real-time PCR targeting specific 16S rRNA genes together with diversity analysis was adopted. The obtained results showed that the SSFCW identified a total of three unknown clusters and two known clusters including Candidatus brocadia and Candidatus kuenenia. The dominant cluster was Candidatus brocadia. The highest diversity levels of ANAMMOX bacteria occurred in autumn (H', 1.21), while the lowest in spring (H', 0.64). The abundance of ANAMMOX bacteria in SSFCW environment ranged from 8.0 x 10(4) to 9.4 x 10(6) copies x g(-1) of fresh weight and the copy number of total bacterial 16S rRNA genes ranged from 7.3 x 10(9) to 9.1 x 10(10) copies x g(-1) of fresh weight during culture cycle. There were significant differences in the ANAMMOX bacteria abundances of different stratum and seasons in SSFCW environment, but the differences in total bacterial abundances were not obvious. In addition, the differences in ANAMMOX bacteria abundances in different stratum and seasons in SSFCW environment were irregular in different culture cycle. According to the distribution characteristics of ANAMMOX bacteria in the wetland, the denitrification effect of SSFCW could be improved by changing the supplying manners of aquaculture wastewater and adjusting the structure of wetland. The research results will provide reference for further optimizing the SSFCW and improving the efficiency of purification.

  4. Bacteriophage cell lysis of Shiga toxin-producing Escherichia coli for top-down proteomic identification of Shiga toxin 1 & 2 using matrix-assisted laser desorption/ionization tandem time-of-light mass spectrometry

    Science.gov (United States)

    RATIONALE: Analysis of bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) often relies upon sample preparation methods that result in cell lysis, e.g. bead-beating. However, Shiga toxin-producing Escherichia coli (STEC) can undergo bacteriophage...

  5. Information processing in bacteria: memory, computation, and statistical physics: a key issues review

    Science.gov (United States)

    Lan, Ganhui; Tu, Yuhai

    2016-05-01

    preserving information, it does not reveal the underlying mechanism that leads to the observed input-output relationship, nor does it tell us much about which information is important for the organism and how biological systems use information to carry out specific functions. To do that, we need to develop models of the biological machineries, e.g. biochemical networks and neural networks, to understand the dynamics of biological information processes. This is a much more difficult task. It requires deep knowledge of the underlying biological network—the main players (nodes) and their interactions (links)—in sufficient detail to build a model with predictive power, as well as quantitative input-output measurements of the system under different perturbations (both genetic variations and different external conditions) to test the model predictions to guide further development of the model. Due to the recent growth of biological knowledge thanks in part to high throughput methods (sequencing, gene expression microarray, etc) and development of quantitative in vivo techniques such as various florescence technology, these requirements are starting to be realized in different biological systems. The possible close interaction between quantitative experimentation and theoretical modeling has made systems biology an attractive field for physicists interested in quantitative biology. In this review, we describe some of the recent work in developing a quantitative predictive model of bacterial chemotaxis, which can be considered as the hydrogen atom of systems biology. Using statistical physics approaches, such as the Ising model and Langevin equation, we study how bacteria, such as E. coli, sense and amplify external signals, how they keep a working memory of the stimuli, and how they use these data to compute the chemical gradient. In particular, we will describe how E. coli cells avoid cross-talk in a heterogeneous receptor cluster to keep a ligand-specific memory. We will also

  6. Information processing in bacteria: memory, computation, and statistical physics: a key issues review.

    Science.gov (United States)

    Lan, Ganhui; Tu, Yuhai

    2016-05-01

    preserving information, it does not reveal the underlying mechanism that leads to the observed input-output relationship, nor does it tell us much about which information is important for the organism and how biological systems use information to carry out specific functions. To do that, we need to develop models of the biological machineries, e.g. biochemical networks and neural networks, to understand the dynamics of biological information processes. This is a much more difficult task. It requires deep knowledge of the underlying biological network-the main players (nodes) and their interactions (links)-in sufficient detail to build a model with predictive power, as well as quantitative input-output measurements of the system under different perturbations (both genetic variations and different external conditions) to test the model predictions to guide further development of the model. Due to the recent growth of biological knowledge thanks in part to high throughput methods (sequencing, gene expression microarray, etc) and development of quantitative in vivo techniques such as various florescence technology, these requirements are starting to be realized in different biological systems. The possible close interaction between quantitative experimentation and theoretical modeling has made systems biology an attractive field for physicists interested in quantitative biology. In this review, we describe some of the recent work in developing a quantitative predictive model of bacterial chemotaxis, which can be considered as the hydrogen atom of systems biology. Using statistical physics approaches, such as the Ising model and Langevin equation, we study how bacteria, such as E. coli, sense and amplify external signals, how they keep a working memory of the stimuli, and how they use these data to compute the chemical gradient. In particular, we will describe how E. coli cells avoid cross-talk in a heterogeneous receptor cluster to keep a ligand-specific memory. We will also

  7. Interspecies interactions result in enhanced biofilm formation by co-cultures of bacteria isolated from a food processing environment

    DEFF Research Database (Denmark)

    Røder, Henriette Lyng; Raghupathi, Prem Krishnan; Herschend, Jakob

    2015-01-01

    examined for multispecies biofilm formation. Eight strains from each sampling site were chosen and all possible combinations of four member co-cultures were tested for enhanced biofilm formation at 15°C and 24°C. In approximately 20% of the multispecies consortia grown at 15°C, the biofilm formation......Bacterial attachment and biofilm formation can lead to poor hygienic conditions in food processing environments. Furthermore, interactions between different bacteria may induce or promote biofilm formation. In this study, we isolated and identified a total of 687 bacterial strains from seven...... different locations in a meat processing environment and evaluated their biofilm formation capability. A diverse group of bacteria was isolated and most were classified as poor biofilm producers in a Calgary biofilm device assay. Isolates from two sampling sites, the wall and the meat chopper, were further...

  8. Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells

    Directory of Open Access Journals (Sweden)

    Wagner Carsten A

    2007-08-01

    Full Text Available Abstract Background Cytotoxic T lymphocytes (CTL and natural killer (NK cells play an essential role in the host defence against intracellular pathogens such as Listeria, and Mycobacteria. The key mediator of bacteria-directed cytotoxicity is granulysin, a 9 kDa protein stored in cytolytic granules together with perforin and granzymes. Granulysin binds to cell membranes and is subsequently taken up via a lipid raft-associated mechanism. In dendritic cells (DC granulysin is further transferred via early endosomes to L. innocua-containing phagosomes were bacteriolysis is induced. In the present study we analysed the role of perforin in granulysin-induced intracellular bacteriolysis in DC. Results We found granulysin-induced lysis of intracellular Listeria significantly increased when perforin was simultaneously present. In pulse-chase experiments enhanced bacteriolysis was observed when perforin was added up to 25 minutes after loading the cells with granulysin demonstrating no ultimate need for simultaneous uptake of granulysin and perforin. The perforin concentration sufficient to enhance granulysin-induced intracellular bacteriolysis did not cause permanent membrane pores in Listeria-challenged DC as shown by dye exclusion test and LDH release. This was in contrast to non challenged DC that were more susceptible to perforin lysis. For Listeria-challenged DC, there was clear evidence for an Ca2+ influx in response to sublytic perforin demonstrating a short-lived change in the plasma membrane permeability. Perforin treatment did not affect granulysin binding, initial uptake or intracellular trafficking to early endosomes. However, enhanced colocalization of granulysin with listerial DNA in presence of perforin was found by confocal laser scanning microscopy. Conclusion The results provide evidence that perforin increases granulysin-mediated killing of intracellular Listeria by enhanced phagosome-endosome fusion triggered by a transient Ca2+ flux.

  9. Isolation, identification and optimization of ethanol producing bacteria from Saccharomyces-based fermentation process of alcohol industries in Iran

    OpenAIRE

    Hoda Ebrahimi; Mojtaba Mohseni

    2013-01-01

    Introduction: Due to the vast growth of world population, consumption of a lot of energy, limited energy supply and rising prices of fuel oil in the future, other alternative energy source is essential. Ethanol is renewable and a safe fuel and it is mainly based on microbial fermentation. The purpose of this study was isolation of high ethanol producing bacteria from the fermentation process of alcohol industries and optimization of growth conditions to be introduced to the industries. Materi...

  10. Deciphering How Pore Formation Causes Strain-Induced Membrane Lysis of Lipid Vesicles.

    Science.gov (United States)

    Jackman, Joshua A; Goh, Haw Zan; Zhdanov, Vladimir P; Knoll, Wolfgang; Cho, Nam-Joon

    2016-02-01

    Pore formation by membrane-active antimicrobial peptides is a classic strategy of pathogen inactivation through disruption of membrane biochemical gradients. It remains unknown why some membrane-active peptides also inhibit enveloped viruses, which do not depend on biochemical gradients. Here, we employ a label-free biosensing approach based on simultaneous quartz crystal microbalance-dissipation and ellipsometry measurements in order to investigate how a pore-forming, virucidal peptide destabilizes lipid vesicles in a surface-based experimental configuration. A key advantage of the approach is that it enables direct kinetic measurement of the surface-bound peptide-to-lipid (P:L) ratio. Comprehensive experiments involving different bulk peptide concentrations and biologically relevant membrane compositions support a unified model that membrane lysis occurs at or above a critical P:L ratio, which is at least several-fold greater than the value corresponding to the onset of pore formation. That is consistent with peptide-induced pores causing additional membrane strain that leads to lysis of highly curved membranes. Collectively, the work presents a new model that describes how peptide-induced pores may destabilize lipid membranes through a membrane strain-related lytic process, and this knowledge has important implications for the design and application of membrane-active peptides.

  11. Modeling cellular lysis in skeletal muscle due to electric shock.

    Science.gov (United States)

    Cela, Carlos J; Lee, Raphael C; Lazzi, Gianluca

    2011-05-01

    High-voltage electrical trauma frequently results in injury patterns that cannot be completely attributed to Joule heating. An electrical-injury model describing cellular lysis damage caused by supraphysiological electric fields is introduced, and used to evaluate the effects of high-voltage electric shock on the skeletal muscle of a human upper limb in a configuration that simulates hand-to-hand contact. A novel multiresolution admittance method, capable of efficiently handling large computational models while maintaining excellent accuracy, was used to perform the numerical computations. Values for the computed current through the arm and the upper limb impedance are reported.

  12. Self-bioremediation of cork-processing wastewaters by (chloro)phenol-degrading bacteria immobilised onto residual cork particles.

    Science.gov (United States)

    del Castillo, I; Hernández, P; Lafuente, A; Rodríguez-Llorente, I D; Caviedes, M A; Pajuelo, E

    2012-04-15

    Cork manufacturing is a traditional industry in Southern Europe, being the main application of this natural product in wine stoppers and insulation. Cork processing begins at boiling the raw material. As a consequence, great volumes of dark wastewaters, with elevated concentrations of chlorophenols, are generated, which must be depurated through costly physicochemical procedures before discarding them into public water courses. This work explores the potential of bacteria, isolated from cork-boiling waters storage ponds, in bioremediation of the same effluent. The bacterial population present in cork-processing wastewaters was analysed by DGGE; low bacterial biodiversity was found. Aerobic bacteria were isolated and investigated for their tolerance against phenol and two chlorophenols. The most tolerant strains were identified by sequencing 16S rDNA. The phenol-degrading capacity was investigated by determining enzyme activities of the phenol-degrading pathway. Moreover, the capacity to form biofilms was analysed in a microtitre plate assay. Finally, the capacity to form biofilms onto the surface of residual small cork particles was evaluated by acridine staining followed by epifluorescence microscopy and by SEM. A low-cost bioremediation system, using phenol-degrading bacteria immobilised onto residual cork particles (a by-product of the industry) is proposed for the remediation of this industrial effluent (self-bioremediation).

  13. Operation of three parallel AN/AO processes to enrich denitrifying phosphorus removing bacteria for low strength wastewater treatment

    Institute of Scientific and Technical Information of China (English)

    XIA Si-qing; LIU Hong-bo

    2006-01-01

    Three parallel anaerobic-anoxic/anaerobic-aerobic (AN/AO) processes were developed to enrich denitrifying phosphorus removal bacteria (DPB) for low strength wastewater treatment. The main body of the parallel AN/AO process consists of an AN(anaerobic-anoxic) process and an AO (anaerobic-aerobic) process. In the AO process, the common phosphorus accumulating organisms (PAOs) was dominate, while in the AN process, DPB was dominate. The volume of anaerobic zone(Vana):anoxic zone(Vano):aerobic zone (Vaer) for the parallel AN/AO process is 1:1:1 in contrast with a Vana:Vaer and Vano,:Vaer of 1:2 and 1:4 for a traditional biological nutrient removal process (BNR). Process 3 excels in the 3 processes on the basis of COD, TN and TP removal. For 4 month operation, the effluent COD concentration of process 3 did not exceed 60 mg/L; the effluent TN concentration of process 3 was lower than 15 mg/L; and the effluent TP concentration of process 3 was lower than 1 mg/L.

  14. Extracellular polymeric substances mediate bioleaching/biocorrosion via interfacial processes involving iron(III) ions and acidophilic bacteria.

    Science.gov (United States)

    Sand, Wolfgang; Gehrke, Tilman

    2006-01-01

    Extracellular polymeric substances seem to play a pivotal role in biocorrosion of metals and bioleaching, biocorrosion of metal sulfides for the winning of precious metals as well as acid rock drainage. For better control of both processes, the structure and function of extracellular polymeric substances of corrosion-causing or leaching bacteria are of crucial importance. Our research focused on the extremophilic bacteria Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans, because of the "simplicity" and knowledge about the interactions of these bacteria with their substrate/substratum and their environment. For this purpose, the composition of the corresponding extracellular polymeric substances and their functions were analyzed. The extracellular polymeric substances of both species consist mainly of neutral sugars and lipids. The functions of the exopolymers seem to be: (i) to mediate attachment to a (metal) sulfide surface, and (ii) to concentrate iron(III) ions by complexation through uronic acids or other residues at the mineral surface, thus, allowing an oxidative attack on the sulfide. Consequently, dissolution of the metal sulfide is enhanced, which may result in an acceleration of 20- to 100-fold of the bioleaching process over chemical leaching. Experiments were performed to elucidate the importance of the iron(III) ions complexed by extracellular polymeric substances for strain-specific differences in oxidative activity for pyrite. Strains of A. ferrooxidans with a high amount of iron(III) ions in their extracellular polymeric substances possess greater oxidation activity than those with fewer iron(III) ions. These data provide insight into the function of and consequently the advantages that extracellular polymeric substances provide to bacteria. The role of extracellular polymeric substances for attachment under the conditions of a space station and resulting effects like biofouling, biocorrosion, malodorous gases, etc. will be discussed.

  15. Weight reduction is associated with increased plasma fibrin clot lysis.

    Science.gov (United States)

    Brzezińska-Kolarz, Beata; Kolarz, Marek; Wałach, Angelika; Undas, Anetta

    2014-11-01

    Obesity is associated with an increased risk of vascular thrombotic events. We sought to investigate how obesity and weight loss affect plasma fibrin clot properties. A total of 29 obese patients were studied before and after 3-month low-fat diet. Plasma fibrin clot parameters, including fibrin clot permeation coefficient (Ks), the lag phase of the turbidity curve, clot lysis time (t 50%), maximum rate of increase in D-dimer levels, and maximum D-dimer concentrations, were determined. Low-fat diet resulted in the reduction of body weight (P < .0001), body mass index (P < .0001), fat mass (P < .0001), total cholesterol (P < .0001), low-density lipoprotein cholesterol (P = .0005), triglycerides (P = .008), and plasminogen activator inhibitor 1 (P = .02), but not in fibrinogen or C-reactive protein. The only change in fibrin clot variables was shorter t 50% (P = .02). Baseline t 50%, but not posttreatment, correlated with waist circumference (r = .44, p = .02). This study demonstrates that weight loss in obese people can increase the efficiency of fibrin clot lysis.

  16. Fe vs. TiO2 Photo-assisted Processes for Enhancing the Solar Inactivation of Bacteria in Water.

    Science.gov (United States)

    Pulgarin, César

    2015-01-01

    Batch solar water disinfection (SODIS) is a known, simple and low-cost water treatment technology. SODIS is based on the synergistic action of temperature increase and light-assisted generation of Reactive Oxygen Species (ROS) on bacteria. ROS are generated via the action of solar photons on i) Natural Organic Matter (NOM), ii) some mineral components of water (Fe oxides or Fe-organic complexes, nitrogen compounds) and iii) endogenous bacteria photosensitizers (e.g. cytochrome). SODIS has proven its effectiveness for remote settlements or urban slums in regions with high incident solar radiation. All of the internal and external simultaneous processes are often driven by photoactive Fe-species present in the cell, as well as in the natural water sources. In SODIS, a temperature of 50 °C is required and due to this temperature dependence, only 1-2 L can be treated at a time. As required exposure time strongly depends on irradiation intensity and temperature, some SODIS households could be overburdened, leading to inadequate treatment and probable bacterial re-growth. This is why TiO(2) photocatalysis and Fe photo-assisted systems (i.e. photo-Fenton reactants) have been considered to enhance the photo-catalytic processes already present in natural water sources when exposed to solar light. Both TiO(2) and Fe-photoassisted processes, when applied to water disinfection aim to improve the performance of solar bacteria inactivation systems by i) enhancing ROS production, ii) making the process independent from the rise in temperature and as a consequence iii) allowing the treatment of larger volumes than 1-2 L of water and iv) prevent bacterial (re)growth, sometimes observed after sole solar treatment.

  17. Isolation, identification and optimization of ethanol producing bacteria from Saccharomyces-based fermentation process of alcohol industries in Iran

    Directory of Open Access Journals (Sweden)

    Hoda Ebrahimi

    2013-01-01

    Full Text Available Introduction: Due to the vast growth of world population, consumption of a lot of energy, limited energy supply and rising prices of fuel oil in the future, other alternative energy source is essential. Ethanol is renewable and a safe fuel and it is mainly based on microbial fermentation. The purpose of this study was isolation of high ethanol producing bacteria from the fermentation process of alcohol industries and optimization of growth conditions to be introduced to the industries. Materials and methods: The samples that were collected from fermentation tanks of alcohol industries were enriched in ZSM medium. To isolate the ethanol producing bacteria, the enriched culture was transferred on RMA agar. Bacterial growth conditions and their effects on ethanol production were optimized based on pH, growth temperature, agitation, fermentation time, initial substrate concentration and carbon and nitrogen sources. In addition, the morphological, physiological and molecular characterizations were investigated for identification of the isolates.Results: Three bacterial isolates ZYM7, ZYM8 and ZYM9 were isolated from fermentation tank. All isolates were able to produce ethanol 5.00, 7.60 and 4.00 gL-1 after 48 hours, respectively. The results demonstrated that all isolates were able to consume most sugars sources specially pentose carbon xylose. The isolate ZYM7 produced 13.00 gL-1 ethanol by consumption of xylose. The results of morphological and physiological characteristics showed that ZYM7 belonged to Lactobacillus sp. and ZYM8 and ZYM9 belonged to Acetobacter sp. Moreover, 16S rRNA sequencing and phylogenetic analyses exhibited that ZYM7 was similar to Lactobacillus rhamnosus with 99% homology and ZYM8 and ZYM9 were similar to Acetobacter pasteurianus with 99 and 98% homology, respectively.Discussion and conclusion: The results showed that that the isolated bacteria were suitable candidates to produce ethanol from raw material enriched with

  18. Vertical distribution of bacteria and intensity of microbiological processes in two stratified gypsum Karst Lakes in Lithuania

    Directory of Open Access Journals (Sweden)

    Krevs A.

    2011-08-01

    Full Text Available Physical-chemical parameters and the vertical distribution of bacteria and organic matter production-destruction processes were studied during midsummer stratification in two karst lakes (Kirkilai and Ramunelis located in northern Lithuania. The lakes were characterized by high sulfate concentrations (369–1248 mg·L-1. The O2/H2S intersection zone formed at 2–3 m depth. In Lake Kirkilai, the highest bacterial densities (up to 8.7 × 106 cell·mL-1 occurred at the O2/H2S intersection zone, whereas in Lake Ramunelis the highest densities were observed in the anoxic hypolimnion (up to 11 × 106 cell·mL-1. Pigment analysis revealed that green sulfur bacteria dominated in the microaerobic–anaerobic water layers in both lakes. The most intensive development of sulfate-reducing bacteria was observed in the anaerobic layer. Photosynthetic production of organic matter was highest in the upper layer. Rates of sulfate reduction reached 0.23 mg S2−·dm3·d-1 in the microaerobic-anaerobic water layer and 1.97 mg S2−·dm3·d-1 in sediments. Karst lakes are very sensitive to organic pollution, because under such impact in the presence of high sulfate amounts, sulfate reduction may become very intensive and, consequently, the increase in hydrogen sulfide and development of sulfur cycle bacteria may reduce the variety of other hydrobionts.

  19. A common, non-optimal phenotypic endpoint in experimental adaptations of bacteriophage lysis time

    Directory of Open Access Journals (Sweden)

    Chantranupong Lynne

    2012-03-01

    Full Text Available Abstract Background Optimality models of evolution, which ignore genetic details and focus on natural selection, are widely used but sometimes criticized as oversimplifications. Their utility for quantitatively predicting phenotypic evolution can be tested experimentally. One such model predicts optimal bacteriophage lysis interval, how long a virus should produce progeny before lysing its host bacterium to release them. The genetic basis of this life history trait is well studied in many easily propagated phages, making it possible to test the model across a variety of environments and taxa. Results We adapted two related small single-stranded DNA phages, ΦX174 and ST-1, to various conditions. The model predicted the evolution of the lysis interval in response to host density and other environmental factors. In all cases the initial phages lysed later than predicted. The ΦX174 lysis interval did not evolve detectably when the phage was adapted to normal hosts, indicating complete failure of optimality predictions. ΦX174 grown on slyD-defective hosts which initially entirely prevented lysis readily recovered to a lysis interval similar to that attained on normal hosts. Finally, the lysis interval still evolved to the same endpoint when the environment was altered to delay optimal lysis interval. ST-1 lysis interval evolved to be ~2 min shorter, qualitatively in accord with predictions. However, there were no changes in the single known lysis gene. Part of ST-1's total lysis time evolution consisted of an earlier start to progeny production, an unpredicted phenotypic response outside the boundaries of the optimality model. Conclusions The consistent failure of the optimality model suggests that constraint and genetic details affect quantitative and even qualitative success of optimality predictions. Several features of ST-1 adaptation show that lysis time is best understood as an output of multiple traits, rather than in isolation.

  20. Does the duration of lysis affect the sensitivity of the in vitro alkaline comet assay?

    Science.gov (United States)

    Enciso, José Manuel; Sánchez, Oscar; López de Cerain, Adela; Azqueta, Amaya

    2015-01-01

    The alkaline comet assay is now the method of choice for measuring different kinds of DNA damage in cells. Several attempts have been made to identify and evaluate the critical points affecting the comet assay outcome, highlighting the requirement of arriving at a standardised protocol in order to be able to compare the results obtained in different laboratories. However, reports on the effect of modifying the time of lysis are lacking. Here we tested different times of lysis (from no lysis to 1 week) in control HeLa cells and HeLa cells treated with different concentrations of methyl methanesulfonate (MMS) or H2O2. We also tested different times of lysis in the comet assay combined with formamidopyrimidine DNA glycosylase (FPG) in untreated and Ro 19-8022 plus light-treated HeLa cells. The same DNA damage levels were detected in the absence of lysis or after 1h of lysis when the standard comet assay was used to detect the MMS- and H2O2-induced lesions; the response increased when longer lysis was used, up to at least 1 week. When FPG was used, a minimum lysis period of 5 min was necessary to allow the enzyme to reach the DNA; the same DNA damage levels were detected after 5 min or 1h of lysis and the response increased up to 24h. In conclusion, the time of lysis can be varied depending on the sensitivity needed in both versions of the assay, and a constant time of lysis should be used if results from different experiments or laboratories are to be compared.

  1. Small-scale extraction of recombinant proteins from bacteria.

    Science.gov (United States)

    Simpson, Richard J

    2010-09-01

    Bacteria are particularly convenient for producing recombinant proteins for purification purposes. To monitor induction as well as the levels of recombinant protein expression, it is important to have a rapid, simple method for estimating bacterial protein expression. This protocol describes the preparation of small-scale bacterial extracts using cell lysis with 0.5% Triton X-100.

  2. Biological variation in tPA-induced plasma clot lysis time

    NARCIS (Netherlands)

    S. Talens (Simone); J.J.M.C. Malfliet (Joyce); G. Rudež (Goran); H.M.H. Spronk (Henri); N.A.H. Janssen (Nicole); P. Meijer (Piet); C. Kluft (Cornelius); M.P.M. de Maat (Moniek); D.C. Rijken (Dingeman)

    2012-01-01

    textabstractHypofibrinolysis is a risk factor for venous and arterial thrombosis, and can be assessed by using a turbidimetric tPA-induced clot lysis time (CLT) assay. Biological variation in clot lysis time may affect the interpretation and usefulness of CLT as a risk factor for thrombosis. Suffici

  3. Cellular lysis in Bacillus subtilis; the affect of multiple extracellular protease deficiencies

    NARCIS (Netherlands)

    Stephenson, K; Bron, S; Harwood, CR

    1999-01-01

    Cellular lysis properties of strains of Bacillus subtilis deficient in the synthesis of extracellular proteases was investigated. In all cases, extracellular protease deficiency was found to increase the extent of cellular lysis of batch cultured strains following the transition to stationary phase,

  4. A Method for Purifying Obligate Intracellular Coxiella burnetii that Employs Digitonin Lysis of Host Cells

    Science.gov (United States)

    Cockrell, Diane C.; Beare, Paul A.; Fischer, Elizabeth R.; Howe, Dale; Heinzen, Robert. A.

    2008-01-01

    Purification of the obligate intracellular bacterium Coxiella burnetii requires physical disruption of infected cells. Here we describe a gentle and safe digitonin lysis procedure to release C. burnetii from infected cells. The purity, yield, and infectivity of digitonin-prepped organisms are comparable to that of organisms purified using cell lysis by sonication. PMID:18242746

  5. On-chip lysis of mammalian cells through a handheld corona device.

    Science.gov (United States)

    Escobedo, C; Bürgel, S C; Kemmerling, S; Sauter, N; Braun, T; Hierlemann, A

    2015-07-21

    On-chip lysis is required in many lab-on-chip applications involving cell studies. In these applications, the complete disruption of the cellular membrane and a high lysis yield is essential. Here, we present a novel approach to lyse cells on-chip through the application of electric discharges from a corona handheld device. The method only requires a microfluidic chip and a low-cost corona device. We demonstrate the effective lysis of BHK and eGFP HCT 116 cells in the sub-second time range using an embedded microelectrode. We also show cell lysis of non-adherent K562 leukemia cells without the use of an electrode in the chip. Cell lysis has been assessed through the use of bright-field microscopy, high-speed imaging and cell-viability fluorescence probes. The experimental results show effective cell lysis without any bubble formation or significant heating. Due to the simplicity of both the components involved and the lysis procedure, this technique offers an inexpensive lysis option with the potential for integration into lab-on-a-chip devices.

  6. Tumor lysis without syndrome in adenocarcinoma of the lung: Case report.

    Science.gov (United States)

    Chopra, Amit; Marak, Creticus; Alappan, Narendra Kumar; Alterman, Daniel; Shim, Chang

    2013-01-01

    Tumor lysis syndrome TLS is commonly seen during the treatment of rapidly proliferating. However TLS is rarely reported in Non-small cell Lung Cancer. This may because of low proliferative rate and chemo-resistant nature of NSCLC. We are reporting a case of tumor lysis without concomitant syndrome in a patient with adenocarcinoma of Lung.

  7. Evidence that Clostridium perfringens theta-toxin induces colloid-osmotic lysis of erythrocytes.

    OpenAIRE

    Harris, R W; Sims, P J; Tweten, R K

    1991-01-01

    Clostridium perfringens theta-toxin was shown to lyse target erythrocytes by a colloid-osmotic mechanism. Analysis showed the onset of lysis of erythrocytes by theta-toxin could be temporarily stabilized with 0.3 M sucrose. Flow cytometry analysis of the size distribution of theta-toxin-treated erythrocytes showed swelling of the erythrocytes prior to lysis.

  8. Tumor lysis syndrome in the emergency department: challenges and solutions

    Directory of Open Access Journals (Sweden)

    Ñamendys-Silva SA

    2015-08-01

    Full Text Available Silvio A Ñamendys-Silva,1,2 Juan M Arredondo-Armenta,1 Erika P Plata-Menchaca,2 Humberto Guevara-García,1 Francisco J García-Guillén,1 Eduardo Rivero-Sigarroa,2 Angel Herrera-Gómez,1 1Department of Critical Care Medicine, Instituto Nacional de Cancerología, 2Department of Critical Care Medicine, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico Abstract: Tumor lysis syndrome (TLS is the most common oncologic emergency. It is caused by rapid tumor cell destruction and the resulting nucleic acid degradation during or days after initiation of cytotoxic therapy. Also, a spontaneous form exists. The metabolic abnormalities associated with this syndrome include hyperkalemia, hyperphosphatemia, hypocalcemia, hyperuricemia, and acute kidney injury. These abnormalities can lead to life-threatening complications, such as heart rhythm abnormalities and neurologic manifestations. The emergency management of overt TLS involves proper fluid resuscitation with crystalloids in order to improve the intravascular volume and the urinary output and to increase the renal excretion of potassium, phosphorus, and uric acid. With this therapeutic strategy, prevention of calcium phosphate and uric acid crystal deposition within renal tubules is achieved. Other measures in the management of overt TLS are prescription of hypouricemic agents, renal replacement therapy, and correction of electrolyte imbalances. Hyperkalemia should be treated quickly and aggressively as its presence is the most hazardous acute complication that can cause sudden death from cardiac arrhythmias. Treatment of hypocalcemia is reserved for patients with electrocardiographic changes or symptoms of neuromuscular irritability. In patients who are refractory to medical management of electrolyte abnormalities or with severe cardiac and neurologic manifestations, early dialysis is recommended.Keywords: tumor lysis syndrome, emergency department, emergency

  9. [Influence of ionizing radiation of low intensity on the processes of reproduction, aging and dying off of Escherichia coli bacteria].

    Science.gov (United States)

    Morozov, I I; Petin, V G; Morozova, G V

    2002-01-01

    The influence of 60Co gamma-ray irradiation of low intensity (0.1-0.4, 0.76 x 10(3) microGy/h) on the processes of reproduction, aging and dying off of E. coli B/r and E. coli BS-1 bacteria have been investigated. It was shown that the reproduction of this bacteria strains was not dependent on the dose rate in the range 0.1-0.4 microGy/h. It was shown in comparison with the irradiated E. coli B/r cells dynamics of the aging and dying off of the irradiated E. coli BS-1 is decreased in the process of prolonged (about 190 days) irradiation with a dose rate of 0.76 x 10(3) microGy/h. It is proposed the relationship between the revealed phenomenon of the decrease in the intensity of the irradiated E. coli BS-1 cell aging and dying and the Vavilov-Cerenkov emission.

  10. Interspecies interactions result in enhanced biofilm formation by co-cultures of bacteria isolated from a food processing environment.

    Science.gov (United States)

    Røder, Henriette L; Raghupathi, Prem K; Herschend, Jakob; Brejnrod, Asker; Knøchel, Susanne; Sørensen, Søren J; Burmølle, Mette

    2015-10-01

    Bacterial attachment and biofilm formation can lead to poor hygienic conditions in food processing environments. Furthermore, interactions between different bacteria may induce or promote biofilm formation. In this study, we isolated and identified a total of 687 bacterial strains from seven different locations in a meat processing environment and evaluated their biofilm formation capability. A diverse group of bacteria was isolated and most were classified as poor biofilm producers in a Calgary biofilm device assay. Isolates from two sampling sites, the wall and the meat chopper, were further examined for multispecies biofilm formation. Eight strains from each sampling site were chosen and all possible combinations of four member co-cultures were tested for enhanced biofilm formation at 15 °C and 24 °C. In approximately 20% of the multispecies consortia grown at 15 °C, the biofilm formation was enhanced when comparing to monospecies biofilms. Two specific isolates (one from each location) were found to be present in synergistic combinations with higher frequencies than the remaining isolates tested. This data provides insights into the ability of co-localized isolates to influence co-culture biofilm production with high relevance for food safety and food production facilities.

  11. Tumor lysis syndrome in a patient with metastatic colon cancer after treatment with oxaliplatin and 5-Fu

    Directory of Open Access Journals (Sweden)

    Ruo-Han Tseng

    2016-12-01

    Full Text Available Tumor lysis syndrome in solid tumors is a rare occurrence, with a poor prognosis. We present the case of a patient of recurrent colon cancer who received chemotherapy with FOLFOX regimen (lencovorin, fluorouracil, and oxaliplatin with subsequent tumor lysis. We present a recurrent rectal cancer patient suffered from tumor lysis syndrome after salvage FOLFOX regimen. After treat with CVVH with improved conscious status. In this case report, we had review the tumor lysis in solid tumor.

  12. Dynamics of hydrogen-producing bacteria in a repeated batch fermentation process using lake sediment as inoculum.

    Science.gov (United States)

    Romano, Stefano; Paganin, Patrizia; Varrone, Cristiano; Tabacchioni, Silvia; Chiarini, Luigi

    2014-02-01

    In this study, we evaluated the effectiveness of lake sediment as inoculum for hydrogen production through dark fermentation in a repeated batch process. In addition, we investigated the effect of heat treatment, applied to enrich hydrogen-producing bacteria, on the bacterial composition and metabolism. Denaturing gradient gel electrophoresis and molecular cloning, both performed using the 16S rDNA gene as target gene, were used to monitor the structure of the bacterial community. Hydrogen production and bacterial metabolism were analysed via gas chromatography and high-performance liquid chromatography. Both treated and non-treated inocula were able to produce high amounts of hydrogen. However, statistical analysis showed a clear difference in their bacterial composition and metabolism. The heat treatment favoured the growth of different Clostridia sp., in particular of Clostridium bifermentans, allowing the production of a constant amount of hydrogen over prolonged time. These cultures showed both butyrate and ethanol fermentation types. Absence of heat treatment allowed species belonging to the genera Bacillus, Sporolactobacillus and Massilia to outgrow Clostridia sp. with a reduction in hydrogen production and a significant metabolic change. Our data indicate that lake sediment harbours bacteria that can efficiently produce hydrogen over prolonged fermentation time. Moreover, we could show that the heat treatment stabilizes the bacterial community composition and the hydrogen production.

  13. State estimation of a batch hydrogen production process using the photosynthetic bacteria Rhodobacter capsulatus

    Energy Technology Data Exchange (ETDEWEB)

    Obeid, Jamila; Magnin, Jean-Pierre [Grenoble University, LEPMI, UMR 5631 (CNRS-INPG-UJF), Laboratoire d' Electrochimie et de Physico chimie des Materiaux et Interfaces, BP 75, 38402 St. Martin d' Heres (France); Flaus, Jean-Marie; Adrot, Olivier [Grenoble University, G-SCOP UMR 272 (CNRS-INPG-UJF), Laboratoire des Sciences pour la Conception, l' Optimisation et la Production, 46, avenue Felix Viallet, 38031 Grenoble (France); Willison, John C. [Laboratoire de Chimie et Biologie des Metaux (UMR 5249 CEA-CNRS-UJF), iRTSV/LCBM, CEA-Grenoble, 38054 Grenoble (France)

    2010-10-15

    This paper addresses the problem of estimating the states of an anaerobic photosynthetic process used for biohydrogen production by the photosynthetic bacterium Rhodobacter capsulatus. The process is described by a non-linear, time-discrete model and the state estimation is solved using an observer based on the Moving-Horizon State Estimation Method (MHSE). This approach is based on the minimization of a criterion (a non-linear function), in this case, the difference between the estimated output and the measured output of the system over a considered time horizon, where the solution is computed by using a numerical interval method. The observer was successfully applied to hydrogen production by R. capsulatus strain B10 in a batch process. (author)

  14. APPLICATION OF RESPIROMETRIC TESTS FOR ASSESSMENT OF METHANOGENIC BACTERIA ACTIVITY IN WASTEWATER SLUDGE PROCESSING

    Directory of Open Access Journals (Sweden)

    Małgorzata Cimochowicz-Rybicka

    2013-07-01

    Full Text Available Production of a methane-rich gas (‘biogas’ is contemporary popular sludges processing technology which allows to generate thermal and/or electric energy. Formal requirements issued by the European Union to promote so called renewable energy resources made these process more attractive leading to its application in WWTPs which were designed based on different sludge handling processes. Authors (as active design engineers noted that dimensioning sludge digestion chamber is usually based on SRT assessment without any emphasis on sludge characteristics. Bio-mass characteristics and the estimation of its activity with respect to methane production are of great importance, from both scientific and practical points of view, as anaerobic digestion appears to be one of crucial processes in municipal wastewater handling and disposal. The authors propose respirometric tests to estimate a biomass potential to produce ‘a biogas’ and several years’ laboratory and full scale experience proved its usefulness and reliability both as a measurement and a design tool applicable in sludge handling. Dimensioning method proposed by authors, allows to construct and optimize operation of digestion chambers based on a methanogenic activity.

  15. Local lysis in acute basilar artery occlusions: a case report

    Energy Technology Data Exchange (ETDEWEB)

    Karnik, R.; Perneczky, G.; Ammerer, H.P.; Brenner, H.; Slany, J. (Krankenanstalt der Stadt Wien Rudolfstiftung (Austria))

    1984-01-01

    Thrombosis of the basilar artery is not a rare disease, and the mortality is reported to be 60 to 80%. Present standard therapy with heparin infusions yields poor results. The high risk of intracerebral haemorrhage prohibits systemic fibrinolytic therapy. Due to these facts and good experience in our department with the use of local intracoronary lysis in acute myocardial infarction, the method of local thrombolysis was applied in a case of acute basiliar artery thrombosis. Fibrinolytic therapy was started via an angiography catheter placed in the vertebral artery in a 28 year-old woman with hemiplegia and severe brain stem symptoms. The patient received 200,000 IU streptokinase within 2 hours and subsequently 300,000 IU urokinase within 10 hours. The vessel re-opened completely. The neurological symptoms decreased during the following weeks. Based on this experience and according to rare reports in the literature we believe local low-dose thrombolysis to be a causal therapy promising success for acute thrombosis of the basilar artery. This therapy can be carried out in every medical centre able to perform selective angiography and experienced in the administration of fibrinolytic drugs.

  16. [Acute renal failure in patients with tumour lysis sindrome].

    Science.gov (United States)

    Poskurica, Mileta; Petrović, Dejan; Poskurica, Mina

    2016-01-01

    `Hematologic malignancies (leukemia, lymphoma, multiple myeloma, et al.), as well as solid tumours (renal, liver, lung, ovarian, etc.), can lead to acute or chronic renal failure.The most common clinical manifestation is acute renal failure within the tumour lysis syndrome (TLS). It is characterized by specific laboratory and clinical criteria in order to prove that kidney disorders result from cytolysis of tumour cells after chemotherapy regimen given, although on significantly fewer occasions it is likely to occur spontaneously or after radiotherapy. Essentially, failure is the disorder of functionally conserved kidney or of kidney with varying degrees of renal insufficiency, which render the kidney impaired and unable to effectively eliminate the end products of massive cytolysis and to correct the resulting disorders: hyperuricemia, hyperkalemia, hypocalcaemia, hyperphosphatemia, and others. The risk of TLS depends on tumour size, proliferative potential of malignant cells, renal function and the presence of accompanying diseases and disorders. Hydration providing adequate diuresis and administration of urinary suppressants (allopurinol, febuxostat) significantly reduce the risk of developing TLS. If prevention of renal impairment isn't possible, the treatment should be supplemented with hemodynamic monitoring and pharmacological support, with the possible application of recombinant urate-oxidase enzyme (rasburicase). Depending on the severity of azotemia and hydroelectrolytic disorders, application of some of the methods of renal replacement therapy may be considered.

  17. The Importance of Endospore-Forming Bacteria Originating from Soil for Contamination of Industrial Food Processing

    Directory of Open Access Journals (Sweden)

    Marc Heyndrickx

    2011-01-01

    Full Text Available Specific endospore formers have become important contaminants in industrial food processing. The direct or indirect soil route of contamination or dispersal is the start of events or processes in the agrofood chain that eventually leads to important problems or concerns for food safety and/or quality. Three important food sectors are discussed in this paper. In the dairy sector, Bacillus cereus, the most important pathogen or spoilage organism in this sector, and Clostridium tyrobutyricum, the most important spoiler in certain cheeses, both contaminate pasteurized milk through the faecal and/or (at least for B. cereus the direct soil route. In the fruit juice industry, Alicyclobacillus acidoterrestris, present on raw fruits, has become a major quality-target organism. In the ready-to-eat food sector, B. cereus and other aerobic endospore formers are introduced via vegetables, fruits, or herbs and spices, while anaerobic spore formers like nonproteolytic Clostridium botulinum and Clostridium estertheticum pose safety and spoilage risks in chilled packaged foods, respectively.

  18. Ecology of Indigenous Lactic Acid Bacteria along Different Winemaking Processes of Tempranillo Red Wine from La Rioja (Spain

    Directory of Open Access Journals (Sweden)

    Lucía González-Arenzana

    2012-01-01

    Full Text Available Ecology of the lactic acid bacteria (LAB during alcoholic fermentation (AF and spontaneous malolactic fermentation (MLF of Tempranillo wines from four wineries of La Rioja has been studied analyzing the influence of the winemaking method, processing conditions, and geographical origin. Five different LAB species were isolated during AF, while, during MLF, only Oenococcus oeni was detected. Although the clonal diversity of O. oeni strains was moderate, mixed populations were observed, becoming at least one strain with distinct PFGE profile the main responsible for MLF. Neither the winemaking method nor the cellar situation was correlated with the LAB diversity. However, processing conditions influenced the total number of isolates and the percentage of each isolated species and strains. The winemaking method could cause that genotypes found in semicarbonic maceration did not appear in other wineries. Four genotypes of O. oeni were isolated in more than one of the rest wineries. These four together with other dominant strains might be included in a future selection process.

  19. Comparison of indicator bacteria inactivation by the ultraviolet and the ultraviolet/hydrogen peroxide disinfection processes in humic waters.

    Science.gov (United States)

    Teksoy, Arzu; Alkan, Ufuk; Eleren, Sevil Çalışkan; Topaç, Burcu Şengül; Sağban, Fatma Olcay Topaç; Başkaya, Hüseyin Savaş

    2011-12-01

    The aim of the present study was to evaluate responses of potential indicator bacteria (i.e. Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis) to the ultraviolet (UV) radiation and the UV/hydrogen peroxide (H₂O₂) disinfection processes of surface waters with different qualities in terms of humic content. The UV and the UV/H₂O₂ processes were applied to waters containing various concentrations of fulvic acid in order to inactivate E. coli, P. aeruginosa and B. subtilis spores. Three fulvic acid (0, 2 and 6 mg l(-1)) and four H₂O₂ (0, 10, 25 and 50 mg l(-1)) concentrations were used. Results showed that the k values of E. coli, P. aeruginosa and B. subtilis spores varied between 2.22 and 4.00, 1.73 and 3.58, and 1.40 and 1.86, respectively, in all test conditions. The sensitivity of the test organisms followed a decreasing order of E. coli > P. aeruginosa > B. subtilis. Results of the study indicated that the blocking effect of fulvic acid for the UV light was diminished by using H₂O₂ in combination with the UV radiation. Findings of the present study strongly suggested that the UV/H₂O₂ process was significantly effective on the inactivation of E. coli and P. aeruginosa in humic waters, whereas it induced little or no apparent contribution to the disinfection efficiency of B. subtilis spores.

  20. Inactivation and regrowth of multidrug resistant bacteria in urban wastewater after disinfection by solar-driven and chlorination processes.

    Science.gov (United States)

    Fiorentino, Antonino; Ferro, Giovanna; Alferez, María Castro; Polo-López, Maria Inmaculada; Fernández-Ibañez, Pilar; Rizzo, Luigi

    2015-07-01

    Solar disinfection and solar-driven advanced oxidation processes (AOPs) (namely H2O2/sunlight, TiO2/sunlight, H2O2/TiO2/sunlight, solar photo-Fenton) were evaluated in the inactivation of indigenous antibiotic-resistant bacteria (ARB) in real urban wastewater. A multidrug resistant (MDR) Escherichia coli strain isolated from the effluent of the biological process of an urban wastewater treatment plant was the target ARB. The higher inactivation rates (residual density under detection limit, 2 CFUm L(-1)) were achieved with H2O2/TiO2/sunlight (cumulative energy per unit of volume (QUV) in the range 3-5 kJ L(-1), depending on H2O2/TiO2 ratio) and H2O2/sunlight (QUV of 8 kJ L(-1)) processes. All investigated processes did not affect antibiotic resistance of survived colonies. Moreover, H2O2/sunlight was compared with conventional chlorination process to evaluate bacterial regrowth potential and particularly the proportion of indigenous MDR E. coli with respect to total indigenous E. coli population. Chlorination (1.0 mg Cl2 L(-1)) was more effective than H2O2/sunlight (50 mg H2O2 L(-1)) to achieve total inactivation of MDR E. coli (15 min Vs 90 min) but less effective in controlling their regrowth (24 h Vs 48 h). Interestingly, the percentage of MDR E. coli in H2O2/sunlight treated samples decreased as incubation time increased; the opposite was observed for chlorinated samples.

  1. Spontaneous Tumor Lysis Syndrome and Secondary Thrombotic Thrombocytopenic Purpura in Early Stage Colorectal Cancer

    Institute of Scientific and Technical Information of China (English)

    Saad Usmani; Joel Appel; Zainab Shahid; Husain Saleh

    2008-01-01

    @@ Acute tumor lysis syndrome (ATLS) is a well-described oncological emergency that is usually associated with hematological malignan-cies complicated by treatment. It is typically related to a high tumor burden, rapidly growing and chemosensitive malignancies.

  2. Evaluation of conventional castaneda and lysis centrifugation blood culture techniques for diagnosis of human brucellosis.

    Science.gov (United States)

    Mantur, Basappa G; Mangalgi, Smita S

    2004-09-01

    We investigated the role of the lysis centrifugation blood culture technique over the conventional Castaneda technique for the diagnosis of human brucellosis. The lysis centrifugation technique has been found to be more sensitive in both acute (20% higher sensitivity; P < 0.00001) and chronic (40% higher sensitivity; P = 0.087) forms of brucellosis. The major advantage of lysis centrifugation was in the mean detection time, which was only 2.4 days in acute and 2.7 days in chronic cases, with 103 out of 110 (93.6%) and 17 out of 20 (85%) cultures from acute and chronic brucellosis, respectively, detected before the conventional culture was positive. Our results confirmed the potential usefulness of the lysis technique in diagnosis and institution of appropriate antibiotic therapy.

  3. Fate of Pathogenic Bacteria Associated with Production of Pickled Sausage by Using a Cold Fill Process.

    Science.gov (United States)

    Gaydos, Nelson J; Cutter, Catherine N; Campbell, Jonathan A

    2016-10-01

    Preservation by pickling has been used for many years to extend the shelf life of various types of food products. By storing meat products in a brine solution containing an organic acid, salt, spices, as well as other preservatives, the pH of the product is reduced, thus increasing the safety and shelf life of the product. Pickling may involve the use of heated brines to further add to the safety of the food product. When precooked, ready-to-eat (RTE) sausages are pickled with a heated brine solution, the process is referred to as hot filling. However, hot filling has been shown to affect the clarity of the brine, making the product cloudy and unappealing to consumers. Because of the potential quality defects caused by higher temperatures associated with hot fill pickling, cold fill pickling, which uses room temperature brine, is preferred by some pickled sausage manufacturers. Because little information exists on the safety of cold fill, pickled sausages, a challenge study was designed using a brine solution (5% acetic acid and 5% salt at 25°C) to pickle precooked, RTE sausages inoculated with a pathogen cocktail consisting of Salmonella Typhimurium, Salmonella Senftenberg, Salmonella Montevideo, Listeria monocytogenes , and Staphylococcus aureus . All pathogens were reduced ~6.80 log CFU/g in 72 h when enumerated on nonselective media. On selective media, Salmonella and L. monocytogenes decreased 6.33 and 6.35 log CFU/g in 12 h, respectively whereas S. aureus was reduced 6.80 log CFU/g in 24 h. Sausages experienced significant (P ≤ 0.05) decreases in pH over the 28 days of storage, whereas no significant differences were observed in water activity (P =0.1291) or salt concentration of the sausages (P =0.1445) or brine (P =0.3180). The results of this experiment demonstrate that cold fill pickling can effectively reduce and inhibit bacterial pathogens.

  4. Tumor lysis syndrome following trastuzumab and pertuzumab for metastatic breast cancer: a case report.

    OpenAIRE

    2016-01-01

    Background Tumor lysis syndrome is a rare and potentially fatal complication of oncologic treatments, especially in solid tumors. To the best of our knowledge, tumor lysis syndrome has never been reported after trastuzumab and pertuzumab combination therapy. Knowledge of risk factors and active prevention proceedings is of utmost importance to avoid fatal outcomes. Case presentation We present the case of a chemo-naive 58-year-old Belgian woman developing hypovolemic shock and multiple organ ...

  5. Polypeptone induces dramatic cell lysis in ura4 deletion mutants of fission yeast.

    Directory of Open Access Journals (Sweden)

    Yuzy Matsuo

    Full Text Available Polypeptone is widely excluded from Schizosaccharomyces pombe growth medium. However, the reasons why polypeptone should be avoided have not been documented. Polypeptone dramatically induced cell lysis in the ura4 deletion mutant when cells approached the stationary growth phase, and this phenotype was suppressed by supplementation of uracil. To determine the specificity of this cell lysis phenotype, we created deletion mutants of other genes involved in de novo biosynthesis of uridine monophosphate (ura1, ura2, ura3, and ura5. Cell lysis was not observed in these gene deletion mutants. In addition, concomitant disruption of ura1, ura2, ura3, or ura5 in the ura4 deletion mutant suppressed cell lysis, indicating that cell lysis induced by polypeptone is specific to the ura4 deletion mutant. Furthermore, cell lysis was also suppressed when the gene involved in coenzyme Q biosynthesis was deleted. This is likely because Ura3 requires coenzyme Q for its activity. The ura4 deletion mutant was sensitive to zymolyase, which mainly degrades (1,3-beta-D glucan, when grown in the presence of polypeptone, and cell lysis was suppressed by the osmotic stabiliser, sorbitol. Finally, the induction of cell lysis in the ura4 deletion mutant was due to the accumulation of orotidine-5-monophosphate. Cell wall integrity was dramatically impaired in the ura4 deletion mutant when grown in the presence of polypeptone. Because ura4 is widely used as a selection marker in S. pombe, caution needs to be taken when evaluating phenotypes of ura4 mutants.

  6. Pseudomonas aeruginosa Pore-Forming Exolysin and Type IV Pili Cooperate To Induce Host Cell Lysis

    Science.gov (United States)

    Basso, Pauline; Ragno, Michel; Elsen, Sylvie; Reboud, Emeline; Golovkine, Guillaume; Bouillot, Stephanie; Huber, Philippe; Lory, Stephen; Faudry, Eric

    2017-01-01

    ABSTRACT   Clinical strains of Pseudomonas aeruginosa lacking the type III secretion system genes employ a toxin, exolysin (ExlA), for host cell membrane disruption. Here, we demonstrated that ExlA export requires a predicted outer membrane protein, ExlB, showing that ExlA and ExlB define a new active two-partner secretion (TPS) system of P. aeruginosa. In addition to the TPS signals, ExlA harbors several distinct domains, which include one hemagglutinin domain, five arginine-glycine-aspartic acid (RGD) motifs, and a C-terminal region lacking any identifiable sequence motifs. However, this C-terminal region is important for the toxic activity, since its deletion abolishes host cell lysis. Using lipid vesicles and eukaryotic cells, including red blood cells, we demonstrated that ExlA has a pore-forming activity which precedes cell membrane disruption of nucleated cells. Finally, we developed a high-throughput cell-based live-dead assay and used it to screen a transposon mutant library of an ExlA-producing P. aeruginosa clinical strain for bacterial factors required for ExlA-mediated toxicity. The screen resulted in the identification of proteins involved in the formation of type IV pili as being required for ExlA to exert its cytotoxic activity by promoting close contact between bacteria and the host cell. These findings represent the first example of cooperation between a pore-forming toxin of the TPS family and surface appendages in host cell intoxication. PMID:28119472

  7. Application of culture culture-independent molecular biology based methods to evaluate acetic acid bacteria diversity during vinegar processing.

    Science.gov (United States)

    Ilabaca, Carolina; Navarrete, Paola; Mardones, Pamela; Romero, Jaime; Mas, Albert

    2008-08-15

    Acetic acid bacteria (AAB) are considered fastidious microorganisms because they are difficult to isolate and cultivate. Different molecular approaches were taken to detect AAB diversity, independently of their capacity to grow in culture media. Those methods were tested in samples that originated during traditional vinegar production. Bacterial diversity was assessed by analysis of 16S rRNA gene, obtained by PCR amplifications of DNA extracted directly from the acetification container. Bacterial composition was analyzed by RFLP-PCR of 16S rRNA gene, Temporal Temperature Gradient Gel Electrophoresis (TTGE) separation of amplicons containing region V3-V5 of 16S rRNA gene and cloning of those amplicons. TTGE bands and clones were grouped based on their electrophoretic pattern similarity and sequenced to be compared with reference strains. The main microorganism identified in vinegar was Acetobacter pasteurianus, which at the end of the acetification process was considered to be the only microorganism present. The diversity was the highest at 2% acetic acid, where indefinite species of Gluconacetobacter xylinus/europaeus/intermedius were also present.

  8. Screening of bacteria for self-healing of concrete cracks and optimization of the microbial calcium precipitation process.

    Science.gov (United States)

    Zhang, J L; Wu, R S; Li, Y M; Zhong, J Y; Deng, X; Liu, B; Han, N X; Xing, F

    2016-08-01

    A novel high-throughput strategy was developed to determine the calcium precipitation activity (CPA) of mineralization bacteria used for self-healing of concrete cracks. A bacterial strain designated as H4 with the highest CPA of 94.8 % was screened and identified as a Bacillus species based on 16S rDNA sequence and phylogenetic tree analysis. Furthermore, the effects of certain influential factors on the microbial calcium precipitation process of H4 were evaluated. The results showed that lactate and nitrate are the best carbon and nitrogen sources, with optimal concentrations of approximately 25 and 18 mM, respectively. The H4 strain is able to maintain a high CPA in the pH range of 9.5-11.0, and a suitable initial spore concentration is 4.0 × 10(7) spores/ml. Moreover, an ambient Ca(2+) concentration greater than 60 mM resulted in a serious adverse impact not only on the CPA but also on the growth of H4, suggesting that the maintenance of the Ca(2+) concentration at a low level is necessary for microbial self-healing of concrete cracks.

  9. Potency of microfiltration membrane process in purifying broccoli (Brassica oleracea L.) fermented by lactic acid bacteria (LAB) as functional food

    Science.gov (United States)

    Susilowati, Agustine; Aspiyanto, Maryati, Yati; Melanie, Hakiki; Lotulung, Puspa D.

    2017-01-01

    Purifying broccoli (Brassica oleracea L.) fermented by Lactic Acid Bacteria (LAB) using mixture of L. bulgaricus, S. thermopillus, L. acidophillusand Bifidobacteriumbifidum and fructooligosaccharides (FOS) as carbon source have been performed to recover biomass concentrate for probiotic and antioxidant. Purification of fermented broccoli was conducted through microfiltration (MF) membrane of 0.15 µm at stirrer rotation speed 400 rpm, room temperature and pressure 40 psia for 30 minutes. Fermented broccoli produced via fermentation process with fermentation time 0 (initial) and 48 hours, and LAB concentration 10% and 20% (v/v) represented as biomass of A, B, C and D. The experimental result showed that based on selectivity of total organic acids, separating optimization was achieved at biomass D (fermentation time 48 hours and mixed LAB culture concentration 20%). Concentrate composition produced in this condition were total acids 6.04%, total solids 24.31%, total polyphenol 0.0252%, reducing sugar 68.25 mg/mL, total sugars 30.89 mg/mL, and dissolved protein 28.54 mg/mL with pH 3.94. In this condition, recovery of biomass concentrate of D for total acids 5.64 folds, total solids 1.82 folds, total polyphenol 3.03 folds, reducing sugar 1.16 folds, total sugars 1.19 folds, and dissolved protein 0.67 folds compared with feed (initial process). Identification of monomer of biomass concentrate D as polyphenol derivatives at T2,01 and T3.01 gave monomer with molecular weight (MW) 192.78 Dalton (Da.), and monomer with MW 191.08, 191.49 and 192.07 Da., while lactic acid derivatives showed MW 251.13, 251.6 and 252.14, and monomer with MW 250.63, 252.14 and 254.22 Da.

  10. Identification of Lactic Acid Bacteria in Fruit Pulp Processing Byproducts and Potential Probiotic Properties of Selected Lactobacillus Strains

    Science.gov (United States)

    Garcia, Estefânia F.; Luciano, Winnie A.; Xavier, Danilo E.; da Costa, Whyara C. A.; de Sousa Oliveira, Kleber; Franco, Octávio L.; de Morais Júnior, Marcos A.; Lucena, Brígida T. L.; Picão, Renata C.; Magnani, Marciane; Saarela, Maria; de Souza, Evandro L.

    2016-01-01

    This study aimed to identify lactic acid bacteria (LAB) in byproducts of fruit (Malpighia glabra L., Mangifera indica L., Annona muricata L., and Fragaria vesca L.) pulp processing. Fifty strains of LAB were identified using matrix-assisted laser desorption/ionization–time of flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequence (16S rRNA) analysis. Species belonging to Lactobacillus genus were the predominant LAB in all fruit pulp processing byproducts. The average congruency between the MALDI-TOF MS and 16S rRNA in LAB species identification reached 86%. Isolates of L. plantarum, L. brevis, L. pentosus, L. lactis and L. mesenteroides were identified with 100% congruency. MALDI-TOF MS and 16S rRNA analysis presented 86 and 100% efficiency of LAB species identification, respectively. Further, five selected Lactobacillus strains (L. brevis 59, L. pentosus 129, L. paracasei 108, L. plantarum 49, and L. fermentum 111) were evaluated for desirable probiotic-related properties and growth behavior on two different cultivation media. The exposure to pH 2.0 sharply decreased the counts of the different Lactobacillus strains after a 1 or 2 h incubation, while varied decreases were noted after 3 h of exposure to pH 3.0. Overall, the exposure to pH 5.0 and to bile salts (0.15, 0.30, and 1.00%) did not decrease the counts of the Lactobacillus strains. All tested Lactobacillus strains presented inhibitory activity against Staphylococcus aureus, Salmonella Typhimurium, Salmonella Enteritidis, Listeria monocytogenes and Escherichia coli, and presented variable susceptibility to different antibiotics. The selected Lactobacillus strains presented satisfactory and reproducible growth behavior. In conclusion, MALDI-TOF MS and 16S rRNA analysis revealed high efficiency and congruency for LAB species identification, and the selected Lactobacillus strains may be candidates for further investigation of novel probiotic strains. PMID:27625647

  11. Bioaugmentation of anaerobic sludge digestion with iron-reducing bacteria: process and microbial responses to variations in hydraulic retention time.

    Science.gov (United States)

    Baek, Gahyun; Kim, Jaai; Shin, Seung Gu; Lee, Changsoo

    2016-01-01

    Although anaerobic digestion (AD) is a widely used option to manage waste activated sludge (WAS), there are some drawbacks related to its slow reaction rate and low energy productivity. This study examined an anaerobic WAS digester, augmented with an iron-reducing microbial consortium, relative to changes in microbial community structure and process performance at decreasing hydraulic retention times (HRTs) of 20 to 10 days. The enhanced methanation performance (approximately 40 % increase in methane yield) by the bioaugmentation was sustained until the HRT was decreased to 12.5 days, under Fe(3+)-rich conditions (ferric oxyhydroxide, 20 mM Fe). Enhanced iron-reducing activity was evidenced by the increased Fe(2+) to total Fe ratio maintained above 50 % during the stable operational phases. A further decrease in HRT to 10 days resulted in a significant performance deterioration, along with a drop in the Fe(2+) to total Fe ratio to <35 %, after four turnovers of operation. Prevailing existence of putative iron-reducing bacteria (IRBs) was identified by denaturing gradient gel electrophoresis (DGGE), with Spirochaetaceae- and Thauera-related organisms being dominant members, and clear dominance shifts among them with respect to decrease in HRT were observed. Lowering HRT led to evident shifts in bacterial community structure likely associated with washout of IRBs, leading to decreases in iron respiration activity and AD performance at a lower HRT. The bacterial community structure shifted dynamically over phases, and the community transitions correlated well with the changes in process performance. Overall, the combined biostimulation and bioaugmentation investigated in this study proved effective for enhanced methane recovery from anaerobic WAS digestion, which suggests an interesting potential for high-rate AD.

  12. Identification of lactic acid bacteria in fruit pulp processing byproducts and potential probiotic properties of selected Lactobacillus strains

    Directory of Open Access Journals (Sweden)

    Estefânia Garcia

    2016-08-01

    Full Text Available This study aimed to identify lactic acid bacteria (LAB in byproducts of fruit (Malpighia glabra L., Mangifera indica L., Annona muricata L. and Fragaria vesca L. pulp processing. Fifty strains of LAB were identified using matrix-assisted laser desorption/ionization–time of flight mass spectrometry (MALDI-TOF MS and 16S rRNA gene sequence (16S rRNA analysis. Species belonging to Lactobacillus genus were the predominant LAB in all fruit pulp processing byproducts. The average congruency between the MALDI-TOF MS and 16S rRNA in LAB species identification reached 86%. Isolates of L. plantarum, L. brevis, L. pentosus, L. lactis and L. mesenteroides were identified with 100% congruency. MALDI-TOF MS and 16S rRNA analysis presented 86% and 100% efficiency of LAB species identification, respectively. Further, five selected Lactobacillus strains (L. brevis 59, L. pentosus 129, L. paracasei 108, L. plantarum 49 and L. fermentum 111 were evaluated for desirable probiotic-related properties and growth behavior on two different cultivation media. The exposure to pH 2.0 sharply decreased the counts of the different Lactobacillus strains after a 1 or 2 h incubation, while varied decreases were noted after 3 h of exposure to pH 3.0. Overall, the exposure to pH 5.0 and to bile salts (0.15, 0.30 and 1.00% did not decrease the counts of the Lactobacillus strains. All tested Lactobacillus strains presented inhibitory activity against Staphylococcus aureus, Salmonella Typhimurium, Salmonella Enteritidis, Listeria monocytogenes and Escherichia coli, and presented variable susceptibility to different antibiotics. The selected Lactobacillus strains presented satisfactory and reproducible growth behavior. In conclusion, MALDI-TOF MS and 16S rRNA analysis revealed high efficiency and congruency for LAB species identification, and the selected Lactobacillus strains may be candidates for further investigation of novel probiotic strains.

  13. Identification of Lactic Acid Bacteria in Fruit Pulp Processing Byproducts and Potential Probiotic Properties of Selected Lactobacillus Strains.

    Science.gov (United States)

    Garcia, Estefânia F; Luciano, Winnie A; Xavier, Danilo E; da Costa, Whyara C A; de Sousa Oliveira, Kleber; Franco, Octávio L; de Morais Júnior, Marcos A; Lucena, Brígida T L; Picão, Renata C; Magnani, Marciane; Saarela, Maria; de Souza, Evandro L

    2016-01-01

    This study aimed to identify lactic acid bacteria (LAB) in byproducts of fruit (Malpighia glabra L., Mangifera indica L., Annona muricata L., and Fragaria vesca L.) pulp processing. Fifty strains of LAB were identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequence (16S rRNA) analysis. Species belonging to Lactobacillus genus were the predominant LAB in all fruit pulp processing byproducts. The average congruency between the MALDI-TOF MS and 16S rRNA in LAB species identification reached 86%. Isolates of L. plantarum, L. brevis, L. pentosus, L. lactis and L. mesenteroides were identified with 100% congruency. MALDI-TOF MS and 16S rRNA analysis presented 86 and 100% efficiency of LAB species identification, respectively. Further, five selected Lactobacillus strains (L. brevis 59, L. pentosus 129, L. paracasei 108, L. plantarum 49, and L. fermentum 111) were evaluated for desirable probiotic-related properties and growth behavior on two different cultivation media. The exposure to pH 2.0 sharply decreased the counts of the different Lactobacillus strains after a 1 or 2 h incubation, while varied decreases were noted after 3 h of exposure to pH 3.0. Overall, the exposure to pH 5.0 and to bile salts (0.15, 0.30, and 1.00%) did not decrease the counts of the Lactobacillus strains. All tested Lactobacillus strains presented inhibitory activity against Staphylococcus aureus, Salmonella Typhimurium, Salmonella Enteritidis, Listeria monocytogenes and Escherichia coli, and presented variable susceptibility to different antibiotics. The selected Lactobacillus strains presented satisfactory and reproducible growth behavior. In conclusion, MALDI-TOF MS and 16S rRNA analysis revealed high efficiency and congruency for LAB species identification, and the selected Lactobacillus strains may be candidates for further investigation of novel probiotic strains.

  14. Taxonomic structure of the yeasts and lactic acid bacteria microbiota of pineapple (Ananas comosus L. Merr.) and use of autochthonous starters for minimally processing.

    Science.gov (United States)

    Di Cagno, Raffaella; Cardinali, Gainluigi; Minervini, Giovanna; Antonielli, Livio; Rizzello, Carlo Giuseppe; Ricciuti, Patrizia; Gobbetti, Marco

    2010-05-01

    Pichia guilliermondii was the only identified yeast in pineapple fruits. Lactobacillus plantarum and Lactobacillus rossiae were the main identified species of lactic acid bacteria. Typing of lactic acid bacteria differentiated isolates depending on the layers. L. plantarum 1OR12 and L. rossiae 2MR10 were selected within the lactic acid bacteria isolates based on the kinetics of growth and acidification. Five technological options, including minimal processing, were considered for pineapple: heating at 72 degrees C for 15 s (HP); spontaneous fermentation without (FP) or followed by heating (FHP), and fermentation by selected autochthonous L. plantarum 1OR12 and L. rossiae 2MR10 without (SP) or preceded by heating (HSP). After 30 days of storage at 4 degrees C, HSP and SP had a number of lactic acid bacteria 1000 to 1,000,000 times higher than the other processed pineapples. The number of yeasts was the lowest in HSP and SP. The Community Level Catabolic Profiles of processed pineapples indirectly confirmed the capacity of autochthonous starters to dominate during fermentation. HSP and SP also showed the highest antioxidant activity and firmness, the better preservation of the natural colours and were preferred for odour and overall acceptability.

  15. Evaluation of cell lysis procedures and use of a micro fluidic system for an automated DNA-based cell identification in interplanetary missions

    Science.gov (United States)

    Hall, J. A.; Felnagle, E.; Fries, M.; Spearing, S.; Monaco, L.; Steele, A.

    2006-12-01

    A Modular Assay System for Solar System Exploration (MASSE) is being developed to include sample handling, pre-treatment, separation and analysis of biological target compounds by both DNA and protein microarrays. To better design sensitive and accurate initial upstream sample handling of the MASSE instrument, experiments investigating the sensitivity and potential extraction bias of commercially available DNA extraction kits between classes of environmentally relevant prokaryotes such as gram-negative bacteria ( Escherichia coli), gram-positive bacteria ( Bacillus megatarium), and Archaea ( Haloarcula marismortui) were performed. For extractions of both planktonic cultures and spiked Mars simulated regolith, FTA ® paper demonstrated the highest sensitivity, with detection as low as ˜1×10 1 cells and ˜3.3×10 2 cells, respectively. In addition to the highest sensitivity, custom modified application of FTA ® paper extraction protocol is the simplest in terms of incorporation into MASSE and displayed little bias in sensitivity with respect to prokaryotic cell type. The implementation of FTA paper for environmental microbiology investigations appears to be a viable and effective option potentially negating the need for other pre-concentration steps such as filtration and negating concerns regarding extraction efficiency of cells. In addition to investigations on useful technology for upstream sample handling in MASSE, we have also evaluated the potential for μTAS to be employed in the MASSE instrument by employing proprietary lab-on-a-chip development technology to investigate the potential for microfluidic cell lysis of different prokaryotic cells employing both chemical and biological lysis agents. Real-time bright-field microscopy and quantitative PMT detection indicated that that gram positive, gram negative and archaeal cells were effectively lyzed in a few seconds using the microfluidic chip protocol developed. This included employing a lysis buffer with

  16. Acid resistance of methanogenic bacteria in a two-stage anaerobic process treating high concentration methanol Wastewater

    Institute of Scientific and Technical Information of China (English)

    ZHOU Xuefei; REN Nanqi

    2007-01-01

    In this study,the two-stage upflow anaerobic sludge blanket(UASB)system and batch experiments were employed to evaluate the performance of anaerobic digestion for the treatment of high concentration methanol wastewater.The acid resistance of granular sludge and methanogenic bacteria and their metabolizing activity were investigated.The results show that the pH of the first UASB changed from 4.9 to 5.8 and 5.5 to 6.2 for the second reactor.Apparently,these were not the advisable pH levels that common metha nogenic bacteria could accept.The methanogenic bacteria of the system,viz.Methanosarcina barkeri,had some acid resistance and could still degrade methanol at pH 5.0.If the methanogenic bacteria were trained further,their acid resistance would be improved somewhat.Granular sludge of the system could protect the methanogenic bacteria within its body against the impact of the acidic environment and make them degrade methanol at pH 4.5.The performance of granular sludge was attributed to its structure,bacteria species,and the distribution of bacterium inside the granule.

  17. Study on processing highly effective Acidophilic bacteria starter culture and Acidophilic bacteria milk powder with new technology%新工艺制备高效嗜酸菌发酵剂和嗜酸菌奶粉的研究

    Institute of Scientific and Technical Information of China (English)

    方景泉; 徐欣

    2013-01-01

    Two different strains of acidophilic bacteria were incubated by different methods and through a series contrast tests,we found the best method for incubating.Then concentrated the bacteria with high sPeed centrifuge and put it in 10% skim milk solution with protective agent D,after vacuum freeze drying the viable count of Lactobacillus acidophilus exist in the starter culture were 8.3 ×1010g-1and its survival rate could be 68.8%.We can process Acidophilic bacteria milk powder when mixed it together with milk powder.%分别使两株嗜酸菌在脱脂乳水解液培养基和脱脂乳培养基中生长,通过一系列的对比实验,以及使菌体浓缩,浓菌体悬浮在添加保护剂D质量分数为10%脱脂乳中,经冷冻干燥后,所得高效嗜酸菌发酵剂中的细胞数达8.3×1010 g-1.其在真空冷冻干燥后的存活率最高可达68.8%.将此高效发酵剂与奶粉混合即可制得嗜酸菌奶粉.

  18. Control of Listeria spp. by competitive-exclusion bacteria in floor drains of a poultry processing plant.

    Science.gov (United States)

    Zhao, Tong; Podtburg, Teresa C; Zhao, Ping; Schmidt, Bruce E; Baker, David A; Cords, Bruce; Doyle, Michael P

    2006-05-01

    In previous studies workers determined that two lactic acid bacterium isolates, Lactococcus lactis subsp. lactis C-1-92 and Enterococcus durans 152 (competitive-exclusion bacteria [CE]), which were originally obtained from biofilms in floor drains, are bactericidal to Listeria monocytogenes or inhibit the growth of L. monocytogenes both in vitro and in biofilms at 4 to 37 degrees C. We evaluated the efficacy of these isolates for reducing Listeria spp. contamination of floor drains of a plant in which fresh poultry is processed. Baseline assays revealed that the mean numbers of Listeria sp. cells in floor drains sampled on six different dates (at approximately biweekly intervals) were 7.5 log(10) CFU/100 cm(2) for drain 8, 4.9 log(10) CFU/100 cm(2) for drain 3, 4.4 log(10) CFU/100 cm(2) for drain 2, 4.1 log(10) CFU/100 cm(2) for drain 4, 3.7 log(10) CFU/100 cm(2) for drain 1, and 3.6 log(10) CFU/100 cm(2) for drain 6. The drains were then treated with 10(7) CE/ml in an enzyme-foam-based cleaning agent four times in 1 week and twice a week for the following 3 weeks. In samples collected 1 week after CE treatments were applied Listeria sp. cells were not detectable (samples were negative as determined by selective enrichment culture) for drains 4 and 6 (reductions of 4.1 and 3.6 log(10) CFU/100 cm(2), respectively), and the mean numbers of Listeria sp. cells were 3.7 log(10) CFU/100 cm(2) for drain 8 (a reduction of 3.8 log(10) CFU/100 cm(2)), culture; a reduction of 3.3 log(10) CFU/100 cm(2)), and 2.6 log(10) CFU/100 cm(2) for drain 3 (a reduction of 2.3 log(10) CFU/100 cm(2)). However, the aerobic plate counts for samples collected from floor drains before, during, and after CE treatment remained approximately the same. The results indicate that application of the two CE can greatly reduce the number of Listeria sp. cells in floor drains at 3 to 26 degrees C in a facility in which fresh poultry is processed.

  19. Arthroscopic lysis of adhesions for the stiff total knee: results after failed manipulation.

    Science.gov (United States)

    Tjoumakaris, Fotios Paul; Tucker, Bradfords Chofield; Post, Zachary; Pepe, Matthew David; Orozco, Fabio; Ong, Alvin C

    2014-05-01

    Arthrofibrosis after total knee arthroplasty (TKA) is a potentially devastating complication, resulting in loss of motion and function and residual pain. For patients in whom aggressive physical therapy and manipulation under anesthesia fail, lysis of adhesions may be the only option to rescue the stiff TKA. The purpose of this study is to report the results of arthroscopic lysis of adhesions after failed manipulation for a stiff, cruciate-substituting TKA. This retrospective study evaluated patients who had undergone arthroscopic lysis of adhesions for arthrofibrosis after TKA between 2007 and 2011. Minimum follow-up was 12 months (average, 31 months). Average total range of motion of patients in this series was 62.3°. Average preoperative flexion contracture was 16° and average flexion was 78.6°. Statistical analysis was performed using Student's t test. Pre- to postoperative increase in range of motion was significant (Psurgery.

  20. Lysis of a Single Cyanobacterium for Whole Genome Amplification

    Directory of Open Access Journals (Sweden)

    Richard N. Zare

    2013-08-01

    Full Text Available Bacterial species from natural environments, exhibiting a great degree of genetic diversity that has yet to be characterized, pose a specific challenge to whole genome amplification (WGA from single cells. A major challenge is establishing an effective, compatible, and controlled lysis protocol. We present a novel lysis protocol that can be used to extract genomic information from a single cyanobacterium of Synechocystis sp. PCC 6803 known to have multilayer cell wall structures that resist conventional lysis methods. Simple but effective strategies for releasing genomic DNA from captured cells while retaining cellular identities for single-cell analysis are presented. Successful sequencing of genetic elements from single-cell amplicons prepared by multiple displacement amplification (MDA is demonstrated for selected genes (15 loci nearly equally spaced throughout the main chromosome.

  1. The effects of differential extraction conditions on the premature lysis of spermatozoa.

    Science.gov (United States)

    Hennekens, Catherine M; Cooper, Elyse S; Cotton, Robin W; Grgicak, Catherine M

    2013-05-01

    The purpose of this study was to determine the effect Proteinase K, sodium dodecyl sulfate (SDS), incubation times, and temperatures had on differential extraction efficiencies and the premature lysis of spermatozoa. The effect was measured using Quantifiler® Duo and Identifiler™ PCR Amplification kits, where the resultant male and female DNA concentrations and their ratios within the nonsperm- and sperm fractions (SFs) were determined. Comparisons between expected and observed ratios illustrate the quantity of female DNA in the SF increased when Proteinase K was absent during the initial incubation. Additionally, there is no indication of simultaneous sperm and epithelial cell lysis in the absence of DTT at Proteinase K concentrations ranging from 10 to 300 μg/mL. All other conditions exhibited minimal variation in DNA concentration. Therefore, despite the various protocols used for the differential lysis of cell mixtures encountered in casework, the method is robust and successful at most conditions.

  2. Preventing acute renal failure is crucial during acute tumor lysis syndrome

    Directory of Open Access Journals (Sweden)

    Darmon Michael

    2007-01-01

    Full Text Available Tumour Lysis syndrome (TLS is characterized by the massive destruction of tumoral cells and the release in the extracellular space of their content. While TLS may occur spontaneously before treatment, it usually develops shortly after the initiation of cytotoxic chemotherapy. These metabolites can overwhelm the homeostatic mechanisms and cause hyperuricemia, hyperkalemia, hyperphosphatemia and hypocalcemia. Moreover, TLS may lead to an acute renal failure (ARF. In addition to the hospital mortality induced by the acute renal failure itself, development of an ARF may preclude optimal cancer treatment. Therefore, prevention of the acute renal failure during acute tumor lysis syndrome is mandatory. The objective of this review is to describe pathophysiological mechanisms leading to acute tumor lysis syndrome, clinical and biological consequences of this syndrome and to provide up-to-date guidelines to ensure prevention and prompt management of this syndrome.

  3. Lysis-deficient phages as novel therapeutic agents for controlling bacterial infection

    Directory of Open Access Journals (Sweden)

    Kempashanaiah Nanjundappa

    2011-08-01

    Full Text Available Abstract Background Interest in phage therapy has grown over the past decade due to the rapid emergence of antibiotic resistance in bacterial pathogens. However, the use of bacteriophages for therapeutic purposes has raised concerns over the potential for immune response, rapid toxin release by the lytic action of phages, and difficulty in dose determination in clinical situations. A phage that kills the target cell but is incapable of host cell lysis would alleviate these concerns without compromising efficacy. Results We developed a recombinant lysis-deficient Staphylococcus aureus phage P954, in which the endolysin gene was rendered nonfunctional by insertional inactivation. P954, a temperate phage, was lysogenized in S. aureus strain RN4220. The native endolysin gene on the prophage was replaced with an endolysin gene disrupted by the chloramphenicol acetyl transferase (cat gene through homologous recombination using a plasmid construct. Lysogens carrying the recombinant phage were detected by growth in presence of chloramphenicol. Induction of the recombinant prophage did not result in host cell lysis, and the phage progeny were released by cell lysis with glass beads. The recombinant phage retained the endolysin-deficient genotype and formed plaques only when endolysin was supplemented. The host range of the recombinant phage was the same as that of the parent phage. To test the in vivo efficacy of the recombinant endolysin-deficient phage, immunocompromised mice were challenged with pathogenic S. aureus at a dose that results in 80% mortality (LD80. Treatment with the endolysin-deficient phage rescued mice from the fatal S. aureus infection. Conclusions A recombinant endolysin-deficient staphylococcal phage has been developed that is lethal to methicillin-resistant S. aureus without causing bacterial cell lysis. The phage was able to multiply in lytic mode utilizing a heterologous endolysin expressed from a plasmid in the propagation host

  4. Rhizosphere Bacteria

    Directory of Open Access Journals (Sweden)

    N.V. Feoktistova

    2016-06-01

    Full Text Available The review deals with the analysis of modern literature data on rhizosphere bacteria and their role in plant life. The structure of rhizosphere has been characterized. The role of plants as the centers of formation of microbial communities has been shown. Data on the main groups of microorganisms inhabiting the rhizosphere have been provided. The associative relationship between rhizobacteria and partner plants has been investigated. The modern concept of holobiont defined as the whole host plant organism and microorganisms associated with it has been reviewed. The role of rhizobacteria in the processes of nitrogen fixation has been discussed in detail. The mechanisms of direct stimulation of plant growth by biosynthesis of phytohormones, improvement of phosphorus and nitrogen nutrition, increase in resistance to stress, and stimulation mediated by antagonism against pathogenic microorganisms have been analyzed. The criteria for selection of rhizobacteria for practical purposes have been discussed.

  5. Tumor lysis syndrome following endoscopic radiofrequency interstitial thermal ablation of colorectal liver metastases.

    LENUS (Irish Health Repository)

    Barry, B D

    2012-02-03

    Radiofrequency interstitial thermal ablation (RITA) provides a palliative option for patients suffering from metastatic liver disease. This procedure can be performed using a laparoscopic approach with laparoscopic ultrasound used to position the RITA probe. We describe a case of laparoscopic RITA performed for colorectal liver metastasis that was complicated by tumor lysis syndrome (TLS) following treatment. We consider RITA to be a safe procedure, as supported by the literature, but where intracorporal tumor lysis is the treatment goal we believe that the systemic release of tumor products can overwhelm the excretory capacity; therefore, TLS is an inevitable consequence in some patients.

  6. Severe acute tumor lysis syndrome in patients with germ-cell tumors

    Directory of Open Access Journals (Sweden)

    Guilherme Alvarenga Feres

    2008-01-01

    Full Text Available Germ-cell tumors are a high-proliferative type of cancer that may evolve to significant bulky disease. Tumor lysis syndrome is rarely reported in this setting. The reports of three patients with germ-cell tumors who developed severe acute tumor lysis syndrome following the start of their anticancer therapy are presented. All patients developed renal dysfunction and multiorgan failure. Patients with extensive germ-cell tumors should be kept on close clinical and laboratory monitoring. Physicians should be aware of this uncommon but severe complication and consider early admission to the intensive care unit for the institution of measures to prevent acute renal failure.

  7. Quantitative fluorescent in-situ hybridization: a hypothesized competition mode between two dominant bacteria groups in hydrogen-producing anaerobic sludge processes.

    Science.gov (United States)

    Huang, C-L; Chen, C-C; Lin, C-Y; Liu, W-T

    2009-01-01

    Two hydrogen-producing continuous flow stirred tank reactors (CSTRs) fed respectively with glucose and sucrose were investigated by polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE) and fluorescent in-situ hybridization (FISH). The substrate was fed in a continuous mode decreased from hydraulic retention time (HRT) 10 hours to 6, 5, 4, 3, and 2 hours. Quantitative fluorescent in-situ hybridization (FISH) observations further demonstrated that two morphotypes of bacteria dominated both microbial communities. One was long rod bacteria which can be targeted either by Chis150 probe designed to hybridize the gram positive low G + C bacteria or the specific oligonucleotide probe Lg10-6. The probe Lg10-6, affiliated with Clostridium pasteurianum, was designed and then checked with other reference organisms. The other type, unknown group, which cannot be detected by Chis150 was curved rod bacteria. Notably, the population ratios of the two predominant groups reflected the different operational performance of the two reactors, such as hydrogen producing rates, substrate turnover rates and metabolites compositions. Therefore, a competition mode of the two dominant bacteria groups was hypothesized. In the study, 16S rRNA-based gene library of hydrogen-producing microbial communities was established. The efficiency of hydrogen yields was correlated with substrates (glucose or sucrose), HRT, metabolites compositions (acetate, propionate, butyrate and ethanol), thermal pre-treatment (seed biomass was heated at 100 degrees C for 45 minutes), and microbial communities in the bioreactor, not sludge sources (municipal sewage sludge, alcohol-processing sludge, or bean-processing sludge). The designed specific oligonucleotide probe Lg10-6 also provides us a useful and fast molecular tool to screen hydrogen-producing microbial communities in the future research.

  8. Complement lysis activity in autologous plasma is associated with lower viral loads during the acute phase of HIV-1 infection.

    Directory of Open Access Journals (Sweden)

    Michael Huber

    2006-11-01

    Full Text Available BACKGROUND: To explore the possibility that antibody-mediated complement lysis contributes to viremia control in HIV-1 infection, we measured the activity of patient plasma in mediating complement lysis of autologous primary virus. METHODS AND FINDINGS: Sera from two groups of patients-25 with acute HIV-1 infection and 31 with chronic infection-were used in this study. We developed a novel real-time PCR-based assay strategy that allows reliable and sensitive quantification of virus lysis by complement. Plasma derived at the time of virus isolation induced complement lysis of the autologous virus isolate in the majority of patients. Overall lysis activity against the autologous virus and the heterologous primary virus strain JR-FL was higher at chronic disease stages than during the acute phase. Most strikingly, we found that plasma virus load levels during the acute but not the chronic infection phase correlated inversely with the autologous complement lysis activity. Antibody reactivity to the envelope (Env proteins gp120 and gp41 were positively correlated with the lysis activity against JR-FL, indicating that anti-Env responses mediated complement lysis. Neutralization and complement lysis activity against autologous viruses were not associated, suggesting that complement lysis is predominantly caused by non-neutralizing antibodies. CONCLUSIONS: Collectively our data provide evidence that antibody-mediated complement virion lysis develops rapidly and is effective early in the course of infection; thus it should be considered a parameter that, in concert with other immune functions, steers viremia control in vivo.

  9. Microphotographs of cyanobacteria documenting the effects of various cell-lysis techniques

    Science.gov (United States)

    Rosen, Barry H.; Loftin, Keith A.; Smith, Christopher E.; Lane, Rachael F.; Keydel, Susan P.

    2011-01-01

    Cyanotoxins are a group of organic compounds biosynthesized intracellularly by many species of cyanobacteria found in surface water. The United States Environmental Protection Agency has listed cyanotoxins on the Safe Drinking Water Act's Contaminant Candidate List 3 for consideration for future regulation to protect public health. Cyanotoxins also pose a risk to humans and other organisms in a variety of other exposure scenarios. Accurate and precise analytical measurements of cyanotoxins are critical to the evaluation of concentrations in surface water to address the human health and ecosystem effects. A common approach to total cyanotoxin measurement involves cell membrane disruption to release the cyanotoxins to the dissolved phase followed by filtration to remove cellular debris. Several methods have been used historically, however no standard protocols exist to ensure this process is consistent between laboratories before the dissolved phase is measured by an analytical technique for cyanotoxin identification and quantitation. No systematic evaluation has been conducted comparing the multiple laboratory sample processing techniques for physical disruption of cell membrane or cyanotoxins recovery. Surface water samples collected from lakes, reservoirs, and rivers containing mixed assemblages of organisms dominated by cyanobacteria, as well as laboratory cultures of species-specific cyanobacteria, were used as part of this study evaluating multiple laboratory cell-lysis techniques in partnership with the U.S. Environmental Protection Agency. Evaluated extraction techniques included boiling, autoclaving, sonication, chemical treatment, and freeze-thaw. Both treated and untreated samples were evaluated for cell membrane integrity microscopically via light, epifluorescence, and epifluorescence in the presence of a DNA stain. The DNA stain, which does not permeate live cells with intact membrane structures, was used as an indicator for cyanotoxin release into the

  10. [Detection of Yersinia Enterocolitica Bacteriophage PhiYe-F10 Lysis Spectrum and Analysis of the Relationship between Lysis Ability and Virulence Gene of Yersinia Enterocolitica].

    Science.gov (United States)

    Zha, Tao; Liang, Junrong; Xiao, Yuchun; Jing, Huaiqi

    2016-03-01

    To determine the lysis spectrum of Yersinia enterocolitica bacteriophage phiYe-F10 and to analyze the relationship between the lysis ability of phiYe-F10 and the virulence gene of Yersinia enterocolitica. To observe the lysis ability of the phage phiYe-F10 to the different Yersinia strains with the double-layer technique. The strains used in this study including 213 of Yersinia enterocolitica and 36 of Yersinia pseudotuberculosis and 1 of Yersinia pestis. The virulence genes of these Yersinia enterocolitica (attachment invasion locus (ail) and enterotoxin (ystA, ystB) and yersinia adhesin A (yadA), virulence factor (virF), specific gene for lipopolysaccharide O-side chain of serotype O : 3 (rfbc) were all detected. Among the 213 Yersinia enterocolitica, 84 strains were O : 3 serotype (78 strains with rfbc gene), 10 were serotype O : 5, 13 were serotype O : 8, 34 were serotype O : 9 and 72 were other serotypes. Of these, 77 were typical pathogenic Yersinia enterocolitica harboring with virulence plasmid (ail+, ystA+, ystB-, yadA+, virF+), and 15 were pathogenic bacterial strains deficiency virulence plasmid (ail+, ystA+, ystB-, yadA-, virF-) and the rest 121 were non pathogenic genotype strains. PhiYe-F10 lysed the 71 serotype O : 3 Yersinia enterocolitica strains which were all carried with rfbc+, including 52 pathogenic Yersinia enterocolitica, 19 nonpathogenic Y. enterocolitica. The phiYe-F10 can not lysed serotype O : 5, O : 9 and other serotype Y. enterocolitica, the lysis rate of serotype O : 3 was as high as 84.5%. The phiYe-F10 can not lysed Yersinia pseudotuberculosis and Yersinia pestis. Yersinia phage phiYe-F10 is highly specific for serotype O : 3 Yersinia enterocolitic at 25 degrees C, which showed a typical narrow lysis spectrum. Phage phiYe-F10 can lysed much more pathogenic Y. enterocolitica than nonpathogenic Y. enterocolitica.

  11. Characteristic Time Scales of Transport Processes for Chemotactic Bacteria in Groundwater: Analysis of Pore-scale to Field-scale Experimental Data

    Science.gov (United States)

    Ford, R. M.

    2010-12-01

    Many processes contribute to the transport of microorganisms in groundwater environments. One process of interest is chemotaxis, whereby motile bacteria are able to detect and swim toward increasing concentrations of industrial hydrocarbons that they perceive as food sources. By enabling bacteria to migrate to the sources of pollutants that they degrade, chemotaxis has the potential to enhance bioremediation efforts, especially in less permeable zones where contamination may persist. To determine the field conditions under which chemotaxis might be exploited in a bioremediation scheme requires an understanding of the characteristic time scales in the system. We defined a dimensionless chemotaxis number that compares the time over which a bacterial population is exposed to a chemical gradient to the time required for a bacterial population to migrate a significant distance in response to a chemical gradient. The exposure time and the response time are dependent upon the experimental conditions and properties of the bacteria and chemical attractant. Experimental data was analyzed for a range of groundwater flow rates over a wide scope of experimental systems including a single-pore with NAPL source, a microfluidic channel with and without a porous matrix, a laboratory column, a bench-scale microcosm and a field-scale study. Chemical gradients were created transverse to the flow direction. Distributions of chemotactic and nonchemotactic bacteria were compared to determine the extent of migration due to chemotaxis. Under some conditions at higher flow rates, the effect of chemotaxis was diminished to the point of not being detected. The goal of the study was to determine a critical value for the dimensionless chemotaxis number (which is independent of scale) that can be used as a design criterion to ascertain a priori the conditions under which a chemotactic response will impact bacterial transport relative to other processes such as advection and dispersion.

  12. Imaging secondary metabolism of Streptomyces sp. Mg1 during cellular lysis and colony degradation of competing Bacillus subtilis.

    Science.gov (United States)

    Barger, Sarah R; Hoefler, B Chris; Cubillos-Ruiz, Andrés; Russell, William K; Russell, David H; Straight, Paul D

    2012-10-01

    Soil streptomycetes are saprotrophic bacteria that secrete numerous secondary metabolites and enzymes for extracellular functions. Many streptomycetes produce antibiotics thought to protect vegetative mycelia from competing organisms. Here we report that an organism isolated from soil, Streptomyces sp. Mg1, actively degrades colonies and causes cellular lysis of Bacillus subtilis when the organisms are cultured together. We predicted that the inhibition and degradation of B. subtilis colonies in this competition depends upon a combination of secreted factors, including small molecule metabolites and enzymes. To begin to unravel this complex competitive phenomenon, we use a MALDI imaging mass spectrometry strategy to map the positions of metabolites secreted by both organisms. In this report, we show that Streptomyces sp. Mg1 produces the macrolide antibiotic chalcomycin A, which contributes to inhibition of B. subtilis growth in combination with other, as yet unidentified factors. We suggest that efforts to understand competitive and cooperative interactions between bacterial species benefit from assays that pair living organisms and probe the complexity of metabolic exchanges between them.

  13. An optimised direct lysis method for gene expression studies on low cell numbers.

    Science.gov (United States)

    Le, Anh Viet-Phuong; Huang, Dexing; Blick, Tony; Thompson, Erik W; Dobrovic, Alexander

    2015-08-05

    There is increasing interest in gene expression analysis of either single cells or limited numbers of cells. One such application is the analysis of harvested circulating tumour cells (CTCs), which are often present in very low numbers. A highly efficient protocol for RNA extraction, which involves a minimal number of steps to avoid RNA loss, is essential for low input cell numbers. We compared several lysis solutions that enable reverse transcription (RT) to be performed directly on the cell lysate, offering a simple rapid approach to minimise RNA loss for RT. The lysis solutions were assessed by reverse transcription quantitative polymerase chain reaction (RT-qPCR) in low cell numbers isolated from four breast cancer cell lines. We found that a lysis solution containing both the non-ionic detergent (IGEPAL CA-630, chemically equivalent to Nonidet P-40 or NP-40) and bovine serum albumin (BSA) gave the best RT-qPCR yield. This direct lysis to reverse transcription protocol outperformed a column-based extraction method using a commercial kit. This study demonstrates a simple, reliable, time- and cost-effective method that can be widely used in any situation where RNA needs to be prepared from low to very low cell numbers.

  14. Erythrocyte lysis in isotonic solution of ammonium chloride: Theoretical modelling and experimental verification

    NARCIS (Netherlands)

    Chernyshev, A.V.; Tarasov, P.A.; Semianov, K.A.; Nekrasov, V.M.; Hoekstra, A.G.; Maltsev, V.P.

    2008-01-01

    A mathematical model of erythrocyte lysis in isotonic solution of ammonium chloride is presented in frames of a statistical approach. The model is used to evaluate several parameters of mature erythrocytes (volume, surface area, hemoglobin concentration, number of anionic exchangers on membrane, ela

  15. An ultra-high temperature flow-through capillary device for bacterial spore lysis.

    Science.gov (United States)

    Hukari, Kyle W; Patel, Kamlesh D; Renzi, Ronald F; West, Jay A A

    2010-08-01

    Rapid and specific characterization of bacterial endospores is dependent on the ability to rupture the cell wall to enable analysis of the intracellular components. In particular, bacterial spores from the bacillus genus are inherently robust and very difficult to lyze or solubilize. Standard protocols for spore inactivation include chemical treatment, sonication, pressure, and thermal lysis. Although these protocols are effective for the inactivation of these agents, they are less well suited for sample preparation for analysis using proteomic and genomic approaches. To overcome this difficulty, we have designed a simple capillary device to perform thermal lysis of bacterial spores. Using this device, we were able to super heat (195 degrees C) an ethylene glycol lysis buffer to perform rapid flow-through rupture and solubilization of bacterial endospores. We demonstrated that the lysates from this preparation method are compatible with CGE as well as DNA amplification analysis. We further demonstrated the flow-through lysing device could be directly coupled to a miniaturized electrophoresis instrument for integrated sample preparation and analysis. In this arrangement, we were enabled to perform sample lysis, fluorescent dye labeling, and protein electrophoresis analysis of bacterial spores in less than 10 min. The described sample preparation device is rapid, simple, inexpensive, and easily integratable with various microfluidic devices.

  16. Viral lysis and microzooplankton grazing of phytoplankton throughout the Southern Ocean

    NARCIS (Netherlands)

    Evans, C.; Brussaard, C.P.D.

    2012-01-01

    We investigated microzooplankton grazing and viral lysis of the pico- and nanophytoplankton community in the Southern Ocean during the "Sensitivity of Sub-Antarctic Zone Waters to Global Change" cruise to the Australian sector (January-February 2007) and the Antarctic-XXIV/3 (February-March 2008) ex

  17. Hypochlorite- and hypobromite-mediated radical formation and its role in cell lysis

    DEFF Research Database (Denmark)

    Hawkins, C L; Brown, B E; Davies, Michael Jonathan

    2001-01-01

    Activated leukocytes generate the potent oxidants HOCl and HOBr via the formation of H(2)O(2) and the release of peroxidase enzymes (myeloperoxidase, eosinophil peroxidase). HOCl and HOBr are potent microbiocidal agents, but excessive or misplaced production can cause tissue damage and cell lysis...

  18. Prevention and treatment of tumor lysis syndrome, and the efficacy and role of rasburicase

    Directory of Open Access Journals (Sweden)

    Alakel N

    2017-02-01

    Full Text Available Nael Alakel,1 Jan Moritz Middeke,1 Johannes Schetelig,1,2 Martin Bornhäuser1 1Department of Internal Medicine I, University Hospital Carl Gustav Carus at the Technische Universitaet Dresden, Dresden, 2German Bone Marrow Donor Center DKMS, Tübigen, Germany Abstract: Tumor lysis syndrome (TLS is a potentially life-threatening condition that occurs in oncologic and hematologic patients with large tumor burden, either due to cytotoxic therapy or, less commonly, spontaneously because of massive tumor cell lysis. TLS is clinically characterized by acute renal failure, hyperuricemia, hyperkalemia, hyperphosphatemia, and hypocalcemia. While limited options are available for treating TLS, identifying patients at high risk for developing TLS and prevention in high-risk patients remain an important aspect in the treatment of cancer patients. In general, treatment of TLS consists of intensive hydration, stimulation of diuresis, and, more specifically, in the use of allopurinol and rasburicase. Rasburicase, a recombinant urate oxidase, rapidly and effectively reduces hyperuricemia, which subsequently significantly decreases the risk of acute renal failure and other clinical manifestations of TLS. For this review, a comprehensive literature search using the term “tumor lysis syndrome” and/or “rasburicase” was performed considering articles listed in MEDLINE. Incidence, prevention, and therapy of TLS with a special focus on the role of rasburicase are discussed. We evaluated 120 relevant articles including 35 case reports, 32 clinical trials, and 14 meta-analyses. Keywords: rasburicase, tumor lysis syndrome, hyperuricemia, acute kidney injury

  19. Analysis of Electric Fields inside Microchannels and Single Cell Electrical Lysis with a Microfluidic Device

    Directory of Open Access Journals (Sweden)

    Tofy Mussivand

    2013-06-01

    Full Text Available Analysis of electric fields generated inside the microchannels of a microfluidic device for electrical lysis of biological cells along with experimental verification are presented. Electrical lysis is the complete disintegration of cell membranes, due to a critical level of electric fields applied for a critical duration on a biological cell. Generating an electric field inside a microchannel of a microfluidic device has many advantages, including the efficient utilization of energy and low-current requirement. An ideal microchannel model was compared with a practical microchannel model using a finite element analysis tool that suggests that the overestimation error can be over 10%, from 2.5 mm or smaller, in the length of a microchannel. Two analytical forms are proposed to reduce this overestimation error. Experimental results showed that the high electric field is confined only inside the microchannel that is in agreement with the simulation results. Single cell electrical lysis was conducted with a fabricated microfluidic device. An average of 800 V for seven seconds across an 8 mm-long microchannel with the dimension of 100 μm × 20 μm was required for lysis, with electric fields exceeding 100 kV/m and consuming 300 mW.

  20. Genetically Determined Variation in Lysis Time Variance in the Bacteriophage φX174.

    Science.gov (United States)

    Baker, Christopher W; Miller, Craig R; Thaweethai, Tanayott; Yuan, Jeffrey; Baker, Meghan Hollibaugh; Joyce, Paul; Weinreich, Daniel M

    2016-04-07

    Researchers in evolutionary genetics recently have recognized an exciting opportunity in decomposing beneficial mutations into their proximal, mechanistic determinants. The application of methods and concepts from molecular biology and life history theory to studies of lytic bacteriophages (phages) has allowed them to understand how natural selection sees mutations influencing life history. This work motivated the research presented here, in which we explored whether, under consistent experimental conditions, small differences in the genome of bacteriophage φX174 could lead to altered life history phenotypes among a panel of eight genetically distinct clones. We assessed the clones' phenotypes by applying a novel statistical framework to the results of a serially sampled parallel infection assay, in which we simultaneously inoculated each of a large number of replicate host volumes with ∼1 phage particle. We sequentially plated the volumes over the course of infection and counted the plaques that formed after incubation. These counts served as a proxy for the number of phage particles in a single volume as a function of time. From repeated assays, we inferred significant, genetically determined heterogeneity in lysis time and burst size, including lysis time variance. These findings are interesting in light of the genetic and phenotypic constraints on the single-protein lysis mechanism of φX174. We speculate briefly on the mechanisms underlying our results, and we discuss the potential importance of lysis time variance in viral evolution.

  1. Genetically Determined Variation in Lysis Time Variance in the Bacteriophage φX174

    Directory of Open Access Journals (Sweden)

    Christopher W. Baker

    2016-04-01

    Full Text Available Researchers in evolutionary genetics recently have recognized an exciting opportunity in decomposing beneficial mutations into their proximal, mechanistic determinants. The application of methods and concepts from molecular biology and life history theory to studies of lytic bacteriophages (phages has allowed them to understand how natural selection sees mutations influencing life history. This work motivated the research presented here, in which we explored whether, under consistent experimental conditions, small differences in the genome of bacteriophage φX174 could lead to altered life history phenotypes among a panel of eight genetically distinct clones. We assessed the clones’ phenotypes by applying a novel statistical framework to the results of a serially sampled parallel infection assay, in which we simultaneously inoculated each of a large number of replicate host volumes with ∼1 phage particle. We sequentially plated the volumes over the course of infection and counted the plaques that formed after incubation. These counts served as a proxy for the number of phage particles in a single volume as a function of time. From repeated assays, we inferred significant, genetically determined heterogeneity in lysis time and burst size, including lysis time variance. These findings are interesting in light of the genetic and phenotypic constraints on the single-protein lysis mechanism of φX174. We speculate briefly on the mechanisms underlying our results, and we discuss the potential importance of lysis time variance in viral evolution.

  2. A simple and novel modification of comet assay for determination of bacteriophage mediated bacterial cell lysis.

    Science.gov (United States)

    Khairnar, Krishna; Sanmukh, Swapnil; Chandekar, Rajshree; Paunikar, Waman

    2014-07-01

    The comet assay is the widely used method for in vitro toxicity testing which is also an alternative to the use of animal models for in vivo testing. Since, its inception in 1984 by Ostling and Johansson, it is being modified frequently for a wide range of application. In spite of its wide applicability, unfortunately there is no report of its application in bacteriophages research. In this study, a novel application of comet assay for the detection of bacteriophage mediated bacterial cell lysis was described. The conventional methods in bacteriophage research for studying bacterial lysis by bacteriophages are plaque assay method. It is time consuming, laborious and costly. The lytic activity of bacteriophage devours the bacterial cell which results in the release of bacterial genomic material that gets detected by ethidium bromide staining method by the comet assay protocol. The objective of this study was to compare efficacy of comet assay with different assay used to study phage mediated bacterial lysis. The assay was performed on culture isolates (N=80 studies), modified comet assay appear to have relatively higher sensitivity and specificity than other assay. The results of the study showed that the application of comet assay can be an economical, time saving and less laborious alternative to conventional plaque assay for the detection of bacteriophage mediated bacterial cell lysis.

  3. Electrochemical lysis at the stage of endoresection for large posterior intraocular tumors

    Directory of Open Access Journals (Sweden)

    Yu. A. Belyy

    2012-01-01

    Full Text Available Purpose: to design the new combined technique of endoresection with intraoperative intraocular electrochemical lysis at the tumor destruction stage for large posterior intraocular tumors.Methods: 3 patients (3 eyes with large choroidal melanomas t3N0M0 (tumor thickness — 8-10 mm, base diameter — 13-15 mm, juxtapapillary localization. Mean age was 55.4 years old. Endoresection with intraoperational intraocular electrochemical lysis of the tumor was performed. Electrochemical lysis was performed with use of the technical unit ECU 300 (Soering, Germany and the original method of combined intratumoral positioning of two platinum electrodes: anode and cathode.Results: the tumor was removal completely in all 3 cases. the anatomical retinal reattachment was reached in all patients. Sclera was safe in all 3 cases. Visual acuity was not changed (NLP. At the place of the removal tumor a surgical choroidal coloboma without pigmentation all over scleral bed and periphery was shown in all cases in distant postoperative period (from 1.5 to 3 years. No local recurrences or metastasis were revealed in all patients.Conclusion: Further investigations in clinical group are necessarily to determinate the real possibilities of the combined method and the indications for endoresection with intraoperative intraocular electrochemical lysis for large intraocular tumors. 

  4. Processing of humic-rich riverine dissolved organic matter by estuarine bacteria: effects of predegradation and inorganic nutrients

    DEFF Research Database (Denmark)

    Asmala, E.; Autio, R.; Kaartokallio, H.

    2014-01-01

    The bioavailability of predegraded dissolved organic matter (DOM) from a humic-rich, boreal river to estuarine bacteria from the Baltic Sea was studied in 39-day bioassays. The river waters had been exposed to various degrees of bacterial degradation by storing them between 0 and 465 days in dark...... of predegradation and the inorganic nutrient status in the receiving estuary has consequences to carbon cycling and will determine the amount of terrestrial-derived DOC being ultimately assimilated into marine food webs...

  5. Fractalkine expression induces endothelial progenitor cell lysis by natural killer cells.

    Directory of Open Access Journals (Sweden)

    Dilyana Todorova

    Full Text Available BACKGROUND: Circulating CD34(+ cells, a population that includes endothelial progenitors, participate in the maintenance of endothelial integrity. Better understanding of the mechanisms that regulate their survival is crucial to improve their regenerative activity in cardiovascular and renal diseases. Chemokine-receptor cross talk is critical in regulating cell homeostasis. We hypothesized that cell surface expression of the chemokine fractalkine (FKN could target progenitor cell injury by Natural Killer (NK cells, thereby limiting their availability for vascular repair. METHODOLOGY/PRINCIPAL FINDINGS: We show that CD34(+-derived Endothelial Colony Forming Cells (ECFC can express FKN in response to TNF-α and IFN-γ inflammatory cytokines and that FKN expression by ECFC stimulates NK cell adhesion, NK cell-mediated ECFC lysis and microparticles release in vitro. The specific involvement of membrane FKN in these processes was demonstrated using FKN-transfected ECFC and anti-FKN blocking antibody. FKN expression was also evidenced on circulating CD34(+ progenitor cells and was detected at higher frequency in kidney transplant recipients, when compared to healthy controls. The proportion of CD34(+ cells expressing FKN was identified as an independent variable inversely correlated to CD34(+ progenitor cell count. We further showed that treatment of CD34(+ circulating cells isolated from adult blood donors with transplant serum or TNF-α/IFN-γ can induce FKN expression. CONCLUSIONS: Our data highlights a novel mechanism by which FKN expression on CD34(+ progenitor cells may target their NK cell mediated killing and participate to their immune depletion in transplant recipients. Considering the numerous diseased contexts shown to promote FKN expression, our data identify FKN as a hallmark of altered progenitor cell homeostasis with potential implications in better evaluation of vascular repair in patients.

  6. Method and apparatus for iterative lysis and extraction of algae

    Energy Technology Data Exchange (ETDEWEB)

    Chew, Geoffrey; Boggs, Tabitha; Dykes, Jr., H. Waite H.; Doherty, Stephen J.

    2015-12-01

    A method and system for processing algae involves the use of an ionic liquid-containing clarified cell lysate to lyse algae cells. The resulting crude cell lysate may be clarified and subsequently used to lyse algae cells. The process may be repeated a number of times before a clarified lysate is separated into lipid and aqueous phases for further processing and/or purification of desired products.

  7. Primary structure and functional analysis of the lysis genes of Lactobacillus gasseri bacteriophage phi adh.

    Science.gov (United States)

    Henrich, B; Binishofer, B; Bläsi, U

    1995-01-01

    The lysis genes of the Lactobacillus gasseri bacteriophage phi adh were isolated by complementation of a lambda Sam mutation in Escherichia coli. Nucleotide sequencing of a 1,735-bp DNA fragment revealed two adjacent coding regions of 342 bp (hol) and 951 bp (lys) in the same reading frame which appear to belong to a common transcriptional unit. Proteins corresponding to the predicted gene products, holin (12.9 kDa) and lysin (34.7 kDa), were identified by in vitro and in vivo expression of the cloned genes. The phi adh holin is a membrane-bound protein with structural similarity to lysis proteins of other phage, known to be required for the transit of murein hydrolases through the cytoplasmic membrane. The phi adh lysin shows homology with mureinolytic enzymes encoded by the Lactobacillus bulgaricus phage mv4, the Streptococcus pneumoniae phage Cp-1, Cp-7, and Cp-9, and the Lactococcus lactis phage phi LC3. Significant homology with the N termini of known muramidases suggests that phi adh lysin acts by a similar catalytic mechanism. In E. coli, the phi adh lysin seems to be associated with the total membrane fraction, from which it can be extracted with lauryl sarcosinate. Either one of the phi adh lysis proteins provoked lysis of E. coli when expressed along with holins or lysins of phage lambda or Bacillus subtilis phage phi 29. Concomitant expression of the combined holin and lysin functions of phi adh in E. coli, however, did not result in efficient cell lysis. PMID:7836307

  8. Optimization of a filter-lysis protocol to purify rat testicular homogenates for automated spermatid counting.

    Science.gov (United States)

    Pacheco, Sara E; Anderson, Linnea M; Boekelheide, Kim

    2012-01-01

    Quantifying testicular homogenization-resistant spermatid heads (HRSH) is a powerful indicator of spermatogenesis. These counts have traditionally been performed manually using a hemocytometer, but this method can be time consuming and biased. We aimed to develop a protocol to reduce debris for the application of automated counting, which would allow for efficient and unbiased quantification of rat HRSH. We developed a filter-lysis protocol that effectively removes debris from rat testicular homogenates. After filtering and lysing the homogenates, we found no statistical differences between manual (classic and filter-lysis) and automated (filter-lysis) counts using 1-way analysis of variance with Bonferroni's multiple comparison test. In addition, Pearson's correlation coefficients were calculated to compare the counting methods, and there was a strong correlation between the classic manual counts and the filter-lysis manual (r = 0.85, P = .002) and the filter-lysis automated (r = 0.89, P = .0005) counts. We also tested the utility of the automated method in a low-dose exposure model known to decrease HRSH. Adult Fischer 344 rats exposed to 0.33% 2,5-hexanedione in the drinking water for 12 weeks demonstrated decreased body (P = .02) and testes (P = .002) weights. In addition, there was a significant reduction in the number of HRSH per testis (P = .002) when compared to controls. A filterlysis protocol was optimized to purify rat testicular homogenates for automated HRSH counts. Automated counting systems yield unbiased data and can be applied to detect changes in the testis after low-dose toxicant exposure.

  9. Gene expression correlates with process rates quantified for sulfate- and Fe(III-reducing bacteria in U(VI-contaminated sediments

    Directory of Open Access Journals (Sweden)

    Denise M Akob

    2012-08-01

    Full Text Available Though iron- and sulfate-reducing bacteria are well known for mediating uranium(VI reduction in contaminated subsurface environments, quantifying the in situ activity of the microbial groups responsible remains a challenge. The objective of this study was to demonstrate the use of quantitative molecular tools that target mRNA transcripts of key genes related to Fe(III and sulfate reduction pathways in order to monitor these processes during in situ U(VI remediation in the subsurface. Expression of the Geobacteraceae-specific citrate synthase gene (gltA and the dissimilatory (bisulfite reductase gene (dsrA, were correlated with the activity of iron- or sulfate-reducing microorganisms, respectively, under stimulated bioremediation conditions in microcosms of sediments sampled from the U.S. Department of Energy’s Oak Ridge Integrated Field Research Challenge (OR-IFRC site at Oak Ridge, Tennessee. In addition, Geobacteraceae-specific gltA and dsrA transcript levels were determined in parallel with the predominant electron acceptors present in moderately and highly contaminated subsurface sediments from the OR-IFRC. Phylogenetic analysis of the cDNA generated from dsrA mRNA, sulfate-reducing bacteria-specific 16S rRNA, and gltA mRNA identified activity of specific microbial groups. Active sulfate reducers were members of the Desulfovibrio, Desulfobacterium, and Desulfotomaculum genera. Members of the subsurface Geobacter clade, closely related to uranium-reducing Geobacter uraniireducens and Geobacter daltonii, were the metabolically-active iron-reducers in biostimulated microcosms and in situ core samples. Direct correlation of transcripts and process rates demonstrated evidence of competition between the functional guilds in subsurface sediments. We further showed that active populations of Fe(III-reducing bacteria and sulfate-reducing bacteria are present in OR-IFRC sediments and are good potential targets for in situ bioremediation.

  10. 蔬菜腌制及其乳酸菌技术的研究进展%Pickling Process of Vegetables and its Lactic Acid Bacteria Technology

    Institute of Scientific and Technical Information of China (English)

    吴祖芳; 赵永威; 翁佩芳

    2012-01-01

    Pickled vegetables processing is an important part of the vegetable processing industry. With the gradual deepening developing of lactic acid bacteria research and new technologies of agricultural products processing, one of the great research interests-was focused on the study of pickled vegetables process. In this paper, the microbial diversity, metabolite of lactic bacteria and flavor compound formation, the functional properties evaluations as well as the ferment starter application of pickled vegetables were elucidated among the domestic and foreign research progress. This can provide the references for technology development and research of pickled vegetables processing.%蔬菜腌制加工是蔬菜加工产业中的重要组成部分,随着农产品加工新技术及乳酸菌研究的逐步深入和发展,腌制蔬菜也成为一个新的研究热点.作者分析了国内外有关蔬菜腌制过程中微生态、乳酸菌的代谢及其蔬菜腌制风味发生机制、腌制蔬菜功能特性评价以及发酵剂开发等方面的研究进展,为蔬菜腌制技术的发展等提供参考.

  11. Meat Processing Plant Microbiome and Contamination Patterns of Cold-Tolerant Bacteria Causing Food Safety and Spoilage Risks in the Manufacture of Vacuum-Packaged Cooked Sausages.

    Science.gov (United States)

    Hultman, Jenni; Rahkila, Riitta; Ali, Javeria; Rousu, Juho; Björkroth, K Johanna

    2015-10-01

    Refrigerated food processing facilities are specific man-made niches likely to harbor cold-tolerant bacteria. To characterize this type of microbiota and study the link between processing plant and product microbiomes, we followed and compared microbiota associated with the raw materials and processing stages of a vacuum-packaged, cooked sausage product affected by a prolonged quality fluctuation with occasional spoilage manifestations during shelf life. A total of 195 samples were subjected to culturing and amplicon sequence analyses. Abundant mesophilic psychrotrophs were detected within the microbiomes throughout the different compartments of the production plant environment. However, each of the main genera of food safety and quality interest, e.g., Leuconostoc, Brochothrix, and Yersinia, had their own characteristic patterns of contamination. Bacteria from the genus Leuconostoc, commonly causing spoilage of cold-stored, modified-atmosphere-packaged foods, were detected in high abundance (up to >98%) in the sausages studied. The same operational taxonomic units (OTUs) were, however, detected in lower abundances in raw meat and emulsion (average relative abundance of 2%±5%), as well as on the processing plant surfaces (processing plant surfaces but to a lesser extent (food safety concerns related to their resilient existence on surfaces.

  12. Dual-species biofilm formation by Escherichia coli O157:H7 and environmental bacteria isolated from fresh-cut processing facilities.

    Science.gov (United States)

    Liu, Nancy T; Nou, Xiangwu; Lefcourt, Alan M; Shelton, Daniel R; Lo, Y Martin

    2014-02-03

    Biofilm formation is a mechanism adapted by many microorganisms that enhances the survival in stressful environments. In food processing facilities, foodborne bacterial pathogens, which many are poor biofilm formers, could potentially take advantage of this protective mechanism by interacting with other strong biofilm producers. The objective of this study was to determine the influence of bacteria native to fresh produce processing environments on the incorporation of Escherichia coli O157:H7 in biofilms. Bacteria strains representing 13 Gram-negative species isolated from two fresh produce processing facilities in a previous study were tested for forming dual-species biofilms with E. coli O157:H7. Strong biofilm producing strains of Burkholderia caryophylli and Ralstonia insidiosa exhibited 180% and 63% increase in biofilm biomass, and significant thickening of the biofilms (B. caryophylli not tested), when co-cultured with E. coli O157:H7. E. coli O157:H7 populations increased by approximately 1 log in dual-species biofilms formed with B. caryophylli or R. insidiosa. While only a subset of environmental isolates with strong biofilm formation abilities increased the presence of E. coli O157:H7 in biofilms, all tested E. coli O157:H7 exhibited higher incorporation in dual-species biofilms with R. insidiosa. These observations support the notion that E. coli O157:H7 and specific strong biofilm producing bacteria interact synergistically in biofilm formation, and suggest a route for increased survival potential of E. coli O157:H7 in fresh produce processing environments.

  13. Abundance and diversity of ammonia-oxidizing archaea and bacteria on biological activated carbon in a pilot-scale drinking water treatment plant with different treatment processes.

    Science.gov (United States)

    Kasuga, Ikuro; Nakagaki, Hirotaka; Kurisu, Futoshi; Furumai, Hiroaki

    2010-01-01

    The effects of different placements of rapid sand filtration on nitrification performance of BAC treatment in a pilot-scale plant were evaluated. In this plant, rapid sand filtration was placed after ozonation-BAC treatment in Process (A), while it preceded ozonation-BAC treatment in Process (B). Analysis of amoA genes of ammonia-oxidizing archaea (AOA) and bacteria (AOB) combined with nitrification potential test was conducted. BAC from Process (A) demonstrated slightly higher nitrification potential at every sampling occasion. This might be due to higher abundances of AOB on BAC from Process (A) than those on BAC from Process (B). However, AOA rather than AOB could be predominant ammonia-oxidizers in BAC treatment regardless of the position of rapid sand filtration. The highest nitrification potential was observed for BAC from both processes in February when the highest abundances of AOA-amoA and AOB-amoA genes were detected. Since rapid sand filtration was placed after BAC treatment in Process (A), residual aluminum concentration in BAC influent was higher in Process (A). However, adverse effects of aluminum on nitrification activity were not observed. These results suggest that factors other than aluminum concentration in different treatment processes could possibly have some influence on abundances of ammonia-oxidizing microorganisms on BAC.

  14. Big bacteria

    DEFF Research Database (Denmark)

    Schulz, HN; Jørgensen, BB

    2001-01-01

    A small number of prokaryotic species have a unique physiology or ecology related to their development of unusually large size. The biomass of bacteria varies over more than 10 orders of magnitude, from the 0.2 mum wide nanobacteria to the largest cells of the colorless sulfur bacteria......, Thiomargarita namibiensis, with a diameter of 750 mum. All bacteria, including those that swim around in the environment, obtain their food molecules by molecular diffusion. Only the fastest and largest swimmers known, Thiovulum majus, are able to significantly increase their food supply by motility...... and by actively creating an advective flow through the entire population. Diffusion limitation generally restricts the maximal size of prokaryotic cells and provides a selective advantage for mum-sized cells at the normally low substrate concentrations in the environment. The largest heterotrophic bacteria...

  15. Anaerobic bacteria

    Science.gov (United States)

    Brook I, Goldstein EJ. Diseases caused by non-spore forming anaerobic bacteria. In: Goldman L, Schafer AI, eds. Goldman's Cecil Medicine . 25th ed. Philadelphia, PA: Elsevier Saunders; 2015:chap 297. Stedman's Online ...

  16. DNA isolation protocols affect the detection limit of PCR approaches of bacteria in samples from the human gastrointestinal tract

    NARCIS (Netherlands)

    Zoetendal, E.G.; Ben-Amor, K.; Akkermans, A.D.L.; Abee, T.; Vos, de W.M.

    2001-01-01

    A major concern in molecular ecological studies is the lysis efficiency of different bacteria in a complex ecosystem. We used a PCR-based 16S rDNA approach to determine the effect of two DNA isolation protocols (i.e. the bead beating and Triton-X100 method) on the detection limit of seven feces-asso

  17. The Use of Lactic Acid Bacteria Starter Cultures during the Processing of Fermented Cereal-based Foods in West Africa: A Review.

    Science.gov (United States)

    Soro-Yao, Amenan Anastasie; Brou, Kouakou; Amani, Georges; Thonart, Philippe; Djè, Koffi Marcelin

    2014-12-01

    Lactic acid bacteria (LAB) are the primary microorganisms used to ferment maize-, sorghum- or millet-based foods that are processed in West Africa. Fermentation contributes to desirable changes in taste, flavour, acidity, digestibility and texture in gruels (ogi, baca, dalaki), doughs (agidi, banku, komé) or steam-cooked granulated products (arraw, ciacry, dégué). Similar to other fermented cereal foods that are available in Africa, these products suffer from inconsistent quality. The use of LAB starter cultures during cereal dough fermentation is a subject of increasing interest in efforts to standardise this step and guaranty product uniformity. However, their use by small-scale processing units or small agro-food industrial enterprises is still limited. This review aims to illustrate and discuss major issues that influence the use of LAB starter cultures during the processing of fermented cereal foods in West Africa.

  18. Characterization of cell lysis in Pseudomonas putida induced upon expression of heterologous killing genes

    DEFF Research Database (Denmark)

    Ronchel, M.C.; Molina, L.; Witte, A.;

    1998-01-01

    Active biological containment systems are based on the controlled expression of killing genes. These systems are of interest for the Pseudomonadaceae because of the potential applications of these microbes as bioremediation agents and biopesticides, The physiological effects that lead to cell death...... upon the induction of expression of two different heterologous killing genes in nonpathogenic Pseudomonas putida KT2440 derivatives have been analyzed, P. putida CMC4 and CMC12 carry in their chromosomes a fusion of the PAl-04/03 promoter to the Escherichia coli gef gene and the phi X174 lysis gene E......, respectively. Expression of the killing genes is controlled by the LacI protein, whose expression is initiated from the XylS-dependent Pm promoter. Under induced conditions, killing of P. putida CMC12 cells mediated by phi X174 lysis protein E was faster than that observed for P. putida CMC4, for which the Gef...

  19. Spontaneous Tumor Lysis Syndrome in the Setting of B-Cell Lymphoma

    Directory of Open Access Journals (Sweden)

    Mateusz Opyrchal

    2010-01-01

    Full Text Available Tumor lysis syndrome (TLS presenting in absence of chemotherapy is a rare occurrence. One of the true oncological emergencies, it can lead to significant morbidity and mortality. TLS is a phenomena usually associated with tumor cell death after treatment. The etiology of the spontaneous TLS is not well understood, which complicates the diagnosis. TLS is well known to oncologists but physicians outside of this specialty have little or no experience with this condition. Early recognition and treatment are the keys to limiting the sequela of the condition. Spontaneous tumor lysis syndrome is rare but presents added risks to the patient because of the potential for delayed diagnosis and no benefit of pretreatment. Diagnosis may be further delayed because this may be the first symptom of underlying malignancy. Therefore, it is imperative that all clinicians are familiar with the syndrome to assure timely recognition.

  20. Feasibility of Wide-Area Decontamination of Bacillus anthracis Spores Using a Germination-Lysis Approach

    Science.gov (United States)

    2011-11-16

    ethanol showed disinfection within 15 min  5% pH-adjusted bleach was effective within 15 min (100% lysis)  3% H2O2 was effective after 30 min (100...508394 Lawrence Livermore National Laboratory LLNL-PRES- 4 Disinfectant >6 Log Reduction on Materials (EPA, 2010a,b; Wood et al., 2011...Target Disinfectant Concentration (ppm) OSHA PEL (ppm) CDC IDLH (ppm) pH-amended bleach (sodium hypochlorite) Stainless Steel, Glass

  1. [Tumor lysis syndrome in a pregnancy complicated with acute lymphoblastic leukemia].

    Science.gov (United States)

    Álvarez-Goris, M P; Sánchez-Zamora, R; Torres-Aguilar, A A; Briones Garduño, J C

    2016-04-01

    Acute leukemia is rare during pregnancy, affects about 1 in 75,000 pregnancies, of all leukemias diagnosed only 28% are acute lymphoblastic leukemia, this is a risk factor to develop spontaneous tumor lysis syndrome, it's a oncologic complication potentially deadly if the prophylactic treatment its avoided. Cases of acute lymphoblastic leukemia associated with pregnancy has been poorly documented in the literature the association of these two entities to pregnancy is the first report published worldwide, so the information is limited.

  2. Prevention and treatment of tumor lysis syndrome, and the efficacy and role of rasburicase

    Science.gov (United States)

    Alakel, Nael; Middeke, Jan Moritz; Schetelig, Johannes; Bornhäuser, Martin

    2017-01-01

    Tumor lysis syndrome (TLS) is a potentially life-threatening condition that occurs in oncologic and hematologic patients with large tumor burden, either due to cytotoxic therapy or, less commonly, spontaneously because of massive tumor cell lysis. TLS is clinically characterized by acute renal failure, hyperuricemia, hyperkalemia, hyperphosphatemia, and hypocalcemia. While limited options are available for treating TLS, identifying patients at high risk for developing TLS and prevention in high-risk patients remain an important aspect in the treatment of cancer patients. In general, treatment of TLS consists of intensive hydration, stimulation of diuresis, and, more specifically, in the use of allopurinol and rasburicase. Rasburicase, a recombinant urate oxidase, rapidly and effectively reduces hyperuricemia, which subsequently significantly decreases the risk of acute renal failure and other clinical manifestations of TLS. For this review, a comprehensive literature search using the term “tumor lysis syndrome” and/or “rasburicase” was performed considering articles listed in MEDLINE. Incidence, prevention, and therapy of TLS with a special focus on the role of rasburicase are discussed. We evaluated 120 relevant articles including 35 case reports, 32 clinical trials, and 14 meta-analyses. PMID:28203093

  3. A novel, compact disk-like centrifugal microfluidics system for cell lysis and sample homogenization.

    Science.gov (United States)

    Kido, Horacio; Micic, Miodrag; Smith, David; Zoval, Jim; Norton, Jim; Madou, Marc

    2007-07-01

    In this paper, we present the design and characterization of a novel platform for mechanical cell lysis of even the most difficult to lyse cell types on a micro or nanoscale (maximum 70 microL total volume). The system incorporates a machined plastic circular disk assembly, magnetic field actuated microfluidics, centrifugal cells and tissue homogenizer and centrifugation system. The mechanism of tissue disruption of this novel cell homogenization apparatus derives from the relative motion of ferromagnetic metal disks and grinding matrices in a liquid medium within individual chambers of the disk in the presence of an oscillating magnetic field. The oscillation of the ferromagnetic disks or blades produces mechanical impaction and shear forces capable of disrupting cells within the chamber both by direct action of the blade and by the motion of the surrounding lysis matrix, and by motion induced vortexing of buffer fluid. Glass beads or other grinding media are integrated into each lysis chamber within the disk to enhance the transfer of energy from the oscillating metal blade to the cells. The system also achieves the centrifugal elimination of solids from each liquid sample and allows the elution of clarified supernatants via siphoning into a collection chamber fabricated into the plastic disk assembly. This article describes system design, implementation and validation of proof of concept on two samples--Escherichia coli and Saccharomyces cerevisiae representing model systems for cells that are easy and difficult to lyse, respectively.

  4. Molecular identification of potential denitrifying bacteria and use of D-optimal mixture experimental design for the optimization of denitrification process.

    Science.gov (United States)

    Ben Taheur, Fadia; Fdhila, Kais; Elabed, Hamouda; Bouguerra, Amel; Kouidhi, Bochra; Bakhrouf, Amina; Chaieb, Kamel

    2016-04-01

    Three bacterial strains (TE1, TD3 and FB2) were isolated from date palm (degla), pistachio and barley. The presence of nitrate reductase (narG) and nitrite reductase (nirS and nirK) genes in the selected strains was detected by PCR technique. Molecular identification based on 16S rDNA sequencing method was applied to identify positive strains. In addition, the D-optimal mixture experimental design was used to optimize the optimal formulation of probiotic bacteria for denitrification process. Strains harboring denitrification genes were identified as: TE1, Agrococcus sp LN828197; TD3, Cronobacter sakazakii LN828198 and FB2, Pedicoccus pentosaceus LN828199. PCR results revealed that all strains carried the nirS gene. However only C. sakazakii LN828198 and Agrococcus sp LN828197 harbored the nirK and the narG genes respectively. Moreover, the studied bacteria were able to form biofilm on abiotic surfaces with different degree. Process optimization showed that the most significant reduction of nitrate was 100% with 14.98% of COD consumption and 5.57 mg/l nitrite accumulation. Meanwhile, the response values were optimized and showed that the most optimal combination was 78.79% of C. sakazakii LN828198 (curve value), 21.21% of P. pentosaceus LN828199 (curve value) and absence (0%) of Agrococcus sp LN828197 (curve value).

  5. Squamous cell carcinoma complicating an hereditary epidermo-lysis bullosa; Carcinome spinocellulaire compliquant une epidermolyse bulleuse hereditaire

    Energy Technology Data Exchange (ETDEWEB)

    Mseddi, M.; Turki, H.; Marrekchi, S.; Abdelmaksoud, W.; Masmoudi, A.; Bouassida, S.; Zahaf, A. [Centre Hospitalier Universitaire Hedi Chaker, Service de Dermatologie, Sfax (Tunisia)

    2004-08-01

    The dystrophic form of hereditary epidermo-lysis bullosa is associated with an increased frequency of squamous cell carcinoma. We report a new case. An 18-year-old patient, carrying a Hallopeau Siemens hereditary epidermo-lysis bullosa, presented a subcutaneous nodular lesion, for 1 year that ulcerated and budded with inguinal lymphadenopathy. The histological study ted to the conclusion of a well differentiated squamous cell carcinoma. The patient was treated surgically. Tumor and metastatic lymph nodes were excised. A radiotherapy was decided but the postoperative course was fatal due to an infection and to a deterioration of her general condition. Squamous cell carcinoma frequently occurs on the cicatricial lesion of hereditary epidermo-lysis bullosa and usually affects males with recessive hereditary epidermo-lysis bullosa. Metastases are frequent, precocious and multiple. The treatment may be surgical. The particularities of our observation are the young age of patient and the localization. (author)

  6. Effect of brine marination on survival and growth of spoilage and pathogenic bacteria during processing and subsequent storage of ready-to-eat shrimp (Pandalus borealis)

    DEFF Research Database (Denmark)

    Mejlholm, Ole; Devitt, Tina D.; Dalgaard, Paw

    2012-01-01

    and growth during processing and subsequent storage of brined and drained shrimp in modified atmosphere packaging (MAP). Different brines with (i) acetic and lactic acids (AL) or (ii) benzoic, citric and sorbic acids (BCS) were studied. V. parahaemolyticus was inactivated in brine AL without shrimp whereas...... of the examined pathogens was able to grow at 7°C in brined and drained MAP shrimp that resembled commercial products. However, reducing the concentration of acetic and lactic acids by 50% resulted in relatively fast growth of L. monocytogenes in brined and drained MAP shrimp at 7°C. Growth of S. aureus......The effect of brine marination at chill temperatures on survival and growth of spoilage and pathogenic bacteria during processing and subsequent storage of ready-to-eat cold water shrimp was studied. Survival and growth of Lactobacillus sakei, Listeria monocytogenes, Salmonella, Staphylococcus...

  7. Isolation and Identification of Concrete Environment Bacteria

    Science.gov (United States)

    Irwan, J. M.; Anneza, L. H.; Othman, N.; Husnul, T.; Alshalif, A. F.

    2016-07-01

    This paper presents the isolation and molecular method for bacteria identification through PCR and DNA sequencing. Identification of the bacteria species is required in order to fully utilize the bacterium capability for precipitation of calcium carbonate in concrete. This process is to enable the addition of suitable catalyst according to the bacterium enzymatic pathway that is known through the bacteria species used. The objective of this study is to isolate, enriched and identify the bacteria species. The bacteria in this study was isolated from fresh urine and acid mine drainage water, Kota Tinggi, Johor. Enrichment of the isolated bacteria was conducted to ensure the bacteria survivability in concrete. The identification of bacteria species was done through polymerase chain reaction (PCR) and rRDNA sequencing. The isolation and enrichment of the bacteria was done successfully. Whereas, the results for bacteria identification showed that the isolated bacteria strains are Bacillus sp and Enterococus faecalis.

  8. Lipopolysaccharides in diazotrophic bacteria.

    Science.gov (United States)

    Serrato, Rodrigo V

    2014-01-01

    Biological nitrogen fixation (BNF) is a process in which the atmospheric nitrogen (N2) is transformed into ammonia (NH3) by a select group of nitrogen-fixing organisms, or diazotrophic bacteria. In order to furnish the biologically useful nitrogen to plants, these bacteria must be in constant molecular communication with their host plants. Some of these molecular plant-microbe interactions are very specific, resulting in a symbiotic relationship between the diazotroph and the host. Others are found between associative diazotrophs and plants, resulting in plant infection and colonization of internal tissues. Independent of the type of ecological interaction, glycans, and glycoconjugates produced by these bacteria play an important role in the molecular communication prior and during colonization. Even though exopolysaccharides (EPS) and lipochitooligosaccharides (LCO) produced by diazotrophic bacteria and released onto the environment have their importance in the microbe-plant interaction, it is the lipopolysaccharides (LPS), anchored on the external membrane of these bacteria, that mediates the direct contact of the diazotroph with the host cells. These molecules are extremely variable among the several species of nitrogen fixing-bacteria, and there are evidences of the mechanisms of infection being closely related to their structure.

  9. Lipopolysaccharides in diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Rodrigo Vassoler Serrato

    2014-09-01

    Full Text Available Biological nitrogen fixation is a process in which the atmospheric nitrogen (N2 is transformed into ammonia (NH3 by a select group of nitrogen-fixing organisms, or diazotrophic bacteria. In order to furnish the biologically useful nitrogen to plants, these bacteria must be in constant molecular communication with their host plants. Some of these molecular plant-microbe interactions are very specific, resulting in a symbiotic relationship between the diazotroph and the host. Others are found between associative diazotrophs and plants, resulting in plant infection and colonization of internal tissues. Independent of the type of ecological interaction, glycans and glycoconjugates produced by these bacteria play an important role in the molecular communication prior and during colonization. Even though exopolysaccharides (EPS and lipochitooligosaccharides (LCO produced by diazotrophic bacteria and released onto the environment have their importance in the microbe-plant interaction, it is the lipopolysaccharides (LPS, anchored on the external membrane of these bacteria, that mediates the direct contact of the diazotroph with the host cells. These molecules are extremely variable among the several species of nitrogen fixing-bacteria, and there are evidences of the mechanisms of infection being closely related to their structure.

  10. Effect of inoculation on strawberry fermentation and acetification processes using native strains of yeast and acetic acid bacteria.

    Science.gov (United States)

    Hidalgo, C; Torija, M J; Mas, A; Mateo, E

    2013-05-01

    The aim of this work was to analyze the microbiota involved in the traditional vinegar elaboration of strawberry fruit during a spontaneous and inoculated process. In the spontaneous processes, low biodiversity was detected in both alcoholic fermentation (AF) and acetification. Nevertheless, a strain of Saccharomyces cerevisiae and of Acetobacter malorum were selected and tested as starter cultures in the inoculation study. The inoculated processes with these strains were compared with another spontaneous process, yielding a significant reduction in time for AF with a total imposition of the S. cerevisiae strain. The resulting strawberry wine was acetified in different containers (glass and wood) yielding an initial imposition of the A. malorum inoculated strain, although displacement by Gluconacetobacter species was observed in the wood barrels.

  11. Screening large numbers of recombinant plasmids: modifications and additions to alkaline lysis for greater efficiency

    Institute of Scientific and Technical Information of China (English)

    XU Yibing; N.V. CHANDRASEKHARAN; Daniel L. SIMMONS

    2006-01-01

    Selecting bacteria transformed with recombinant plasmid is a laborious step in gene cloning experiments. This selection process is even more tedious when large numbers of clones need to be screened. We describe here modifications to the ultra fast plasmid preparation method described previously by Law and Crickmore. The modified method is coupled to an efficient PCR step to rapidly determine orientation of the inserts. Compared to traditional methods of analysis requiring growth of overnight cultures, plasmid isolation and restriction enzyme digestion to determine orientation this procedure allows for the analysis and storage of a large number of recombinants within a few hours.

  12. Constraints on mechanisms and rates of anaerobic oxidation of methane by microbial consortia: process-based modeling of ANME-2 archaea and sulfate reducing bacteria interactions

    Directory of Open Access Journals (Sweden)

    B. Orcutt

    2008-11-01

    Full Text Available Anaerobic oxidation of methane (AOM is the main process responsible for the removal of methane generated in Earth's marine subsurface environments. However, the biochemical mechanism of AOM remains elusive. By explicitly resolving the observed spatial arrangement of methanotrophic archaea and sulfate reducing bacteria found in consortia mediating AOM, potential intermediates involved in the electron transfer between the methane oxidizing and sulfate reducing partners were investigated via a consortium-scale reaction transport model that integrates the effect of diffusional transport with thermodynamic and kinetic controls on microbial activity. Model simulations were used to assess the impact of poorly constrained microbial characteristics such as minimum energy requirements to sustain metabolism and cell specific rates. The role of environmental conditions such as the influence of methane levels on the feasibility of H2, formate and acetate as intermediate species, and the impact of the abundance of intermediate species on pathway reversal were examined. The results show that higher production rates of intermediates via AOM lead to increased diffusive fluxes from the methane oxidizing archaea to sulfate reducing bacteria, but the build-up of the exchangeable species can cause the energy yield of AOM to drop below that required for ATP production. Comparison to data from laboratory experiments shows that under the experimental conditions of Nauhaus et al. (2007, none of the potential intermediates considered here is able to support metabolic activity matching the measured rates.

  13. Direct identification of microorganisms from positive blood cultures using the lysis-filtration technique and matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS): a multicentre study.

    Science.gov (United States)

    Farina, Claudio; Arena, Fabio; Casprini, Patrizia; Cichero, Paola; Clementi, Massimo; Cosentino, Marina; Degl'Innocenti, Roberto; Giani, Tommaso; Luzzaro, Francesco; Mattei, Romano; Mauri, Carola; Nardone, Maria; Rossolini, Gian Maria; Serna Ortega, Paula Andrea; Vailati, Francesca

    2015-04-01

    Microbial identification from blood cultures is essential to institute optimal antibiotic therapy and improve survival possibilities. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied to identify bacteria and yeasts from positive blood cultures broths. The aim of this multicentre study was to evaluate the reliability of the lysis-filtration technique associated with MALDI-TOF MS to directly identify microorganisms from 765 positive blood cultures collected in six Italian hospitals. Overall, 675/765 (78.1%) blood isolates were correctly identified at the species level, with significant differences between Gram-negative and Gram-positive bacteria (92.6%, and 69.8%, respectively). Some difficulties arise in identifying Streptococcus pneumoniae, Staphylococcus aureus, yeasts and anaerobes. The lysis-filtration protocol is a suitable procedure in terms of performance in identifying microorganisms, but it is quite expensive and technically time-consuming since the time of filtration is not regular for all the samples. The application of the MALDI-TOF MS technique to the direct microbial identification from positive blood cultures is a very promising approach, even if more experience must be gained to minimize errors and costs.

  14. Evaluation of the removal of indicator bacteria from domestic sludge processed by Autothermal Thermophilic Aerobic Digestion (ATAD).

    Science.gov (United States)

    Piterina, Anna V; Bartlett, John; Pembroke, Tony J

    2010-09-01

    The degradation of sludge solids in an insulated reactor during Autothermal Thermophilic Aerobic Digestion (ATAD) processing results in auto-heating, thermal treatment and total solids reduction, however, the ability to eliminate pathogenic organisms has not been analysed under large scale process conditions. We evaluated the ATAD process over a period of one year in a two stage, full scale Irish ATAD plant established in Killarney and treating mixed primary and secondary sludge, by examining the sludge microbiologically at various stages during and following ATAD processing to determine its ability to eliminate indicator organisms. Salmonella spp. (pathogen) and fecal-coliform (indicator) densities were well below the limits used to validate class A biosolids in the final product. Enteric pathogens present at inlet were deactivated during the ATAD process and were not detected in the final product using both traditional microbial culture and molecular phylogenetic techniques. A high DNase activity was detected in the bulk sludge during the thermophilic digestion stage which may be responsible for the rapid turn over of DNA from lysed cells and the removal of mobile DNA. These results offer assurance for the safe use of ATAD sludge as a soil supplement following processing.

  15. Evaluation of the Removal of Indicator Bacteria from Domestic Sludge Processed by Autothermal Thermophilic Aerobic Digestion (ATAD

    Directory of Open Access Journals (Sweden)

    Tony J. Pembroke

    2010-09-01

    Full Text Available The degradation of sludge solids in an insulated reactor during Autothermal Thermophilic Aerobic Digestion (ATAD processing results in auto-heating, thermal treatment and total solids reduction, however, the ability to eliminate pathogenic organisms has not been analysed under large scale process conditions. We evaluated the ATAD process over a period of one year in a two stage, full scale Irish ATAD plant established in Killarney and treating mixed primary and secondary sludge, by examining the sludge microbiologically at various stages during and following ATAD processing to determine its ability to eliminate indicator organisms. Salmonella spp. (pathogen and fecal-coliform (indicator densities were well below the limits used to validate class A biosolids in the final product. Enteric pathogens present at inlet were deactivated during the ATAD process and were not detected in the final product using both traditional microbial culture and molecular phylogenetic techniques. A high DNase activity was detected in the bulk sludge during the thermophilic digestion stage which may be responsible for the rapid turn over of DNA from lysed cells and the removal of mobile DNA. These results offer assurance for the safe use of ATAD sludge as a soil supplement following processing.

  16. Potential of lactic acid bacteria to reduce the growth ofFusarium culmorum in the malting process.

    Science.gov (United States)

    Liske, R; Niessen, L; Vogel, R

    2000-03-01

    25 microorganisms were used for tested for their potential to reduce growth of toxigenic and gushing-activeFusarium culmorum strains during the malting process. Twelve isolates were found to substantially decrease the growth ofF. culmorum under the conditions applied.

  17. Lactic acid bacteria and natural antimicrobials to improve the safety and shelf-life of minimally processed sliced apples and lamb's lettuce.

    Science.gov (United States)

    Siroli, Lorenzo; Patrignani, Francesca; Serrazanetti, Diana I; Tabanelli, Giulia; Montanari, Chiara; Gardini, Fausto; Lanciotti, Rosalba

    2015-05-01

    Outbreaks of food-borne disease associated with the consumption of fresh and minimally processed fruits and vegetables have increased dramatically over the last few years. Traditional chemical sanitizers are unable to completely eradicate or kill the microorganisms on fresh produce. These conditions have stimulated research to alternative methods for increasing food safety. The use of protective cultures, particularly lactic acid bacteria (LAB), has been proposed for minimally processed products. However, the application of bioprotective cultures has been limited at the industrial level. From this perspective, the main aims of this study were to select LAB from minimally processed fruits and vegetables to be used as biocontrol agents and then to evaluate the effects of the selected strains, alone or in combination with natural antimicrobials (2-(E)-hexenal/hexanal, 2-(E)-hexenal/citral for apples and thyme for lamb's lettuce), on the shelf-life and safety characteristics of minimally processed apples and lamb's lettuce. The results indicated that applying the Lactobacillus plantarum strains CIT3 and V7B3 to apples and lettuce, respectively, increased both the safety and shelf-life. Moreover, combining the selected strains with natural antimicrobials produced a further increase in the shelf-life of these products without detrimental effects on the organoleptic qualities.

  18. Leukocyte Lysis and Cytokine Induction by the Human Sexually Transmitted Parasite Trichomonas vaginalis

    Science.gov (United States)

    Mercer, Frances; Diala, Fitz Gerald I.; Chen, Yi-Pei; Molgora, Brenda M.; Ng, Shek Hang; Johnson, Patricia J.

    2016-01-01

    Trichomonas vaginalis (Tv) is an extracellular protozoan parasite that causes the most common non-viral sexually transmitted infection: trichomoniasis. While acute symptoms in women may include vaginitis, infections are often asymptomatic, but can persist and are associated with medical complications including increased HIV susceptibility, infertility, pre-term labor, and higher incidence of cervical cancer. Heightened inflammation resulting from Tv infection could account for these complications. Effective cellular immune responses to Tv have not been characterized, and re-infection is common, suggesting a dysfunctional adaptive immune response. Using primary human leukocyte components, we have established an in vitro co-culture system to assess the interaction between Tv and the cells of the human immune system. We determined that in vitro, Tv is able to lyse T-cells and B-cells, showing a preference for B-cells. We also found that Tv lysis of lymphocytes was mediated by contact-dependent and soluble factors. Tv lysis of monocytes is far less efficient, and almost entirely contact-dependent. Interestingly, a common symbiont of Tv, Mycoplasma hominis, did not affect cytolytic activity of the parasite, but had a major impact on cytokine responses. M. hominis enabled more diverse inflammatory cytokine secretion in response to Tv and, of the cytokines tested, Tv strains cleared of M. hominis induced only IL-8 secretion from monocytes. The quality of the adaptive immune response to Tv is therefore likely influenced by Tv symbionts, commensals, and concomitant infections, and may be further complicated by direct parasite lysis of effector immune cells. PMID:27529696

  19. Sorafenib induced tumor lysis syndrome in an advanced hepatocellular carcinoma patient

    Institute of Scientific and Technical Information of China (English)

    Wu-Shiung Huang; Chang-Hsu Yang

    2009-01-01

    A 55-year-old male patient with hepatitis B-related liver cirrhosis was found to have advanced hepatocellular carcinoma. His AFP was initially 9828 mg/L and rapidly dropped to 5597 mg/L in ten days after oral sorafenib treatment. However, he developed acute renal failure, hyperkalemia, and hyperuricemia 30 d after receiving the sorafenib treatment. Tumor lysis syndrome was suspected and intensive hemodialysis was performed. Despite intensive hemodialysis and other supportive therapy, he developed multiple organ failure (liver, renal, and respiratory failure) and metabolic acidosis. The patient expired 13 d after admission.

  20. Optimization of Electrical Methods for Sub -surface Monitoring of Biological Contamination: From Micro-scale to Macroscopic one through Sub-micrometric Topographic and Electrochemical Studies of Oxydation/Reduction Processes Provoked by Bacteria

    Science.gov (United States)

    Dhahri, S.; Marliere, C.

    2012-12-01

    The presence of biological matter (bacteria) in deep geological sites for storage of, for instance, radioactive elements or groundwater in aquifers was clearly proved. That biomass triggers physical and chemical processes which greatly modify the durability and the sustainability of the storage sites. These processes, mainly from oxidative/reductive reactions, are poorly understood. This is mainly due to the fact that former studies were done at the macroscopic level far away from the micrometric scale where relevant processes induced by bacteria take place. Investigations at microscopic level are needed. Thus, we developed an experimental set -up based on the combined use of optical microscopy (epifluorescence and transmission), atomic force microscopy (AFM) and scanning electro -chemical microscopy (SECM) in order to get simultaneous information on topographic and electro -chemical processes at different length scales. The first highly sensitive step was to use AFM and optical microscopy with biological samples in liquid environment: We will present a new, non -perturbative method for imaging bacteria in their natural liquid environment using AFM. No immobilization protocol, neither chemical nor mechanical, is needed, contrary to what has been regarded till now as essential. Furthermore we were able to follow the natural gliding movements of bacteria, directly proving their living state during the AFM investigation: we thus directly prove the low impact of these breakthrough AFM observations on the native behavior of the bacteria. The second delicate step was to combine AFM and optical measurements with electrical ones. We mounted a new experimental set-up coupling real -time (i) monitoring of optical properties as the optical density (OD) evolution related to bulk bacterial growth in liquid or as the counting of number of bacteria adhering on the surface of the sample as well and (ii) electrical and electrochemical measurements. We thus will present results on

  1. Ecophysiology of the anammox bacteria

    NARCIS (Netherlands)

    Kartal, Mustafa Boran

    2008-01-01

    Anaerobic ammonium oxidizing (anammox) bacteria oxidize ammonium to dinitrogen gas with nitrite as the electron acceptor. These bacteria are the key players in the global nitrogen cycle, responsible for the most of nitrogen production in natural ecosystems. The anammox process is also a cost-effecti

  2. Hydrogen production from food processing waste by anaerobic bacteria; Kenkisei saikin ni yoru shokuhin kako haikibutsu kara no suiso seisei

    Energy Technology Data Exchange (ETDEWEB)

    Mizuno, O.; Ohara, T.; Noike, T. [Tohoku University, Sendai (Japan)

    1997-08-22

    To produce hydrogen from bean-curd refuse discharged from bean-curd production process, effects of solid concentration on the hydrogen production activity and the final amount of hydrogen production are clarified using hydrogen production sludge obtained from a soybean silo with hydrogen explosion. Decomposition characteristics of bean-curd refuse by hydrogen fermentation are also investigated. Hydrogen contents of fermentation gas produced from decomposed bean-curd refuse were between 54 and 78%. It was found that bean-curd refuse can be used for hydrogen fermentation. The final amounts of hydrogen production were between 0.014 and 0.020 m{sup 3}centre dotkgVS{sup -1}, which was not greatly affected by the solid concentration. Even under high solid concentration conditions, sufficient hydrogen was produced. The hydrogen production activity was affected by the solid concentration. During the hydrogen production process, concentrations of alcohol and volatile fatty acid increased with decreasing the concentration of soluble sugars in the liquid phase. Acetate, propionate, n-butyrate and ethanol were the main metabolic products. 10 refs., 5 figs., 2 tabs.

  3. Constraints on mechanisms and rates of anaerobic oxidation of methane by microbial consortia: process-based modeling of ANME-2 archaea and sulfate reducing bacteria interactions

    Directory of Open Access Journals (Sweden)

    B. Orcutt

    2008-05-01

    Full Text Available Anaerobic oxidation of methane (AOM is the main process responsible for the removal of methane generated in Earth's marine subsurface environments. However, the biochemical mechanism of AOM remains elusive. By explicitly resolving the observed spatial arrangement of methanotrophic archaea and sulfate reducing bacteria found in consortia mediating AOM, potential intermediates involved in the electron transfer between the methane oxidizing and sulfate reducing partners were investigated via a consortium-scale reaction transport model that integrates the effect of diffusional transport with thermodynamic and kinetic controls on microbial activity. Model simulations were used to assess the impact of poorly constrained microbial characteristics such as minimum energy requirements to sustain metabolism, substrate affinity and cell specific rates. The role of environmental conditions such as the influence of methane levels on the feasibility of H2, formate and acetate as intermediate species, and the impact of the abundance of intermediate species on pathway reversal was examined. The results show that higher production rates of intermediates via AOM lead to increased diffusive fluxes from the methane oxidizing archaea to sulfate reducing bacteria, but the build-up of the exchangeable species causes the energy yield of AOM to drop below that required for ATP production. Comparison to data from laboratory experiments shows that under the experimental conditions of Nauhaus et al. (2007, neither hydrogen nor formate is exchanged fast enough between the consortia partners to achieve measured rates of metabolic activity, but that acetate exchange might support rates that approach those observed.

  4. Variability in bacteria and virus-like particle abundances during purging of unconfined aquifers.

    Science.gov (United States)

    Roudnew, Ben; Lavery, Trish J; Seymour, Justin R; Jeffries, Thomas C; Mitchell, James G

    2014-01-01

    Standard methodologies for sampling the physicochemical conditions of groundwater recommend purging a bore for three bore volumes to avoid sampling the stagnant water within a bore and instead gain samples representative of the aquifer. However, there are currently no methodological standards addressing the amount of purging required to gain representative biological samples to assess groundwater bacterial and viral abundances. The objective of this study was to examine how bacterial and viral abundances change during the purging of bore volumes. Six bores infiltrating into unconfined aquifers were pumped for five or six bore volumes each and bacteria and virus-like particles (VLPs) were enumerated from each bore volume using flow cytometry. In examination of the individual bores trends in bacterial abundances were observed to increase, decrease, or remain constant with each purged bore volume. Furthermore, triplicates taken at each bore volume indicated substantial variations in VLP and bacterial abundances that are often larger than the differences between bore volumes. This indicates a high level of small scale heterogeneity in microbial community abundance in groundwater samples, and we suggest that this may be an intrinsic feature of bore biology. The heterogeneity observed may be driven by bottom up processes (variability in the distribution of organic and inorganic nutrients), top-down processes (grazing and viral lysis), physical heterogeneities in the bore, or technical artifacts associated with the purging process. We suggest that a more detailed understanding of the ecology underpinning this variability is required to adequately describe the microbiological characteristics of groundwater ecosystems.

  5. Exploring the use of natural antimicrobial agents and pulsed electric fields to control spoilage bacteria during a beer production process.

    Science.gov (United States)

    Galvagno, M A; Gil, G R; Iannone, L J; Cerrutti, P

    2007-01-01

    Different natural antimicrobials affected viability of bacterial contaminants isolated at critical steps during a beer production process. In the presence of 1 mg/ml chitosan and 0.3 mg/ml hops, the viability of Escherichia coli in an all malt barley extract wort could be reduced to 0.7 and 0.1% respectively after 2 hour- incubation at 4 degrees C. The addition of 0.0002 mg/ml nisin, 0.1 mg/ml chitosan or 0.3 mg/ml hops, selectively inhibited growth of Pediococcus sp. in more than 10,000 times with respect to brewing yeast in a mixed culture. In the presence of 0.1 mg ml chitosan in beer, no viable cells of the thermoresistant strain Bacillus megaterium were detected. Nisin, chitosan and hops increased microbiological stability during storage of a local commercial beer inoculated with Lactobacillus plantarum or Pediococcus sp. isolated from wort. Pulsed Electric Field (PEF) (8 kV/cm, 3 pulses) application enhanced antibacterial activity of nisin and hops but not that of chitosan. The results herein obtained suggest that the use of these antimicrobial compounds in isolation or in combination with PEF would be effective to control bacterial contamination during beer production and storage.

  6. STUDIES ON THE BACTERIOPHAGE OF D'HERELLE : III. SOME OF THE FACTORS DETERMINING THE NUMBER AND SIZE OF PLAQUES OF BACTERIAL LYSIS ON AGAR.

    Science.gov (United States)

    Bronfenbrenner, J J; Korb, C

    1925-09-30

    The experiments reported above confirm the fact that lytic principle is distributed in active solution in a state of indivisible units. This permits its quantitative evaluation by serial dilution, as well as by plating on agar. The latter method, however, often gives readings considerably lower than those obtained by the broth dilution method of titration. By varying the concentration of agar it has been possible to show that the discrepancy is due to adsorption of the lytic agent on agar. When the concentration of the latter is increased from 0.3 per cent to 2.5 per cent the number of plaques of lysis is reduced more than 100 times. At the same time the average size of the plaques also decreases approximately to one-tenth of the original. The size, as well as the number of plaques, has been found to depend also on the condition of the culture employed in titration. Thus, when the culture exposed to the action of lytic agent is composed of young susceptible bacteria, the greater the concentration of bacteria, the smaller the plaques. When the culture is composed partly of young and partly of old susceptible bacteria, both the size and the number of the plaques are diminished with the increase in the relative concentration of old bacteria. On the other hand, presence in the culture of resistant bacteria does not affect either the size or the number of the plaques so long as the relative concentration of susceptible bacteria in the culture is sufficient to allow formation of them. The plaques appearing in the presence of a high concentration of resistant variants in the culture are relatively indistinct owing to overgrowth. Under carefully controlled conditions the size of plaques is found to be determined by the character of the lytic filtrate. Thus in the case of lytic agents which act upon more than one bacterial species the size of the plaques remains constant, irrespective of the bacterial substratum used for the production of the active filtrate.

  7. Revisiting bistability in the lysis/lysogeny circuit of bacteriophage lambda.

    Directory of Open Access Journals (Sweden)

    Michael Bednarz

    Full Text Available The lysis/lysogeny switch of bacteriophage lambda serves as a paradigm for binary cell fate decision, long-term maintenance of cellular state and stimulus-triggered switching between states. In the literature, the system is often referred to as "bistable." However, it remains unclear whether this term provides an accurate description or is instead a misnomer. Here we address this question directly. We first quantify transcriptional regulation governing lysogenic maintenance using a single-cell fluorescence reporter. We then use the single-cell data to derive a stochastic theoretical model for the underlying regulatory network. We use the model to predict the steady states of the system and then validate these predictions experimentally. Specifically, a regime of bistability, and the resulting hysteretic behavior, are observed. Beyond the steady states, the theoretical model successfully predicts the kinetics of switching from lysogeny to lysis. Our results show how the physics-inspired concept of bistability can be reliably used to describe cellular phenotype, and how an experimentally-calibrated theoretical model can have accurate predictive power for cell-state switching.

  8. High-k Dielectric Passivation: Novel Considerations Enabling Cell Specific Lysis Induced by Electric Fields.

    Science.gov (United States)

    Wassermann, Klemens J; Barth, Sven; Keplinger, Franz; Noehammer, Christa; Peham, Johannes R

    2016-08-24

    A better understanding of the electrodynamic behavior of cells interacting with electric fields would allow for novel scientific insights and would lead to the next generation of cell manipulation, diagnostics, and treatment. Here, we introduce a promising electrode design by using metal oxide high-k dielectric passivation. The thermally generated dielectric passivation layer enables efficient electric field coupling to the fluid sample comprising cells while simultaneously decoupling the electrode ohmically from the electrolyte, allowing for better control and adjustability of electric field effects due to reduced electrochemical reactions at the electrode surface. This approach demonstrates cell-size specific lysis with electric fields in a microfluidic flow-through design resulting in 99.8% blood cell lysis at 6 s exposure without affecting the viability of Gram-positive and Gram-negative bacterial spike-ins. The advantages of this new approach can support next-generation investigations of electrodynamics in biological systems and their exploitation for cell manipulation in multiple fields of medicine, life science, and industry.

  9. [Study of a lysis medium stabilizing microfilaments and microtubules in vitro and in vivo].

    Science.gov (United States)

    Foucault, G; Raymond, M N; Coffe, G; Pudles, J

    1984-01-01

    Determination of experimental conditions which allow the evaluation of the variations in the ratio of non polymerized and polymerized forms of actin and tubulin during the reorganization of the cytoskeletal cell system is of most valuable importance. In order to prepare cell homogenates which would reflect the in vivo situation, we tested in vitro a lysis medium which stabilized both microfilaments and microtubules, which were determined by DNase inhibition assays and colchicine binding assays respectively. This lysis medium containing 10 mM potassium phosphate, 1mM magnesium chloride, 5 mM EGTA, 1 M hexylene glycol, 1% Triton X-100, pH 6.4, used at 4 degrees C a) diffused rapidly into the cells; b) did not denature actin and tubulin; c) did not displace the equilibrium between non polymerized and polymerized forms of actin and tubulin, allowing biochemical assays on cell homogenates; d) blocked the evolution of the cytoskeletal system and permitted structural studies; e) and allowed the decoration of microfilaments by heavy meromyosin.

  10. [Effectiveness of locoregional rt-PA lysis in acute leg and arm vein thrombosis].

    Science.gov (United States)

    Perkmann, R; Neuhauser, B; Bodner, G; Tauscher, T; Fraedrich, G

    2001-06-01

    39 patients suffering from a thrombosis of the peripheral venous system were treated with a loco-regional lysis, using rt-PA. Two cycles of 40 mg rt-PA a day were applicated by means of a special drainage-management, using perforans veins. During lytic therapy, 20,000 to 30,000 IE of unfractionized heparin were additionally administered. Laboratory work including aPTT and fibrinogen measurement was performed every 8 hours. Additionally a phlebography was performed after 24 hours. Patients received an anticoagulative therapy using sintrom or marcumar the following 3 months. We obtained a successful thrombolysis without any major complications in 90%. Minor complications included 3 peripheral pulmonary embolisms. Duplex sonographic and plethysmographic follow up was performed in 25 patients one year after operation. In two patients with ankle edema insufficient valves at the popliteal vein were found with both diagnostic modalities. 23 patients showed no signs of insufficient valves neither clinically nor at duplex sonography and plethysmography. The locoregional lysis appears to be an effective method for the treatment of acute peripheral vein thrombosis.

  11. Plasminogen associates with phosphatidylserine-exposing platelets and contributes to thrombus lysis under flow.

    Science.gov (United States)

    Whyte, Claire S; Swieringa, Frauke; Mastenbroek, Tom G; Lionikiene, Ausra S; Lancé, Marcus D; van der Meijden, Paola E J; Heemskerk, Johan W M; Mutch, Nicola J

    2015-04-16

    The interaction of plasminogen with platelets and their localization during thrombus formation and fibrinolysis under flow are not defined. Using a novel model of whole blood thrombi, formed under flow, we examine dose-dependent fibrinolysis using fluorescence microscopy. Fibrinolysis was dependent upon flow and the balance between fibrin formation and plasminogen activation, with tissue plasminogen activator-mediated lysis being more efficient than urokinase plasminogen activator-mediated lysis. Fluorescently labeled plasminogen radiates from platelet aggregates at the base of thrombi, primarily in association with fibrin. Hirudin attenuates, but does not abolish plasminogen binding, denoting the importance of fibrin. Flow cytometry revealed that stimulation of platelets with thrombin/convulxin significantly increased the plasminogen signal associated with phosphatidylserine (PS)-exposing platelets. Binding was attenuated by tirofiban and Gly-Pro-Arg-Pro amide, confirming a role for fibrin in amplifying plasminogen binding to PS-exposing platelets. Confocal microscopy revealed direct binding of plasminogen and fibrinogen to different platelet subpopulations. Binding of plasminogen and fibrinogen co-localized with PAC-1 in the center of spread platelets. In contrast, PS-exposing platelets were PAC-1 negative, and bound plasminogen and fibrinogen in a protruding "cap." These data show that different subpopulations of platelets harbor plasminogen by diverse mechanisms and provide an essential scaffold for the accumulation of fibrinolytic proteins that mediate fibrinolysis under flow.

  12. Predicting the concentration of verotoxin-producing Escherichia coli bacteria during processing and storage of fermented raw-meat sausages.

    Science.gov (United States)

    Quinto, E J; Arinder, P; Axelsson, L; Heir, E; Holck, A; Lindqvist, R; Lindblad, M; Andreou, P; Lauzon, H L; Marteinsson, V Þ; Pin, C

    2014-05-01

    A model to predict the population density of verotoxigenic Escherichia coli (VTEC) throughout the elaboration and storage of fermented raw-meat sausages (FRMS) was developed. Probabilistic and kinetic measurement data sets collected from publicly available resources were completed with new measurements when required and used to quantify the dependence of VTEC growth and inactivation on the temperature, pH, water activity (aw), and concentration of lactic acid. Predictions were compared with observations in VTEC-contaminated FRMS manufactured in a pilot plant. Slight differences in the reduction of VTEC were predicted according to the fermentation temperature, 24 or 34°C, with greater inactivation at the highest temperature. The greatest reduction was observed during storage at high temperatures. A population decrease greater than 6 decimal logarithmic units was observed after 66 days of storage at 25°C, while a reduction of only ca. 1 logarithmic unit was detected at 12°C. The performance of our model and other modeling approaches was evaluated throughout the processing of dry and semidry FRMS. The greatest inactivation of VTEC was predicted in dry FRMS with long drying periods, while the smallest reduction was predicted in semidry FMRS with short drying periods. The model is implemented in a computing tool, E. coli SafeFerment (EcSF), freely available from http://www.ifr.ac.uk/safety/EcoliSafeFerment. EcSF integrates growth, probability of growth, and thermal and nonthermal inactivation models to predict the VTEC concentration throughout FRMS manufacturing and storage under constant or fluctuating environmental conditions.

  13. Phages of lactic acid bacteria: The role of genetics in understanding phage-host interactions and their co-evolutionary processes

    Energy Technology Data Exchange (ETDEWEB)

    Mahony, Jennifer, E-mail: j.mahony@ucc.ie [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Ainsworth, Stuart; Stockdale, Stephen [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Sinderen, Douwe van, E-mail: d.vansinderen@ucc.ie [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Alimentary Pharmabiotic Centre, Biosciences Institute, University College Cork, Western Road, Cork (Ireland)

    2012-12-20

    Dairy fermentations are among the oldest food processing applications, aimed at preservation and shelf-life extension through the use of lactic acid bacteria (LAB) starter cultures, in particular strains of Lactococcus lactis, Streptococcus thermophilus, Lactobacillus spp. and Leuconostoc spp. Traditionally this was performed by continuous passaging of undefined cultures from a finished fermentation to initiate the next fermentation. More recently, consumer demands on consistent and desired flavours and textures of dairy products have led to a more defined approach to such processes. Dairy (starter) companies have responded to the need to define the nature and complexity of the starter culture mixes, and dairy fermentations are now frequently based on defined starter cultures of low complexity, where each starter component imparts specific technological properties that are desirable to the product. Both mixed and defined starter culture approaches create the perfect environment for the proliferation of (bacterio)phages capable of infecting these LAB. The repeated use of the same starter cultures in a single plant, coupled to the drive towards higher and consistent production levels, increases the risk and negative impact of phage infection. In this review we will discuss recent advances in tracking the adaptation of phages to the dairy industry, the advances in understanding LAB phage-host interactions, including evolutionary and genomic aspects.

  14. Phages of lactic acid bacteria: the role of genetics in understanding phage-host interactions and their co-evolutionary processes.

    Science.gov (United States)

    Mahony, Jennifer; Ainsworth, Stuart; Stockdale, Stephen; van Sinderen, Douwe

    2012-12-20

    Dairy fermentations are among the oldest food processing applications, aimed at preservation and shelf-life extension through the use of lactic acid bacteria (LAB) starter cultures, in particular strains of Lactococcus lactis, Streptococcus thermophilus, Lactobacillus spp. and Leuconostoc spp. Traditionally this was performed by continuous passaging of undefined cultures from a finished fermentation to initiate the next fermentation. More recently, consumer demands on consistent and desired flavours and textures of dairy products have led to a more defined approach to such processes. Dairy (starter) companies have responded to the need to define the nature and complexity of the starter culture mixes, and dairy fermentations are now frequently based on defined starter cultures of low complexity, where each starter component imparts specific technological properties that are desirable to the product. Both mixed and defined starter culture approaches create the perfect environment for the proliferation of (bacterio)phages capable of infecting these LAB. The repeated use of the same starter cultures in a single plant, coupled to the drive towards higher and consistent production levels, increases the risk and negative impact of phage infection. In this review we will discuss recent advances in tracking the adaptation of phages to the dairy industry, the advances in understanding LAB phage-host interactions, including evolutionary and genomic aspects.

  15. All-in-one nanowire-decorated multifunctional membrane for rapid cell lysis and direct DNA isolation.

    KAUST Repository

    So, Hongyun

    2014-11-24

    This paper describes a handheld device that uses an all-in-one membrane for continuous mechanical cell lysis and rapid DNA isolation without the assistance of power sources, lysis reagents, and routine centrifugation. This nanowire-decorated multifunctional membrane was fabricated to isolate DNA by selective adsorption to silica surface immediately after disruption of nucleus membranes by ultrasharp tips of nanowires for a rapid cell lysis, and it can be directly assembled with commercial syringe filter holders. The membrane was fabricated by photoelectrochemical etching to create microchannel arrays followed by hydrothermal synthesis of nanowires and deposition of silica. The proposed membrane successfully purifies high-quality DNA within 5 min, whereas a commercial purification kit needs more than an hour.

  16. The susceptibility to cytotoxic T lymphocyte mediated lysis of chemically induced sarcomas from immunodeficient and normal mice

    DEFF Research Database (Denmark)

    Svane, I M; Engel, A M; Thomsen, Allan Randrup;

    1997-01-01

    tested for susceptibility to cytolysis by virus specific cytotoxic T cells. Tumour cells originating from tumours induced in immunocompetent C.B.-17 mice presented virus antigen more efficiently than tumour cells from immunodeficient SCID mice. No significant difference in virus antigen presentation...... was found between tumours from nude and nu/+ BALB/c mice. The sensitivity of target cells from the individual tumours to cytotoxic T lymphocyte (CTL) mediated lysis correlated negatively with their sensitivity to natural killer (NK) cell mediated lysis. There was a positive correlation between...... the sensitivity to CTL mediated lysis and surface expression of the MHC class I molecule Ld of the tumour cells. Tumour cells incapable of in vitro presentation of viral antigen to specific cytotoxic T cells originated from tumours known from previous experiments to be readily accepted after transplantation...

  17. Rapid method for DNA extraction from the honey bee Apis mellifera and the parasitic bee mite Varroa destructor using lysis buffer and proteinase K.

    Science.gov (United States)

    Issa, M R C; Figueiredo, V L C; De Jong, D; Sakamoto, C H; Simões, Z L P

    2013-10-22

    We developed a rapid method for extraction of DNA from honey bees, Apis mellifera, and from the parasitic bee mite, Varroa destructor. The advantages include fast processing and low toxicity of the substances that are utilized. We used lysis buffer with nonionic detergents to lyse cell walls and proteinase K to digest proteins. We tested whole thorax, thoracic muscle mass, legs, and antennae from individual bees; the mites were processed whole (1 mite/sample). Each thorax was incubated whole, without cutting, because exocuticle color pigment darkened the extraction solution, interfering with PCR results. The procedure was performed with autoclaved equipment and laboratory gloves. For each sample, we used 100 µL lysis buffer (2 mL stock solution of 0.5 M Tris/HCl, pH 8.5, 10 mL stock solution of 2 M KCl, 500 µL solution of 1 M MgCl2, 2 mL NP40, and 27.6 g sucrose, completed to 200 mL with bidistilled water and autoclaved) and 2 µL proteinase K (10 mg/mL in bidistilled water previously autoclaved, as proteinase K cannot be autoclaved). Tissues were incubated in the solutions for 1-2 h in a water bath (62°-68 °C) or overnight at 37 °C. After incubation, the tissues were removed from the extraction solution (lysis buffer + proteinase K) and the solution heated to 92 °C for 10 min, for proteinase K inactivation. Then, the solution with the extracted DNA was stored in a refrigerator (4°-8 °C) or a freezer (-20 °C). This method does not require centrifugation or phenol/chloroform extraction. The reduced number of steps allowed us to sample many individuals/day. Whole mites and bee antennae were the most rapidly processed. All bee tissues gave the same quality DNA. This method, even using a single bee antenna or a single mite, was adequate for extraction and analysis of bee genomic and mitochondrial DNA and mite genomic DNA.

  18. Contribution of midgut bacteria to blood digestion and egg production in aedes aegypti (diptera: culicidae (L.

    Directory of Open Access Journals (Sweden)

    Pimenta Paulo FP

    2011-06-01

    Full Text Available Abstract Background The insect gut harbors a variety of microorganisms that probably exceed the number of cells in insects themselves. These microorganisms can live and multiply in the insect, contributing to digestion, nutrition, and development of their host. Recent studies have shown that midgut bacteria appear to strengthen the mosquito's immune system and indirectly enhance protection from invading pathogens. Nevertheless, the physiological significance of these bacteria for mosquitoes has not been established to date. In this study, oral administration of antibiotics was employed in order to examine the contribution of gut bacteria to blood digestion and fecundity in Aedes aegypti. Results The antibiotics carbenicillin, tetracycline, spectinomycin, gentamycin and kanamycin, were individually offered to female mosquitoes. Treatment of female mosquitoes with antibiotics affected the lysis of red blood cells (RBCs, retarded the digestion of blood proteins and reduced egg production. In addition, antibiotics did not affect the survival of mosquitoes. Mosquito fertility was restored in the second gonotrophic cycle after suspension of the antibiotic treatment, showing that the negative effects of antibiotics in blood digestion and egg production in the first gonotrophic cycle were reversible. Conclusions The reduction of bacteria affected RBC lysis, subsequently retarded protein digestion, deprived mosquito from essential nutrients and, finally, oocyte maturation was affected, resulting in the production of fewer viable eggs. These results indicate that Ae. aegypti and its midgut bacteria work in synergism to digest a blood meal. Our findings open new possibilities to investigate Ae. aegypti-associated bacteria as targets for mosquito control strategies.

  19. Overexpressions of Lambda Phage Lysis Genes and Biosynthetic Genes of Poly-β-hydroxybutyrate in Recombinant E.coli

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    A plasmid (pTU9) containing the lambda (λ) phage lysis genes S(-)RRz and the biosynthetic genes phbCAB of poly-β-hydroxybutyrate (PHB) was constructed and transformed into E.coli JM109. Cultured in Luria-Bertani (LB) medium with 20 g/L glucose, E.coli JM109 (pTU9) could accumulate PHB in cells up to 40% (g PHB per g dry cells). A chelating agent EDTA was applied to induce a complete cell lysis and PHB granules were released. This method has a potential application in PHB separation.

  20. A biological process for the reclamation of flue gas desulfurization gypsum using mixed sulfate-reducing bacteria with inexpensive carbon sources.

    Science.gov (United States)

    Kaufman, E N; Little, M H; Selvaraj, P

    1997-01-01

    A combined chemical and biological process for the recycling of flue gas desulfurization (FGD) gypsum into calcium carbonate and elemental sulfur is demonstrated. In this process, a mixed culture of sulfate-reducing bacteria (SRB) utilizes inexpensive carbon sources, such as sewage digest or synthesis gas, to reduce FGD gypsum to hydrogen sulfide. The sulfide is then oxidized to elemental sulfur via reaction with ferric sulfate, and accumulating calcium ions are precipitated as calcium carbonate using carbon dioxide. Employing anaerobically digested municipal sewage sludge (AD-MSS) medium as a carbon source, SRBs in serum bottles demonstrated an FGD gypsum reduction rate of 8 mg/L/h (10(9) cells)(-1). A chemostat with continuous addition of both AD-MSS media and gypsum exhibited sulfate reduction rates as high as 1.3 kg FGD gypsum/m(3)d. The increased biocatalyst density afforded by cell immobilization in a columnar reactor allowed a productivity of 152 mg SO(4) (-2)/Lh or 6.6 kg FGD gypsum/m(3)d. Both reactors demonstrated 100% conversion of sulfate, with 75-100% recovery of elemental sulfur and chemical oxygen demand utilization as high as 70%. Calcium carbonate was recovered from the reactor effluent on precipitation using carbon dioxide. It was demonstrated that SRBs may also use synthesis gas (CO, H(2), and CO(2) in the reduction of gypsum, further decreasing process costs. The formation of two marketable products-elemental sulfur and calcium carbonate-from FGD gypsum sludge, combined with the use of a low-cost carbon source and further improvements in reactor design, promises to offer an attractive alternative to the landfilling of FGD gypsum.

  1. Factors involved in the regulation of early enzyme synthesis and lysis of Escherichia coli B infected with bacteriophage T4D

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, S.

    1976-01-01

    The capability of T4/sup +/-infected Escherichia coli cells to display handling-induced lysis may implicate the activity of a known, untested gene, or a heretofore undiscovered gene. This gene product may be involved in the normal lytic process; it may be a new phage-induced phospholipase or a late function that activates a latent E. coli phospholipase. Another possibility is that the late function is involved in membrane biosynthesis or modification. Alternatively, the required late protein(s) may have some other role that only secondarily affects the cell's permeability barrier. Whatever the case may be, this work is of practical interest since chilling and centrifugation of infected cells are very common procedures and researchers should be aware of any handling-induced damage.

  2. In black south africans from rural and urban communities, the 4G/5G PAI-1 polymorphism influences PAI-1 activity, but not plasma clot lysis time

    NARCIS (Netherlands)

    Z. de Lange (Zelda); D.C. Rijken (Dingeman); T. Hoekstra (Tiny); K.R. Conradie (Karin); J.C. Jerling (Johann); M. Pieters (Marlien)

    2013-01-01

    textabstractData on genetic and environmental factors influencing PAI-1 levels and their consequent effect on clot lysis in black African populations are limited. We identified polymorphisms in the promoter area of the PAI-1 gene and determined their influence on PAI-1act levels and plasma clot lysi

  3. Entropy Production Rate Changes in Lysogeny/Lysis Switch Regulation of Bacteriophage Lambda*

    Institute of Scientific and Technical Information of China (English)

    DING Hui; LUO Liao-Fu; LIN Hao

    2011-01-01

    According to the chemical kinetic model of lysogeny/lysis switch in Escherichia coli (E. coli) intected by bacteriophage λ, the entropy production rates of steady states are calculated. The results show that the lysogenic state has lower entropy production rate than lytic state, which provides an explanation on why the lysogenic state of λ phage is so stable. We also notice that the entropy production rates of both lysogenic state and lytic state are lower than that of saddle-point and bifurcation state, which is consistent with the principle of minimum entropy production for living organism in nonequilibrium stationary state. Subsequently, the relations between CI and Cro degradation rates at two bifurcations and the changes of entropy production rate with CI and Cro degradation are deduced. The theory and method can be used to calculate entropy change in other molecular network.

  4. LYSIS OF BACTERIAL PROTOPLASTS AND SPHEROPLASTS BY STAPHYLOCOCCAL ALPHA-TOXIN AND STREPTOLYSIN S.

    Science.gov (United States)

    BERNHEIMER, A W; SCHWARTZ, L L

    1965-05-01

    Bernheimer, Alan W. (New York University School of Medicine, New York, N.Y.), and Lois L. Schwartz. Lysis of bacterial protoplasts and spheroplasts by staphylococcal alpha-toxin and streptolysin S. J. Bacteriol. 89:1387-1392. 1965.-Protoplasts of Bacillus megaterium, Sarcina lutea, and Streptococcus pyogenes, and spheroplasts of Escherichia coli were lysed by staphylococcal alpha-toxin, whereas spheroplasts of Vibrio metschnikovii and V. comma were not. In the spectrum of its lytic action, streptolysin S qualitatively resembled staphylococcal alpha-toxin except for failure to lyse S. pyogenes protoplasts. In contrast to the two foregoing agents, streptolysin O did not lyse protoplasts and spheroplasts. The observations are interpreted in relation to similarities and differences in lipid composition of bacterial and mammalian cell membranes.

  5. Abundance and diversity of ammonia-oxidizing archaea and bacteria on granular activated carbon and their fates during drinking water purification process.

    Science.gov (United States)

    Niu, Jia; Kasuga, Ikuro; Kurisu, Futoshi; Furumai, Hiroaki; Shigeeda, Takaaki; Takahashi, Kazuhiko

    2016-01-01

    Ammonia is a precursor to trichloramine, which causes an undesirable chlorinous odor. Granular activated carbon (GAC) filtration is used to biologically oxidize ammonia during drinking water purification; however, little information is available regarding the abundance and diversity of ammonia-oxidizing archaea (AOA) and bacteria (AOB) associated with GAC. In addition, their sources and fates in water purification process remain unknown. In this study, six GAC samples were collected from five full-scale drinking water purification plants in Tokyo during summer and winter, and the abundance and community structure of AOA and AOB associated with GAC were studied in these two seasons. In summer, archaeal and bacterial amoA genes on GACs were present at 3.7 × 10(5)-3.9 × 10(8) gene copies/g-dry and 4.5 × 10(6)-4.2 × 10(8) gene copies/g-dry, respectively. In winter, archaeal amoA genes remained at the same level, while bacterial amoA genes decreased significantly for all GACs. No differences were observed in the community diversity of AOA and AOB from summer to winter. Phylogenetic analysis revealed high AOA diversity in group I.1a and group I.1b in raw water. Terminal-restriction fragment length polymorphism analysis of processed water samples revealed that AOA diversity decreased dramatically to only two OTUs in group I.1a after ozonation, which were identical to those detected on GAC. It suggests that ozonation plays an important role in determining AOA diversity on GAC. Further study on the cell-specific activity of AOA and AOB is necessary to understand their contributions to in situ nitrification performance.

  6. Prediction of recurrent venous thromboembolism by clot lysis time: a prospective cohort study.

    Directory of Open Access Journals (Sweden)

    Ludwig Traby

    Full Text Available Venous thromboembolism (VTE is a chronic disease, which tends to recur. Whether an abnormal fibrinolytic system is associated with an increased risk of VTE is unclear. We assessed the relationship between fibrinolytic capacity (reflected by clot lysis time [CLT] and risk of recurrent VTE. We followed 704 patients (378 women; mean age 48 yrs with a first unprovoked VTE for an average of 46 months after anticoagulation withdrawal. Patients with natural coagulation inhibitor deficiency, lupus anticoagulant, cancer, homozygosity for factor V Leiden or prothrombin mutation, or requirement for indefinite anticoagulation were excluded. Study endpoint was symptomatic recurrent VTE. For measurement of CLT, a tissue factor-induced clot was lysed by adding tissue-type plasminogen activator. Time between clot formation and lysis was determined by measuring the turbidity. 135 (19% patients had recurrent VTE. For each increase in CLT of 10 minutes, the crude relative risk (RR of recurrence was 1.13 (95% CI 1.02-1.25; p = 0.02 and was 1.08 (95% CI 0.98-1.20; p = 0.13 after adjustment for age and sex. For women only, the adjusted RR was 1.14 (95% CI, 0.91-1.42, p = 0.22 for each increase in CLT of 10 minutes. CLT values in the 4(th quartile of the female patient population, as compared to values in the 1(st quartile, conferred a risk of recurrence of 3.28 (95% CI, 1.07-10.05; p = 0.04. No association between CLT and recurrence risk was found in men. Hypofibrinolysis as assessed by CLT confers a moderate increase in the risk of recurrent VTE. A weak association between CLT and risk of recurrence was found in women only.

  7. Stabilizing additives added during cell lysis aid in the solubilization of recombinant proteins.

    Directory of Open Access Journals (Sweden)

    David J Leibly

    Full Text Available Insoluble recombinant proteins are a major issue for both structural genomics and enzymology research. Greater than 30% of recombinant proteins expressed in Escherichia coli (E. coli appear to be insoluble. The prevailing view is that insolubly expressed proteins cannot be easily solubilized, and are usually sequestered into inclusion bodies. However, we hypothesize that small molecules added during the cell lysis stage can yield soluble protein from insoluble protein previously screened without additives or ligands. We present a novel screening method that utilized 144 additive conditions to increase the solubility of recombinant proteins expressed in E. coli. These selected additives are natural ligands, detergents, salts, buffers, and chemicals that have been shown to increase the stability of proteins in vivo. We present the methods used for this additive solubility screen and detailed results for 41 potential drug target recombinant proteins from infectious organisms. Increased solubility was observed for 80% of the recombinant proteins during the primary and secondary screening of lysis with the additives; that is 33 of 41 target proteins had increased solubility compared with no additive controls. Eleven additives (trehalose, glycine betaine, mannitol, L-Arginine, potassium citrate, CuCl(2, proline, xylitol, NDSB 201, CTAB and K(2PO(4 solubilized more than one of the 41 proteins; these additives can be easily screened to increase protein solubility. Large-scale purifications were attempted for 15 of the proteins using the additives identified and eight (40% were prepared for crystallization trials during the first purification attempt. Thus, this protocol allowed us to recover about a third of seemingly insoluble proteins for crystallography and structure determination. If recombinant proteins are required in smaller quantities or less purity, the final success rate may be even higher.

  8. Sustainable microbial water quality monitoring programme design using phage-lysis and multivariate techniques.

    Science.gov (United States)

    Nnane, Daniel Ekane

    2011-11-15

    Contamination of surface waters is a pervasive threat to human health, hence, the need to better understand the sources and spatio-temporal variations of contaminants within river catchments. River catchment managers are required to sustainably monitor and manage the quality of surface waters. Catchment managers therefore need cost-effective low-cost long-term sustainable water quality monitoring and management designs to proactively protect public health and aquatic ecosystems. Multivariate and phage-lysis techniques were used to investigate spatio-temporal variations of water quality, main polluting chemophysical and microbial parameters, faecal micro-organisms sources, and to establish 'sentry' sampling sites in the Ouse River catchment, southeast England, UK. 350 river water samples were analysed for fourteen chemophysical and microbial water quality parameters in conjunction with the novel human-specific phages of Bacteroides GB-124 (Bacteroides GB-124). Annual, autumn, spring, summer, and winter principal components (PCs) explained approximately 54%, 75%, 62%, 48%, and 60%, respectively, of the total variance present in the datasets. Significant loadings of Escherichia coli, intestinal enterococci, turbidity, and human-specific Bacteroides GB-124 were observed in all datasets. Cluster analysis successfully grouped sampling sites into five clusters. Importantly, multivariate and phage-lysis techniques were useful in determining the sources and spatial extent of water contamination in the catchment. Though human faecal contamination was significant during dry periods, the main source of contamination was non-human. Bacteroides GB-124 could potentially be used for catchment routine microbial water quality monitoring. For a cost-effective low-cost long-term sustainable water quality monitoring design, E. coli or intestinal enterococci, turbidity, and Bacteroides GB-124 should be monitored all-year round in this river catchment.

  9. 大米乳酸菌发酵降镉工艺优化%Process optimization for lactic acid bacteria fermentation to reduce cadmium in rice

    Institute of Scientific and Technical Information of China (English)

    刘也嘉; 林亲录; 肖冬梅; 张珺; 林利忠

    2016-01-01

    为研究乳酸发酵对大米重金属镉的脱除效果,该试验以镉超标大米为原料,采用大米发酵液进行发酵降镉,通过 Plackett-Burman(PB)设计、最陡爬坡试验设计和 Box-Behnken 设计对发酵降镉工艺进行优化。结果表明:PB 试验筛选出的对大米发酵降镉效果显著影响的因素分别为加水量、强化菌种添加量和发酵温度;根据最陡爬坡确定的各因素逼近区域进行响应面试验设计,得出最佳降镉工艺为加水量120%、强化菌种添加量0.08‰、发酵温度32℃,其他发酵条件还包括米粉发酵液添加量2%、食盐添加量0.8%、以及有效降镉时间22~26 h,就镉质量分数为0.52 mg/kg的早米,此条件下降镉率为79.24%。研究结果为乳酸发酵降镉、缓解大米镉超标问题提供参考。%A kind of rice noodle fermentation broth from Changde provides strains to remove the heavy metal cadmium in rice. And the main microbial flora in this fermented liquid is plant lactobacillus. Plant lactobacillus can break down the rice protein to make cadmium free from combining state and then move out from rice. The bacterial cell membrane can also remove cadmium from rice by the effect of adsorption. Plackett-Burman design (PBD), Steepest ascent experiment (SAE), and Box-Behnken design (BBD) were used to optimize the process of this experiment. The results showed that, main factors which obviously affected the rates of cadmium removal in rice screened by PBD were the additive quantity of water, the amount of strengthening bacteria and the fermentation temperature. We also adopted the response surface design of experiment to find the optimum fermentation conditions by using SAE that is a high efficient screening experiment method. The scopes of 3 main factors were respectively that, the additive quantity of water was from 110%-130%, the amount of strengthening bacteria was from 0.5‰ to 0.9‰ and the fermentation temperature was

  10. Viruses, bacteria, and heterotrophic nanoflagellates in Laptev Sea plankton

    Science.gov (United States)

    Kopylov, A. I.; Sazhin, A. F.; Zabotkina, E. A.; Romanenko, A. V.; Romanova, N. D.; Makarevich, P. R.; Wenger, M. P.

    2016-11-01

    The paper considers the concentrations and functional characteristics of viruses, bacteria, and heterotrophic nanoflagellates determined for the first time in the Laptev Sea in August-September, 2014. The abundance of bacteria, viruses, and heterotrophic nanoflagellates varied from 110.1 × 103 to 828.4 × 103 cells/mL, from 384.2 × 103 to 2932.8 × 103 particles/mL, and from 108 to 651 cells/mL, respectively. The daily bacterioplankton production varied from 4.2 × 103 to 381.7 × 103 cells/mL, with an average of 117.6 × 103 cells/mL. Electron transmission microscopy has for the first time shown that the frequency of visibly infected bacterial cells varied from 0.2 to 2.0% (0.8% on average) of NB. The average virus-induced mortality of bacteria was 6.3% of bacterioplankton production, with variations ranging from 1.4 to 16.9%. Grazing on bacteria by heterotrophic nanoflagellates contributed more to bacteria mortality than virus-induced bacterial lysis. By grazing on bacteria, heterotrophic nanoflagellates consumed large quantities of viruses located on the surface and inside bacterial cells.

  11. Inhibitors of histone deacetylase 1 reverse the immune evasion phenotype to enhance T-cell mediated lysis of prostate and breast carcinoma cells.

    Science.gov (United States)

    Gameiro, Sofia R; Malamas, Anthony S; Tsang, Kwong Y; Ferrone, Soldano; Hodge, James W

    2016-02-16

    The clinical promise of cancer immunotherapy relies on the premise that the immune system can recognize and eliminate tumor cells identified as non-self. However, tumors can evade host immune surveillance through multiple mechanisms, including epigenetic silencing of genes involved in antigen processing and immune recognition. Hence, there is an unmet clinical need to develop effective therapeutic strategies that can restore tumor immune recognition when combined with immunotherapy, such as immune checkpoint blockade and therapeutic cancer vaccines. We sought to examine the potential of clinically relevant exposure of prostate and breast human carcinoma cells to histone deacetylase (HDAC) inhibitors to reverse tumor immune escape to T-cell mediated lysis. Here we demonstrate that prostate (LNCAP) and breast (MDA-MB-231) carcinoma cells are more sensitive to T-cell mediated lysis in vitro after clinically relevant exposure to epigenetic therapy with either the pan-HDAC inhibitor vorinostat or the class I HDAC inhibitor entinostat. This pattern of immunogenic modulation was observed against a broad range of tumor-associated antigens, such as CEA, MUC1, PSA, and brachyury, and associated with augmented expression of multiple proteins involved in antigen processing and tumor immune recognition. Genetic and pharmacological inhibition studies identified HDAC1 as a key determinant in the reversal of carcinoma immune escape. Further, our findings suggest that the observed reversal of tumor immune evasion is driven by a response to cellular stress through activation of the unfolded protein response. This offers the rationale for combining HDAC inhibitors with immunotherapy, including therapeutic cancer vaccines.

  12. Comparison of the efficiency of different methods for the lysis of cells in lab-on-chip systems

    NARCIS (Netherlands)

    Röser, T.; Drese, K.S.; Fütterer, X.; Germar, F. von; Hansen-Hagge, T.E.; Ritzi, M.; Bullema, J.E.; Bolt, P.J.

    2007-01-01

    Quantitative or qualitative examination of DNA has enormous impact on medical, forensic, or genealogical, analysis. The macro as well as micro world knows a panel of different methods for cell lysis that has to happen prior to the purification of the DNA. To our knowledge the efficiency of those pro

  13. Complicated Tumor Lysis Syndrome after CVVH Treatment in a Renal Transplantation Patient: One Case Report and Literature Review

    Institute of Scientific and Technical Information of China (English)

    Zengbo Liu; Li Li; Jianhui Yang; Zhishuang Han; Zengyi Ma; Aimei Feng

    2009-01-01

    @@ Introduction Tumor lysis syndrome (TLS) is a potentially lethal emergency caused by lysed tumor cells, and it frequently occurs in tumors of hematologic origin. Up until now, there has been only one known report published overseas about TLS resulting from post-transplant lymphoproliferative disorder (PTLD)[1].

  14. Symbiotic interaction of endophytic bacteria with arbuscular mycorrhizal fungi and its antagonistic effect on Ganoderma boninense.

    Science.gov (United States)

    Sundram, Shamala; Meon, Sariah; Seman, Idris Abu; Othman, Radziah

    2011-08-01

    Endophytic bacteria (Pseudomonas aeruginosa UPMP3 and Burkholderia cepacia UMPB3), isolated from within roots of oil palm (Elaeis guineensis Jacq.) were tested for their presymbiotic effects on two arbuscular mcorrhizal fungi, Glomus intraradices UT126 and Glomus clarum BR152B). These endophytic bacteria were also tested for antagonistic effects on Ganoderma boninense PER 71, a white wood rot fungal pathogen that causes a serious disease in oil palm. Spore germination and hyphal length of each arbuscular mycorrhizal fungal (AMF) pairing with endophytic bacteria was found to be significantly higher than spores plated in the absence of bacteria. Scanning electron microscopy (SEM) showed that the endophytic bacteria were scattered, resting or embedded on the surface hyaline layer or on the degraded walls of AMF spores, possibly feeding on the outer hyaline spore wall. The antagonistic effect of the endophytic bacteria was expressed as severe morphological abnormalities in the hyphal structures of G. boninense PER 71. The effects of the endophytic bacteria on G. boninense PER 71 hyphal structures were observed clearly under SEM. Severe inter-twisting, distortion, lysis and shriveling of the hyphal structures were observed. This study found that the effect of endophytic bacteria on G. intraradices UT126 and G. clarum BR152B resembled that of a mycorrhiza helper bacteria (MHB) association because the association significantly promoted AMF spore germination and hyphal length. However, the endophytic bacteria were extremely damaging to G. boninense PER 71.

  15. Bacteria and viruses of the ice-free aquatic area of the Barents Sea at the beginning of polar night.

    Science.gov (United States)

    Shirokolobova, T I; Zhichkin, A P; Venger, M P; Vodopyanova, V V; Moiseev, D V

    2016-07-01

    The most massive components of the microplankton were studied in the open sea waters for the first time at the end of the autumn season. It has been found that abundance of the virio- and bacterioplankton exceeded that observed in winter in the coastal zone. Against the background of a relatively uniform distribution of bacteria, the viral abundance and the lysis-mediated bacterioplankton death rate reached the maximum values in the most cold and salty waters of the northern sea areas.

  16. [BPO-Specific, complement-dependant cell-lysis of differently sensitized sheep red cells: evaluation of haptenic groups and their influence on IgM and IgG-induced lysis (author's transl)].

    Science.gov (United States)

    Wiedermann, G; Stemberger, H; Förster, O; Müller, M

    1976-04-01

    Sheep erythrocytes were coated with bencylpenicilloyl-(BPO)groups. Different incubation periods resulted in erythrocyte preparations with different hapten density. Complement dependent lysis induced by IgM or IgG antibodies was studied with the cell preparations. The calculation of hapten density on the erythrocyte surface was not possible by direct measurement of coupled radioactive BPO since more than 90% of radioactive material was found in the soluble supernatant after osmotic cell lysis and less than 10% was fixed to the cellular membrane. Measurement of membrane bound immunologically relevant BPO-groups was achieved, therefore, by comparison of the inhibitory capacity of the test cells with that of a standard cell preparation. The latter consisted of tannic acid treated erythrocytes coated with protein complexed radioactive BPO. Surface hapten density of the different target cell preparations varied between 1.9 x 10(5) and 4.8 10(5) BPO-groups per cell depending on the time of incubation. Complement dependent antibody mediated cell lysis was significantly reduced by reduction of haptenic sites per target cell, IgG induced lysis being much more affected than hemolysis induced by IgM antibodies. Statistical calculations led to the conclusion that 18,000 protein islets per cell bearing 4 or more BPO-groups are not sufficient for hemolysis induced by IgG antibodies. 48,000 protein islets with this hapten density are necessary for "optimal" sensitization. IgG antibodies must be apparently bound to the cell surface in bivalent form.

  17. Trophic interactions between viruses, bacteria and nanoflagellates under various nutrient conditions and simulated climate change.

    Science.gov (United States)

    Bouvy, M; Bettarel, Y; Bouvier, C; Domaizon, I; Jacquet, S; Le Floc'h, E; Montanié, H; Mostajir, B; Sime-Ngando, T; Torréton, J P; Vidussi, F; Bouvier, T

    2011-07-01

    Population dynamics in the microbial food web are influenced by resource availability and predator/parasitism activities. Climatic changes, such as an increase in temperature and/or UV radiation, can also modify ecological systems in many ways. A series of enclosure experiments was conducted using natural microbial communities from a Mediterranean lagoon to assess the response of microbial communities to top-down control [grazing by heterotrophic nanoflagellates (HNF), viral lysis] and bottom-up control (nutrients) under various simulated climatic conditions (temperature and UV-B radiations). Different biological assemblages were obtained by separating bacteria and viruses from HNF by size fractionation which were then incubated in whirl-Pak bags exposed to an increase of 3°C and 20% UV-B above the control conditions for 96 h. The assemblages were also provided with an inorganic and organic nutrient supply. The data show (i) a clear nutrient limitation of bacterial growth under all simulated climatic conditions in the absence of HNF, (ii) a great impact of HNF grazing on bacteria irrespective of the nutrient conditions and the simulated climatic conditions, (iii) a significant decrease in burst size (BS) (number of intracellular lytic viruses per bacterium) and a significant increase of VBR (virus to bacterium ratio) in the presence of HNF, and (iv) a much larger temperature effect than UV-B radiation effect on the bacterial dynamics. These results show that top-down factors, essentially HNF grazing, control the dynamics of the lagoon bacterioplankton assemblage and that short-term simulated climate changes are only a secondary effect controlling microbial processes.

  18. Prolonged clot lysis time increases the risk of a first but not recurrent venous thrombosis.

    Science.gov (United States)

    Karasu, Alev; Baglin, Trevor P; Luddington, Roger; Baglin, Caroline A; van Hylckama Vlieg, Astrid

    2016-03-01

    The role of the fibrinolytic system in the development of venous thrombosis (VT) is unclear. We studied the risk of first and recurrent VT associated with reduced fibrinolysis, as measured by clot lysis time (CLT). We also studied the relationship between CLT and thrombin generation to determine if any relationship between CLT and VT was affected by thrombin generation. Analyses were performed in the Thrombophilia Hypercoagulability Environmental risk for Venous Thromboembolism Study, a two-centre population-based case-control study, including 579 patients and 338 controls, with patients followed from the event to determine incidence of recurrent VT. Hypofibrinolysis was associated with a 1·8-fold increased risk of a first VT [95% confidence interval (CI) 1·2-2·7]. Adjustment for sex, age, study location and Endogenous Thrombin Potential (ETP) did not change the result. The risk of VT was 2·9-fold increased when the 90th percentiles of prolonged CLT and high ETP were combined, with the highest risk for unprovoked first events (Odds Ratio = 4·2, 95% CI 1·3-13·5). In the follow-up study the Hazard Ratio for a recurrent VT associated with hypofibrinolysis was 1·5 (95% CI 0·9-2·6). A weak dose response effect was observed in relation to prolongation of CLT and recurrent VT. Although hypofibrinolysis constitutes a risk factor for a first VT, an association with recurrence is, at best, weak.

  19. Spontaneous acute tumor lysis syndrome in a DBA/1J mouse: a case report and review.

    Science.gov (United States)

    Lovelace, Karen; vanGessel, Yvonne; Asher, Ludmila V; Vogel, Peter

    2003-01-01

    Spontaneous acute tumor lysis syndrome (ATLS) was diagnosed in a 10-month-old female DBA/1J sentinel mouse with leukemic lymphoma. The mouse was unable to maintain balance and died shortly after being observed rolling around in its cage. Disseminated neoplastic disease, including a large cranial mediastinal mass, enlarged lymph nodes and splenomegaly, was present at necropsy. Histopathologic examination revealed widespread massive necrosis of lymphoblastic tumor cells, and widely disseminated microemboli composed of nuclear and cytoplasmic cell debris. Although ATLS is widely recognized as an oncologic emergency in humans, acute lesions of ATLS have not been described. The mechanical obstruction of capillary beds by microemboli originating from disintegrating necrotic tumor cells was the likely cause of clinical signs and death in this mouse. We propose that similar microemboli may contribute to the pathogenesis of the acute renal failure and other clinical signs associated with ATLS in humans. Recognition of spontaneous ATLS in laboratory animals is especially important in studies that assess the efficacy and/or toxicity of anticancer treatments, where early deaths due to ATLS might mistakenly be attributed to a direct test article effect.

  20. Screening of plants acting against Heterometrus laoticus scorpion venom activity on fibroblast cell lysis.

    Science.gov (United States)

    Uawonggul, Nunthawun; Chaveerach, Arunrat; Thammasirirak, Sompong; Arkaravichien, Tarinee; Chuachan, Chattong; Daduang, Sakda

    2006-01-16

    The aqueous extracts of 64 plant species, listed as animal- or insect-bite antidotes in old Thai drug recipes were screened for their activity against fibroblast cell lysis after Heterometrus laoticus scorpion venom treatment. The venom was preincubated with plant extract for 30 min and furthered treated to confluent fibroblast cells for 30 min. More than 40% efficiency (test/control) was obtained from cell treatment with venom preincubated with extracts of Andrographis paniculata Nees (Acanthaceae), Barringtonia acutangula (L.) Gaertn. (Lecythidaceae), Calamus sp. (Palmae), Clinacanthus nutans Lindau (Acanthaceae), Euphorbia neriifolia L. (Euphorbiaceae), Ipomoea aquatica Forssk (Convolvulaceae), Mesua ferrea L. (Guttiferae), Passiflora laurifolia L. (Passifloraceae), Plectranthus amboinicus (Lour.) Spreng. (Labiatae), Ricinus communis L. (Euphorbiaceae), Rumex sp. (Polygonaceae) and Sapindus rarak DC. (Sapindaceae), indicating that they had a tendency to be scorpion venom antidotes. However, only Andrographis paniculata and Barringtonia acutangula extracts provided around 50% viable cells from extract treatments without venom preincubation. These two plant extracts are expected to be scorpion venom antidotes with low cytotoxicity.

  1. Rasburicase represents a new tool for hyperuricemia in tumor lysis syndrome and in gout

    Directory of Open Access Journals (Sweden)

    Lisa Cammalleri, Mariano Malaguarnera

    2007-01-01

    Full Text Available Hyperuricemia is a feature of several pathologies and requires an appropriate and often early treatment, owing to the severe consequences that it may cause. A rapid and massive raise of uric acid, during tumor lysis syndrome (TLS, and also a lower and chronic hyperuricemia, as in gout, mainly damage the kidney. To prevent or treat these consequences, a new therapeutic option is represented by rasburicase, a recombinant form of an enzyme, urate oxidase. This enzyme converts hypoxanthine and xanthine into allantoin, a more soluble molecule, easily cleared by kidney. The several types of urate oxidase have followed each other, with progressive reduction of adverse reactions. The most important among them are allergenicity and the development of antibodies which compromise their effectiveness. Nevertheless, a limit of rasburicase's use remains its cost, which obliges to a judicious choice to prevent TLS in high risk patients with cancer and in case of allergy or impossibility to take allopurinol orally both in TLS and in gout. A large body of evidence confirms the efficacy and safety of rasburicase, even in comparison to the standard drugs used in the aforementioned pathologies.

  2. Ag nanoparticles generated using bio-reduction and -coating cause microbial killing without cell lysis.

    Science.gov (United States)

    Gade, Aniket; Adams, Joshua; Britt, David W; Shen, Fen-Ann; McLean, Joan E; Jacobson, Astrid; Kim, Young-Cheol; Anderson, Anne J

    2016-04-01

    Cost-effective "green" methods of producing Ag nanoparticles (NPs) are being examined because of the potential of these NPs as antimicrobials. Ag NPs were generated from Ag ions using extracellular metabolites from a soil-borne Pythium species. The NPs were variable in size, but had one dimension less than 50 nm and were biocoated; aggregation and coating changed with acetone precipitation. They had dose-dependent lethal effects on a soil pseudomonad, Pseudomonas chlororaphis O6, and were about 30-fold more effective than Ag(+) ions. A role of reactive oxygen species in cell death was demonstrated by use of fluorescent dyes responsive to superoxide anion and peroxide accumulation. Also mutants of the pseudomonad, defective in enzymes that protect against oxidative stress, were more sensitive than the wild type strain; mutant sensitivity differed between exposure to Ag NPs and Ag(+) ions demonstrating a nano-effect. Imaging of bacterial cells treated with the biocoated Ag NPs revealed no cell lysis, but there were changes in surface properties and cell height. These findings support that biocoating the NPs results in limited Ag release and yet they retained potent antimicrobial activity.

  3. Physical lysis only (PLO) methods suitable as rapid sample pretreatment for qPCR assay.

    Science.gov (United States)

    Wang, Xiaofang; Lee, Byung-Tae; Son, Ahjeong

    2014-10-01

    Quantitative PCR (qPCR) enables rapid and sensitive gene quantification and is widely used in genomics, such as biological, medical, environmental, and food sciences. However, sample pretreatment requires the use of conventional DNA extraction kits which are time-consuming and labor intensive. In this study, we investigated four physical lysis only (PLO) methods which are rapid and could serve as alternatives to conventional DNA extraction kits. These PLO methods are bead mill, heating, sonication, and freeze-thaw. Using ethidium bromide-based assay, their performance was evaluated and compared. The effects of cell debris and its removal were also investigated. Bead mill method without cell debris removal appeared to yield the best qPCR results among the four PLO methods. In addition, bead mill method also performed better than conventional DNA extraction kits. It is probably due to the substantial loss of DNA material during the extensive purification of the conventional DNA extraction kits. The bead mill method has been demonstrated to successfully quantify 10(2) to 10(7) copies of the PAH-RHDα gene of Pseudomonas putida.

  4. Ultrastructural study on the morphological changes in indigenous bacteria of mucous layer and chyme throughout the rat intestine.

    Science.gov (United States)

    Mantani, Youhei; Ito, Eri; Nishida, Miho; Yuasa, Hideto; Masuda, Natsumi; Qi, Wang-Mei; Kawano, Junichi; Yokoyama, Toshifumi; Hoshi, Nobuhiko; Kitagawa, Hiroshi

    2015-09-01

    Indigenous bacteria in the alimentary tract are exposed to various bactericidal peptides and digestive enzymes, but the viability status and morphological changes of indigenous bacteria are unclear. Therefore, the present study aimed to ultrastructurally clarify the degeneration and viability status of indigenous bacteria in the rat intestine. The majority of indigenous bacteria in the ileal mucous layer possessed intact cytoplasm, but the cytoplasm of a few bacteria contained vacuoles. The vacuoles were more frequently found in bacteria of ileal chyme than in those of ileal mucous layer and were found in a large majority of bacteria in both the mucous layer and chyme throughout the large intestine. In the dividing bacteria of the mucous layer and chyme throughout the intestine, the ratio of area occupied by vacuoles was almost always less than 10%. Lysis or detachment of the cell wall in the indigenous bacteria was more frequently found in the large intestine than in the ileum, whereas bacterial remnants, such as cell walls, were distributed almost evenly throughout the intestine. In an experimental control of long-time-cultured Staphylococcus epidermidis on agar, similar vacuoles were also found, but cell-wall degeneration was never observed. From these findings, indigenous bacteria in the mucous layer were ultrastructurally confirmed to be the source of indigenous bacteria in the chyme. Furthermore, the results suggested that indigenous bacteria were more severely degenerated toward the large intestine and were probably degraded in the intestine.

  5. Viral lysis of Phaeocystis pouchetii: implications for algal population dynamics and heterotrophic C, N and P cycling

    DEFF Research Database (Denmark)

    Haaber, Jakob Brandt Borup; Middelboe, Mathias

    2009-01-01

    and phosphorus mineralization. Lysis of P. pouchetii by PpV had strong positive effects on bacterial and HNF abundance, and the mass balance of C, N and P suggested an efficient transfer of organic material from P. pouchetii to bacterial and HNF biomass through viral lysis. At the same time, the degradation of P....... pouchetii lysates was associated with significant regeneration of inorganic N and P resulting in 148 microg N l(-1) and 7 microg P l(-1), corresponding to 78% and 26% of lysate N and P being mineralized to NH(4)(+) and PO(4)(3-), respectively. These results showed that the turnover of viral lysates...... in the microbial food web was associated with significant N and P mineralization, supporting the current view that viral lysates can be an important source of inorganic nutrients in marine systems. In the presence of R. salina, the generated NH(4)(+) supported 11% of the observed R. salina growth. Regrowth...

  6. Sulfolobus Turreted Icosahedral Virus c92 Protein Responsible for the Formation of Pyramid-Like Cellular Lysis Structures

    DEFF Research Database (Denmark)

    Snyder, Jamie C; Brumfield, Susan K; Peng, Nan;

    2011-01-01

    Host cells infected by Sulfolobus turreted icosahedral virus (STIV) have been shown to produce unusual pyramid-like structures on the cell surface. These structures represent a virus-induced lysis mechanism that is present in Archaea and appears to be distinct from the holin/endolysin system...... described for DNA bacteriophages. This study investigated the STIV gene products required for pyramid formation in its host Sulfolobus solfataricus. Overexpression of STIV open reading frame (ORF) c92 in S. solfataricus alone is sufficient to produce the pyramid-like lysis structures in cells. Gene...... disruption of c92 within STIV demonstrates that c92 is an essential protein for virus replication. Immunolocalization of c92 shows that the protein is localized to the cellular membranes forming the pyramid-like structures....

  7. Sulfolobus turreted icosahedral virus c92 protein responsible for the formation of pyramid-like cellular lysis structures.

    Science.gov (United States)

    Snyder, Jamie C; Brumfield, Susan K; Peng, Nan; She, Qunxin; Young, Mark J

    2011-07-01

    Host cells infected by Sulfolobus turreted icosahedral virus (STIV) have been shown to produce unusual pyramid-like structures on the cell surface. These structures represent a virus-induced lysis mechanism that is present in Archaea and appears to be distinct from the holin/endolysin system described for DNA bacteriophages. This study investigated the STIV gene products required for pyramid formation in its host Sulfolobus solfataricus. Overexpression of STIV open reading frame (ORF) c92 in S. solfataricus alone is sufficient to produce the pyramid-like lysis structures in cells. Gene disruption of c92 within STIV demonstrates that c92 is an essential protein for virus replication. Immunolocalization of c92 shows that the protein is localized to the cellular membranes forming the pyramid-like structures.

  8. Acute spontaneous tumor lysis in anaplastic large T-cell lymphoma presenting with hyperuricemic acute renal failure.

    Science.gov (United States)

    Hsu, Hsiang-Hao; Huang, Chiu-Ching

    2004-01-01

    Acute spontaneous tumor lysis (ASTL) syndrome, an extremely rare disease, requires prompt recognition and aggressive management because it is fulminant at its outset, associated with severe metabolic derangement, and potentially reversible. We describe an unusual case in which spontaneous tumor lysis occurred in anaplastic large T-cell lymphoma associated with acute uric acid nephropathy, persistent oliguria, and shock. This case contrasts markedly with previously reported cases of ASTL syndrome, which developed mainly in the pathologic type of Burkitt lymphoma. To our knowledge, this is the first reported occurrence of ASTL syndrome associated with anaplastic large T-cell type lymphoma. This report also chronicles our successful experience with continuous renal replacement therapy in the presence of compromised hemodynamic status.

  9. Sampling bacteria with a laser

    Science.gov (United States)

    Schwarzwälder, Kordula; Rutschmann, Peter

    2014-05-01

    Water quality is a topic of high interest and it's getting more and more important due to climate change and the implementation of European Water Framework Directive (WFD). One point of interest here is the inflow of bacteria into a river caused by combined sewer overflows which lead untreated wastewater including bacteria directly into a river. These bacteria remain in the river for a certain time, they settle down and can be remobilised again. In our study we want to investigate these processes of sedimentation and resuspension and use the results for the development of a software module coupled with the software Flow3D. Thereby we should be able to simulate and therefore predict the water quality influenced by combined sewer overflows. Hence we need to get information about the bacteria transport and fate. We need to know about the size of the bacteria or of the bacteria clumps and the size of the particles the bacteria are attached to. The agglomerates lead to different characteristics and velocities of settlement. The timespan during this bacteria can be detected in the bulk phase depends on many factors like the intensity of UV light, turbidity of the water, the temperature of the water, if there are grazers and a lot more. The size, density and composition of the agglomerates is just a part of all these influencing factors, but it is extremely difficult to differ between the other effects if we have no information about the simple sedimentation in default of these basic information. However we have a big problem getting the data. The chaining between bacteria or bacteria and particles is not too strong, so filtering the water to get a sieving curve may destroy these connections. We did some experiments similar to PIV (particle image velocimetry) measurements and evaluated the pictures with a macro written for the software ImageJ. Doing so we were able to get the concentration of bacteria in the water and collect information about the size of the bacteria. We

  10. Lisis de adherencias epidurales (Parte 2): Técnica intervencionista Percutaneous epidutal lysis of adhesions (2): Interventional technique

    OpenAIRE

    Racz, G.; Flores, J. C.

    2013-01-01

    This technique consists in the use of catheters and special needles to introduce solutions into difficult access sites in order to treat fibrosis or chronic inflammation conditions. Solutions such as hyaluronidase and hypertonic saline solution can be used for the lysis of adhesions caused by the pathology. This paper refers to different alternatives to correctly apply the technique at different spine levels. The technique has evolved from the conventional Tuhoy solution needles with some neu...

  11. Determining Human Clot Lysis Time (in vitro with Plasminogen/Plasmin from Four Species (Human, Bovine, Goat, and Swine

    Directory of Open Access Journals (Sweden)

    Omaira Cañas Bermúdez

    2015-05-01

    Full Text Available Cardiovascular disease is the leading cause of death worldwide, including failures in the plasminogen/plasmin system which is an important factor in poor lysis of blood clots. This article studies the fibrinolytic system in four species of mammals, and it identifies human plasminogen with highest thrombolysis efficiency. It examines plasminogen from four species (human, bovine, goat, and swine and identifies the most efficient one in human clot lysis in vitro. All plasminogens were identically purified by affinity chromatography. Human fibrinogen was purified by fractionation with ethanol. The purification of both plasminogen and fibrinogen was characterized by one-dimensional SDS-PAGE (10%. Human clot formation in vitro and its dissolution by plasminogen/plasmin consisted of determining lysis time from clot formation to its dilution. Purification of proteins showed greater than 95% purity, human plasminogen showed greater ability to lyse clot than animal plasminogen. The article concludes that human plasminogen/plasmin has the greatest catalysis and efficiency, as it dissolves human clot up to three times faster than that of irrational species.

  12. Comparison of clot lysis activity and biochemical properties of originator tenecteplase (Metalyse(®)) with those of an alleged biosimilar.

    Science.gov (United States)

    Kliche, Werner; Krech, Ingo; Michel, Martin C; Sangole, Nishant V; Sathaye, Sadhana

    2014-01-01

    The bioengineered tissue plasminogen activator tenecteplase is an important treatment modality of acute myocardial infarction recommended by international guidelines. Following introduction of originator tenecteplase (brand names Metalyse(®) and TNKase(®)), a "biosimilar" tenecteplase became available for commercial use in India under the brand name Elaxim(®) in the absence of Indian biosimilar guidelines which came into force from September 15th, 2012. Based on a report of biochemical and fibrinolytical differences between Metalyse and Elaxim, we have systematically compared them in a range of routine quality testing assays. As compared to Metalyse, Elaxim exhibited less clot lysis activity and contained less of the two-chain form of tenecteplase. Even upon full in vitro conversion to the two-chain form Elaxim exhibited less clot lysis activity. This was linked to differences in sialic acid content and glycosylation pattern with Elaxim exhibiting less bi- and more tetra-antennary glycosylation, leading to a different charge heterogeneity profile. Regarding purity, Elaxim contained more tenecteplase aggregates and, in contrast to Metalyse, considerable amounts of Chinese hamster ovary cell protein. Taken together these data demonstrate that Metalyse and Elaxim differ considerably in clot lysis activity and biochemical properties. These data question whether Elaxim indeed can be considered a "biosimilar" of Metalyse, i.e., whether and to which extent the clinical efficacy and safety properties of Metalyse can be extrapolated to Elaxim in the absence of comparative clinical data.

  13. Comparison of clot lysis activity and biochemical properties of originator tenecteplase (Metalyse® with those of an alleged biosimilar

    Directory of Open Access Journals (Sweden)

    Werner eKliche

    2014-02-01

    Full Text Available The bioengineered tissue plasminogen activator tenecteplase is an important treatment modality of acute myocardial infarction recommended by international guidelines. Following introduction of originator tenecteplase (brand names Metalyse® and TNKase®, a ‘biosimilar’ tenecteplase became available for commercial use in India under the brand name Elaxim® in the absence of Indian biosimilar guidelines which came into force from September 15th, 2012. Based on a report of biochemical and fibrinolytical differences between Metalyse and Elaxim, we have systematically compared them in a range of routine quality testing assays. As compared to Metalyse, Elaxim exhibited less clot lysis activity and contained less of the two-chain form of tenecteplase. Even upon full in vitro conversion to the two-chain form Elaxim exhibited less clot lysis activity. This was linked to differences in sialic acid content and glycosylation pattern with Elaxim exhibiting less bi- and more tetra-antennary glycosylation, leading to different charge heterogeneity profile. Regarding purity, Elaxim contained more tenecteplase aggregates and, in contrast to Metalyse, considerable amounts of Chinese hamster ovary cell protein. Taken together these data demonstrate that Metalyse and Elaxim differ considerably in clot lysis activity and biochemical properties. These data question whether Elaxim indeed can be considered a ‘biosimilar’ of Metalyse, i.e. whether and to which extent the clinical efficacy and safety properties of Metalyse can be extrapolated to Elaxim in the absence of comparative clinical data.

  14. RNA integrity as a quality indicator during the first steps of RNP purifications : A comparison of yeast lysis methods

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    Jansen Ralf-Peter

    2004-10-01

    Full Text Available Abstract Background The completion of several genome-sequencing projects has increased our need to assign functions to newly identified genes. The presence of a specific protein domain has been used as the determinant for suggesting a function for these new genes. In the case of proteins that are predicted to interact with mRNA, most RNAs bound by these proteins are still unknown. In yeast, several protocols for the identification of protein-protein interactions in high-throughput analyses have been developed during the last years leading to an increased understanding of cellular proteomics. If any of these protocols or similar approaches shall be used for the identification of mRNA-protein complexes, the integrity of mRNA is a critical factor. Results We compared the effect of different lysis protocols on RNA integrity. We report dramatic differences in RNA stability depending on the method used for yeast cell lysis. Glass bead milling and French Press lead to degraded mRNAs even in the presence of RNase inhibitors. Thus, they are not suitable to purify intact mRNP complexes or to identify specific mRNAs bound to proteins. Conclusion We suggest a novel protocol, grinding deep-frozen cells, for the preparation of protein extracts that contain intact RNAs, as lysis method for the purification of mRNA-protein complexes from yeast cells.

  15. Lysis of pig endothelium by IL-2 activated human natural killer cells is inhibited by swine and human major histocompatibility complex (MHC) class I gene products.

    Science.gov (United States)

    Itescu, S; Artrip, J H; Kwiatkowski, P A; Wang, S F; Minanov, O P; Morgenthau, A S; Michler, R E

    1997-01-01

    We have previously described a form of xenograft rejection, mediated by natural killer (NK) cells, occurring in pig-to-primate organ transplants beyond the period of antibody-mediated hyperacute rejection. In this study, two distinct NK activation pathways were identified as mechanisms of pig aortic endotheliual cell (PAEC) lysis by human NK cells. Using an antibody-dependent cellular cytotoxicity (ADCC) assay, a progressive increase in human NK lysis of PAEC was observed following incubation with human IgG at increasing serum titer. In the absence of IgG, a second mechanism of PAEC lysis by human NK cells was observed following activation with IL-2. IL-2 activation of human NK cells increased lysis of PAEC by over 3-fold compared with ADCC. These results indicate that IL-2 activation of human NK cells induces significantly higher levels of lytic activity than does conventional ADCC involving IgG and FcRIII. We next investigated the role of MHC class I molecules in the regulation of NK lysis following IL-2 activation. PAEC expression of SLA class I molecules was increased by up to 75% by treatment with human TNFa. Following treatment with TNFa at 1 u/ml, IL-2 activated human NK lysis of PAEC was inhibited at every effector:target (E:T) ratio tested. Maximal effect occurred at an E:T ratio of 10:1, with TNFa inhibiting specific lysis by 59% (p < 0.01). Incubation with an anti-SLA class I Mab, but not IgG isotype control, abrogated the protective effects of TNFa on NK lysis of PAEC, suggesting direct inhibitory effects of SLA class I molecules on human NK function. To investigate whether human MHC class I molecules might have similar effects on human NK lysis of PAEC, further experiments were performed using a soluble peptide derived from the alpha-helical region of HLA-B7. Incubation with the HLA-B7 derived peptide significantly reduced the IL-2 activated NK lytic activity against PAEC in a dose-dependent fashion. Maximal effect occurred at a concentration of 10 mg

  16. Rapid start-up of the anammox process: Effects of five different sludge extracellular polymeric substances on the activity of anammox bacteria.

    Science.gov (United States)

    Guo, Jianbo; Wang, Sihui; Lian, Jing; Ngo, Huu Hao; Guo, Wenshan; Liu, Yunman; Song, Yuanyuan

    2016-11-01

    This study investigated the rapid start-up of the anaerobic ammonium oxidation (anammox) strategy by inoculating different biomass ratios of denitrifying granular sludge and anammox bacteria. The results demonstrated that two reactors (R1 and R2) were rapidly and successfully started-up on days 25 and 28, respectively, with nitrogen removal rates (NRRs) of 0.70kg/(m(3)·d) and 0.72kg/(m(3)·d) at biomass ratios of 10:1 (R1) and 50:1 (R2). The explanation for rapid start-up was found by examining the effect of five different sludge extracellular polymeric substances (EPS) on the activity of anammox bacteria in the batch experiments. Batch experiments results first demonstrated that the denitrification sludge EPS (DS-EPS) enhanced the anammox bacteria activity the most, and NO2(-)-N, NH4(+)-N removal rates were 1.88- and 1.53-fold higher than the control with optimal DS-EPS volume of 10mL. The rapid start-up strategy makes possible the application of anammox to practical engineering.

  17. Myxoma virus infection promotes NK lysis of malignant gliomas in vitro and in vivo.

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    Henry Ogbomo

    Full Text Available Myxoma virus (MYXV is a well-established oncolytic agent against different types of tumors. MYXV is also known for its immunomodulatory properties in down-regulating major histocompatibility complex (MHC I surface expression (via the M153R gene product, a viral E3-ubiquitin ligase and suppressing T cell killing of infected target cells. MHC I down-regulation, however, favors NK cell activation. Brain tumors including gliomas are characterized by high MHC I expression with impaired NK activity. We thus hypothesized that MYXV infection of glioma cells will promote NK cell-mediated recognition and killing of gliomas. We infected human gliomas with MYXV and evaluated their susceptibility to NK cell-mediated cytotoxicity. MYXV enhanced NK cell-mediated killing of glioma cells (U87 cells, MYXV vs. Mock: 51.73% vs. 28.63%, P = .0001, t test; U251 cells, MYXV vs. Mock: 40.4% vs. 20.03%, P .0007, t test. Using MYXV M153R targeted knockout (designated vMyx-M153KO to infect gliomas, we demonstrate that M153R was responsible for reduced expression of MHC I on gliomas and enhanced NK cell-mediated antiglioma activity (U87 cells, MYXV vs. vMyx-M153KO: 51.73% vs. 25.17%, P = .0002, t test; U251 cells, MYXV vs. vMyx-M153KO: 40.4% vs. 19.27, P = .0013, t test. Consequently, NK cell-mediated lysis of established human glioma tumors in CB-17 SCID mice was accelerated with improved mouse survival (log-rank P = .0072. These results demonstrate the potential for combining MYXV with NK cells to effectively kill malignant gliomas.

  18. Myxoma virus infection promotes NK lysis of malignant gliomas in vitro and in vivo.

    Science.gov (United States)

    Ogbomo, Henry; Zemp, Franz J; Lun, Xueqing; Zhang, Jiqing; Stack, Danuta; Rahman, Masmudur M; McFadden, Grant; Mody, Christopher H; Forsyth, Peter A

    2013-01-01

    Myxoma virus (MYXV) is a well-established oncolytic agent against different types of tumors. MYXV is also known for its immunomodulatory properties in down-regulating major histocompatibility complex (MHC) I surface expression (via the M153R gene product, a viral E3-ubiquitin ligase) and suppressing T cell killing of infected target cells. MHC I down-regulation, however, favors NK cell activation. Brain tumors including gliomas are characterized by high MHC I expression with impaired NK activity. We thus hypothesized that MYXV infection of glioma cells will promote NK cell-mediated recognition and killing of gliomas. We infected human gliomas with MYXV and evaluated their susceptibility to NK cell-mediated cytotoxicity. MYXV enhanced NK cell-mediated killing of glioma cells (U87 cells, MYXV vs. Mock: 51.73% vs. 28.63%, P = .0001, t test; U251 cells, MYXV vs. Mock: 40.4% vs. 20.03%, P .0007, t test). Using MYXV M153R targeted knockout (designated vMyx-M153KO) to infect gliomas, we demonstrate that M153R was responsible for reduced expression of MHC I on gliomas and enhanced NK cell-mediated antiglioma activity (U87 cells, MYXV vs. vMyx-M153KO: 51.73% vs. 25.17%, P = .0002, t test; U251 cells, MYXV vs. vMyx-M153KO: 40.4% vs. 19.27, P = .0013, t test). Consequently, NK cell-mediated lysis of established human glioma tumors in CB-17 SCID mice was accelerated with improved mouse survival (log-rank P = .0072). These results demonstrate the potential for combining MYXV with NK cells to effectively kill malignant gliomas.

  19. Single-dose rasburicase 6 mg in the management of tumor lysis syndrome in adults.

    Science.gov (United States)

    McDonnell, Anne M; Lenz, Kristi L; Frei-Lahr, Debra A; Hayslip, John; Hall, Philip D

    2006-06-01

    Rasburicase is currently approved at a dosage of 0.15-0.2 mg/kg once/day for 5 days in pediatric patients with cancer to lower plasma uric acid concentrations and manage tumor lysis syndrome (TLS). Information on rasburicase dosing in adults is limited, with some data on using rasburicase as a single dose instead of multiple daily doses. Therefore, we evaluated the efficacy of a single dose of rasburicase for preventing or managing TLS in adults. We collected retrospective data for 11 adults with hematologic malignancies who received a single 6-mg dose of rasburicase. All patients received intravenous hydration with urinary alkalinization and allopurinol; however, due to adverse reactions, two patients received short courses of allopurinol. Only patients at high risk for TLS (e.g., large tumor burden, increasing uric acid concentration) or those with TLS received rasburicase. The single dose of rasburicase 6 mg resulted in a median 0.0773-mg/kg dose (range 0.0232-0.1361 mg/kg). The single 6-mg dose rapidly lowered uric acid concentrations in 10 of the 11 patients. The median uric acid concentration of 11.7 mg/dl (range 7.4-17.4 mg/dl) declined to 2.0 mg/dl (range 0.5-15.4 mg/dl) within a day after rasburicase administration (p=0.022). In these 10 patients, uric acid concentrations remained low despite subsequent chemotherapy, and none required additional rasburicase doses. The only patient who did not respond to the single 6-mg rasburicase dose was a morbidly obese man (259 kg, body mass index 87 kg/m2) who subsequently responded to an additional dose of rasburicase 12 mg. These results warrant further investigation of a single 6-mg dose of rasburicase in adults with TLS or at high-risk for developing TLS.

  20. Enhancement of antibody-dependent mechanisms of tumor cell lysis by a targeted activator of complement.

    Science.gov (United States)

    Imai, Masaki; Ohta, Rieko; Varela, Juan C; Song, Hongbin; Tomlinson, Stephen

    2007-10-01

    Complement inhibitors expressed on tumor cells provide a hindrance to the therapeutic efficacy of some monoclonal antibodies (mAb). We investigated a novel strategy to overwhelm complement inhibitor activity and amplify complement activation on tumor cells. The C3-binding domain of human complement receptor 2 (CR2; CD21) was linked to the complement-activating Fc region of human IgG1 (CR2-Fc), and the ability of the construct to target and amplify complement deposition on tumor cells was investigated. CR2 binds C3 activation fragments, and CR2-Fc targeted tumor cells by binding to C3 initially deposited by a tumor-specific antibody. Complement deposition on Du145 cells (human prostate cancer cell line) and anti-MUC1 mAb-mediated complement-dependent lysis of Du145 cells were significantly enhanced by CR2-Fc. Anti-MUC1 antibody-dependent cell-mediated cytotoxicity of Du145 by human peripheral blood mononuclear cells was also significantly enhanced by CR2-Fc in both the presence and the absence of complement. Radiolabeled CR2-Fc targeted to s.c. Du145 tumors in nude mice treated with anti-MUC1 mAb, validating the targeting strategy in vivo. A metastatic model was used to investigate the effect of CR2-Fc in a therapeutic paradigm. Administration of CR2-Fc together with mAb therapy significantly improved long-term survival of nude mice challenged with an i.v. injection of EL4 cells. The data show that CR2-Fc enhances the therapeutic efficacy of antibody therapy, and the construct may provide particular benefits under conditions of limiting antibody concentration or low tumor antigen density.

  1. Human leukocyte antigen E contributes to protect tumor cells from lysis by natural killer cells.

    Science.gov (United States)

    Lo Monaco, Elisa; Tremante, Elisa; Cerboni, Cristina; Melucci, Elisa; Sibilio, Leonardo; Zingoni, Alessandra; Nicotra, Maria Rita; Natali, Pier Giorgio; Giacomini, Patrizio

    2011-09-01

    The nonclassic class I human leukocyte antigen E (HLA-E) molecule engages the inhibitory NKG2A receptor on several cytotoxic effectors, including natural killer (NK) cells. Its tissue distribution was claimed to be wider in normal than in neoplastic tissues, and surface HLA-E was undetectable in most tumor cell lines. Herein, these issues were reinvestigated taking advantage of HLA-E-specific antibodies, immunohistochemistry, and biochemical methods detecting intracellular and surface HLA-E regardless of conformation. Contrary to published evidence, HLA-E was detected in a few normal epithelia and in a large fraction (approximately 1/3) of solid tumors, including those derived from HLA-E-negative/low-normal counterparts. Remarkably, HLA-E was detected in 30 of 30 tumor cell lines representative of major lymphoid and nonlymphoid lineages, and in 11 of 11, it was surface-expressed, although in a conformation poorly reactive with commonly used antibodies. Coexpression of HLA-E and HLA class I ligand donors was not required for surface expression but was associated with NKG2A-mediated protection from lysis by the cytotoxic cell line NKL and polyclonal NK cells from healthy donors, as demonstrated by antibody-mediated relief of protection in 10% to 20% of the tested target-effector combinations. NKG2A-mediated protection of additional targets became evident on NK effector blocking with antibodies to activating receptors (DNAM-1, natural cytotoxicity receptors, and NKG2D). Thus, initial evidence that the long-elusive HLA-E molecule is enhanced by malignant transformation and is functional in tumor cells is presented here, although its importance and precise functional role remain to be addressed in the context of a general understanding of the NK ligand-receptor network.

  2. Human Leukocyte Antigen E Contributes to Protect Tumor Cells from Lysis by Natural Killer Cells

    Directory of Open Access Journals (Sweden)

    Elisa Lo Monaco

    2011-09-01

    Full Text Available The nonclassic class I human leukocyte antigen E (HLA-E molecule engages the inhibitory NKG2A receptor on several cytotoxic effectors, including natural killer (NK cells. Its tissue distribution was claimed to be wider in normal than in neoplastic tissues, and surface HLA-E was undetectable in most tumor cell lines. Herein, these issues were reinvestigated taking advantage of HLA-E-specific antibodies, immunohistochemistry, and biochemical methods detecting intracellular and surface HLA-E regardless of conformation. Contrary to published evidence, HLA-E was detected in a few normal epithelia and in a large fraction (approximately 1/3 of solid tumors, including those derived from HLA-E-negative/low-normal counterparts. Remarkably, HLA-E was detected in 30 of 30 tumor cell lines representative of major lymphoid and nonlymphoid lineages, and in 11 of 11, it was surface-expressed, although in a conformation poorly reactive with commonly used antibodies. Coexpression of HLA-E and HLA class I ligand donors was not required for surface expression but was associated with NKG2A-mediated protection from lysis by the cytotoxic cell line NKL and polyclonal NK cells from healthy donors, as demonstrated by antibody-mediated relief of protection in 10% to 20% of the tested target-effector combinations. NKG2A-mediated protection of additional targets became evident on NK effector blocking with antibodies to activating receptors (DNAM-1, natural cytotoxicity receptors, and NKG2D. Thus, initial evidence that the long-elusive HLA-E molecule is enhanced by malignant transformation and is functional in tumor cells is presented here, although its importance and precise functional role remain to be addressed in the context of a general understanding of the NK ligand-receptor network.

  3. Human Leukocyte Antigen E Contributes to Protect Tumor Cells from Lysis by Natural Killer Cells12

    Science.gov (United States)

    Monaco, Elisa Lo; Tremante, Elisa; Cerboni, Cristina; Melucci, Elisa; Sibilio, Leonardo; Zingoni, Alessandra; Nicotra, Maria Rita; Natali, Pier Giorgio; Giacomini, Patrizio

    2011-01-01

    The nonclassic class I human leukocyte antigen E (HLA-E) molecule engages the inhibitory NKG2A receptor on several cytotoxic effectors, including natural killer (NK) cells. Its tissue distribution was claimed to be wider in normal than in neoplastic tissues, and surface HLA-E was undetectable in most tumor cell lines. Herein, these issues were reinvestigated taking advantage of HLA-E-specific antibodies, immunohistochemistry, and biochemical methods detecting intracellular and surface HLA-E regardless of conformation. Contrary to published evidence, HLA-E was detected in a few normal epithelia and in a large fraction (approximately 1/3) of solid tumors, including those derived from HLA-E-negative/low-normal counterparts. Remarkably, HLA-E was detected in 30 of 30 tumor cell lines representative of major lymphoid and nonlymphoid lineages, and in 11 of 11, it was surface-expressed, although in a conformation poorly reactive with commonly used antibodies. Coexpression of HLA-E and HLA class I ligand donors was not required for surface expression but was associated with NKG2A-mediated protection from lysis by the cytotoxic cell line NKL and polyclonal NK cells from healthy donors, as demonstrated by antibody-mediated relief of protection in 10% to 20% of the tested target-effector combinations. NKG2A-mediated protection of additional targets became evident on NK effector blocking with antibodies to activating receptors (DNAM-1, natural cytotoxicity receptors, and NKG2D). Thus, initial evidence that the long-elusive HLA-E molecule is enhanced by malignant transformation and is functional in tumor cells is presented here, although its importance and precise functional role remain to be addressed in the context of a general understanding of the NK ligand-receptor network. PMID:21969815

  4. P2X7 receptors mediate resistance to toxin-induced cell lysis.

    Science.gov (United States)

    Schoenauer, Roman; Atanassoff, Alexander P; Wolfmeier, Heidi; Pelegrin, Pablo; Babiychuk, Eduard B; Draeger, Annette

    2014-05-01

    In the majority of cells, the integrity of the plasmalemma is recurrently compromised by mechanical or chemical stress. Serum complement or bacterial pore-forming toxins can perforate the plasma membrane provoking uncontrolled Ca(2+) influx, loss of cytoplasmic constituents and cell lysis. Plasmalemmal blebbing has previously been shown to protect cells against bacterial pore-forming toxins. The activation of the P2X7 receptor (P2X7R), an ATP-gated trimeric membrane cation channel, triggers Ca(2+) influx and induces blebbing. We have investigated the role of the P2X7R as a regulator of plasmalemmal protection after toxin-induced membrane perforation caused by bacterial streptolysin O (SLO). Our results show that the expression and activation of the P2X7R furnishes cells with an increased chance of surviving attacks by SLO. This protective effect can be demonstrated not only in human embryonic kidney 293 (HEK) cells transfected with the P2X7R, but also in human mast cells (HMC-1), which express the receptor endogenously. In addition, this effect is abolished by treatment with blebbistatin or A-438079, a selective P2X7R antagonist. Thus blebbing, which is elicited by the ATP-mediated, paracrine activation of the P2X7R, is part of a cellular non-immune defense mechanism. It pre-empts plasmalemmal damage and promotes cellular survival. This mechanism is of considerable importance for cells of the immune system which carry the P2X7R and which are specifically exposed to toxin attacks.

  5. Chemotaxis toward phytoplankton drives organic matter partitioning among marine bacteria.

    Science.gov (United States)

    Smriga, Steven; Fernandez, Vicente I; Mitchell, James G; Stocker, Roman

    2016-02-09

    The microenvironment surrounding individual phytoplankton cells is often rich in dissolved organic matter (DOM), which can attract bacteria by chemotaxis. These "phycospheres" may be prominent sources of resource heterogeneity in the ocean, affecting the growth of bacterial populations and the fate of DOM. However, these effects remain poorly quantified due to a lack of quantitative ecological frameworks. Here, we used video microscopy to dissect with unprecedented resolution the chemotactic accumulation of marine bacteria around individual Chaetoceros affinis diatoms undergoing lysis. The observed spatiotemporal distribution of bacteria was used in a resource utilization model to map the conditions under which competition between different bacterial groups favors chemotaxis. The model predicts that chemotactic, copiotrophic populations outcompete nonmotile, oligotrophic populations during diatom blooms and bloom collapse conditions, resulting in an increase in the ratio of motile to nonmotile cells and in the succession of populations. Partitioning of DOM between the two populations is strongly dependent on the overall concentration of bacteria and the diffusivity of different DOM substances, and within each population, the growth benefit from phycospheres is experienced by only a small fraction of cells. By informing a DOM utilization model with highly resolved behavioral data, the hybrid approach used here represents a new path toward the elusive goal of predicting the consequences of microscale interactions in the ocean.

  6. Potential role of bacteria packaging by protozoa in the persistence and transmission of pathogenic bacteria.

    Science.gov (United States)

    Denoncourt, Alix M; Paquet, Valérie E; Charette, Steve J

    2014-01-01

    Many pathogenic bacteria live in close association with protozoa. These unicellular eukaryotic microorganisms are ubiquitous in various environments. A number of protozoa such as amoebae and ciliates ingest pathogenic bacteria, package them usually in membrane structures, and then release them into the environment. Packaged bacteria are more resistant to various stresses and are more apt to survive than free bacteria. New evidence indicates that protozoa and not bacteria control the packaging process. It is possible that packaging is more common than suspected and may play a major role in the persistence and transmission of pathogenic bacteria. To confirm the role of packaging in the propagation of infections, it is vital that the molecular mechanisms governing the packaging of bacteria by protozoa be identified as well as elements related to the ecology of this process in order to determine whether packaging acts as a Trojan Horse.

  7. Potential role of bacteria packaging by protozoa in the persistence and transmission of pathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Alix M Denoncourt

    2014-05-01

    Full Text Available Many pathogenic bacteria live in close association with protozoa. These unicellular eukaryotic microorganisms are ubiquitous in various environments. A number of protozoa such as amoebae and ciliates ingest pathogenic bacteria, package them usually in membrane structures, and then release them into the environment. Packaged bacteria are more resistant to various stresses and are more apt to survive than free bacteria. New evidence indicates that protozoa and not bacteria control the packaging process. It is possible that packaging is more common than suspected and may play a major role in the persistence and transmission of pathogenic bacteria. To confirm the role of packaging in the propagation of infections, it is vital that the molecular mechanisms governing the packaging of bacteria by protozoa be identified as well as elements related to the ecology of this process in order to determine whether packaging acts as a Trojan Horse.

  8. Modeling the fate of glucosinolates during thermal processing of Brassica vegetables

    NARCIS (Netherlands)

    Sarvan, I.; Verkerk, R.; Dekker, M.

    2012-01-01

    Glucosinolates are secondary metabolites of Brassicavegetables that have been associated with health benefits. The concentrations of these compounds are strongly affected by processing of the vegetables. Various mechanisms are responsible for these changes: Lysis of plant cells and compartments, dif

  9. Polymer/bacteria composite nanofiber non-wovens by electrospinning of living bacteria protected by hydrogel microparticles.

    Science.gov (United States)

    Gensheimer, Marco; Brandis-Heep, Astrid; Agarwal, Seema; Thauer, Rudolf K; Greiner, Andreas

    2011-03-10

    Physically crosslinked PVA-hydrogel microparticles are utilized for encapsulation of E. coli and M. luteus. The bacteria survive dry storage or treatment with bacteria-hostile organic solvents significantly better than unprotected bacteria as proven by culture-test experiments. The bacteria-protecting PVA microparticles are available for standard polymer-solution-processing techniques, as exemplarily shown by co-electrospinning of living bacteria encapsulated in dry PVA-hydrogel microparticles together with PVB-, PLLA-, and PCL-form organic solvents.

  10. Electron microscope observation of Salmonella Pullorum transmembrane tunnel mediated by the bacteriophage lysis protein E%噬菌体裂解E蛋白介导鸡白痢沙门菌跨膜孔道的电镜观察

    Institute of Scientific and Technical Information of China (English)

    郭荣显; 耿士忠; 焦红梅; 焦新安; 潘志明; 陈义芳; 安树敏

    2014-01-01

    Bacteriophage lysis protein E can lead to a fusion of the inner and outer membranes to form a specific transmembrane tunnel in Salmonella cytoderm. In this study, scanning electron microscopy ( SEM) and transmission electron microscopy ( TEM) were used to observe the characteristic of the transmembrane tunnel. Bacterial ghosts were prepared by inducing expression of the bacteriophage lysis gene E in Salmonella Pullorum S06004 harboring plasmid pBBR1MCS2-E. The pre-treatment of samples were observed by SEM and TEM. After induced in 42 ℃, the vast majority of Salmonella Pullorum recombinants could form a transmembrane tunnel at the polar sites of bacteria with a diameter varying in the range of 200~400 nm. Bacterial ghosts show the loss of cytoplasmic material and maintain the integrity of the outer membrane structure. The cell surface wrinkled obviously. This research provides an efficient morphological evidence for Salmonella ghosts as vaccine candidate, which is mediated by the bacteriophage lysis protein E.%应用电镜对噬菌体裂解 E蛋白介导沙门菌内外膜融合而形成的特异性跨膜孔道进行超微结构观察。42℃诱导携带温控表达质粒的重组鸡白痢沙门菌S06004(pBBR1MCS2-E)表达E基因以制备鸡白痢沙门菌菌影;样品经前处理,扫描电镜和透射电镜观察其形态结构变化。重组鸡白痢沙门菌经温度诱导后,绝大多数发生裂解,在两极形成直径介于200~400 nm之间的跨膜孔道,细菌整体呈空泡状,并保持完整的外膜结构,但细胞表面发生明显的褶皱。本研究为噬菌体E蛋白裂解沙门菌制备新型灭活疫苗的研究提供了形态学依据。

  11. Immune-gene therapy for renal cancer: chimeric receptor-mediated lysis of tumor cells

    NARCIS (Netherlands)

    M.E.M. Weijtens (Mo)

    2001-01-01

    textabstractThe immune system serves as a protective system against infectious agents such as bacteria, viruses and parasites. Foreign molecules (antigens) can be recognized by the immune system and induce an immune response resulting in destruction and elimination of the pathogens. In addition to i

  12. Bleach vs. Bacteria

    Science.gov (United States)

    ... Articles | Inside Life Science Home Page Bleach vs. Bacteria By Sharon Reynolds Posted April 2, 2014 Your ... hypochlorous acid to help kill invading microbes, including bacteria. Researchers funded by the National Institutes of Health ...

  13. Antibacterial Activity and Action Mechanism of the Essential Oil from Enteromorpha linza L. against Foodborne Pathogenic Bacteria.

    Science.gov (United States)

    Patra, Jayanta Kumar; Baek, Kwang-Hyun

    2016-03-21

    Foodborne illness and disease caused by foodborne pathogenic bacteria is continuing to increase day by day and it has become an important topic of concern among various food industries. Many types of synthetic antibacterial agents have been used in food processing and food preservation; however, they are not safe and have resulted in various health-related issues. Therefore, in the present study, essential oil from an edible seaweed, Enteromorpha linza (AEO), was evaluated for its antibacterial activity against foodborne pathogens, along with the mechanism of its antibacterial action. AEO at 25 mg/disc was highly active against Bacillus cereus (12.3-12.7 mm inhibition zone) and Staphylococcus aureus (12.7-13.3 mm inhibition zone). The minimum inhibitory concentration and minimum bactericidal concentration values of AEO ranged from 12.5-25 mg/mL. Further investigation of the mechanism of action of AEO revealed its strong impairing effect on the viability of bacterial cells and membrane permeability, as indicated by a significant increase in leakage of 260 nm absorbing materials and K⁺ ions from the cell membrane and loss of high salt tolerance. Taken together, these data suggest that AEO has the potential for use as an effective antibacterial agent that functions by impairing cell membrane permeability via morphological alternations, resulting in cellular lysis and cell death.

  14. Bacteria and lignin degradation

    Institute of Scientific and Technical Information of China (English)

    Jing LI; Hongli YUAN; Jinshui YANG

    2009-01-01

    Lignin is both the most abundant aromatic (phenolic) polymer and the second most abundant raw material.It is degraded and modified by bacteria in the natural world,and bacteria seem to play a leading role in decomposing lignin in aquatic ecosystems.Lignin-degrading bacteria approach the polymer by mechanisms such as tunneling,erosion,and cavitation.With the advantages of immense environmental adaptability and biochemical versatility,bacteria deserve to be studied for their ligninolytic potential.

  15. The detrimental influence of bacteria (E. coli, Shigella and Salmonella) on the degradation of organic compounds (and vice versa) in TiO2 photocatalysis and near-neutral photo-Fenton processes under simulated solar light.

    Science.gov (United States)

    Moncayo-Lasso, Alejandro; Mora-Arismendi, Luis Enrique; Rengifo-Herrera, Julián Andrés; Sanabria, Janeth; Benítez, Norberto; Pulgarin, César

    2012-05-01

    TiO2 photocatalytic and near-neutral photo-Fenton processes were tested under simulated solar light to degrade two models of natural organic matter - resorcinol (R) (which should interact strongly with TiO2 surfaces) and hydroquinone (H) - separately or in the presence of bacteria. Under similar oxidative conditions, inactivation of Escherichia coli, Shigella sonnei and Salmonella typhimurium was carried out in the absence and in the presence of 10 mg L(-1) of R and H. The 100% abatement of R and H by using a TiO2 photocatalytic process in the absence of bacteria was observed in 90 min for R and in 120 min for H, while in the presence of microorganisms abatement was only of 55% and 35% for R and H, respectively. Photo-Fenton reagent at pH 5.0 completely removed R and H in 40 min, whereas in the presence of microorganisms their degradation was of 60% to 80%. On the other hand, 2 h of TiO2 photocatalytic process inactivated S. typhimurium and E. coli cells in three and six orders of magnitude, respectively, while S. sonnei was completely inactivated in 10 min. In the presence of R or H, the bacterial inactivation via TiO2 photocatalysis was significantly decreased. With photo-Fenton reagent at pH 5 all the microorganisms tested were completely inactivated in 40 min of simulated solar light irradiation in the absence of organics. When R and H were present, bacterial photo-Fenton inactivation was less affected. The obtained results suggest that in both TiO2 and iron photo-assisted processes, there is competition between organic substances and bacteria simultaneously present for generated reactive oxygen species (ROS). This competition is most important in heterogeneous systems, mainly when there are strong organic-TiO2 surface interactions, as in the resorcinol case, suggesting that bacteria-TiO2 interactions could play a key role in photocatalytic cell inactivation processes.

  16. 乳酸菌在干辣椒发酵中的应用%The Appl ication of Lactic Acid Bacteria in Fermentation Process of Dry Red Pepper

    Institute of Scientific and Technical Information of China (English)

    万春秋; 刘德忠

    2015-01-01

    The orthogonal test is conducted by taking dry pepper as the raw material,adding lactic acid bacteria to produce chili paste.The results show that the optimal production conditions are as follows:the salt concentration of 10%,the inoculum size of 1 .0%,the fermentation temperature of 25 ℃ and the fermentation time of 20 days.Then the scale validation is carried out in the factory,the experimental results show that adding lactic acid bacteria not only shortens the fermentation time,but also the color of product is brighter,the aroma is stronger,the freshness is higher and the stability is better.%文章阐述了以干辣椒为原料,添加乳酸菌发酵生产辣椒酱,通过正交试验结果表明:最佳生产条件为食盐浓度10%,接种量1.0%,发酵温度25℃,发酵时间20天。然后在工厂进行了规模化验证,实验表明:添加乳酸菌发酵技术不仅缩短发酵的时间,且其产品的色泽更鲜艳、香味更浓郁、鲜度更高、稳定性更好。

  17. 生鲜调理鱼香肉丝在销售中的细菌变化%Changes of bacteria on fresh prepared fried shredded pork with sweet and sour sauce during the process of sales

    Institute of Scientific and Technical Information of China (English)

    李苗云; 朱应举; 张建威; 张秋会; 赵改名; 柳艳霞

    2011-01-01

    In order to protect the interests of consumers and enhance the health quality of prepared food, the bacteria changes of fresh prepared fried shredded pork with Sweet and sour sauce at different seasons and sale times were reported in the trial. The results showed that the microorganisms in prepared meat products during the processing of sales increased regularly, and in spring the micro-organisms were significantly higher than in winter, In spring, the total number of bacteria have reached to 7. 191 9 lg(CFU/g)after 9 h on the sales, and made a great impact on fresh prepared fried shredded pork with Sweet and sour sauce. In addition, Pseudomonas, Intestinal bacteria, Lactic acid bacteria increased with the extension of the sales' times,but The proportion of them had short odds. Therefore, they were the main microorganisms in fresh prepared fried shredded pork with Sweet and sour sauce.%对不同季节和不同销售时间生鲜调理鱼香肉丝中的细菌变化进行研完.结果表明,生鲜调理鱼香肉丝中微生物在销售过程中呈上升趋势,而且春季微生物含量明显高于冬季,春季销售9h时细菌总数已达到7.1919 1g(CFU/g),对生鲜调理鱼香肉丝的安全性影响较大;假单胞菌、肠道菌、乳酸菌在销售过程中随着销售时间的延长不断增加,但三者所占比例变化不大,是生鲜调理鱼香肉丝中的主要微生物.

  18. Influence of Legionella pneumophila and other water bacteria on the survival and growth of Acanthamoeba polyphaga.

    Science.gov (United States)

    Anacarso, I; Guerrieri, E; Bondi, M; de Niederhäusern, S; Iseppi, R; Sabia, C; Contri, M; Borella, P; Messi, P

    2010-10-01

    We investigated in solid medium, in water microcosm co-cultures and by light and transmission electron microscopy the influence of Legionella pneumophila Lp-1, Pseudomonas aeruginosa ATCC 27853, Burkholderia cepacia ATCC 25416 and Pseudomonas fluorescens SSD35 on the growth and survival of Acanthamoeba polyphaga. The infection with L. pneumophila was microscopically characterized by the presence of few bacteria inside protozoa at 4th h, and by the beginning of disruptive effects in late phase of trial. In water microcosm studies, performed at different temperature, the more significant interactions were observed at 30°C. In these conditions, L. pneumophila caused a marked reduction in trophozoite and cyst counts from the 4th day until the end of incubation (11 days). B. cepacia showed, by microscopic observation, few and generally single rods within protozoan phagosomes and caused a light reduction of trophozoite viability and cyst formation in co-cultures. A more invasive type of endocytosis, characterized by an early invasion with the presence of a high bacteria number inside amoebae, was observed for Pseudomonas strains. P. fluorescens produced a violent lysis of the host, whereas P. aeruginosa did not cause lysis or suffering. These results underline that water bacteria other than legionella are capable of intracellular survival in Acanthamoeba, influencing the protozoa viable cycle.

  19. Intracellular Bacteria in Protozoa

    Science.gov (United States)

    Görtz, Hans-Dieter; Brigge, Theo

    Intracellular bacteria in humans are typically detrimental, and such infections are regarded by the patients as accidental and abnormal. In protozoa it seems obvious that many bacteria have coevolved with their hosts and are well adapted to the intracellular way of life. Manifold interactions between hosts and intracellular bacteria are found, and examples of antibacterial resistance of unknown mechanisms are observed. The wide diversity of intracellular bacteria in protozoa has become particularly obvious since they have begun to be classified by molecular techniques. Some of the bacteria are closely related to pathogens; others are responsible for the production of toxins.

  20. Pitfalls, prevention, and treatment of hyperuricemia during tumor lysis syndrome in the era of rasburicase (recombinant urate oxidase

    Directory of Open Access Journals (Sweden)

    Andrea Pession

    2008-03-01

    Full Text Available Andrea Pession, Fraia Melchionda, Claudia CastelliniOncologia Ematologia Pediatrica “Lalla Seràgnoli”, Clinica Pediatrica, Università degli Studi di Bologna, Bologna, ItalyAbstract: Along with hydration and urinary alkalinization, allopurinol has been the standard agent for the management of hyperuricemia in patients with a high tumor burden at risk of tumor lysis syndrome; however, this agent often fails to prevent and treat this complication effectively. Rasburicase (recombinant urate oxidase has been shown to be effective in reducing uric acid and preventing uric acid accumulation in patients with hematologic malignancies with hyperuricemia or at high risk of developing it. Rasburicase acts at the end of the purine catabolic pathway and, unlike allopurinol, does not induce accumulation of xanthine or hypoxanthine. Its rapid onset of action and the ability to lower pre-existing elevated uric acid levels are the advantages of rasburicase over allopurinol. Rasburicase represents an effective alternative to allopurinol to promptly reduce uric acid levels, improve patient’s electrolyte status, and reverse renal insufficiency. The drug, initially studied in pediatric patients with acute lymphoblastic leukemia and aggressive non-Hodgkin lymphoma, seems to show comparable benefit in adults with similar lymphoid malignancies or at high risk of tumor lysis syndrome. Current and future trials will evaluate alternative doses and different schedules of rasburicase to maintain its efficacy while reducing its cost. The review provides a comprehensive and detailed review of pathogenesis, laboratory, and clinical presentation of TLS together with clinical studies already performed both in pediatric and adult patients.Keywords: tumor lysis syndrome, urate oxidase, rasburicase, allopurinol, uric acid

  1. 肿瘤溶解综合征的诊治%Diagnosis and treatment of tumor lysis syndrome

    Institute of Scientific and Technical Information of China (English)

    冯顺乔

    2015-01-01

    Tumor lysis syndrome(tumor lysis syndrome,TLS) is a common severe early complication of malignant disease in blood system.It also can occur in other malignancies.TLS is due to the large number of tumor cells dissolved in the short term,and release the intracellular metabolites,which cause a group of metabolic disorders syndromes.Its clinical features are hyperuricemia, hyperkalemia,hyperphosphatemia,hypocalcemia,acute renal failure.Positive prevention.Early diagnosis and effective treatment can prevent serious complications.In this paper,we review the relevant literature to explore pathological mechanism,epidemiology, risk factors,clinical diagnosis,prevention and treatment of TLS.%肿瘤溶解综合征(Tumor lysis syndrome,TLS)是血液系统恶性病中较常见的早期严重并发症,也可发生于其他恶性肿瘤。TLS是由于肿瘤细胞短期内大量溶解,释放细胞内代谢产物,引起代谢紊乱的一组综合征,临床特点为高尿酸血症、高血钾、高血磷、低血钙、急性肾功能衰竭。积极预防、早期诊断和有效的治疗可避免严重并发症的发生。本文复习相关文献以探讨TLS发生的病理机制、流行病学特征、高危因素、临床诊断、预防及治疗。

  2. Severe Tumor Lysis Syndrome and Acute Pulmonary Edema Requiring Extracorporeal Membrane Oxygenation Following Initiation of Chemotherapy for Metastatic Alveolar Rhabdomyosarcoma.

    Science.gov (United States)

    Sanford, Ethan; Wolbrink, Traci; Mack, Jennifer; Rowe, R Grant

    2016-05-01

    We present an 8-year-old male with metastatic alveolar rhabdomyosarcoma (ARMS) who developed precipitous cardiopulmonary collapse with severe tumor lysis syndrome (TLS) 48 hr after initiation of chemotherapy. Despite no detectable pulmonary metastases, acute hypoxemic respiratory failure developed, requiring extracorporeal membrane oxygenation (ECMO). Although TLS has been reported in disseminated ARMS, this singular case of life-threatening respiratory deterioration developing after initiation of chemotherapy presented unique therapeutic dilemmas. We review the clinical aspects of this case, including possible mechanisms of respiratory failure, and discuss the role of ECMO utilization in pediatric oncology.

  3. Can the development and autolysis of lactic acid bacteria influence the cheese volatile fraction? The case of Grana Padano.

    Science.gov (United States)

    Lazzi, Camilla; Povolo, Milena; Locci, Francesco; Bernini, Valentina; Neviani, Erasmo; Gatti, Monica

    2016-09-16

    In this study, the relationship between the dynamics of the growth and lysis of lactic acid bacteria in Grana Padano cheese and the formation of the volatile flavor compounds during cheese ripening was investigated. The microbial dynamics of Grana Padano cheeses that were produced in two different dairies were followed during ripening. The total and cultivable lactic microflora, community composition as determined by length heterogeneity-PCR (LH-PCR), and extent of bacterial lysis using an intracellular enzymatic activity assay were compared among cheeses after 2, 6 and 13months of ripening in two dairies. The evolution of whole and lysed microbiota was different between the two dairies. In dairy 2, the number of total cells was higher than that in dairy 1 in all samples, and the number of cells that lysed during ripening was lower. In addition, at the beginning of ripening (2months), the community structure of the cheese from dairy 2 was more complex and was composed of starter lactic acid bacteria (Lactobacillus helveticus and Lactobacillus delbrueckii) and NSLAB, possibly arising from raw milk, including Lactobacillus rhamnosus/Lactobacillus casei and Pediococcus acidilactici. On the other hand, the cheese from dairy 1 that ripened for 2months was mainly composed of the SLAB L. helveticus and L. delbrueckii. An evaluation of the free-DNA fraction through LH-PCR identified those species that had a high degree of lysis. Data on the dynamics of bacterial growth and lysis were evaluated with respect to the volatile profile and the organic acid content of the two cheeses after 13months of ripening, producing very different results. Cheese from dairy 1 showed a higher content of free fatty acids, particularly those deriving from milk fat lipolysis, benzaldehyde and organic acids, such as pGlu and citric. In contrast, cheese from dairy 2 had a greater amount of ketones, alcohols, hydrocarbons, acetic acid and propionic acid. Based on these results, we can conclude that

  4. Fibrinolytic efficacy of Amediplase, Tenecteplase and scu-PA in different external plasma clot lysis models: sensitivity to the inhibitory action of thrombin activatable fibrinolysis inhibitor (TAFI).

    Science.gov (United States)

    Guimarães, Ana H C; Barrett-Bergshoeff, Marrie M; Criscuoli, Marco; Evangelista, Stefano; Rijken, Dingeman C

    2006-09-01

    In this study, the in-vitro fibrinolytic efficacy of Tenecteplase, Amediplase and scu-PA was investigated in different external lysis models by measuring the lysis of human plasma clots after the addition of the plasminogen activators (PAs) to the surrounding plasma. The effect of TAFI was examined for each PA by neutralising TAFIa with potato carboxypeptidase inhibitor (PCI). The lytic efficacy of Amediplase was lower than that of Tenecteplase at low PA concentrations but slightly higher at therapeutic concentrations. The activity of scu-PA was clearly lower than that of either Tenecteplase or Amediplase. The TAFI system inhibited external clot lysis mediated by all the PAs when thrombomodulin was present in the model. In the therapeutic range (5-10 mug/ml) however, the TAFIa effect was negligible for both Amediplase and Tenecteplase. At lower PA concentrations the effect of TAFI on Amediplase was slightly stronger than that on Tenecteplase. Under static conditions the lysis rates were lower than with stirring. The role of TAFI was similar under both conditions. In conclusion, at therapeutic concentrations Amediplase was slightly more active than Tenecteplase and scu-PA under all conditions used. Therefore, Amediplase might possibly be a more potent thrombolytic agent at these concentrations and increase the efficacy of thrombolysis. The potential of TAFI for inhibiting thrombolytic therapy is probably low. However in conditions where the local PA concentrations are sub-optimal TAFI might affect the lysis rate.

  5. Organic and Inorganic Nitrogen Impact Chlorella variabilis Productivity and Host Quality for Viral Production and Cell Lysis.

    Science.gov (United States)

    Cheng, Yu-Shen; Labavitch, John; VanderGheynst, Jean S

    2015-05-01

    Microalgae have been proposed as a potential feedstock for biofuel production; however, cell disruption is usually required for collection and utilization of cytoplasmic polysaccharides and lipids. Virus infection might be one approach to disrupt the cell wall. The concentration of yeast extract and presence of KNO3 in algae cultivation media were investigated to observe their effects on Chlorella variabilis NC64A physiology and composition and the subsequent effect on production of Chlorella virus and disruption of infected cells. Cytoplasmic starch accumulation increased from 5% to approximately 35% of the total dry weight when yeast extract decreased from 1 to 0.25 g L(-1). When cells were cultured with the lowest nitrogen levels, the total polysaccharide accounted for more than 50% of the cell wall, which was 1.7 times higher than the content in cells cultured with the highest nitrogen levels. The C/N ratio of the algal biomass decreased by a factor of approximately 2 when yeast extract increased from 0.25 to 1 g L(-1). After virus infection, cells with a low C/N ratio produced a 7.6 times higher burst size than cells with a high C/N ratio, suggesting that the nitrogen content in C. variabilis has a large influence on viral production and cell lysis. The results have implications on management of nitrogen for both the synthesis of products from algae and product recovery via viral lysis.

  6. Staphylococcal α-hemolysin is neurotoxic and causes lysis of brain cells in vivo and in vitro.

    Science.gov (United States)

    Dahlberg, Daniel; Mariussen, Espen; Goverud, Ingeborg Løstegaard; Tønjum, Tone; Mæhlen, Jan; Antal, Ellen-Ann; Hassel, Bjørnar

    2015-05-01

    Formation of a bacterial brain abscess entails loss of brain cells and formation of pus. The mechanisms behind the cell loss are not fully understood. Staphylococcus aureus, a common cause of brain abscesses, produces various exotoxins, including α-hemolysin, which is an important factor in brain abscess formation. α-Hemolysin may cause cytolysis by forming pores in the plasma membrane of various eukaryotic cells. However, whether α-hemolysin causes lysis of brain cells is not known. Nor is it known whether α-hemolysin in the brain causes cell death through pore formation or by acting as a chemoattractant, recruiting leukocytes and causing inflammation. Here we show that α-hemolysin injected into rat brain causes cell damage and edema formation within 30 min. Cell damage was accompanied by an increase in extracellular concentrations of zinc, GABA, glutamate, and other amino acids, indicating plasma membrane damage, but leukocytic infiltration was not seen 0.5-12h after α-hemolysin injection. This was in contrast to injection of S. aureus, which triggered extensive infiltration with neutrophils within 8h. In vitro, α-hemolysin caused concentration-dependent lysis of isolated nerve endings and cultured astrocytes. We conclude that α-hemolysin contributes to the cell death inherent in staphylococcal brain abscess formation as a pore-forming neurotoxin.

  7. Antigen-Specific Inhibition of High-Avidity T Cell Target Lysis by Low-Avidity T Cells via Trogocytosis

    Directory of Open Access Journals (Sweden)

    Brile Chung

    2014-08-01

    Full Text Available Current vaccine conditions predominantly elicit low-avidity cytotoxic T lymphocytes (CTLs, which are non-tumor-cytolytic but indistinguishable by tetramer staining or enzyme-linked immunospot from high-avidity CTLs. Using CTL clones of high or low avidity for melanoma antigens, we show that low-avidity CTLs can inhibit tumor lysis by high-avidity CTLs in an antigen-specific manner. This phenomenon operates in vivo: high-avidity CTLs control tumor growth in animals but not in combination with low-avidity CTLs specific for the same antigen. The mechanism involves stripping of specific peptide-major histocompatibility complexes (pMHCs via trogocytosis by low-avidity melanoma-specific CTLs without degranulation, leading to insufficient levels of specific pMHC on target cell surface to trigger lysis by high-avidity CTLs. As such, peptide repertoire on the cell surface is dynamic and continually shaped by interactions with T cells. These results describe immune regulation by low-avidity T cells and have implications for vaccine design.

  8. Fibrinolytic enzyme production by newly isolated Bacillus cereus SRM-001 with enhanced in-vitro blood clot lysis potential.

    Science.gov (United States)

    Narasimhan, Manoj Kumar; Chandrasekaran, Muthukumaran; Rajesh, Mathur

    2015-01-01

    The discovery of plasmin-like microbial fibrinolytic enzymes having high specificity and negligible side effects is crucial for thrombolytic therapy. Herein, we report one such extra-cellular fibrinolytic enzyme producing Bacillus cereus SRM-001 isolated from the blood-laden soil of a chicken dump yard. The potency of the enzyme was established with fibrin plate assay and in-vitro blood clot lysis assay. The shake-flask operating parameters and media composition were optimized for maximizing the productivity of the enzyme. The operating parameters, pH 7, 37°C, 1% inoculum volume and 24 h inoculum age, were found to be the optimum. The levels of media components, corn flour (0.3% w/v), soyabean powder (1.9% w/v) and MnSO4 (11.5 mM) were optimized by statistical analysis using Box-Behnken design derived RSM. This resulted in an almost 1.8 fold increase in fibrinolytic enzyme productivity. The 3D response surface plots showed soyabean powder and MnSO4 to be the key ingredients for enhancing the enzyme productivity, whereas corn flour had a marginal effect. The in-vitro blood clot lysis assay conducted at near physiological pH 7 at 37°C showed the enzyme to be a potential therapeutic thrombolytic agent.

  9. Segmentation of Bacteria Image Based on Level Set Method

    Institute of Scientific and Technical Information of China (English)

    WANG Hua; CHEN Chun-xiao; HU Yong-hong; YANG Wen-ge

    2008-01-01

    In biology ferment engineering, accurate statistics of the quantity of bacte-ria is one of the most important subjects. In this paper, the quantity of bacteria which was observed traditionally manuauy can be detected automatically. Image acquisition and pro-cessing system is designed to accomplish image preprocessing, image segmentation and statistics of the quantity of bacteria. Segmentation of bacteria images is successfully real-ized by means of a region-based level set method and then the quantity of bacteria is com-puted precisely, which plays an important role in optimizing the growth conditions of bac-teria.

  10. Effects of bacterially produced precipitates on the metabolism of sulfate reducing bacteria during the bio-treatment process of copper-containing wastewater

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A large volume of bacterially produced precipitates are generated during the bio-treatment of heavy metal wastewater.The composition of the bacterially produced precipitates and its effects on sulfate reducing bacteria (SRB) in copper-containing waste stream were evaluated in this study.The elemental composition of the microbial precipitate was studied using electrodispersive X-ray spectroscopy (EDX),and it was found that the ratio of S:Cu was 1.12.Combining with the results of copper distribution in the SRB metabolism culture,which was analyzed by the sequential extraction procedure,copper in the precipitates was determined as covellite (CuS).The bacterially produced precipitates caused a decrease of the sulfate reduction rate,and the more precipitates were generated,the lower the sulfate reduction rate was.The particle sizes of bacterially generated covellite were ranging from 0.03 to 2 m by particles size distribution (PSD) analysis,which was smaller than that of the SRB cells.Transmission electron microscopy (TEM) analysis showed that the microbial covellite was deposited on the surface of the cell.The effects of the microbial precipitate on SRB metabolism were found to be weakened by increasing the precipitation time and adding microbial polymeric substances in later experiments.These results provided direct evidence that the SRB activity was inhibited by the bacterially produced covellite,which enveloped the bacterium and thus affected the metabolism of SRB on mass transfer.

  11. Study on the processing of chaenoneles lagenoria juice fermented by lactic acid bacteria%木瓜乳酸发酵饮料的工艺研究

    Institute of Scientific and Technical Information of China (English)

    谢主兰; 吴文龙; 雷晓凌

    2001-01-01

    Using chaenoneles lagenoria as main material to extract its juice to be fermented by lactic acid bacteria, the optimum fermenting technical condition was determined by orthogonal experiment. The result showed that the optimum density of chaenoneles lagenoria juice 50% ~ 60%, the optimum scale of L. bulgaricus: Str. thermophilus 1: 1, inoculation 4%, fermenting temperature 42℃ ,the first fermenting time 6~ 8 hours. The fermented drinks was stable,delicious,with optimum sweetness and sourness.%对以木瓜为原料提取的木瓜浆进行乳酸发酵,采用正交试验法优选出最佳发酵条件。结果表明,木瓜浆浓度50%~60%,保加利亚乳杆菌:嗜热链球菌最适比例为1:1,接种量4%,发酵温度42℃,前发酵时间6~8h,研制的木瓜乳酸发酵饮料组织均匀,口感细腻,甜酸适度。

  12. 玉竹沙参乳酸菌饮料的工艺研究%Process of Lactic acid Bacteria Beverage of Polygonatum and Adenophora

    Institute of Scientific and Technical Information of China (English)

    王媛; 梁彦

    2016-01-01

    Polygonatum,adenophora and fresh milk are used as raw materials. After sterilization,milk was inoculated with starter cultures for lactic acid fermentation,turning into solidified yogurt,then developing activated lactic acid bacteria beverage. Through single factor experiments and orthogonal experiments,optimal volumes of polygonatum, adenophora,sugar and composite stabilizer were determined: polygonatum of 9%,adenophora of 7%,sugar of 13%and stabilizer of 0.20%.%以玉竹、沙参以及鲜牛乳为主要原料,牛乳经杀菌后接种发酵剂进行乳酸发酵制成凝固型酸奶,经过调配制成活性乳酸菌饮料。通过单因素实验和正交试验,确定出玉竹浆、沙参浆、蔗糖以及复合稳定剂的最适添加量,玉竹浆添加量9%,沙参浆添加量7%,蔗糖添加量13%,复合稳定剂添加量0.20%。

  13. Genomics of Probiotic Bacteria

    Science.gov (United States)

    O'Flaherty, Sarah; Goh, Yong Jun; Klaenhammer, Todd R.

    Probiotic bacteria from the Lactobacillus and Bifidobacterium species belong to the Firmicutes and the Actinobacteria phylum, respectively. Lactobacilli are members of the lactic acid bacteria (LAB) group, a broadly defined family of microorganisms that ferment various hexoses into primarily lactic acid. Lactobacilli are typically low G + C gram-positive species which are phylogenetically diverse, with over 100 species documented to date. Bifidobacteria are heterofermentative, high G + C content bacteria with about 30 species of bifidobacteria described to date.

  14. 氯离子对含砷金精矿细菌氧化过程的影响研究%Research on the Chloride Ions’ Effect on the Bacteria Pre-oxidation Process of Arsenic-containing Gold Concentrates

    Institute of Scientific and Technical Information of China (English)

    杨玮; 刘瑞强; 任允超

    2014-01-01

    研究了不同Cl-浓度下含砷金精矿的细菌预氧化过程,试验结果表明:TCJ菌能够适应水源地的水质变化满足生产要求,在目前的生产条件下,TCJ菌耐受Cl-浓度的临界值是2.7 g/L;在不同的原料和浸出体系条件下,细菌对Cl-的适应性和耐受度是不同的,适宜的Cl-浓度在一定程度上有利于细菌氧化,过高的Cl-浓度会抑制细菌活性,影响预氧化效果。%The paper mainly studies the pre-oxidation process of bacteria of Arsenic-containing gold concentrates under different chloride ion concentrations.The results show that TCJ bacteria can adapt to the changes of water quality in water-head sources to meet the production requirements.On the present producing conditions,the critical value of TCJ bacterial tolerance concentration of chloride ions is 2.7 g/L.The experiment shows that the adaptability and the extent of tolerance of bacteria on Cl-is different in different raw materials and leaching system.Appropriate chloride ion is conducive to bacterial oxidation to some extent,while high chloride ions will inhibit bacterial activity and affect the pre-oxidation effect.

  15. Investigation of the Efficiencies of Bioaerosol Samplers for Collecting Aerosolized Bacteria Using a Fluorescent Tracer. I: Effects of Non-sampling Processes on Bacterial Culturability

    NARCIS (Netherlands)

    Zhao, Y.; Aarnink, A.J.A.; Doornenbal, P.; Huynh, T.T.T.; Groot Koerkamp, P.W.G.; Jong, de M.C.M.; Landman, W.J.M.

    2011-01-01

    By sampling aerosolized microorganisms, the efficiency of a bioaerosol sampler can be calculated depending on its ability both to collect microorganisms and to preserve their culturability during a sampling process. However, those culturability losses in the non-sampling processes should not be coun

  16. Survey on the community and dynamics of lactic acid bacteria in Grana Padano cheese.

    Science.gov (United States)

    Santarelli, Marcela; Bottari, Benedetta; Lazzi, Camilla; Neviani, Erasmo; Gatti, Monica

    2013-12-01

    Grana Padano (GP) is a Protected Designation of Origin cheese made with raw milk and natural whey culture (NWC) that is characterised by a long ripening period. In this study, six GP productions were considered in order to evaluate the trend of microbial dynamics and compare lactic acid bacteria (LAB) population levels in cheeses during the entire cheese-making process. To reach this goal, for each GP production, samples of vat raw milk, NWC and cheeses at 48h, 2, 6, 9 and 13 months were subjected to plate counts and direct counts by fluorescence microscopy, as well as amplicon length heterogeneity-PCR (LH-PCR). Statistical analysis was applied to the results and ecological indices were estimated. It was demonstrated that the LAB able to grow in the cheese-environment conditions could arise from both raw milk and NWC. Starter lactobacilli (SLAB) from NWC were the main species present during acidification, and non-starter LAB (NSLAB), mainly from milk but also from NWC, were able to grow after brining and they dominated during ripening. The peak areas of LH-PCR profiles were used to determine ecological indices during manufacture and ripening. Among cheese ecosystems with different ageing times, diversity, Evenness and Richness were different, with highest bacterial growth and diversity occurring in cheese ripening at 2 months. At this time point, which seemed to be a crucial moment for GP microbial evolution, cell lysis of both SLAB and NSLAB was also observed. Sampling modality and statistical analysis gave greater significance to the results used to describe the microbiological characteristics of a cheese recognised worldwide.

  17. The choice of fermentation strains of lactic acid bacteria in conserving process%保藏过程中乳酸菌发酵菌种的选择

    Institute of Scientific and Technical Information of China (English)

    杜磊; 袁超; 杜杨

    2011-01-01

    Lactobacillus bulgaricus and Streptococcus thermophilus are very primary in yogurt production. They play an important role in yogurt production. But they are easily being subject to degenerate activity in refrigeration so that the activity of preserved zymogen will consumedly debase. And the degenerating activity affected the zymogen's property. The Lactobacillus bulgaricus, Streptococcus thermophilus and their mixture were subcultured. Contrast the experimental data,it was found that mixing the two kinds of lactobacillus was the best way to conserve without exceptional case. It can greatly reduce the change of the lactic acid bacteria activity during the conserving time.%保加利亚乳杆菌和嗜热链球菌是酸奶生产过程的主要发酵菌种,在酸奶制作过程中起着极其重要的作用。但是由于菌种在其保藏过程中极易出现活性退化的现象,使得保藏后的菌种活性大大降低,影响了菌种的使用特性。分别对单独的保加利亚乳杆菌、嗜热链球菌及这两种菌的混合菌种进行菌种在保藏期间的传代实验,通过对实验数据的分析比较可得出,在没有特殊要求的情况下,乳酸菌菌种的保藏最好是使用保加刺亚乳杆菌和嗜热链球菌的混合菌种.这样可以大大降低乳酸菌菌种在保藏期内的特性变化。

  18. Chryseobacterium indologenes, novel mannanase-producing bacteria

    Directory of Open Access Journals (Sweden)

    Surachai Rattanasuk

    2009-10-01

    Full Text Available Mannanase is a mannan degrading enzyme which is produced by microorganisms, including bacteria. This enzyme can be used in many industrial processes as well as for improving the quality of animal feeds. The aim of the present study was toscreen and characterize the mannanase-producing bacteria. Two genera of bacteria were isolated from Thai soil samples,fermented coconut, and fertilizer. Screening was carried out on agar plates containing mannan stained with iodine solution.The bacteria were identified by partial 16S rRNA gene sequence, biochemical test and morphology, respectively. The mannanase activity was determined by zymogram and DNS method. Two strains of bacteria with mannanase activity were identified as Bacillus and Chryseobacterium. This is the first report of mannanase-producing Chryseobacterium.

  19. Nitrogen acquisition in Agave tequilana from degradation of endophytic bacteria

    OpenAIRE

    Beltran-Garcia, Miguel J.; White, JR; Prado, Fernanda M; Prieto, Katia R.; Yamaguchi, Lydia F.; Torres, Monica S.; Kato, Massuo J.; Medeiros, Marisa H. G.; Di Mascio,Paolo

    2014-01-01

    Plants form symbiotic associations with endophytic bacteria within tissues of leaves, stems, and roots. It is unclear whether or how plants obtain nitrogen from these endophytic bacteria. Here we present evidence showing nitrogen flow from endophytic bacteria to plants in a process that appears to involve oxidative degradation of bacteria. In our experiments we employed Agave tequilana and its seed-transmitted endophyte Bacillus tequilensis to elucidate organic nitrogen transfer from 15N-labe...

  20. Daratumumab-mediated lysis of primary multiple myeloma cells is enhanced in combination with the human anti-KIR antibody IPH2102 and lenalidomide

    DEFF Research Database (Denmark)

    Nijhof, I. S.; Lammerts van Bueren, J. J.; van Kessel, B.;

    2015-01-01

    killer cell inhibitory receptors with the human monoclonal anti-KIR antibody IPH2102, next to activation of natural killer cells with the immune modulatory drug lenalidomide. In 4-hour antibody-dependent cell-mediated cytotoxicity assays, IPH2102 did not induce lysis of multiple myeloma cell lines......RIIa-131R allele, who bind IgG1 with lower affinity than patients carrying the FcgammaRIIIa-158V allele or the FcgammaRIIa-131H allele. Finally, a further synergistically improved myeloma cell lysis with the daratumumab-IPH2102 combination was observed by adding lenalidomide, which suggests that more...

  1. Very High Throughput Electrical Cell Lysis and Extraction of Intracellular Compounds Using 3D Carbon Electrodes in Lab-on-a-Chip Devices

    Directory of Open Access Journals (Sweden)

    Philippe Renaud

    2012-08-01

    Full Text Available Here we present an electrical lysis throughput of 600 microliters per minute at high cell density (108 yeast cells per ml with 90% efficiency, thus improving the current common throughput of one microliter per minute. We also demonstrate the extraction of intracellular luciferase from mammalian cells with efficiency comparable to off-chip bulk chemical lysis. The goal of this work is to develop a sample preparation module that can act as a stand-alone device or be integrated to other functions already demonstrated in miniaturized devices, including sorting and analysis, towards a true lab-on-a-chip.

  2. 大黄4种饮片水提液体外肠道菌群转化比较研究%Comparison of transformation of four processed rhubarb aqueous extracts in intestinal bacteria in vitro

    Institute of Scientific and Technical Information of China (English)

    宋瑞; 田媛; 张尊建

    2012-01-01

    目的:研究比较大黄4种饮片水提液在大鼠肠道菌群中的代谢转化情况.方法:在厌氧条件下,分别将大黄4种饮片水提液与大鼠肠道菌试液在37℃温孵,采用HPLC-DAD,HPLC-MS/MS对可被肠道菌群代谢转化的成分及代谢产物进行定性分析,并根据各母体化合物在不同孵育时间的的色谱峰面积变化分析其代谢转化趋势.结果:蒽醌类化合物、没食子酰葡萄糖苷类化合物、萘苷类化合物能在肠道菌群的作用下发生代谢转化.其中,蒽醌类化合物占有绝对优势,在发现的17个代谢物中初步推测有12个由蒽醌类化合物转化而来.实验还发现,随着孵育时间的增加,不同饮同中同一成分代谢转化的趋势不尽相同.结论:炮制改变了大黄饮片所含成分的组合及比例关系,对它们的肠道菌群转化产生了影响.%Objective: To compare the metabolic transformation of four processed rhubarb aqueous extracts in rat intestinal bacteria in vitro. Method: Rat intestinal bacteria test solution and each of four processed rhubarb aqueous extracts were incubated under anaerobic conditions at 37 ℃. High-performance liquid chromatography with diode-array detection (HPLC-DAD) and tandem mass spectrometry (HPLC-MS/MS) was used for the qualitative analysis on the components that can be bio-transformed by rat intestinal bacteria as well as the trend of metabolic transformation of each parent compounds according to the changes in chromatographic peak areas in different incubation times. Result: Anthraquinones, glucose gallates and naphthalenes glucosides could be bio-transformed by rat intestinal bacteria. Of them, anthraquinones were undoubtedly the most prevalent parent compounds, as 12 out of the 17 metabolites were tentatively assigned as metabolites transformed from anthraquinones. Besides, it was also found that each parent compound in four processed rhubarb extract were diverse from each other with the incubation time

  3. Functional bacteria and process metabolism of the Denitrifying Sulfur conversion-associated Enhanced Biological Phosphorus Removal (DS-EBPR) system: An investigation by operating the system from deterioration to restoration.

    Science.gov (United States)

    Guo, Gang; Wu, Di; Hao, Tianwei; Mackey, Hamish Robert; Wei, Li; Wang, Haiguang; Chen, Guanghao

    2016-05-15

    A sulfur conversion-associated Enhanced Biological Phosphorus (P) Removal (EBPR) system is being developed to cater for the increasing needs to treat saline/brackish wastewater resulting from seawater intrusion into groundwater and sewers and frequent use of sulfate coagulants during drinking water treatment, as well as to meet the demand for eutrophication control in warm climate regions. However, the major functional bacteria and metabolism in this emerging biological nutrient removal system are still poorly understood. This study was thus designed to explore the functional microbes and metabolism in this new EBPR system by manipulating the deterioration, failure and restoration of a lab-scale system. This was achieved by changing the mixed liquor suspended solids (MLSS) concentration to monitor and evaluate the relationships among sulfur conversion (including sulfate reduction and sulfate production), P removal, variation in microbial community structures, and stoichiometric parameters. The results show that the stable Denitrifying Sulfur conversion-associated EBPR (DS-EBPR) system was enriched by sulfate-reducing bacteria (SRB) and sulfide-oxidizing bacteria (SOB). These bacteria synergistically participated in this new EBPR process, thereby inducing an appropriate level of sulfur conversion crucial for achieving a stable DS-EBPR performance, i.e. maintaining sulfur conversion intensity at 15-40 mg S/L, corresponding to an optimal sludge concentration of 6.5 g/L. This range of sulfur conversion favors microbial community competition and various energy flows from internal polymers (i.e. polysulfide or elemental sulfur (poly-S(2-)/S(0)) and poly-β-hydroxyalkanoates (PHA)) for P removal. If this range was exceeded, the system might deteriorate or even fail due to enrichment of glycogen-accumulating organisms (GAOs). Four methods of restoring the failed system were investigated: increasing the sludge concentration, lowering the salinity or doubling the COD

  4. Optimization of process conditions of dry longan pulp fermented by complex lactic acid bacteria%龙眼果浆复合乳酸菌发酵工艺优化

    Institute of Scientific and Technical Information of China (English)

    赖婷; 张名位; 刘磊; 张瑞芬; 邓媛元; 池建伟

    2016-01-01

    Longan (Dimocarpus longan Lour.) is a subtropical fruit, and it contains many nutritional and functional components such as carbohydrates, fiber, and phenolics. However, the fresh longan fruit is susceptible to spoilage and has a short storage life, so it is necessary to develop a new product of longan by fermentation. The purpose of this study was to obtain the complex lactic acid bacteria being suitable for fermentation of dry longan pulp, and the optimizaiton of process conditions of fermentation. The effect of seven lactic acid bacteria (Lactobacillus acidophilus,Leuconostoc mesenteroides, Lactobacillus casei,Pediococcus pentosaceus,Lactobacillus plantarum subsp·Plantarum,Lactobacillus delbrueckii subsp. lactis ) fermentations on the Lactobacillus colonies, the pH value, the acidity and the content of reducing sugar and free phenolic was analyzed. The result showed that the content of reducing sugar was consumed and the content of free phenolic was increased in seven lactic acid bacteria fermentations in dry longan pulp.Leuconostoc mesenteroides and Lactobacillus plantarumsubsp·Plantarum were selected by comparing the effect of seven lactic acid bacteria fermention. The conditions of longan pulp fermented by complex lactic acid bacteria were optimized through the single factor experiment, and factors included solid-liquid ratio, fermentation temperature, fermentation time, complex lactobacillus inoculation, complex lactobacillus matching shift. The result showed that solid-liquid ratio, fermentation temperature, fermentation time, and complex lactobacillus inoculation were significant influence factors in dry longan pulp fermented by complex lactic acid bacteria. And the suitable conditions in dry longan pulp fermented by complex lactic acid bacteria were solid-liquid ratio1:7 (g/mL), fermentation temperature 35℃, fermentation time 48 h, complex lactobacillus inoculation 1%. Complex lactobacillus matching shift could not significantly effect on the fuzzy

  5. Dual-species biofilms formation by Escherichia coli O157:H7 and environmental bacteria isolated from fresh-cut processing plants

    Science.gov (United States)

    Biofilm formation is a mechanism adapted by many microorganisms that enhances the survival in stressful environments. In food processing facilities, bacterial strains with strong biofilm forming capacities are more likely to survive the daily cleaning and disinfection. Foodborne bacterial pathogens,...

  6. Comparison of BACTEC MYCO/F LYTIC and WAMPOLE ISOLATOR 10 (lysis-centrifugation) systems for detection of bacteremia, mycobacteremia, and fungemia in a developing country.

    Science.gov (United States)

    Archibald, L K; McDonald, L C; Addison, R M; McKnight, C; Byrne, T; Dobbie, H; Nwanyanwu, O; Kazembe, P; Reller, L B; Jarvis, W R

    2000-08-01

    In less-developed countries, studies of bloodstream infections (BSI) have been hindered because of the difficulty and costs of culturing blood for bacteria, mycobacteria, and fungi. During two study periods (study period I [1997] and study period II [1998]), we cultured blood from patients in Malawi by using the BACTEC MYCO/F LYTIC (MFL), ISOLATOR 10 (Isolator), Septi-Chek AFB (SC-AFB), and Septi-Chek bacterial (SC-B) systems. During study period I, blood was inoculated at 5 ml into an MFL bottle, 10 ml into an Isolator tube for lysis and centrifugation, and 10 ml into an SC-B bottle. Next, 0.5-ml aliquots of Isolator concentrate were inoculated into an SC-AFB bottle and onto Middlebrook 7H11 agar slants, chocolate agar slants, and Inhibitory Mold Agar (IMA) slants. During study period II, the SC-B and chocolate agar cultures were discontinued. MFL growth was detected by fluorescence caused by shining UV light (lambda = 365 nm) onto the indicator on the bottom of the bottle. During study period I, 251 blood cultures yielded 44 bacterial isolates. For bacteremia, the MFL was similar to the Isolator concentrate on chocolate agar (34 of 44 versus 27 of 44; P, not significant [NS]), but more sensitive than the SC-B bottle (34 of 44 versus 24 of 44; P = 0.05). For both study periods combined, 486 blood cultures yielded 37 mycobacterial and 13 fungal isolates. For mycobacteremia, the sensitivities of the MFL and Isolator concentrate in the SC-AFB bottle were similar (30 of 37 versus 29 of 37; P, NS); the MFL bottle was more sensitive than the concentrate on Middlebrook agar (30 of 37 versus 15 of 37; P = 0.002). For fungemia, the MFL bottle was as sensitive as the SC-B bottle or Isolator concentrate on chocolate agar or IMA slants. We conclude that the MFL bottle, inoculated with just 5 ml of blood and examined under UV light, provides a sensitive and uncomplicated method for comprehensive detection of BSI in less-developed countries.

  7. Sulfate Reducing Bacteria Processing of Chromium-containing Waste Water Research Present Situation%硫酸盐还原菌处理含铬废水的研究现状

    Institute of Scientific and Technical Information of China (English)

    安文辉; 马斯然; 王磊峰

    2012-01-01

    Containing Cr (VI) of wastewater is a kind of difficult processing and hazard of heavy metal waste water, biological treatment of heavy metal wastewater has the characteristics of high efficiency low cost, sulfate reducing bacteria (SRB) is a kind of heavy metals to strong resistance of the anaerobic bacteria, including Cr (VI) wastewater treatment has the advantage. The paper introduced the SRB Cr-containing wastewater treatment using the related research and new technology.%含Cr(VI)的废水是一类难处理且危害大的重金属废水,生物处理重金属废水具有效率商成本低的特点,硫酸盐还原菌(SRB)是一类对重金属有较强抗性的厌氧菌,在含Cr(VI)废水处理方面具有优势。文章介绍了目前利用SRB处理含铬废水的相关研究及新工艺。

  8. How honey kills bacteria

    NARCIS (Netherlands)

    P.H.S. Kwakman; A.A. te Velde; L. de Boer; D. Speijer; C.M.J.E. Vandenbroucke-Grauls; S.A.J. Zaat

    2010-01-01

    With the rise in prevalence of antibiotic-resistant bacteria, honey is increasingly valued for its antibacterial activity. To characterize all bactericidal factors in a medical-grade honey, we used a novel approach of successive neutralization of individual honey bactericidal factors. All bacteria t

  9. Identification, characterization, and cloning of TIP-B1, a novel protein inhibitor of tumor necrosis factor-induced lysis.

    Science.gov (United States)

    Berleth, E S; Nadadur, S; Henn, A D; Eppolito, C; Shiojiri, S; Gurtoo, H L; Ehrke, M J; Mihich, E

    1999-11-01

    Some cancer cells evade elimination by virtue of their insensitivity to agents that induce apoptosis. Conversely, the side effects of anticancer agents could be diminished if normal cells were more resistant. To further elucidate the factors that contribute to the susceptibility of a cell to apoptosis, these investigations were designed to identify proteins isolated from cells exposed to low concentrations of tumor necrosis factor (TNF) that, when incubated with normally TNF-sensitive cells, protect these cells from TNF-induced cytotoxicity. TIP-B1, a novel protein, has been identified, purified, and characterized from cytosolic extracts of TNF-treated human fibroblasts. The approximately 27 kDa pI-4.5 TIP-B1 protein is unique based on both the sequence of three internal peptides (comprising 51 amino acids) and the nucleotide sequence of the corresponding 783-bp cDNA partial clone. Western blot analyses using polyclonal antisera raised against both the purified native TIP-B1 and the approximately 14 kDa product of the cDNA partial TIP-B1 clone, as well as Northern blot analyses using the cDNA insert as a probe, indicate that TIP-B1 may belong to a family of proteins that are expressed in a number of cell lines from diverse tissues. TNF-sensitive cells, when exposed to 4-10 microg/ml concentrations of TIP-B1 prior to the addition of TNF, are completely protected from TNF-induced lysis. Furthermore, TIP-B1 protects cells from apoptotic lysis induced by TNF. Preincubation of TIP-B1 with TNF does not affect the ability of TNF to induce lysis. Moreover, TIP-B1 does not seem to interfere with the interactions between TNF and the TNF receptors, based on a preliminary flow cytometric analysis of the cellular binding of biotinylated TNF. On the basis of these characteristics, TIP-B1 is not a soluble TNF receptor, an anti-TNF antibody, nor a protease that degrades TNF; yet TIP-B1 functions when added exogenously to cells. These characteristics, its novel sequence, and its

  10. Metallization of bacteria cells

    Institute of Scientific and Technical Information of China (English)

    黎向锋; 李雅芹; 蔡军; 张德远

    2003-01-01

    Bacteria cells with different standard shapes are well suited for use as templates for the fabrication of magnetic and electrically conductive microstructures. In this paper, metallization of bacteria cells is demonstrated by an electroless deposition technique of nickel-phosphorus initiated by colloid palladium-tin catalyst on the surfaces of Citeromyces matritensis and Bacillus cereus. The activated and metallized bacteria cells have been characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray diffraction analysis (XRD). Results showed that both Citeromyces matritensis and Bacillus cereus had no deformation in shape after metallization; the metallized films deposited on the surfaces of bacteria cells are homogeneous in thickness and noncrystalline in phase structure. The kinetics of colloid palladium-tin solution and electroless plating on bacteria cells is discussed.

  11. Cell wall elongation mode in Gram-negative bacteria is determined by peptidoglycan architecture.

    Science.gov (United States)

    Turner, Robert D; Hurd, Alexander F; Cadby, Ashley; Hobbs, Jamie K; Foster, Simon J

    2013-01-01

    Cellular integrity and morphology of most bacteria is maintained by cell wall peptidoglycan, the target of antibiotics essential in modern healthcare. It consists of glycan strands, cross-linked by peptides, whose arrangement determines cell shape, prevents lysis due to turgor pressure and yet remains dynamic to allow insertion of new material, and hence growth. The cellular architecture and insertion pattern of peptidoglycan have remained elusive. Here we determine the peptidoglycan architecture and dynamics during growth in rod-shaped Gram-negative bacteria. Peptidoglycan is made up of circumferentially oriented bands of material interspersed with a more porous network. Super-resolution fluorescence microscopy reveals an unexpected discontinuous, patchy synthesis pattern. We present a consolidated model of growth via architecture-regulated insertion, where we propose only the more porous regions of the peptidoglycan network that are permissive for synthesis.

  12. Complement-Dependent Lysis of Influenza A Virus-Infected Cells by Broadly Cross-Reactive Human Monoclonal Antibodies ▿

    Science.gov (United States)

    Terajima, Masanori; Cruz, John; Co, Mary Dawn T.; Lee, Jane-Hwei; Kaur, Kaval; Wilson, Patrick C.; Ennis, Francis A.

    2011-01-01

    We characterized human monoclonal antibodies (MAbs) cloned from influenza virus-infected patients and from influenza vaccine recipients by complement-dependent lysis (CDL) assay. Most MAbs active in CDL were neutralizing, but not all neutralizing MAbs can mediate CDL. Two of the three stalk-specific neutralizing MAbs tested were able to mediate CDL and were more cross-reactive to temporally distant H1N1 strains than the conventional hemagglutination-inhibiting and neutralizing MAbs. One of the stalk-specific MAbs was subtype cross-reactive to H1 and H2 hemagglutinins, suggesting a role for stalk-specific antibodies in protection against influenza illness, especially by a novel viral subtype which can cause pandemics. PMID:21994454

  13. High-intensity focused ultrasound sonothrombolysis: the use of perfluorocarbon droplets to achieve clot lysis at reduced acoustic power.

    Science.gov (United States)

    Pajek, Daniel; Burgess, Alison; Huang, Yuexi; Hynynen, Kullervo

    2014-09-01

    The purpose of this study was to evaluate use of intravascular perfluorocarbon droplets to reduce the sonication power required to achieve clot lysis with high-intensity focused ultrasound. High-intensity focused ultrasound with droplets was initially applied to blood clots in an in vitro flow apparatus, and inertial cavitation thresholds were determined. An embolic model for ischemic stroke was used to illustrate the feasibility of this technique in vivo. Recanalization with intravascular droplets was achieved in vivo at 24 ± 5% of the sonication power without droplets. Recanalization occurred in 71% of rabbits that received 1-ms pulsed sonications during continuous intravascular droplet infusion (p = 0.041 vs controls). Preliminary experiments indicated that damage was confined to the ultrasonic focus, suggesting that tolerable treatments would be possible with a more tightly focused hemispheric array that allows the whole focus to be placed inside of the main arteries in the human brain.

  14. Nitric oxide hinders antibody clearance from the surface of Trypanoplasma borreli and increases susceptibility to complement-mediated lysis.

    Science.gov (United States)

    Forlenza, Maria; Nakao, Miki; Wibowo, Indra; Joerink, Maaike; Arts, Joop A J; Savelkoul, Huub F J; Wiegertjes, Geert F

    2009-10-01

    Trypanoplasma borreli is an extracellular blood parasite of carp belonging to the same Order (Kinetoplastida) as African trypanosomes. These mammalian parasites have developed different strategies to evade the host immune system including antigenic variation, immunosuppression and clearance of surface-bound antibodies. The latter mechanism allows trypanosomes to use their swimming movement to cause surface-bound antibodies to 'sail' and accumulate at the posterior end of the parasite, to be internalized via the flagellar pocket and be degraded. There is no evidence that T. borreli shows antigenic variation, but during the late phases of infection NO-mediated immunosuppression is observed. High levels of nitric oxide (NO) lead to extensive tissue nitration whereas the parasite itself is not affected. Therefore, the induction of NO has thus far been considered a parasite-driven response with immunosuppressive effects. In the present study, we show that the induction of NO, particularly during the early phase of T. borreli infections, should be re-considered an effective part of the host immune response. We show that T. borreli rapidly removes surface-bound IgM. In addition, moderate concentrations of NO, by hindering surface antibody clearance, maintain high the concentrations of membrane-bound IgM, thereby favoring antibody-dependent complement-mediated parasite lysis. We performed a comprehensive quantitative gene expression analysis of in total seven different complement factors involved in all three activation pathways, differentiating between 1 and 4 isoforms for each complement gene. Our gene expression analysis supports an important role for antibody-dependent complement-mediated lysis of T. borreliin vivo. To our knowledge, NO-dependent inhibition of antibody clearance from the surface of kinetoplastid parasites has not been investigated. Our data support a role for NO as an important player in host-parasite interactions, not only as immune suppressor (late

  15. Rasburicase in the management of tumor lysis: an evidence-based review of its place in therapy

    Directory of Open Access Journals (Sweden)

    Dinnel J

    2015-01-01

    Full Text Available Jennifer Dinnel,1 Bonny L Moore,1 Brent M Skiver,1 Prithviraj Bose1,2 1Department of Internal Medicine, Virginia Commonwealth University, 2VCU Massey Cancer Center, Richmond, VA, USA Abstract: Tumor lysis syndrome (TLS is a potentially life-threatening complication of cancer therapy characterized by two or more of the following laboratory abnormalities: hyperuricemia, hyperkalemia, hypocalcemia, and hyperphosphatemia, with resultant end-organ damage, eg, renal failure, seizures, or cardiac arrhythmias. High-risk patients include those with highly proliferative cancers and/or large tumor burdens, particularly in the setting of highly effective chemotherapy, among other risk factors. Before 2002, antihyperuricemic drug therapy was limited to allopurinol, a xanthine oxidase inhibitor. Rasburicase, a recombinant urate oxidase, was approved by the US Food and Drug Administration for children in 2002 and adults in 2009, ushering in a new era in TLS therapy. We attempted to critically appraise the available evidence supporting the perceived benefits of rasburicase in the management of TLS. A Medline search yielded 98 relevant articles, including 26 retrospective and 22 prospective studies of rasburicase for the treatment of TLS, which were then evaluated to determine the best available evidence for the effectiveness of rasburicase in terms of disease-oriented, patient-oriented, and economic outcomes. Rasburicase is now a standard of care for patients at high risk of TLS despite continuing debate on the correlation between its profound and rapid lowering of plasma uric acid levels with hard patient outcomes, eg, need for renal replacement therapy and mortality. Rasburicase is dramatically effective in lowering plasma uric acid levels. The mortality and cost-effectiveness benefits of this expensive drug remain to be conclusively proven, and well designed, randomized controlled trials are needed to answer these fundamentally important questions. Keywords

  16. Modelling and predicting the simultaneous growth of Listeria monocytogenes and psychrotolerant lactic acid bacteria in processed seafood and mayonnaise-based seafood salads

    DEFF Research Database (Denmark)

    Mejlholm, Ole; Dalgaard, Paw

    2015-01-01

    A new combined model for Listeria monocytogenes and psychrotolerant Lactobacillus spp. was constructed and evaluated for processed seafood and mayonnaise-based seafood salads. The new model was constructed by combining existing cardinal parameter models for L. monocytogenes and Lactobacillus spp....... using the classical Jameson effect to model microbial interaction. Maximum population density (MPD) values of L. monocytogenes were accurately predicted in processed seafood with a known initial cell concentration of Lactobacillus spp. For these experiments, average MPD values of 4.5 and 4.3 log (cfu....../g) were observed and predicted, respectively for L. monocytogenes. In seafood salads, growth of L. monocytogenes continued at a reduced growth rate after Lactobacillus sakei had reached their MPD. This growth pattern was successfully described by an expanded version of the classical Jameson effect model...

  17. An integrated direct loop-mediated isothermal amplification microdevice incorporated with an immunochromatographic strip for bacteria detection in human whole blood and milk without a sample preparation step.

    Science.gov (United States)

    Lee, Dohwan; Kim, Yong Tae; Lee, Jee Won; Kim, Do Hyun; Seo, Tae Seok

    2016-05-15

    We have developed an integrated direct loop-mediated isothermal amplification (Direct LAMP) microdevice incorporated with an immunochromatographic strip (ICS) to identify bacteria contaminated in real samples. The Direct LAMP is a novel isothermal DNA amplification technique which does not require thermal cycling steps as well as any sample preparation steps such as cell lysis and DNA extraction for amplifying specific target genes. In addition, the resultant amplicons were colorimetrically detected on the ICS, thereby enabling the entire genetic analysis process to be simplified. The two functional units (Direct LAMP and ICS) were integrated on a single device without use of the tedious and complicated microvalve and tubing systems. The utilization of a slidable plate allows us to manipulate the fluidic control in the microchannels manually and the sequential operation of the Direct LAMP and ICS detection could be performed by switching the slidable plate to each functional unit. Thus, the combination of the direct isothermal amplification without any sample preparation and thermal cycling steps, the ICS based amplicon detection by naked eyes, and the slidable plate to eliminate the microvalves in the integrated microdevice would be an ideal platform for point-of-care DNA diaganotics. On the integrated Direct LAMP-ICS microdevice, we could analyze Staphylococcus aureus (S. aureus) and Escherichia coli O157:H7 (E. coli O157:H7) contaminated in human whole blood or milk at a single-cell level within 1h.

  18. Mexican Trypanosoma cruzi isolates: in vitro susceptibility of epimastigotes to anti-trypanosoma cruzi drugs and metacyclic forms to complement-mediated lysis.

    Science.gov (United States)

    León-Pérez, Floribeth; Gómez-Garcia, Lorena; Alejandre-Aguilar, R; López, R; Monteón, V M

    2007-01-01

    Trypanosoma cruzi has a clonal organization with an ample array of genetic and phenotypic features and probably anaploid constitution. Consequently, the biological behavior, biochemistry, and molecular attributes may be distinctive for each parasite strain in different geographical regions. As far as we know, there is no published information on the susceptibility of Mexican T. cruzi stocks to anti-T. cruzi drugs such as benznidazole and gentian violet, or on its resistance to complement-mediated lysis. We studied 10 Mexican T. cruzi isolates from different geographical areas, such as the pacific coast (Oaxaca, Guerrero, and Nayarit States), central part of Mexico (Guanajuato State), Gulf of Mexico (Veracruz State), and the Yucatan Peninsula (Campeche State). We searched for the natural resistance to drugs in in vitro assay against the 10 Mexican isolates using epimastigote forms and the complement-mediated lysis using metacyclic trypomastigotes insect-derived in three of them (one for each geographic region). In general, we observed high resistance to benznidazole in all the Mexican isolates tested, but in the complement-mediated lysis test, they showed moderate to high susceptibility. Although it is necessary to expand this study by using trypomastigotes and the intracellular form to verify its biological role, we suggest that Mexican T. cruzi parasites may have a variable susceptibility to antibody-mediated lysis and high resistance to benznidazole.

  19. Comparison of Two Cell Lysis Procedures for Recovery of Microcystins in Water Samples from Silver Lake in Dover, Delaware, with Microcystin Producing Cyanobacterial Accumulations

    Science.gov (United States)

    Loftin, Keith A.; Meyer, Michael T.; Rubio, Fernando; Kamp, Lisa; Humphries, Edythe; Whereat, Ed

    2008-01-01

    A collaboration was developed between Abraxis, LLC, the State of Delaware Department of Natural Resources and Environmental Control Division of Water Resources Environmental Laboratory, the University of Delaware, and the United States Geological Survey to investigate the efficacy of the QuikLyse procedure developed by Abraxis, LLC as an alternative cell-lysis technique suitable for use with an existing liquid chromatography/tandem mass spectrometry research method developed at the United States Geological Survey Organic Geochemistry Research Laboratory to analyze cyanotoxins. A comparison of three sequential freeze/thaw cycles versus QuikLyse, a proprietary chemical lysis procedure was conducted on four water samples collected from Silver Lake in Dover, Delaware. Results from the Abraxis Microcystins-DM enzyme-linked immunosorbent assay and liquid chromatography/tandem mass spectrometry were tabulated as a function of the cell lysis technique. Stastical comparison of percent relative standard deviations showed no significant difference (alpha = 0.05) between both cell-lysis techniques when measured by enzyme-linked immunosorbent assay or liquid chromatography/tandem mass spectrometry for three of the four samples.

  20. Antibiotics from predatory bacteria

    Directory of Open Access Journals (Sweden)

    Juliane Korp

    2016-03-01

    Full Text Available Bacteria, which prey on other microorganisms, are commonly found in the environment. While some of these organisms act as solitary hunters, others band together in large consortia before they attack their prey. Anecdotal reports suggest that bacteria practicing such a wolfpack strategy utilize antibiotics as predatory weapons. Consistent with this hypothesis, genome sequencing revealed that these micropredators possess impressive capacities for natural product biosynthesis. Here, we will present the results from recent chemical investigations of this bacterial group, compare the biosynthetic potential with that of non-predatory bacteria and discuss the link between predation and secondary metabolism.

  1. Porphyromonas gingivalis as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells.

    Science.gov (United States)

    Wunsch, Christopher M; Lewis, Janina P

    2015-12-17

    Anaerobic bacteria far outnumber aerobes in many human niches such as the gut, mouth, and vagina. Furthermore, anaerobic infections are common and frequently of indigenous origin. The ability of some anaerobic pathogens to invade human cells gives them adaptive measures to escape innate immunity as well as to modulate host cell behavior. However, ensuring that the anaerobic bacteria are live during experimental investigation of the events may pose challenges. Porphyromonas gingivalis, a Gram-negative anaerobe, is capable of invading a variety of eukaryotic non-phagocytic cells. This article outlines how to successfully culture and assess the ability of P. gingivalis to invade human umbilical vein endothelial cells (HUVECs). Two protocols were developed: one to measure bacteria that can successfully invade and survive within the host, and the other to visualize bacteria interacting with host cells. These techniques necessitate the use of an anaerobic chamber to supply P. gingivalis with an anaerobic environment for optimal growth. The first protocol is based on the antibiotic protection assay, which is largely used to study the invasion of host cells by bacteria. However, the antibiotic protection assay is limited; only intracellular bacteria that are culturable following antibiotic treatment and host cell lysis are measured. To assess all bacteria interacting with host cells, both live and dead, we developed a protocol that uses fluorescent microscopy to examine host-pathogen interaction. Bacteria are fluorescently labeled with 2',7'-Bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM) and used to infect eukaryotic cells under anaerobic conditions. Following fixing with paraformaldehyde and permeabilization with 0.2% Triton X-100, host cells are labeled with TRITC phalloidin and DAPI to label the cell cytoskeleton and nucleus, respectively. Multiple images taken at different focal points (Z-stack) are obtained for temporal

  2. [Darwin and bacteria].

    Science.gov (United States)

    Ledermann D, Walter

    2009-02-01

    As in 2009 the scientific world celebrates two hundreds years from the birthday of Charles Darwin and one hundred and fifty from the publication of The Origin of Species, an analysis of his complete work is performed, looking for any mention of bacteria. But it seems that the great naturahst never took knowledge about its existence, something rather improbable in a time when the discovery of bacteria shook the medical world, or he deliberately ignored them, not finding a place for such microscopic beings into his theory of evolution. But the bacteria badly affected his familiar life, killing scarlet fever one of his children and worsening to death the evolution of tuberculosis of his favourite Annie. Darwin himself could suffer the sickness of Chagas, whose etiological agent has a similar level to bacteria in the scale of evolution.

  3. 余甘子褐色乳酸菌饮料加工工艺研究%Study on the Processing Technology of Brown Emblica Lactic Acid Bacteria Beverage

    Institute of Scientific and Technical Information of China (English)

    杨洋; 高航; 段艳珠; 崔立红; 段玉禄

    2015-01-01

    以脱脂乳粉、余甘子为主要原料,研究了余甘子褐色乳酸菌饮料的制备工艺条件。通过正交试验研究了余甘子褐色乳酸菌饮料产品配方及乳化稳定剂的复配方案。结果表明:余甘子褐色乳酸菌饮料最适配料为白砂糖6.0%、余甘子汁2.0%、蜂蜜1.0%、甜橙香精0.08%。余甘子褐色乳酸菌饮料最佳稳定剂为可溶性大豆多糖0.20%、果胶0.30%、三聚磷酸钠0.05%。制得的余甘子褐色乳酸菌饮料色泽呈黄褐色,具有甜橙特征风味,口感清爽适口,并具有一定的营养保健功能。%skim milk powder and emblic leaflfower fruit are used as raw material and the manufacture process conditions of brown emblica lactic acid bacteria beverage were discussed. Product formula and compositional formulation of emulsion stabilizer were studied by orthogonal experiments. The results showed that the most adapted material formula of brown emblica lactic acid bacteria beverage was as fol ows:white sugar was 6.0%, phyl anthus emblica juice was 2.0%, honey was 1.0%, and an orange lfavour was 0.08%. The best stabilizer compositional formulation was as fol ows:soluble soybean polysaccharide was 0.20%, pectin was 0.30%, and sodium tripolyphosphate was 0.05%. The resulting yel owish-brown color of brown emblica lactic acid bacteria beverage, with characteristic lfavor of orange, fresh taste palatability, and has a certain nutrition and health function.

  4. Label-free bacteria detection using evanescent mode of a suspended core terahertz fiber

    CERN Document Server

    Mazhorova, Anna; Ng, Andy; Chinnappan, Raja; Zourob, Mohammed; Skorobogatiy, Maksim

    2011-01-01

    We propose for the first time an E. coli bacteria sensor based on the evanescent field of the fundamental mode of a suspended-core terahertz fiber. The sensor is capable of E. coli detection at concentrations in the range of 104-109 cfu/ml. The polyethylene fiber features a 150 {\\mu}m core suspended by three deeply sub-wavelength bridges in the center of a 5.1 mm-diameter cladding tube. The fiber core is biofunctionalized with T4 bacteriophages which bind and eventually destroy (lyse) their bacterial target. Using environmental SEM we demonstrate that E. coli is first captured by the phages on the fiber surface. After 25 minutes, most of the bacteria is infected by phages and then destroyed with ~1{\\mu}m-size fragments remaining bound to the fiber surface. The bacteria-binding and subsequent lysis unambiguously correlate with a strong increase of the fiber absorption. This signal allows the detection and quantification of bacteria concentration. Presented bacteria detection method is label-free and it does no...

  5. Production processing of fermented soy-milk by lactic acid bacteria%乳酸菌发酵酸豆乳生产工艺的研究

    Institute of Scientific and Technical Information of China (English)

    方海田; 刘慧燕

    2009-01-01

    以大豆为原料,对生产酸豆乳的工艺进行了研究.纯豆乳经浸泡、磨浆、脱腥后制成,通过正交试验,获得最佳的发酵工艺,豆水比为大豆∶水=1∶ 10.嗜热链球菌∶保加利亚乳杆菌=1∶ 1,85℃杀菌30min,蔗糖的添加量为8%,前发酵温度为40℃,前发酵时间为4h,接种量5%,后发酵温度4℃.%Using soybeans as raw material, production processing of fermented soy-milk was investigated in this paper. Pure soybean milk was obtained by soaking, grinding, removing fishy smell. The optimal fermentation conditions were achieved through orthogonal experiment the ratio of Streptococcus thermophilus to Lactobacillus pasteurization time 30 min, content of sucrose addition 8%, pre-fermentation temperature 40℃, pre-fermentation time 4 h, inoculum 5% and post-fermentation temperature 4℃.

  6. Bacteria-mediated bisphenol A degradation.

    Science.gov (United States)

    Zhang, Weiwei; Yin, Kun; Chen, Lingxin

    2013-07-01

    Bisphenol A (BPA) is an important monomer in the manufacture of polycarbonate plastics, food cans, and other daily used chemicals. Daily and worldwide usage of BPA and BPA-contained products led to its ubiquitous distribution in water, sediment/soil, and atmosphere. Moreover, BPA has been identified as an environmental endocrine disruptor for its estrogenic and genotoxic activity. Thus, BPA contamination in the environment is an increasingly worldwide concern, and methods to efficiently remove BPA from the environment are urgently recommended. Although many factors affect the fate of BPA in the environment, BPA degradation is mainly depended on the metabolism of bacteria. Many BPA-degrading bacteria have been identified from water, sediment/soil, and wastewater treatment plants. Metabolic pathways of BPA degradation in specific bacterial strains were proposed, based on the metabolic intermediates detected during the degradation process. In this review, the BPA-degrading bacteria were summarized, and the (proposed) BPA degradation pathway mediated by bacteria were referred.

  7. The fecal bacteria

    Science.gov (United States)

    Sadowsky, Michael J.; Whitman, Richard L.

    2011-01-01

    The Fecal Bacteria offers a balanced, integrated discussion of fecal bacteria and their presence and ecology in the intestinal tract of mammals, in the environment, and in the food supply. This volume covers their use in examining and assessing water quality in order to offer protection from illnesses related to swimming in or ingesting contaminated water, in addition to discussing their use in engineering considerations of water quality, modeling, monitoring, and regulations. Fecal bacteria are additionally used as indicators of contamination of ready-to-eat foods and fresh produce. The intestinal environment, the microbial community structure of the gut microbiota, and the physiology and genomics of this broad group of microorganisms are explored in the book. With contributions from an internationally recognized group of experts, the book integrates medicine, public health, environmental, and microbiological topics in order to provide a unique, holistic understanding of fecal bacteria. Moreover, it shows how the latest basic science and applied research findings are helping to solve problems and develop effective management strategies. For example, readers will discover how the latest tools and molecular approaches have led to our current understanding of fecal bacteria and enabled us to improve human health and water quality. The Fecal Bacteria is recommended for microbiologists, clinicians, animal scientists, engineers, environmental scientists, food safety experts, water quality managers, and students. It will help them better understand fecal bacteria and use their knowledge to protect human and environmental health. They can also apply many of the techniques and molecular tools discussed in this book to the study of a broad range of microorganisms in a variety of habitats.

  8. The Effects and Revelations of Manufacturing Process on the Health Benefits of Probiotic Bacteria%制造工艺对益生菌健康效应的影响及其启示

    Institute of Scientific and Technical Information of China (English)

    刘茂柯; 唐玉明; 曹晓涵; 任道群; 姚万春; 敖晓琳

    2014-01-01

    近年来的研究表明,益生菌产品的功效往往独立于活菌数存在并在生产加工过程中受到多种因素的影响。通过对相关研究报道的收集与分析,总结了生产环节影响益生菌健康效应的主要因素,并针对产品研发过程中应注意的问题和如何提高益生菌产品的功效提出了一些初步的观点和认识。%Recent researches indicated that the functionality of probiotic was affected by multiple factors during the manufacturing process, while the functionality was independent of viable bacteria number. In this paper, summarizes the main factors influencing the functionality of probiotic in the manufacturing process , and recommends some suggestions forward on the matters needing attention about the probiotic products during which were produced and on how to develop the efficacy of probiotic foods.

  9. Significance of postgrowth processing of ZnO nanostructures on antibacterial activity against gram-positive and gram-negative bacteria.

    Science.gov (United States)

    Mehmood, Shahid; Rehman, Malik A; Ismail, Hammad; Mirza, Bushra; Bhatti, Arshad S

    2015-01-01

    In this work, we highlighted the effect of surface modifications of one-dimensional (1D) ZnO nanostructures (NSs) grown by the vapor-solid mechanism on their antibacterial activity. Two sets of ZnO NSs were modified separately - one set was modified by annealing in an Ar environment, and the second set was modified in O2 plasma. Annealing in Ar below 800°C resulted in a compressed lattice, which was due to removal of Zn interstitials and increased O vacancies. Annealing above 1,000°C caused the formation of a new prominent phase, Zn2SiO4. Plasma oxidation of the ZnO NSs caused an expansion in the lattice due to the removal of O vacancies and incorporation of excess O. Photoluminescence (PL) spectroscopy was employed for the quantification of defects associated with Zn and O in the as-grown and processed ZnO NS. Two distinct bands were observed, one in the ultraviolet (UV) region, due to interband transitions, and other in the visible region, due to defects associated with Zn and O. PL confirmed the surface modification of ZnO NS, as substantial decrease in intensities of visible band was observed. Antibacterial activity of the modified ZnO NSs demonstrated that the surface modifications by Ar annealing limited the antibacterial characteristics of ZnO NS against Staphylococcus aureus. However, ZnO NSs annealed at 1,000°C or higher showed a remarkable antibacterial activity against Escherichia coli. O2 plasma-treated NS showed appreciable antibacterial activity against both E. coli and S. aureus. The minimum inhibition concentration was determined to be 0.5 mg/mL and 1 mg/mL for Ar-annealed and plasma-oxidized ZnO NS, respectively. It was thus proved that the O content at the surface of the ZnO NS was crucial to tune the antibacterial activity against both selected gram-negative (E. coli) and gram-positive (S. aureus) bacterial species.

  10. X-ray-induced lysis of the Fe-CO bond in carbonmonoxy-myoglobin.

    Science.gov (United States)

    Della Longa, Stefano; Arcovito, Alessandro

    2010-11-01

    By using X-ray absorption near edge structure (XANES) spectroscopy, we show that under prolonged exposure to Synchrotron X-rays, at T Fe-heme in carbonmonoxy-myoglobin (MbCO) undergoes a slow two-state transition process. The final spectrum is nearly identical to that of the classical photoproduct (Mb*CO) obtained by UV-visible light illumination at 15 K. By increasing the temperature, the starting spectrum of MbCO is recovered at T > 100 K, demonstrating that the process is reversible and no damage occurred at the heme site in the time course of the experiment. Thus, the overall X-ray-induced process at low temperature is identical to the well-known (light-induced) photolysis of CO-hemeproteins.

  11. Design and application of the method for isolating magnetotactic bacteria

    Institute of Scientific and Technical Information of China (English)

    XIAO Zhijie; LIAN Bin; CHEN Jun; H. Henry Teng

    2007-01-01

    A simple apparatus was designed to effectively isolate magnetotactic bacteria from soils or sediments based on their magnetotaxis. Through a series of processes including sample incubation, MTB harvesting, isolation, purification and identification, several strains of bacteria were isolated from the samples successfully. By Transmission Electron Microscopy (TEM) and Energy-Dispersive X-ray Analysis (EDXA), these bacteria were certificated to be magnetotactic bacteria. The phylogenetic relationship between the isolated magnetic strains and some known magnetotactic bacteria was inferred by the construction of phylogenetic tree based on 16SrDNA sequences. This apparatus has been proven to have the advantages of being inexpensive, simple to assemble, easy to perform and highly efficient to isolate novel magnetotactic bacteria. The research indicated that the combined approach of harvesting MTB by home-made apparatus and the method of plate colony isolation could purify and isolate magnetotactic bacteria effectively.

  12. Anaerobic bacteria in otitis media.

    Science.gov (United States)

    Fulghum, R S; Daniel, H J; Yarborough, J G

    1977-01-01

    Anaerobic bacteria, Peptostrepotococcus intermedius and Propionibacterium acnes, were found in mixed culture specimens from four to ten tested cases of chronic secretory otitis media. These anaerobic bacteria were in a mixed infection flora with aerobic bacteria most often Staphylococcus epidermidis and Cornybacterium sp. which do not fit any established species. The findings of anaerobic bacteria in otitis media is consistent with the sporadic report of the involvement of anaerobic bacteria in otitis media in the literature since 1898.

  13. Olive oil glycero lysis with the immobilized lipase Candida antarctica in a solvent free system

    Energy Technology Data Exchange (ETDEWEB)

    Singh, A. K.; Mukhopadhyay, M.

    2012-11-01

    In the present work, the solvent free lipase glycerolysis of olive oil for the production of monoglyceride (MG) and diglyceride (DG) with an immobilized Lipase B Candida antarctica was studied. The experiments were performed in batch mode by varying different process parameters. The Results showed that the MG and DG yields were dependent on operating conditions such as time, temperature, glycerol/ oil molar ratio, enzyme concentration and the water content in glycerol. The optimum operating time for maximum MG, 26 wt% and DG, 30 wt% production was 3h. The initial reaction rate was studied by varying different process parameters for 1h. The initial reaction rate increased at 30 degree centigrade temperature, 2:1 glycerol/oil molar ratio, 3.5% (w/w) water content in glycerol and 0.015g of enzyme loading. Comparative data for MG and DG yields for different oils and enzyme combinations were presented.

  14. Biological control of potato black scurf by rhizosphere associated bacteria

    Directory of Open Access Journals (Sweden)

    Mohsin Tariq

    2010-06-01

    Full Text Available The present work was carried out to study the potential of plant rhizosphere associated bacteria for the biocontrol of potato black scurf disease caused by Rhizoctonia solani Khun AG-3. A total of twenty-eight bacteria isolated from diseased and healthy potato plants grown in the soil of Naran and Faisalabad, Pakistan were evaluated for their antagonistic potential. Nine bacterial strains were found to be antagonistic in vitro, reduced the fungal growth and caused the lysis of sclerotia of R. solani in dual culture assay as well as in extracellular metabolite efficacy test. The selected antagonistic strains were further tested for the production and efficacy of volatile and diffusible antibiotics, lytic enzymes and siderophores against R. solani. Selected antagonistic bacteria were also characterized for growth promoting attributes i.e., phosphate solubilization, nitrogen fixation and indole acetic acid production. Biocontrol efficacy and percent yield increase by these antagonists was estimated in greenhouse experiment. Statistical analysis showed that two Pseudomonas spp. StT2 and StS3 were the most effective with 65.1 and 73.9 percent biocontrol efficacy, as well as 87.3 and 98.3 percent yield increase, respectively. Potential antagonistic bacterial strain StS3 showed maximum homology to Pseudomonas sp. as determined by 16S rRNA gene sequencing. These results suggest that bacterial isolates StS3 and StT2 have excellent potential to be used as effective biocontrol agents promoting plant growth with reduced disease incidence.

  15. Investigation of low-voltage pulse parameters on electroporation and electrical lysis using a microfluidic device with interdigitated electrodes.

    Science.gov (United States)

    Morshed, Bashir I; Shams, Maitham; Mussivand, Tofy

    2014-03-01

    Electroporation (EP) of biological cells leads to the exchange of materials through the permeabilized cell membrane, while electrical lysis (EL) irreversibly disrupts the cell membrane. We report a microfluidic device to study these two phenomena with low-voltage excitation for lab-on-a-chip (LOC) applications. For systematic study of EP, we have employed a quantification metric: flow Index (FI) of EP. Simulation and experimental results with the microfluidic device containing interdigitated, coplanar, integrated electrodes to electroporate, and rapidly lyse biological cells are presented. H&E stained human buccal cells were subjected to various pulse magnitudes, pulsewidths, and number of pulses. Simulations show that an electric field of 25 kV/cm with a 20 V applied potential produced 1.3 (°)C temperature rise for a 5 s of excitation. For a 20 V pulse-excitation with pulse-widths between 0.5 to 5 s, EL was observed, whereas for lower excitations, only EP was observed. FI of EP is found to be a direct function of pulse magnitudes, pulsewidths, and numbers of pulses. To release DNA from nucleus, excitation-pulses of 5 s were required. Quantification of EP would be useful for systematic study of EP toward optimization with various excitation pulses, while low-voltage requirement and high yield of EP and EL are critical to develop LOC for drug delivery and cell-sample preparation, respectively.

  16. Early postnatal diagnosis of hereditary spherocytosis by combining light microscopy, acidified glycerol lysis test and eosin-5'-maleimide binding assay.

    Science.gov (United States)

    Andres, Oliver; Eber, Stefan; Speer, Christian P

    2015-12-01

    Exact diagnosis of hereditary spherocytosis (HS) is widely considered unreliable around birth. However, early postnatal diagnosis at the beginning of congenital hemolysis may be essential for managing neonatal anemia and hemolytic icterus, identifying those at high risk for severe hyperbilirubinemia, irreversible kernicterus, or sudden need for red cell transfusion. We analyzed 37 blood samples from neonates or infants up to six weeks of life that had been collected in-house or shipped to our laboratory due to suspected red cell membrane disorder. By combining assessment of red cell morphology, acidified glycerol lysis test (AGLT), and eosin-5'-maleimide (EMA) binding assay, we were able to clearly exclude HS in 22 and confirm HS in 10 patients, of which one had undergone red cell transfusion prior to blood sampling. Assessment of red cell morphology and normal test results allowed diagnosis of infantile pyknocytosis or Heinz body anemia in three neonates. Re-evaluation of five patients with inconsistent results of AGLT and EMA binding led to confirmation of HS in two cases. Automated analysis of hematologic parameters revealed elevated proportion of hyperdense cells to be a highly significant indicator for HS in neonatal infants. We showed that assessment of red cell morphology in combination with AGLT and EMA binding assay is a reliable basis for confirming or rejecting suspected diagnosis of HS even in neonates. Our data underline the necessity for blood sampling and laboratory exploration in suspected red cell membrane or enzyme defects at the earliest occasion.

  17. Modified Atkins diet induces subacute selective ragged-red-fiber lysis in mitochondrial myopathy patients.

    Science.gov (United States)

    Ahola, Sofia; Auranen, Mari; Isohanni, Pirjo; Niemisalo, Satu; Urho, Niina; Buzkova, Jana; Velagapudi, Vidya; Lundbom, Nina; Hakkarainen, Antti; Muurinen, Tiina; Piirilä, Päivi; Pietiläinen, Kirsi H; Suomalainen, Anu

    2016-11-01

    Mitochondrial myopathy (MM) with progressive external ophthalmoplegia (PEO) is a common manifestation of mitochondrial disease in adulthood, for which there is no curative therapy. In mice with MM, ketogenic diet significantly delayed progression of the disease. We asked in this pilot study what effects high-fat, low-carbohydrate "modified Atkins" diet (mAD) had for PEO/MM patients and control subjects and followed up the effects by clinical, morphological, transcriptomic, and metabolomic analyses. All of our five patients, irrespective of genotype, showed a subacute response after 1.5-2 weeks of diet, with progressive muscle pain and leakage of muscle enzymes, leading to premature discontinuation of the diet. Analysis of muscle ultrastructure revealed selective fiber damage, especially in the ragged-red-fibers (RRFs), a MM hallmark. Two years of follow-up showed improvement of muscle strength, suggesting activation of muscle regeneration. Our results indicate that (i) nutrition can modify mitochondrial disease progression, (ii) dietary counseling should be part of MM care, (iii) short mAD is a tool to induce targeted RRF lysis, and (iv) mAD, a common weight-loss method, may induce muscle damage in a population subgroup.

  18. Granulocytes of sea anemone Actinia equina (Linnaeus, 1758 body fluid contain and release cytolysins forming plaques of lysis

    Directory of Open Access Journals (Sweden)

    MG Parisi

    2014-01-01

    Full Text Available The Cnidaria phylum includes organisms that are among the most poisonous animals. The exact composition of cnidarian bioactive molecules is not known in detail, but little is known on the cells that produce the toxins. Here we have shown that the presence of cytolysins is not exclusive of nematocysts. A plaque-forming assay was carried out with cell populations extracted from the percoled body fluid showed for the first time that anthozoan granulocytes are able to form plaque of lysis. We have partitioned the total population of free cells into three distinct discrete bands by discontinuous Percoll gradient, and we have identified six small different types cells: morular granulocytes; cells with large or small peripherical granules, granulocytes with irregular shape containing blue and red granules, cells showing one fine red granule of uniform size and, finally, cells with elongated shape and small dispersed granules. Cell lysate of each cellular band resulted cytolytic toward different erythrocytes types. SDS page analysis of the lysate cell fraction showed a predominant of 20 kDa that corresponds to the weight of the cytolytic equinatoxin. The nature of equinatoxins-related activity was demonstrated by inhibition experiments using bovine sphingomyelin.

  19. Real-time monitoring of bioaerosols via cell-lysis by air ion and ATP bioluminescence detection.

    Science.gov (United States)

    Park, Chul Woo; Park, Ji-Woon; Lee, Sung Hwa; Hwang, Jungho

    2014-02-15

    In this study, we introduce a methodology for disrupting cell membranes with air ions coupled with ATP bioluminescence detection for real-time monitoring of bioaerosol concentrations. A carbon fiber ionizer was used to extract ATP from bacterial cells for generating ATP bioluminescence. Our methodology was tested using Staphylococcus epidermidis and Escherichia coli, which were aerosolized with an atomizer, and then indoor bioaerosols were also used for testing the methodology. Bioaerosol concentrations were estimated without culturing which requires several days for colony formation. Correlation equations were obtained for results acquired using our methodology (Relative Luminescent Unit (RLU)/m(3)) and a culture-based (Colony Forming Unit (CFU)/m(3)) method; CFU/m(3)=1.8 × measured RLU/m(3) for S. epidermidis and E. coli, and CFU/m(3)=1.1 × measured RLU/m(3) for indoor bioaerosols under the experimental conditions. Our methodology is an affordable solution for rapidly monitoring bioaerosols due to rapid detection time (cell-lysis time: 3 min; bioluminescence detection time: <1 min) and easy operation.

  20. An unusual presentation of tumor lysis syndrome in a patient with advanced gastric adenocarcinoma: case report and literature review.

    Science.gov (United States)

    Vodopivec, Danica Maria; Rubio, Jose Enrique; Fornoni, Alessia; Lenz, Oliver

    2012-01-01

    Tumor lysis syndrome (TLS) is characterized by hyperuricemia, hyperkalemia, hyperphosphatemia, and secondary hypocalcemia in patients with a malignancy. When these laboratory abnormalities develop rapidly, clinical complications such as cardiac arrhythmias, acute renal failure, seizures, or death may occur. TLS is caused by rapid release of intracellular contents by dying tumor cells, a condition that is expected to be common in hematologic malignancies. However, TLS rarely occurs with solid tumors, and here we present the second chemotherapy-induced TLS in a patient with advanced gastric adenocarcinoma to be reported in the literature. We also provide information regarding the total cases of TLS in solid tumors reported from 1977 to present day. Our methodology involved identifying key articles from existing reviews of the literature and then using search terms from these citations in MEDLINE to find additional publications. We relied on a literature review published in 2003 by Baeksgaard et al., where they gathered all total 45 cases reported from 1977 to 2003. Then, we looked for new reported cases from 2004 to present day. All reports (case reports, brief reports, letters to editor, correspondence, reviews, journals, and short communications) identified through these searches were reviewed and included.

  1. An Unusual Presentation of Tumor Lysis Syndrome in a Patient with Advanced Gastric Adenocarcinoma: Case Report and Literature Review

    Directory of Open Access Journals (Sweden)

    Danica Maria Vodopivec

    2012-01-01

    Full Text Available Tumor lysis syndrome (TLS is characterized by hyperuricemia, hyperkalemia, hyperphosphatemia, and secondary hypocalcemia in patients with a malignancy. When these laboratory abnormalities develop rapidly, clinical complications such as cardiac arrhythmias, acute renal failure, seizures, or death may occur. TLS is caused by rapid release of intracellular contents by dying tumor cells, a condition that is expected to be common in hematologic malignancies. However, TLS rarely occurs with solid tumors, and here we present the second chemotherapy-induced TLS in a patient with advanced gastric adenocarcinoma to be reported in the literature. We also provide information regarding the total cases of TLS in solid tumors reported from 1977 to present day. Our methodology involved identifying key articles from existing reviews of the literature and then using search terms from these citations in MEDLINE to find additional publications. We relied on a literature review published in 2003 by Baeksgaard et al., where they gathered all total 45 cases reported from 1977 to 2003. Then, we looked for new reported cases from 2004 to present day. All reports (case reports, brief reports, letters to editor, correspondence, reviews, journals, and short communications identified through these searches were reviewed and included.

  2. Nd:YAG Laser Lysis of the Fibrinous Membrane and Remnant Substance on the Anterior Surface of Intraocular Lens

    Institute of Scientific and Technical Information of China (English)

    ZhendeLin; ChenjinJin

    1995-01-01

    Purpose:To determine the effects of ND:YAGlaser to disrupt the fibrinous membrane and remnant substance on the anterior surface of intraocular lens.Methods:Nd:YAGlaser was applied on 23cases of fibrinous membrane forma-tion and 8cases of remnant substance on the anterior surface of intaocular lenses(IOL)which had not reponded well to steroid therapy,Eighteen cases were male and 13female.The mean age was49.7years(range,5-78years),The in-terval between IOL implantation and laser therapy was 0.5-30months in the fibrinous membrane cases and 3-10days in the remnant substance cases.The energy applied was0.8-3.0mJ/exp.with2-112exposures.mean follow-up period was3.6months.Results:Complications during therapy included only 2cases of slight iris bleed-ing,Visual acuity after therapy was improved 1line in 16cases,2lines in 11cas-es,3lines in 1case,4lines in 1case and 5lines in 2cases,No post-therapy com-plication was found.Conclusions:Nd:YAG laser lysis is an effective alternative to ramove the fibri-nous membrane and remnant substances on the anterior surface of IOL.Eye Sci-ence1995;11:128-130.

  3. Study of Lactobacillus as Probiotic Bacteria

    Directory of Open Access Journals (Sweden)

    J Nowroozi

    2004-07-01

    Full Text Available Because of inhibitory effect, selected probiotic lactobacilli may be used as biological preservative, so, the aim of this study was to present some data on lactobacillus as probiotic bacteria. Lactic acid bacteria were isolated from sausage. Each isolate of lactobacillus species was identified by biochemical tests and comparing their sugar fermentation pattern. Antibacterial activities were done by an agar spot, well diffusion and blank disk method. Enzyme sensitivity of supernatant fluid and concentrated cell free culture after treatment with α-amylase, lysozyme and trypsin was determined. The isolated bacteria were Lacto. plantarum, Lacto delbruekii, Lacto. acidophilus, Lacto. brevis. The isolated bacteria had strong activity against indicator strains. The antibacterial activity was stable at 100ºC for 10 min and at 56ºC for 30 min, but activity was lost after autoclaving. The maximum production of plantaricin was obtained at 25 - 30ºC at pH 6.5. Because, lactobacilli that used to process sausage fermentation are producing antimicrobial activity with heat stability bacteriocin, so, these bacteria may be considered to be a healthy probiotic diet. Lactobacilli originally isolated from meat products are the best condidates as probiotic bacteria to improve the microbiological safety of these foods.

  4. Small molecule ice recrystallization inhibitors mitigate red blood cell lysis during freezing, transient warming and thawing

    Science.gov (United States)

    Briard, Jennie G.; Poisson, Jessica S.; Turner, Tracey R.; Capicciotti, Chantelle J.; Acker, Jason P.; Ben, Robert N.

    2016-03-01

    During cryopreservation, ice recrystallization is a major cause of cellular damage. Conventional cryoprotectants such as dimethyl sulfoxide (DMSO) and glycerol function by a number of different mechanisms but do not mitigate or control ice recrystallization at concentrations utilized in cryopreservation procedures. In North America, cryopreservation of human red blood cells (RBCs) utilizes high concentrations of glycerol. RBC units frozen under these conditions must be subjected to a time-consuming deglycerolization process after thawing in order to remove the glycerol to increased post-thaw recovery of RBCs. The most potent IRI from this study is also capable of protecting frozen RBCs against the large temperature fluctuations associated with transient warming.

  5. Mycophagous soil bacteria

    NARCIS (Netherlands)

    Rudnick, M.B.

    2015-01-01

    Abstract

    Soil microorganisms evolved several strategies to compete for limited nutrients in soil. Bacteria of the genus Collimonas developed a way to exploit fungi as a source of organic nutrients. This strategy has been termed “mycophagy&r

  6. Antibiotic-Resistant Bacteria.

    Science.gov (United States)

    Longenecker, Nevin E.; Oppenheimer, Dan

    1982-01-01

    A study conducted by high school advanced bacteriology students appears to confirm the hypothesis that the incremental administration of antibiotics on several species of bacteria (Escherichia coli, Staphylococcus epidermis, Bacillus sublitus, Bacillus megaterium) will allow for the development of antibiotic-resistant strains. (PEB)

  7. Bacteria-surface interactions.

    Science.gov (United States)

    Tuson, Hannah H; Weibel, Douglas B

    2013-05-14

    The interaction of bacteria with surfaces has important implications in a range of areas, including bioenergy, biofouling, biofilm formation, and the infection of plants and animals. Many of the interactions of bacteria with surfaces produce changes in the expression of genes that influence cell morphology and behavior, including genes essential for motility and surface attachment. Despite the attention that these phenotypes have garnered, the bacterial systems used for sensing and responding to surfaces are still not well understood. An understanding of these mechanisms will guide the development of new classes of materials that inhibit and promote cell growth, and complement studies of the physiology of bacteria in contact with surfaces. Recent studies from a range of fields in science and engineering are poised to guide future investigations in this area. This review summarizes recent studies on bacteria-surface interactions, discusses mechanisms of surface sensing and consequences of cell attachment, provides an overview of surfaces that have been used in bacterial studies, and highlights unanswered questions in this field.

  8. Mechanism of Action of Melaleuca alternifolia (Tea Tree) Oil on Staphylococcus aureus Determined by Time-Kill, Lysis, Leakage, and Salt Tolerance Assays and Electron Microscopy

    OpenAIRE

    Carson, Christine F.; Mee, Brian J.; Riley, Thomas V

    2002-01-01

    The essential oil of Melaleuca alternifolia (tea tree) has broad-spectrum antimicrobial activity. The mechanisms of action of tea tree oil and three of its components, 1,8-cineole, terpinen-4-ol, and α-terpineol, against Staphylococcus aureus ATCC 9144 were investigated. Treatment with these agents at their MICs and two times their MICs, particularly treatment with terpinen-4-ol and α-terpineol, reduced the viability of S. aureus. None of the agents caused lysis, as determined by measurement ...

  9. Hyperphosphatemia during spontaneous tumor lysis syndrome culminate in severe hyphosphatemia at the time of blast crisis of Phneg CML to acute myelomoncytic leukemia

    Directory of Open Access Journals (Sweden)

    Salomon Ophira

    2012-08-01

    Full Text Available Abstract Extreme swing of phosphor from severe hyperphosphatemia to severe hypophosphatemia in a patient with blast crisis of myeloid origin was the result of imbalance between massive apoptosis of leukemic cells in the context of spontaneous tumor lysis syndrome and massive production of leukemic cells with only 1% of blast in peripheral blood. The mutated p53 protein suggested acting as oncogene in the presented case and possibly affecting phosphor status.

  10. Cloning of xylanase gene of Streptomyces flavogriseus in Escherichia coli and bacteriophage lambda-induced lysis for the release of cloned enzyme.

    Science.gov (United States)

    Srivastava, R; Ali, S S; Srivastava, B S

    1991-03-01

    The xylanase gene of Streptomyces flavogriseus was cloned in pUC8 plasmid and expressed in Escherichia coli lysogenic for lambda cI857. lambda-Induced lysis of E. coli at 42 degrees C allowed efficient release of cloned enzyme activity in extracellular environment. The xylanase gene was located in the 0.8-kb HindIII fragment and coded for 18,000 Mr xylanase.

  11. Biological activity of a red-tide alga--A. tamarense under co-cultured condition with bacteria

    Institute of Scientific and Technical Information of China (English)

    SU Jian-qiang; YU Zhi-ming; TIAN Yun; SONG Xiu-xian; HONG Hua-sheng; ZHENG Tian-ling

    2005-01-01

    The relationship between Alexandrium tamarense(Lebour) Balech, one of red-tide alga, and two strains of marine bacteria,Bacillius megaterium(S7 ) and B. halmapulus(S10) isolated from Xiamen Western Sea, was investigated by evaluating the growth state of A. tamarense and the variation of β-glucosidase activity in co-culture system. The results showed the growth and multiplication of the alga were related with the concentration, genus speciality of the bacteria, and growth stage of the alga itself. The growth of A. tamarense was obviously inhibited by S7 and S10 at high concentration. Either inhibition or promotion contributed much more clearly in earlier than in later stage of the growth of the alga. Furthermore, there was a roughly similar variation trend of the activity of extra-cellular enzyme, β-glucosidase, in the water of the separately co-cultured bacteria S7 and S10 with the alga. The β-glucosidase activity(β-GlcA) rapidly increased during the later algal growth accompanying the increase of the lysis of the alga cells. The obvious inhibition of A. tamarense by marine bacteria at high concentration and evident increase of β-GlcA in co-colture system would help us in better understanding the relationship between red-tide alga and bacteria, and also enlightened us the possible use of bacteria in the bio-control of red-tide.

  12. 混菌发酵制备鸭肉香肠的4艺优化及其氧化控制研究%Process Optimization of Mixed Bacteria Fermentation for Preparation of Duck Sausage and Its Oxidation Control

    Institute of Scientific and Technical Information of China (English)

    李超; 王乃馨; 陈尚龙; 耿中华; 印炎

    2016-01-01

    首先采用单因素试验和 Box-Behnken 试验对混菌发酵制备鸭肉香肠的工艺进行优化,然后研究葡萄籽提取物对其氧化性能的影响,最后制定产品质量标准。实验结果表明:清酒乳杆菌和乳酸片球菌混菌发酵制备鸭肉香肠的最佳工艺条件为菌种配比2∶1、接种量5 mL/kg、发酵温度35℃、发酵时间20.3 h、烘烤温度76℃和烘烤时间5.4 h;在此条件下感官得分为86.3;葡萄籽提取物能明显提高发酵鸭肉香肠的耐氧化性能。%According to the single factor experiment and Box-Behnken design,the process of mixed bacteria fermentation for preparation of duck sausage is firstly optimized in the paper.And then the effect of grape seed extract on oxidation properties of sausage is studied.At last,the product quality standards are established.The results show that the best processing conditions of duck sausage are strains proportion of 2 ∶ 1,inoculation quantity of 5 mL/kg,fermentation temperature of 35 ℃, fermentation time of 20.3 h,baking temperature of 76 ℃ and baking time of 5.4 h.Under the optimum conditions,the sensory score is 86.3.Grape seed extract can significantly increase the oxidation resistance properties of duck sausage.

  13. Demonstration of NK cell-mediated lysis of varicella-zoster virus (VZV)-infected cells: characterization of the effector cells

    Energy Technology Data Exchange (ETDEWEB)

    Tilden, A.B.; Cauda, R.; Grossi, C.E.; Balch, C.M.; Lakeman, A.D.; Whitley, R.J.

    1986-06-01

    Infection with varicella-zoster virus (VZV) rendered RAJI cells more susceptible to lysis by non-adherent blood lymphocytes. At an effector to target ratio of 80:1 the mean percentage of /sup 51/Cr release of VZV-infected RAJI cells was 41 +/- 12%, whereas that of uninfected RAJI cells was 15 +/- 6%. The increased susceptibility to lysis was associated with increased effector to target conjugate formation in immunofluorescence binding assays. The effector cells cytotoxic for VZV-infected RAJI cells were predominantly Leu-11a/sup +/ Leu-4/sup -/ granular lymphocytes as demonstrated by fluorescence-activated cell sorting. The effector cell active against VZV-infected RAJI cells appeared similar to those active against herpes simplex virus (HSV)-infected cells, because in cold target competition experiments the lysis of /sup 51/Cr-labeled VZV-infected RAJI cells was efficiently inhibited by either unlabeled VZV-infected RAJI cells (mean 71% inhibition, 2:1 ratio unlabeled to labeled target) or HSV-infected RAJI cells (mean 69% inhibition) but not by uninfected RAJI cells (mean 10% inhibition). In contrast, competition experiments revealed donor heterogeneity in the overlap between effector cells for VZV- or HSV-infected RAJI vs K-562 cells.

  14. Using Mathematical Modelling to Explore Hypotheses about the Role of Bovine Epithelium Structure in Foot-And-Mouth Disease Virus-Induced Cell Lysis.

    Directory of Open Access Journals (Sweden)

    Kyriaki Giorgakoudi

    Full Text Available Foot-and-mouth disease (FMD is a highly contagious disease of cloven-hoofed animals. FMD virus (FMDV shows a strong tropism for epithelial cells, and FMD is characterised by cell lysis and the development of vesicular lesions in certain epithelial tissues (for example, the tongue. By contrast, other epithelial tissues do not develop lesions, despite being sites of viral replication (for example, the dorsal soft palate. The reasons for this difference are poorly understood, but hypotheses are difficult to test experimentally. In order to identify the factors which drive cell lysis, and consequently determine the development of lesions, we developed a partial differential equation model of FMDV infection in bovine epithelial tissues and used it to explore a range of hypotheses about epithelium structure which could be driving differences in lytic behaviour observed in different tissues. Our results demonstrate that, based on current parameter estimates, epithelial tissue thickness and cell layer structure are unlikely to be determinants of FMDV-induced cell lysis. However, differences in receptor distribution or viral replication amongst cell layers could influence the development of lesions, but only if viral replication rates are much lower than current estimates.

  15. Production of lipase from Pseudomonas gessardii using blood tissue lipid and thereof for the hydrolysis of blood cholesterol and triglycerides and lysis of red blood cells.

    Science.gov (United States)

    Ramani, K; Sekaran, G

    2012-08-01

    The study demonstrates the production of lipase (LIP) from Pseudomonas gessardii using blood tissue lipid as the substrate for the hydrolysis of blood cholesterol and triglycerides. The lipase was purified with the specific activity of 828 U/mg protein and the molecular weight of 56 kDa. The maximum lipase activity was observed at the pH 7.0 and the temperature 37 °C. The amino acid composition of purified lipase was determined by HPLC. The mesoporous activated carbon (MAC) was used for the immobilization of lipase for the repeated use of the enzyme catalyst. The K (m) value of immobilized lipase (MAC-LIP) and the free lipase (LIP) was 0.182 and 1.96 mM, respectively. The V (max) value of MAC-LIP and LIP was 1.33 and 1.26 mM/min, respectively. The MAC and MAC-LIP were characterized by scanning electron microscopy (SEM). The hydrolysis study showed 78 and 100% hydrolysis of triglycerides and cholesterol, respectively, for LIP and 84 and 100% hydrolysis of triglycerides and cholesterol, respectively, for MAC-LIP at the reaction time of 1 h. The effect of lipase on cell wall lysis was carried out on the RBCs of blood plasma. Interestingly, 99.9% lysis of RBCs was observed within 2 h. SEM images and phase contrast microscopy confirmed the lysis of RBCs. This work provides a potential biocatalyst for the hydrolysis of blood cholesterol and triglycerides.

  16. Combined Transfection with EBV-Specific Epitopes and HLA-A2 genes is More Effective than Separate Transfection in Promoting CTL Lysis against Nasopharyngeal Carcinoma

    Institute of Scientific and Technical Information of China (English)

    Weijun Ding; Choylen Fong

    2004-01-01

    To augment specific cytotoxic T lymphocyte (CTL) lysis is a promising strategy for cancer therapy. In this study,we examined the boosting effect of CTLs upon autologous lymphoblastoid B cell lines (LCLs) transfected with diverse plasmids, to explore the possible CTL-based immunotherapy of nasopharyngeal carcinoma (NPC).FCM analysis displayed rather high ratio (>30%) of successfully transfected LCLs by utilizing the DMRIE-C kit. CTL assays demonstrated that substantially higher ratio of CTL specific lysis was observed upon the LCLs transfected with both expression vectors encoding EBV-specific epitopes and their presentation molecule HLA-A2, in contrast with those transfected separately. By transfecting the vector encoding HLA-A2 alone, only the LCLs of HLA-A2+ donors elicited markedly higher CTL lysis. CTL assays also showed that there existed no marked differences upon transfection by either different vectors (pcDNA3, pNGVL3 or pNGVL3-hFlex), or different EBV-derived peptides (LMP2Pep1 or LMP2Pep2), or with or without the doubled DNA sequence encoding peptides. This study indicated a promising immunotherapy strategy on NPC through boosting and eliciting the EBV-specific CTL activation by transferring vectors encoding both EBV-specific epitopes and their presentation molecule HLA-A2 into autologous LCL, the presentation cells of MHC/peptide tetrameric complex.

  17. Combined Transfection with EBV-Specific Epitopes and HLA-A2 genes is More Effective than Separate Transfection in Promoting CTL Lysis against Nasopharyngeal Carcinoma

    Institute of Scientific and Technical Information of China (English)

    WeijunDing; ChoylenFong

    2004-01-01

    To augment specific cytotoxic T lymphocyte (CTL) lysis is a promising strategy for cancer therapy. In this study,we examined the boosting effect of CTLs upon autologous lymphoblastoid B cell lines (LCLs) transfected with diverse plasmids, to explore the possible CTL-based immunotherapy of nasopharyngeal carcinoma (NPC).FCM analysis displayed rather high ratio (>30%) of successfully transfected LCLs by utilizing the DMRIE-C kit. CTL assays demonstrated that substantially higher ratio of CTL specific lysis was observed upon the LCLs transfected with both expression vectors encoding EBV-specific epitopes and their presentation molecule HLA-A2, In contrast with those transfected separately. By transfecting the vector encoding HLA-A2 alone, only the LCLs of HLA-A2+ donors elicited markedly higher CTL lysis. CTL assays also showed that there existed no marked differences upon transfection by either different vectors (pcDNA3, pNGVL3 or pNGVL3-hFIex), or different EBV-derived peptides (LMP2Pep1 or LMP2Pep2), or with or without the doubled DNA sequence encoding peptides. This study indicated a promising immunotherapy strategy on NPC through boosting and eliciting the EBV-specific CTL activation by transferring vectors encoding both EBV-specific epitopes and their presentation molecule HLA-A2 into autologous LCL, the presentation cells of MHC/peptide tetrameric complex.

  18. Reduction of excess sludge in a sequencing batch reactor by lysis-cryptic growth using quick lime for disintegration under low temperature.

    Science.gov (United States)

    Lv, Xiao-Mei; Song, Ju-Sheng; Li, Ji; Zhai, Kun

    2016-10-06

    In the present study, quick-lime-based thermal-alkaline sludge disintegration (SD) under low temperature was combined with cryptic growth to investigate the excess sludge reduction efficiency in the sequencing batch reactor (SBR). The optimized condition of SD was as follows: T = 80℃, pH = 11, t = 180 min, and the SD rate was about 42.1%. With 65.6% of excess sludge disintegrated and returned to the SBR, the system achieved sludge reduction rate of about 40.1%. The lysis-cryptic growth still obtained satisfactory sludge reduction efficiency despite the comparative low SD rate, which suggested that disintegration rate might not be the decisive factor for cryptic-growth-based sludge reduction. Lysis-cryptic growth did not impact the effluent quality, yet the phosphorus removal performance was enhanced, with effluent total phosphorus concentration decreased by 0.3 mg/L (33%). Crystal compounds of calcium phosphate precipitate were detected in the system by Fourier transform infrared spectroscopy and X-ray diffraction, which indicated the phosphorus removal potential of SD using lime. Moreover, endogenous dehydrogenase activity of activated sludge in the lysis-cryptic system was enhanced, which was beneficial for sludge reduction. SD and cryptic growth in the present study demonstrates an economical and effective approach for sludge reduction.

  19. Enhancement of NK cell-mediated lysis of non-small lung cancer cells by nPKC activator, ingenol 3,20 dibenzoate.

    Science.gov (United States)

    Gong, Chenyuan; Yao, Chao; Xu, Zihang; Ni, Zhongya; Zhu, Xiaowen; Wang, Lixin; Yan, Xuewei; Zhou, Wuxiong; Zhu, Shiguo

    2017-03-01

    The IFN-γ production is crucial for NK cell-mediated lysis of cancer cells. Thus increasing the IFN-γ production by NK cells may be an ideal strategy to improve their tumoricidal effect. Since the focus on new drug development has shifted towards natural products, limited information is out there about natural products that enhance the IFN-γ production by NK cells. In this study, through a high-throughput screening, we have identified a natural product ingenol 3,20 dibenzoate (IDB), an activator of tumor suppressor protein kinase C (PKC) isozymes, could increase the IFN-γ production and degranulation by NK cells, especially when NK cells were stimulated by non-small lung cancer (NSCLC) cells. IDB also significantly enhanced the NK cell-mediated lysis of NSCLC cells. Furthermore, PKC inhibitor, sotrastaurin abrogated IDB-induced IFN-γ production, degranulation and cytotoxicity, but did not affect IFN-γ production by NK cells without IDB treatment and NSCLC cell stimulation. The IFN-γ neutralization reversed the IDB-induced enhancement of NK cell mediated killing. In conclusion, our study indicated that IDB enhanced NK cell-mediated lysis of NSCLC cells is dependent on specific PKC mediated IFN-γ production and degranulation. Thus, IDB may have a promising application in clinic for NK cell-based cancer immunotherapy.

  20. Effects of deposit-feeding macrofauna on benthic bacteria, viruses, and protozoa in a silty freshwater sediment.

    Science.gov (United States)

    Wieltschnig, Claudia; Fischer, Ulrike R; Velimirov, Branko; Kirschner, Alexander K T

    2008-07-01

    In microcosm experiments, we simultaneously tested the effects of increased numbers of deposit-feeding macrofauna (chironomids, oligochaetes and cladocerans) on the standing stock, activities and interactions of heterotrophic bacteria, viruses, and bacterivorous protozoa (heterotrophic nanoflagellates and ciliates) in the aerobic layer of a silty littoral freshwater sediment. On average, bacterial secondary production was stimulated between 11 and 29% by all macrofaunal groups compared to control experiments without macrofauna addition. Bacterial standing stock increased significantly by 8 and 13% in case of chironomids and cladocerans, respectively. Oligochaetes and chironomids produced significant negative effects on viral abundance while the results with cladocerans were inconsistent. The addition of oligochaetes and chironomids resulted in a significant decrease by on average 68 and 32% of viral decay rates, respectively, used as a measure of viral production. The calculated contribution of virus-induced lysis to benthic bacterial mortality was low, with 2.8 to 11.8% of bacterial secondary production, and decreased by 39 to 81% after the addition of macrofauna compared to the control. The abundances of heterotrophic nanoflagellates were significantly reduced by 20% by all tested macrofauna groups, while ciliates showed inconsistent results. The importance of heterotrophic nanoflagellate grazing on benthic bacteria was very low (virus-induced cell lysis and protozoan grazing.

  1. Integrated printed circuit board device for cell lysis and nucleic acid extraction.

    Science.gov (United States)

    Marshall, Lewis A; Wu, Liang Li; Babikian, Sarkis; Bachman, Mark; Santiago, Juan G

    2012-11-01

    Preparation of raw, untreated biological samples remains a major challenge in microfluidics. We present a novel microfluidic device based on the integration of printed circuit boards and an isotachophoresis assay for sample preparation of nucleic acids from biological samples. The device has integrated resistive heaters and temperature sensors as well as a 70 μm × 300 μm × 3.7 cm microfluidic channel connecting two 15 μL reservoirs. We demonstrated this device by extracting pathogenic nucleic acids from 1 μL dispensed volume of whole blood spiked with Plasmodium falciparum. We dispensed whole blood directly onto an on-chip reservoir, and the system's integrated heaters simultaneously lysed and mixed the sample. We used isotachophoresis to extract the nucleic acids into a secondary buffer via isotachophoresis. We analyzed the convective mixing action with micro particle image velocimetry (micro-PIV) and verified the purity and amount of extracted nucleic acids using off-chip quantitative polymerase chain reaction (PCR). We achieved a clinically relevant limit of detection of 500 parasites per microliter. The system has no moving parts, and the process is potentially compatible with a wide range of on-chip hybridization or amplification assays.

  2. Small molecule ice recrystallization inhibitors mitigate red blood cell lysis during freezing, transient warming and thawing.

    Science.gov (United States)

    Briard, Jennie G; Poisson, Jessica S; Turner, Tracey R; Capicciotti, Chantelle J; Acker, Jason P; Ben, Robert N

    2016-03-29

    During cryopreservation, ice recrystallization is a major cause of cellular damage. Conventional cryoprotectants such as dimethyl sulfoxide (DMSO) and glycerol function by a number of different mechanisms but do not mitigate or control ice recrystallization at concentrations utilized in cryopreservation procedures. In North America, cryopreservation of human red blood cells (RBCs) utilizes high concentrations of glycerol. RBC units frozen under these conditions must be subjected to a time-consuming deglycerolization process after thawing in order to remove the glycerol to recrystallization inhibitors (IRIs) that are effective cryoprotectants for human RBCs, resulting in 70-80% intact RBCs using only 15% glycerol and slow freezing rates. These compounds are capable of reducing the average ice crystal size of extracellular ice relative to a 15% glycerol control validating the positive correlation between a reduction in ice crystal size and increased post-thaw recovery of RBCs. The most potent IRI from this study is also capable of protecting frozen RBCs against the large temperature fluctuations associated with transient warming.

  3. Bacteria that purify sludge; Des bacteries epuratrices

    Energy Technology Data Exchange (ETDEWEB)

    Peignen-Seraline, P.; Manem, J. [Cirsee, Lyonnaise des Eaux, 92 - Nanterre (France)

    1997-03-01

    Inherent in water purification processes, the formation of sludges is intensively studied. Recently, original bacteria have been observed by searchers: some of them purify water making ``tassels``, others separate them and some of them even participate in the elimination of the first. This research study is described into details and will probably be used in the future at the industrial scale. (O.M.)

  4. Is Your ATM Dispensing Bacteria?

    Science.gov (United States)

    ... news/fullstory_162067.html Is Your ATM Dispensing Bacteria? Study in New York City found most of ... keypads in New York City were covered in bacteria, researchers reported, with most of the microbes coming ...

  5. Genomics of oral bacteria.

    Science.gov (United States)

    Duncan, Margaret J

    2003-01-01

    Advances in bacterial genetics came with the discovery of the genetic code, followed by the development of recombinant DNA technologies. Now the field is undergoing a new revolution because of investigators' ability to sequence and assemble complete bacterial genomes. Over 200 genome projects have been completed or are in progress, and the oral microbiology research community has benefited through projects for oral bacteria and their non-oral-pathogen relatives. This review describes features of several oral bacterial genomes, and emphasizes the themes of species relationships, comparative genomics, and lateral gene transfer. Genomics is having a broad impact on basic research in microbial pathogenesis, and will lead to new approaches in clinical research and therapeutics. The oral microbiota is a unique community especially suited for new challenges to sequence the metagenomes of microbial consortia, and the genomes of uncultivable bacteria.

  6. Manufacture of Probiotic Bacteria

    Science.gov (United States)

    Muller, J. A.; Ross, R. P.; Fitzgerald, G. F.; Stanton, C.

    Lactic acid bacteria (LAB) have been used for many years as natural biopreservatives in fermented foods. A small group of LAB are also believed to have beneficial health effects on the host, so called probiotic bacteria. Probiotics have emerged from the niche industry from Asia into European and American markets. Functional foods are one of the fastest growing markets today, with estimated growth to 20 billion dollars worldwide by 2010 (GIA, 2008). The increasing demand for probiotics and the new food markets where probiotics are introduced, challenges the industry to produce high quantities of probiotic cultures in a viable and stable form. Dried concentrated probiotic cultures are the most convenient form for incorporation into functional foods, given the ease of storage, handling and transport, especially for shelf-stable functional products. This chapter will discuss various aspects of the challenges associated with the manufacturing of probiotic cultures.

  7. Exopolysaccharides from Marine Bacteria

    Institute of Scientific and Technical Information of China (English)

    CHI Zhenming; FANG Yan

    2005-01-01

    Microbial polysaccharides represent a class of important products of growing interest for many sectors of industry. In recent years, there has been a growing interest in isolating new exopolysaccharides (EPSs)-producing bacteria from marine environments, particularly from various extreme marine environments. Many new marine microbial EPSs with novel chemical compositions, properties and structures have been found to have potential applications in fields such as adhesives,textiles, pharmaceuticals and medicine for anti-cancer, food additives, oil recovery and metal removal in mining and industrial waste treatments, etc This paper gives a brief summary of the information about the EPSs produced by marine bacteria,including their chemical compositions, properties and structures, together with their potential applications in industry.

  8. Síndrome de lise tumoral: uma revisão abrangente da literatura Acute tumor lysis syndrome: a comprehensive review

    Directory of Open Access Journals (Sweden)

    Michael Darmon

    2008-09-01

    égias baseadas no risco dos pacientes são necessários para limitar a alta morbidade e mortalidade desta complicação.Tumor lysis syndrome is characterized by the massive destruction of malignant cells and the release in the extra-cellular space of their content. While Tumor lysis syndrome may occur spontaneously before treatment, it usually develops shortly after the initiation of cytotoxic chemotherapy. These metabolites can overwhelm the homeostatic mechanisms with development of hyperuricaemia, hyperkalaemia, hyperphosphataemia, and hypocalcaemia. These biological manifestations may lead to clinical manifestations including, acute kidney injury, seizure, or sudden death that require intensive care. Since clinical tumor lysis syndrome is associated with a poor prognosis both prevention of tumor lysis syndrome and prevention of clinical consequences of tumor lysis syndrome are mandatory. The objective of this review is to describe pathophysiological mechanisms, biological and clinical manifestations of tumor Lysis syndrome, and to provide upto-date guidelines to ensure prevention of tumor lysis syndrome. Review of selected studies on tumor lysis syndrome published at the PubMed database www.pubmed.gov during the last 20 years. Additional references were retrieved from the studies initially selected. Tumor lysis syndrome is a frequent and life-threatening complication of the newly diagnosed malignancies. Preventive measures, including hydration, uricolytic agents, eviction of factors predisposing to acute kidney injury and, in the more severe patients, on prophylactic renal replacement therapy, are required to prevent or limit clinical consequences of Tumor lysis syndrome. However optimal timing and modalities of prevention remains unknown and may be modified by the changing spectrum of patients at risk of tumor lysis syndrome. Development and validation of risk based strategies is required to limit the high morbidity and mortality of this complication.

  9. Cable Bacteria in Freshwater Sediments

    OpenAIRE

    Risgaard-Petersen, Nils; Kristiansen, Michael; Frederiksen, Rasmus B.; Dittmer, Anders Lindequist; Bjerg, Jesper Tataru; Trojan, Daniela; Schreiber, Lars; Damgaard, Lars Riis; Schramm, Andreas; Nielsen, Lars Peter

    2015-01-01

    In marine sediments cathodic oxygen reduction at the sediment surface can be coupled to anodic sulfide oxidation in deeper anoxic layers through electrical currents mediated by filamentous, multicellular bacteria of the Desulfobulbaceae family, the so-called cable bacteria. Until now, cable bacteria have only been reported from marine environments. In this study, we demonstrate that cable bacteria also occur in freshwater sediments. In a first step, homogenized sediment collected from the fre...

  10. Tumorous Stem Mustard Lactic Acid Bacteria Beverage Stability Research and Process Conditions Optimization%茎瘤芥乳酸菌饮料稳定性研究及工艺条件优化

    Institute of Scientific and Technical Information of China (English)

    徐安书; 郭健; 殷永玲

    2015-01-01

    基于茎瘤芥胡萝卜混合汁乳酸菌饮料稳定性研究基础上进行工艺条件优化,进一步研究发酵过程中酸度对感官品质和组织状态的影响,采用四因素三水平正交实验,研究菌种比例、接种量、发酵温度、发酵时间对发酵过程中稳定性的影响,根据沉淀率的大小判定其稳定性,试验结果:保加利亚乳杆菌和嗜热链球菌比例为1∶1,母发酵剂接种量3%,发酵温度37℃,发酵时间12 h,均质压力20 MPa,均质温度在30℃~40℃之间,2次均质,效果更好,最后添加复合稳定剂耐酸羧甲基纤维素钠CMC0.3%+黄原胶0.01%。%Tumorous stem mustard carrot juice lactic acid bacteria beverage stability study on the basis of the optimization of process conditions based on the further study of the effect of acidity and fermentation process on the sensory quality and organizational status, using four factors and three levels orthogonal experiment to study the effect of proportion, strain, inoculation, fermentation temperature and fermentation time on the stability of the fermentation process, according to the precipitation the size of rate to determine its stability, the results show that:the Lactobacillus bulgaricus and Streptococcus thermophilus ratio 1∶1, mother culture inoculation amount 3%, fermentation temperature of 37 ℃, fermentation time 12 h, homogenization pressure 20 MPa, homogeneous temperature between 30 ℃-40 ℃, the better the two homogeneous, effect, finally adding compound stabilizer acid Suo Jiaji cellulose sodium CMC0.3%+0.01%xanthan gum.

  11. 冬瓜生腌过程细菌种群变化及其品质相关性%Bacteria Community Changes and Its Quality Related in Raw Process of Pickled Wax Gourd

    Institute of Scientific and Technical Information of China (English)

    沈锡权; 赵永威; 吴祖芳; 翁佩芳; 卓鸿雁

    2012-01-01

    采用16S rDNA基因克隆文库的方法,对生腌冬瓜腌制体系的微生物多样性、优势种群及其相关品质变化过程进行分析.结果表明;冬瓜生腌开始时优势细菌为肠杆菌属、乳球菌属和魏斯氏茵属;早期阶段优势菌为戊糖片球菌;腌制30 d时优势茵属变成了肠杆菌属和片球菌属;在腌制中后期微生物组成基本稳定,其中片球菌属、魏斯氏茵属和枝芽孢杆菌属为优势菌属.冬瓜生腌制过程中pH值和亚硝酸盐质量分数下降,最低值分别为3.7和0.5 mg/kg,盐度稳定在7.0左右,细菌总数和乳酸茵总数由下降至最后稳定的过程,腌制中后期乳酸菌总数稳定在1.0×107 cfu/mL左右.冬瓜生腌过程检测到的乳球菌、片球菌和枝芽孢杆菌等菌属对维持腌冬瓜质量具有重要意义.%The microbial community diversity, predominant species and its related characters of the product were investigated during the fermentation process of pickled wax gourd using the 16S rDNA clone library. The results showed that in the pickling system Enterobacter, Lacto-coccus and Weissella are the predominant microbial community at earlier phase. After 15 days of fermentation, the dominant population has changed to be Pediococcus pentosaceus, on day 30 Enterobacter and Pediococcus become to be the predominant species. In the later stage of pickling, the community system is stable, and Pediococcus, Weissella and Virgibacillus are the major microbial species. At the same time, the pH value and nitrite concentration of pickled wax gourd samples had been decreasing, its minimum values reached 3. 7 and 0. 5 mg/kg, respectively. The salinity value was finally stable in 7. 0%. The total population of bacteria and lactic acid bacteria (LAB) changed from the high level to stable, the population of LAB was 1. O×07cfu/mL at the later phase of pickled. Some species including Lactococcus, Pediococcus and Virgibacillus detected in the pickled wax gourd

  12. Bacteria in crude oil survived autoclaving and stimulated differentially by exogenous bacteria.

    Directory of Open Access Journals (Sweden)

    Xiao-Cui Gong

    Full Text Available Autoclaving of crude oil is often used to evaluate the hydrocarbon-degrading abilities of bacteria. This may be potentially useful for bioaugmentation and microbial enhanced oil recovery (MEOR. However, it is not entirely clear if "endogenous" bacteria (e.g., spores in/on crude oil survive the autoclaving process, or influence subsequent evaluation of the hydrocarbon-degradation abilities of the "exogenous" bacterial strains. To test this, we inoculated autoclaved crude oil medium with six exogenous bacterial strains (three Dietzia strains, two Acinetobacter strains, and one Pseudomonas strain. The survival of the spore-forming Bacillus and Paenibacillus and the non-spore-forming mesophilic Pseudomonas, Dietzia, Alcaligenes, and Microbacterium was detected using a 16S rRNA gene clone library and terminal restriction fragment length polymorphism (T-RFLP analysis. However, neither bacteria nor bacterial activity was detected in three controls consisting of non-inoculated autoclaved crude oil medium. These results suggest that detection of endogenous bacteria was stimulated by the six inoculated strains. In addition, inoculation with Acinetobacter spp. stimulated detection of Bacillus, while inoculation with Dietzia spp. and Pseudomonas sp. stimulated the detection of more Pseudomonas. In contrast, similar exogenous bacteria stimulated similar endogenous bacteria at the genus level. Based on these results, special emphasis should be applied to evaluate the influence of bacteria capable of surviving autoclaving on the hydrocarbon-degrading abilities of exogenous bacteria, in particular, with regard to bioaugmentation and MEOR. Bioaugmentation and MEOR technologies could then be developed to more accurately direct the growth of specific endogenous bacteria that may then improve the efficiency of treatment or recovery of crude oil.

  13. Bacteriocins of lactic acid bacteria : extending the family

    NARCIS (Netherlands)

    Alvarez-Sieiro, Patricia; Montalbán-López, Manuel; Mu, Dongdong; Kuipers, Oscar P

    2016-01-01

    Lactic acid bacteria (LAB) constitute a heterogeneous group of microorganisms that produce lactic acid as the major product during the fermentation process. LAB are Gram-positive bacteria with great biotechnological potential in the food industry. They can produce bacteriocins, which are proteinaceo

  14. [The Role of Ombrophilic Dissipotrophic Bacteria in Wood Decomposition].

    Science.gov (United States)

    Zaichikova, M V; Berestovskaya, Y Y; Vasil'eva, L V

    2016-01-01

    The summarized experimental data on ombrophilic bacteria isolated from dystrophic waters formed by a mycobacterial community during the process of spruce wood decomposition are presented. It was demonstrated that the ombrophilic microbial community was characterized by wide phylogenetic diversity at the initial stage of spruce wood decomposition by xylotrophic fungi under low mineralization conditions. It was noted that bacteria were able to grow under acidic and ultrafresh conditions and most of them were referred to oligotrophs. It was determined that all isolated ombrophilic bacteria divided into three groups depending on the substrate specifity: saccharolytic, acidotrophic bacteria, and bacteria, which used C1-compounds as the substrate. The position of the ombrophilic bacteria in the trophic chain was determined.

  15. Another turn of the screw in shaving Gram-positive bacteria: Optimization of proteomics surface protein identification in Streptococcus pneumoniae.

    Science.gov (United States)

    Olaya-Abril, Alfonso; Gómez-Gascón, Lidia; Jiménez-Munguía, Irene; Obando, Ignacio; Rodríguez-Ortega, Manuel J

    2012-06-27

    Bacterial surface proteins are of outmost importance as they play critical roles in the interaction between cells and their environment. In addition, they can be targets of either vaccines or antibodies. Proteomic analysis through "shaving" live cells with proteases has become a successful approach for a fast and reliable identification of surface proteins. However, this protocol has not been able to reach the goal of excluding cytoplasmic contamination, as cell lysis is an inherent process during culture and experimental manipulation. In this work, we carried out the optimization of the "shaving" strategy for the Gram-positive human pathogen Streptococcus pneumoniae, a bacterium highly susceptible to autolysis, and set up the conditions for maximizing the identification of surface proteins containing sorting or exporting signals, and for minimizing cytoplasmic contamination. We also demonstrate that cell lysis is an inherent process during culture and experimental manipulation, and that a low level of lysis is enough to contaminate a "surfome" preparation with peptides derived from cytoplasmic proteins. When the optimized conditions were applied to several clinical isolates, we found the majority of the proteins described to induce protection against pneumococcal infection. In addition, we found other proteins whose protection capacity has not been yet tested. In addition, we show the utility of this approach for providing antigens that can be used in serological tests for the diagnosis of pneumococcal disease.

  16. 山西老陈醋发酵过程菌群微生态分析及乳酸菌分离%Analysis of Microbial Community Microecology in Shanxi Aged Vinegar Fermentation Process and Isolation of Lactic Acid Bacteria

    Institute of Scientific and Technical Information of China (English)

    王梦颖; 赵国忠; 赵建新; 张灏; 陈卫

    2016-01-01

    Shanxi aged vinegar is one of Chinese four famous vinegars,with unique flavor,long history and being beneficial to human health,in order to promote the development of Shanxi aged vinegar industry and prevent corruption caused by microorganisms,16S rDNA metagenomics sequencing is used to analyze the microecology of microbial community during the fermentation process of Shanxi aged vinegar,to make bacterial species and the relative abundance of them clearly,which plays a leading role during the fermentation process of Shanxi aged vinegar.Based on the study mentioned above,lactic acid bacteria are isolated and identified,in order to excavate potential microorganism resources in traditional fermented food.%山西老陈醋作为中国四大名醋之一,风味独特,历史悠久,且对人体健康有较好的促进作用。为了促进山西老陈醋发酵工业的发展及对其生产中出现的不可控微生物腐败造成的质量问题进行预防,研究采用16S rDNA 宏基因组的测序手段对山西老陈醋发酵过程中的菌群微生态进行分析,了解陈醋在酒精发酵及醋酸发酵两个重要过程中起主导作用的细菌种类及各种属微生物的相对丰度,并在对主要发酵微生物有一定了解的基础上对其中的乳酸菌进行分离、鉴定,充分挖掘传统发酵食品中具有潜力的微生物资源。

  17. Brotes germinados y bacterias

    OpenAIRE

    García Olmedo, Francisco

    2011-01-01

    Ante la confusión y el revuelo asociados al último incidente causado por una cepa de la bacteria Escherichia coli (E. coli) en Alemania, tal vez no esté de más esta carta para recordar y actualizar escritos míos anteriores aparecidos en Revista de Libros sobre los riesgos alimentarios en general y sobre los peligros de dicho microorganismo en particular. 1 . Aunque es cierto que la proporción de cepas peligrosas de E. coli es quizás inferior a la de delincuentes entre los humanos, exi...

  18. 乳酸菌发酵技术脱除大米粉中镉的工艺优化%Optimization of fermentation process of removal of cadmium in rice powder using lactic acid bacteria

    Institute of Scientific and Technical Information of China (English)

    傅亚平; 廖卢艳; 刘阳; 吴卫国

    2015-01-01

    In recent years, along with our country cadmium pollution in rice is increasingly serious, the safety and quality of rice in China has been under serious threat. Contamination of rice with cadmium also draws great attention because of its serious threat to food safety and its potential harm to human health. Thus, it’s very important and necessary for food industry to find a method to decrease the degree of cadmium pollution in polished rice. Cadmium in rice has been demonstrated that it mainly bonded with protein and little cadmium accumulated in starch and other nutritional ingredients. And processing of soaking, heating, washing can reduce the content of cadmium in rice powder, but the effect of cadmium removal is not obvious. Hence, it also was proved that the amount of heavy metals in rice dropped significantly after fermentation using lactic acid bacteria. Therefore, in order to resolve the problem of“excessive cadmium”polished rice utilization, this paper tried to reduce the content of cadmium in“excessive cadmium”polished rice(cadmium content of 0.6479mg/kg) by lactic acid bacteria and produce safe fermented rice products, and the fermentation conditions were studied and optimized. The mixed strains of Lactobacillus plantarum and Pediococcus pentosaceus(2:1,v/v) were selected as the suitable fermentation strain by comparative test. The effects of rice powder size, fermentation time, fermentation temperature and inoculum size of mixed strains on the removal rate of cadmium in polished rice also were evaluated by single-factor experiment. The results showed that fermentation time, fermentation temperature and inoculum size of mixed strains could significantly affect the removal rate of cadmium(P0.05). Then, fermentation time, fermentation temperature and inoculum size of mixed strains were further researched using a three-variable, three level Box-Benhnken design to optimize the fermentation conditions. The results indicated that the interaction

  19. Collective Sensing-Capacity of Bacteria Populations

    CERN Document Server

    Einolghozati, Arash; Fekri, Faramarz

    2012-01-01

    The design of biological networks using bacteria as the basic elements of the network is initially motivated by a phenomenon called quorum sensing. Through quorum sensing, each bacterium performs sensing the medium and communicating it to others via molecular communication. As a result, bacteria can orchestrate and act collectively and perform tasks impossible otherwise. In this paper, we consider a population of bacteria as a single node in a network. In our version of biological communication networks, such a node would communicate with one another via molecular signals. As a first step toward such networks, this paper focuses on the study of the transfer of information to the population (i.e., the node) by stimulating it with a concentration of special type of a molecules signal. These molecules trigger a chain of processes inside each bacteria that results in a final output in the form of light or fluorescence. Each stage in the process adds noise to the signal carried to the next stage. Our objective is ...

  20. Metabolic engineering of bacteria for ethanol production

    Energy Technology Data Exchange (ETDEWEB)

    Ingram, L.O.; Gomez, P.F.; Lai, X.; Moniruzzaman, M.; Wood, B.E.; Yomano, L.P.; York, S.W. [Univ. of Florida, Gainesville, FL (United States). Dept. of Microbiology and Cell Science

    1998-04-20

    Technologies are available which will allow the conversion of lignocellulose into fuel ethanol using genetically engineered bacteria. Assembling these into a cost-effective process remains a challenge. The authors` work has focused primarily on the genetic engineering of enteric bacteria using a portable ethanol production pathway. Genes encoding Zymomonas mobilis pyruvate decarboxylase and alcohol dehydrogenase have been integrated into the chromosome of Escherichia coli B to produce strain KO11 for the fermentation of hemicellulose-derived syrups. This organism can efficiently ferment all hexose and pentose sugars present in the polymers of hemicellulose. Klebsiella oxytoca M5A1 has been genetically engineered in a similar manner to produce strain P2 for ethanol production from cellulose. This organism has the native ability to ferment cellobiose and cellotriose, eliminating the need for one class of cellulase enzymes. The optimal pH for cellulose fermentation with this organism is near that of fungal cellulases. The general approach for the genetic engineering of new biocatalysts has been most successful with enteric bacteria thus far. However, this approach may also prove useful with gram-positive bacteria which have other important traits for lignocellulose conversion. Many opportunities remain for further improvements in the biomass to ethanol processes.

  1. Beneficial bacteria inhibit cachexia.

    Science.gov (United States)

    Varian, Bernard J; Goureshetti, Sravya; Poutahidis, Theofilos; Lakritz, Jessica R; Levkovich, Tatiana; Kwok, Caitlin; Teliousis, Konstantinos; Ibrahim, Yassin M; Mirabal, Sheyla; Erdman, Susan E

    2016-03-15

    Muscle wasting, known as cachexia, is a debilitating condition associated with chronic inflammation such as during cancer. Beneficial microbes have been shown to optimize systemic inflammatory tone during good health; however, interactions between microbes and host immunity in the context of cachexia are incompletely understood. Here we use mouse models to test roles for bacteria in muscle wasting syndromes. We find that feeding of a human commensal microbe, Lactobacillus reuteri, to mice is sufficient to lower systemic indices of inflammation and inhibit cachexia. Further, the microbial muscle-building phenomenon extends to normal aging as wild type animals exhibited increased growth hormone levels and up-regulation of transcription factor Forkhead Box N1 [FoxN1] associated with thymus gland retention and longevity. Interestingly, mice with a defective FoxN1 gene (athymic nude) fail to inhibit sarcopenia after L. reuteri therapy, indicating a FoxN1-mediated mechanism. In conclusion, symbiotic bacteria may serve to stimulate FoxN1 and thymic functions that regulate inflammation, offering possible alternatives for cachexia prevention and novel insights into roles for microbiota in mammalian ontogeny and phylogeny.

  2. Chemical communication in bacteria

    Science.gov (United States)

    Suravajhala, Srinivasa Sandeep; Saini, Deepak; Nott, Prabhu

    Luminescence in Vibrio fischeri is a model for quorum-sensing-gene-regulation in bacteria. We study luminescence response of V. fischeri to both internal and external cues at the single cell and population level. Experiments with ES114, a wild-type strain, and ainS mutant show that luminescence induction in cultures is not always proportional to cell-density and there is always a basal level of luminescence. At any given concentration of the exogenously added signals, C6-HSL and C8-HSL, luminescence per cell reaches a maximum during the exponential phase and decreases thereafter. We hypothesize that (1) C6-HSL production and LuxR activity are not proportional to cell-density, and (2) there is a shift in equilibrium from C6-HSL to C8-HSL during the later stages of growth of the culture. RT-PCR analysis of luxI and luxR shows that the expression of these genes is maximum corresponding to the highest level of luminescence. The shift in equilibrium is shown by studying competitive binding of C6-HSL and C8-HSL to LuxR. We argue that luminescence is a unicellular behaviour, and an intensive property like per cell luminescence is more important than gross luminescence of the population in understanding response of bacteria to chemical signalling. Funding from the Department of Science and Technology, India is acknowledged.

  3. Sublethal exposure to alpha radiation (223Ra dichloride) enhances various carcinomas' sensitivity to lysis by antigen-specific cytotoxic T lymphocytes through calreticulin-mediated immunogenic modulation.

    Science.gov (United States)

    Malamas, Anthony S; Gameiro, Sofia R; Knudson, Karin M; Hodge, James W

    2016-12-27

    Radium-223 dichloride (Xofigo®; 223Ra) is an alpha-emitting radiopharmaceutical FDA-approved for the treatment of bone metastases in patients with advanced castration-resistant prostate cancer. It is also being examined clinically in patients with breast and lung carcinoma and patients with multiple myeloma. As with other forms of radiation, the aim of 223Ra is to reduce tumor burden by directly killing tumor cells. External beam (photon) and proton radiation have been shown to augment tumor sensitivity to antigen-specific CD8+ cytotoxic T lymphocytes (CTLs). However, little is known about whether treatment with 223Ra can also induce such immunogenic modulation in tumor cells that survive irradiation. We examined these effects in vitro by exposing human prostate, breast, and lung carcinoma cells to sublethal doses of 223Ra. 223Ra significantly enhanced T cell-mediated lysis of each tumor type by CD8+ CTLs specific for MUC-1, brachyury, and CEA tumor antigens. Immunofluorescence analysis revealed that the increase in CTL killing was accompanied by augmented protein expression of MHC-I and calreticulin in each tumor type, molecules that are essential for efficient antigen presentation. Enhanced tumor-cell lysis was facilitated by calreticulin surface translocation following 223Ra exposure. The phenotypic changes observed after treatment appear to be mediated by induction of the endoplasmic reticulum stress response pathway. By rendering tumor cells more susceptible to T cell-mediated lysis, 223Ra may potentially be effective in combination with various immunotherapies, particularly cancer vaccines that are designed to generate and expand patients' endogenous antigen-specific T-cell populations against specific tumor antigens.

  4. Ammonia effect on hydrogenotrophic methanogens and syntrophic acetate oxidizing bacteria

    DEFF Research Database (Denmark)

    Wang, Han; Fotidis, Ioannis; Angelidaki, Irini

    2015-01-01

    Ammonia-rich substrates can cause inhibition on anaerobic digestion process. Syntrophic acetate oxidizing bacteria (SAOB) and hydrogenotrophic methanogens are important for the ammonia inhibitory mechanism on anaerobic digestion. The roles and interactions of SAOB and hydrogenotrophic methanogens...

  5. A novel trifunctional IgG-like bispecific antibody to inhibit HIV-1 infection and enhance lysis of HIV by targeting activation of complement

    Directory of Open Access Journals (Sweden)

    Tomlinson Stephen

    2010-06-01

    Full Text Available Abstract Background The complement system is not only a key component of innate immunity but also provides a first line of defense against invading pathogens, especially for viral pathogens. Human immunodeficiency virus (HIV, however, possesses several mechanisms to evade complement-mediated lysis (CoML and exploit the complement system to enhance viral infectivity. Responsible for this intrinsic resistance against complement-mediated virolysis are complement regulatory membrane proteins derived from the host cell that inherently downregulates complement activation at several stages of the cascade. In addition, HIV is protected from complement-mediated lysis by binding soluble factor H (fH through the viral envelope proteins, gp120 and gp41. Whereas inhibition of complement activity is the desired outcome in the vast majority of therapeutic approaches, there is a broader potential for complement-mediated inhibition of HIV by complement local stimulation. Presentation of the hypothesis Our previous studies have proven that the complement-mediated antibody-dependent enhancement of HIV infection is mediated by the association of complement receptor type 2 bound to the C3 fragment and deposited on the surface of HIV virions. Thus, we hypothesize that another new activator of complement, consisting of two dsFv (against gp120 and against C3d respectively linked to a complement-activating human IgG1 Fc domain ((anti-gp120 × anti-C3d-Fc, can not only target and amplify complement activation on HIV virions for enhancing the efficiency of HIV lysis, but also reduce the infectivity of HIV through blocking the gp120 and C3d on the surface of HIV. Testing the hypothesis Our hypothesis was tested using cell-free HIV-1 virions cultivated in vitro and assessment of virus opsonization was performed by incubating appropriate dilutions of virus with medium containing normal human serum and purified (anti-gp120 × anti-C3d-Fc proteins. As a control group, viruses

  6. Changes in clot lysis levels of reteplase and streptokinase following continuous wave ultrasound exposure, at ultrasound intensities following attenuation from the skull bone

    Directory of Open Access Journals (Sweden)

    Roijer Anders

    2008-08-01

    Full Text Available Abstract Background Ultrasound (US has been used to enhance thrombolytic therapy in the treatment of stroke. Considerable attenuation of US intensity is however noted if US is applied over the temporal bone. The aim of this study was therefore to explore possible changes in the effect of thrombolytic drugs during low-intensity, high-frequency continuous-wave ultrasound (CW-US exposure. Methods Clots were made from fresh venous blood drawn from healthy volunteers. Each clot was made from 1.4 ml blood and left to coagulate for 1 hour in a plastic test-tube. The thrombolytic drugs used were, 3600 IU streptokinase (SK or 0.25 U reteplase (r-PA, which were mixed in 160 ml 0.9% NaCl solution. Continuous-wave US exposure was applied at a frequency of 1 MHz and intensities ranging from 0.0125 to 1.2 W/cm2. For each thrombolytic drug (n = 2, SK and r-PA and each intensity (n = 9 interventional clots (US-exposed, n = 6 were submerged in thrombolytic solution and exposed to CW-US while control clots (also submerged in thrombolytic solution, n = 6 were left unexposed to US. To evaluate the effect on clot lysis, the haemoglobin (Hb released from each clot was measured every 20 min for 1 hour (20, 40 and 60 min. The Hb content (mg released was estimated by spectrophotometry at 540 nm. The difference in effect on clot lysis was expressed as the difference in the amount of Hb released between pairs of US-exposed clots and control clots. Statistical analysis was performed using Wilcoxon's signed rank test. Results Continuous-wave ultrasound significantly decreased the effects of SK at intensities of 0.9 and 1.2 W/cm2 at all times (P 2 and at 1.2 W/cm2, following 40 min exposure at 0.3, 0.6, 0.9 and at 1.2 W/cm2, and following 60 min of exposure at 0.05 0.3, 0.6, 0.9 and at 1.2 W/cm2 (all P Conclusion Increasing intensities of CW-US exposure resulted in increased clot lysis of r-PA-treated blood clots, but decreased clot lysis of SK-treated clots.

  7. Influence of histone deacetylase inhibitors and DNA-methyltransferase inhibitors on the NK cell-mediated lysis of pediatric B-lineage leukemia.

    Directory of Open Access Journals (Sweden)

    Matthias Manuel Pfeiffer

    2013-04-01

    Full Text Available Epigenetic drugs like histone deacetylase inhibitors and DNA methyltransferase inhibitors have been shown to be effective against a variety of tumor entities. Among different molecular anticancer activities of epigenetic active substances, up-regulation of NK cell ligands was described to contribute to an enhanced NK cell-mediated killing of tumor cell lines. So far, no data is available on this effect in childhood acute lymphoblastic leukemia. We investigated the effect of two HDACi (vorinostat, VPA and two DNMTi (azacytidine, decitabine on the viability, expression of NK ligands and NK-susceptibility of the pre-B-cell-ALL cell line MHH-CALL-4. Whereas vorinostat, azacytidine and decitabine directly reduced viability of the cell line, VPA had no direct cytotoxic effect. NKG2D ligands were expressed only at very low levels and not affected by epigenetic treatment. Higher expression was found for the DNAM1 ligands with significant up regulation of CD112 after treatment with VPA (p=0.02. No significant increase in lysis mediated by resting NK cells could be observed, whereas incubation of target cells with decitabine resulted in a significant increase in lysis mediated by IL-2 activated NK cells (p=0.0051, p=0.06 for azacytidine. Vorinostat and VPA could increase the lysis by expanded NK cells which was statistically not significant due to high inter-individual variability. Furthermore, HDACi but not DNMTi reduced the NK mediated lysis of MHH-CALL-4 after incubation of effector cells. In conclusion, there is a synergistic effect between epigenetic drugs and NK cells against MHH-CALL-4 which is not as strong as in other tumor entities. In situations where NK mediated control of leukemia is assumed or wanted, a sophisticated combination of single epigenetic drugs and ex vivo expanded NK cells is needed to maximize the synergistic effect of both treatment strategies and DNMTIs may be preferred based on the direct inhibitory effect of HDACi on NK cell

  8. Nitrogen acquisition in Agave tequilana from degradation of endophytic bacteria.

    Science.gov (United States)

    Beltran-Garcia, Miguel J; White, James F; Prado, Fernanda M; Prieto, Katia R; Yamaguchi, Lydia F; Torres, Monica S; Kato, Massuo J; Medeiros, Marisa H G; Di Mascio, Paolo

    2014-11-06

    Plants form symbiotic associations with endophytic bacteria within tissues of leaves, stems, and roots. It is unclear whether or how plants obtain nitrogen from these endophytic bacteria. Here we present evidence showing nitrogen flow from endophytic bacteria to plants in a process that appears to involve oxidative degradation of bacteria. In our experiments we employed Agave tequilana and its seed-transmitted endophyte Bacillus tequilensis to elucidate organic nitrogen transfer from (15)N-labeled bacteria to plants. Bacillus tequilensis cells grown in a minimal medium with (15)NH4Cl as the nitrogen source were watered onto plants growing in sand. We traced incorporation of (15)N into tryptophan, deoxynucleosides and pheophytin derived from chlorophyll a. Probes for hydrogen peroxide show its presence during degradation of bacteria in plant tissues, supporting involvement of reactive oxygen in the degradation process. In another experiment to assess nitrogen absorbed as a result of endophytic colonization of plants we demonstrated that endophytic bacteria potentially transfer more nitrogen to plants and stimulate greater biomass in plants than heat-killed bacteria that do not colonize plants but instead degrade in the soil. Findings presented here support the hypothesis that some plants under nutrient limitation may degrade and obtain nitrogen from endophytic microbes.

  9. Immunomodulatory properties of probiotic bacteria

    DEFF Research Database (Denmark)

    Fink, Lisbeth Nielsen

    2007-01-01

    Certain lactic acid bacteria (LAB) are part of the commensal intestinal flora and considered beneficial for health, as they compete with pathogens for adhesion sites in the intestine and ferment otherwise indigestible compounds. Another important property of these so-called probiotic bacteria...... with bacteria, and the cytokine pattern induced by specific bacteria resembled the pattern induced in MoDC, except for TNF-alpha and IL-6, which were induced in response to different bacteria in blood DC/monocytes and monocyte-derived DC. Autologous NK cells produced IFN-gamma when cultured with blood DC......, monocytes and monocyte-derived DC and IL-12-inducing bacteria, whereas only DC induced IFN-gamma production in allogeneic T cells. In vitro-generated DC is a commonly used model of tissue DC, but they differ in certain aspects from intestinal DC, which are in direct contact with the intestinal microbiota...

  10. Cable Bacteria in Freshwater Sediments

    DEFF Research Database (Denmark)

    Risgaard-Petersen, Nils; Kristiansen, Michael; Frederiksen, Rasmus

    2015-01-01

    In marine sediments cathodic oxygen reduction at the sediment surface can be coupled to anodic sulfide oxidation in deeper anoxic layers through electrical currents mediated by filamentous, multicellular bacteria of the Desulfobulbaceae family, the so-called cable bacteria. Until now, cable...... bacteria have only been reported from marine environments. In this study, we demonstrate that cable bacteria also occur in freshwater sediments. In a first step, homogenized sediment collected from the freshwater stream Giber Å, Denmark, was incubated in the laboratory. After 2 weeks, pH signatures...... marine cable bacteria, with the genus Desulfobulbus as the closest cultured lineage. The results of the present study indicate that electric currents mediated by cable bacteria could be important for the biogeochemistry in many more environments than anticipated thus far and suggest a common evolutionary...

  11. Acoustofluidic bacteria separation

    Science.gov (United States)

    Li, Sixing; Ma, Fen; Bachman, Hunter; Cameron, Craig E.; Zeng, Xiangqun; Huang, Tony Jun

    2017-01-01

    Bacterial separation from human blood samples can help with the identification of pathogenic bacteria for sepsis diagnosis. In this work, we report an acoustofluidic device for label-free bacterial separation from human blood samples. In particular, we exploit the acoustic radiation force generated from a tilted-angle standing surface acoustic wave (taSSAW) field to separate Escherichia coli from human blood cells based on their size difference. Flow cytometry analysis of the E. coli separated from red blood cells shows a purity of more than 96%. Moreover, the label-free electrochemical detection of the separated E. coli displays reduced non-specific signals due to the removal of blood cells. Our acoustofluidic bacterial separation platform has advantages such as label-free separation, high biocompatibility, flexibility, low cost, miniaturization, automation, and ease of in-line integration. The platform can be incorporated with an on-chip sensor to realize a point-of-care sepsis diagnostic device.

  12. Bacteria, phages and septicemia.

    Directory of Open Access Journals (Sweden)

    Ausra Gaidelyte

    Full Text Available The use of phages is an attractive option to battle antibiotic resistant bacteria in certain bacterial infections, but the role of phage ecology in bacterial infections is obscure. Here we surveyed the phage ecology in septicemia, the most severe type of bacterial infection. We observed that the majority of the bacterial isolates from septicemia patients spontaneously secreted phages active against other isolates of the same bacterial strain, but not to the strain causing the disease. Such phages were also detected in the initial blood cultures, indicating that phages are circulating in the blood at the onset of sepsis. The fact that most of the septicemic bacterial isolates carry functional prophages suggests an active role of phages in bacterial infections. Apparently, prophages present in sepsis-causing bacterial clones play a role in clonal selection during bacterial invasion.

  13. Bacteriophages of methanotrophic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Tyutikow, F.M. (All-Union Research Inst. for Genetics and Selection of Industrial Microorganisms, Moscow, USSR); Bespalova, I.A.; Rebentish, B.A.; Aleksandrushkina, N.N.; Krivisky, A.S.

    1980-10-01

    Bacteriophages of methanotrophic bacteria have been found in 16 out of 88 studied samples (underground waters, pond water, soil, gas and oil installation waters, fermentor cultural fluids, bacterial paste, and rumen of cattle) taken in different geographic zones of the Soviet Union. Altogether, 23 phage strains were isolated. By fine structure, the phages were divided into two types (with very short or long noncontractile tails); by host range and serological properties, they fell into three types. All phages had guanine- and cytosine-rich double-stranded deoxyribonucleic acid consisting of common nitrogen bases. By all of the above-mentioned properties, all phages within each of the groups were completely identical to one another, but differed from phages of other groups.

  14. Freeze-drying of lactic acid bacteria.

    Science.gov (United States)

    Fonseca, Fernanda; Cenard, Stéphanie; Passot, Stéphanie

    2015-01-01

    Lactic acid bacteria are of great importance for the food and biotechnology industry. They are widely used as starters for manufacturing food (e.g., yogurt, cheese, fermented meats, and vegetables) and probiotic products, as well as for green chemistry applications. Freeze-drying or lyophilization is a convenient method for preservation of bacteria. By reducing water activity to values below 0.2, it allows long-term storage and low-cost distribution at suprazero temperatures, while minimizing losses in viability and functionality. Stabilization of bacteria via freeze-drying starts with the addition of a protectant solution to the bacterial suspension. Freeze-drying includes three steps, namely, (1) freezing of the concentrated and protected cell suspension, (2) primary drying to remove ice by sublimation, and (3) secondary drying to remove unfrozen water by desorption. In this chapter we describe a method for freeze-drying of lactic acid bacteria at a pilot scale, thus allowing control of the process parameters for maximal survival and functionality recovery.

  15. Studies on nonidet P40 lysis of murine lymphoid cells. I. Use of cholera toxin and cell surface Ig to determine degree of dissociation of the plasma membrane.

    Science.gov (United States)

    Hart, D A

    1975-09-01

    Lymphoid cells from A/J mice were iodinated (125I) by the lactoperoxidase lysed with the non-ionic detergent NP-40. The plasma membrane glycolipid receptor for cholera toxin and cell surface immunoglobulin were utilized in immune precipitation systems to characterize the degree of dissociation of the plasma membrane under various conditions. It was found that at 0.1% NP-40 and at cell concentration from 5 to 10 times 10(7) cells/ml, lipid-protein and protein-lipid-protein complexes formed in NP-40 which were soluble after centrifugation at 10(5) times G. Column chromatography of 125I-cell lysates on agarose A-0.5 M in 0.1% or 0.5% NP-40/PBS indicated that the majority of iodinated cell surface material existed as aggregates in detergent micelles. The availability of the oligosaccharide moiety of the glycolipid to interact with the cholera toxin was dependent on both the detergent concentration and the cell concentration used for cell lysis. However, the cell surface immunoglobulin was immunoprecipitable under all conditions of lysis tested.

  16. [White blood cell lysis syndrome after autologous peripheral blood stem cell transplantation in the treatment of renal AL amyloidosis. Case report].

    Science.gov (United States)

    Gatica, Antonio; Bertin, Pablo; Tagle, Rodrigo

    2006-06-01

    The treatment of AL amyloidosis was not successful until the advent of myeloablative chemotherapy consisting of high-dose intravenous melphalan followed by autologous peripheral blood stem cell transplantation. This new treatment has achieved better survival rates and, remarkably, it has obtained complete remission. Among patients with renal involvement, achievement of a complete hematological response was associated with a 50% reduction in proteinuria and stable creatinine clearance in more than 2/3 of patients. Despite of these excellent results, this new therapy is associated with significant toxicity, including the development of acute renal failure due to white blood cell lysis syndrome. We report a 59 year-old female with a nephrotic syndrome due to primary amyloidosis successfully treated autologous stem cell transplantation who developed acute renal failure caused by white blood cell lysis syndrome. The patient required treatment with granulocytic colony stimulating factor and intermittent hemofiltration and was discharged 23 days after melphalan administration with a satisfactory renal function and white blood cell count. After one year of follow up, she maintains a good glomerular filtration rate, a proteinuria of less than, 1 g/day and normal hematological values.

  17. Mechanism of action of Melaleuca alternifolia (tea tree) oil on Staphylococcus aureus determined by time-kill, lysis, leakage, and salt tolerance assays and electron microscopy.

    Science.gov (United States)

    Carson, Christine F; Mee, Brian J; Riley, Thomas V

    2002-06-01

    The essential oil of Melaleuca alternifolia (tea tree) has broad-spectrum antimicrobial activity. The mechanisms of action of tea tree oil and three of its components, 1,8-cineole, terpinen-4-ol, and alpha-terpineol, against Staphylococcus aureus ATCC 9144 were investigated. Treatment with these agents at their MICs and two times their MICs, particularly treatment with terpinen-4-ol and alpha-terpineol, reduced the viability of S. aureus. None of the agents caused lysis, as determined by measurement of the optical density at 620 nm, although cells became disproportionately sensitive to subsequent autolysis. Loss of 260-nm-absorbing material occurred after treatment with concentrations equivalent to the MIC, particularly after treatment with 1,8-cineole and alpha-terpineol. S. aureus organisms treated with tea tree oil or its components at the MIC or two times the MIC showed a significant loss of tolerance to NaCl. When the agents were tested at one-half the MIC, only 1,8-cineole significantly reduced the tolerance of S. aureus to NaCl. Electron microscopy of terpinen-4-ol-treated cells showed the formation of mesosomes and the loss of cytoplasmic contents. The predisposition to lysis, the loss of 260-nm-absorbing material, the loss of tolerance to NaCl, and the altered morphology seen by electron microscopy all suggest that tea tree oil and its components compromise the cytoplasmic membrane.

  18. A Simple Method for DNA Extraction from Mature Date Palm Leaves: Impact of Sand Grinding and Composition of Lysis Buffer

    Directory of Open Access Journals (Sweden)

    Mohammad Al Sadoon

    2010-09-01

    Full Text Available Molecular marker techniques have been widely used for cultivar identification of inbred date palms (Phoenix dactylifera L.; Arecaceae and biodiversity conservation. Isolation of highly pure DNA is the prerequisite for PCR amplification and subsequent use such as DNA fingerprinting and sequencing of genes that have recently been developed for barcoding. To avoid problems related to the preservation and use of liquid nitrogen, we examined sterile sand for grinding the date palm leaves. Individual and combined effects of sodium chloride (NaCl, polyvinylpyrrolidone (PVP and lithium chloride (LiCl with the cetyltrimethylammonium bromide (CTAB method for a DNA yield of sufficient purity and PCR amplification were evaluated in this study. Presence of LiCl and PVP alone or together in the lysis buffer did not significantly improve the DNA yield and purity compared with the addition of NaCl. Our study suggested that grinding of date palm leaf with sterile sand and inclusion of NaCl (1.4 M in the lysis buffer without the costly use of liquid nitrogen, PVP and LiCl, provides a DNA yield of sufficient purity, suitable for PCR amplification.

  19. Image-Based Single Cell Profiling: High-Throughput Processing of Mother Machine Experiments

    Science.gov (United States)

    Sachs, Christian Carsten; Grünberger, Alexander; Helfrich, Stefan; Probst, Christopher; Wiechert, Wolfgang; Kohlheyer, Dietrich; Nöh, Katharina

    2016-01-01

    Background Microfluidic lab-on-chip technology combined with live-cell imaging has enabled the observation of single cells in their spatio-temporal context. The mother machine (MM) cultivation system is particularly attractive for the long-term investigation of rod-shaped bacteria since it facilitates continuous cultivation and observation of individual cells over many generations in a highly parallelized manner. To date, the lack of fully automated image analysis software limits the practical applicability of the MM as a phenotypic screening tool. Results We present an image analysis pipeline for the automated processing of MM time lapse image stacks. The pipeline supports all analysis steps, i.e., image registration, orientation correction, channel/cell detection, cell tracking, and result visualization. Tailored algorithms account for the specialized MM layout to enable a robust automated analysis. Image data generated in a two-day growth study (≈ 90 GB) is analyzed in ≈ 30 min with negligible differences in growth rate between automated and manual evaluation quality. The proposed methods are implemented in the software molyso (MOther machine AnaLYsis SOftware) that provides a new profiling tool to analyze unbiasedly hitherto inaccessible large-scale MM image stacks. Conclusion Presented is the software molyso, a ready-to-use open source software (BSD-licensed) for the unsupervised analysis of MM time-lapse image stacks. molyso source code and user manual are available at https://github.com/modsim/molyso. PMID:27661996

  20. Swimming bacteria in liquid crystal

    Science.gov (United States)

    Sokolov, Andrey; Zhou, Shuang; Aranson, Igor; Lavrentovich, Oleg

    2014-03-01

    Dynamics of swimming bacteria can be very complex due to the interaction between the bacteria and the fluid, especially when the suspending fluid is non-Newtonian. Placement of swimming bacteria in lyotropic liquid crystal produces a new class of active materials by combining features of two seemingly incompatible constituents: self-propelled live bacteria and ordered liquid crystals. Here we present fundamentally new phenomena caused by the coupling between direction of bacterial swimming, bacteria-triggered flows and director orientations. Locomotion of bacteria may locally reduce the degree of order in liquid crystal or even trigger nematic-isotropic phase transition. Microscopic flows generated by bacterial flagella disturb director orientation. Emerged birefringence patterns allow direct optical observation and quantitative characterization of flagella dynamics. At high concentration of bacteria we observed the emergence of self-organized periodic texture caused by bacteria swimming. Our work sheds new light on self-organization in hybrid bio-mechanical systems and can lead to valuable biomedical applications. Was supported by the US DOE, Office of Basic Energy Sciences, Division of Materials Science and Engineering, under the Contract No. DE AC02-06CH11357.

  1. Determination of Ammonia Oxidizing Bacteria and Nitrate Oxidizing Bacteria in Wastewater and Bioreactors

    Science.gov (United States)

    Francis, Somilez Asya

    2014-01-01

    The process of water purification has many different physical, chemical, and biological processes. One part of the biological process is the task of ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB). Both play critical roles in the treatment of wastewater by oxidizing toxic compounds. The broad term is nitrification, a naturally occurring process that is carried out by AOB and NOB by using oxidation to convert ammonia to nitrite and nitrite to nitrate. To monitor this biological activity, bacterial staining was performed on wastewater contained in inoculum tanks and biofilm samples from bioreactors. Using microscopy and qPCR, the purpose of this experiment was to determine if the population of AOB and NOB in wastewater and membrane bioreactors changed depending on temperature and hibernation conditions to determine the optimal parameters for AOB/NOB culture to effectively clean wastewater.

  2. Chromium(VI) Bioremoval by Pseudomonas Bacteria: Role of Microbial Exudates for Natural Attenuation and Biotreatment of Cr(VI) Contamination

    Energy Technology Data Exchange (ETDEWEB)

    N Mercan Dogan; C Kantar; S Gulcan; C Dodge; B Coskun Yilmaz; M Ali Mazmanci

    2011-12-31

    Laboratory batch and column experiments were conducted to investigate the role of microbial exudates, e.g., exopolymeric substance (EPS) and alginic acid, on microbial Cr(VI) reduction by two different Pseudomonas strains (P. putida P18 and P. aeuroginosa P16) as a method for treating subsurface environment contaminated with Cr(VI). Our results indicate that microbial exudates significantly enhanced microbial Cr(VI) reduction rates by forming less toxic and highly soluble organo-Cr(III) complexes despite the fact Cr(III) has a very low solubility under the experimental conditions studied (e.g., pH 7). The formation of soluble organo-Cr(III) complexes led to the protection of the cells and chromate reductases from inactivation. In systems with no organic ligands, soluble organo-Cr(III) end products were formed between Cr(III) and the EPS directly released by bacteria due to cell lysis. Our results also provide evidence that cell lysis played an important role in microbial Cr(VI) reduction by Pseudomonas bacteria due to the release of constitutive reductases that intracellularly and/or extracellularly catalyzed the reduction of Cr(VI) to Cr(III). The overall results highlight the need for incorporation of the release and formation of organo-Cr(III) complexes into reactive transport models to more accurately design and monitor in situ microbial remediation techniques for the treatment of subsurface systems contaminated with Cr(VI).

  3. Chromium(VI) bioremoval by pseudomonas bacteria: role of microbial exudates for natural attenuation and biotreatment of Cr(VI) contamination

    Energy Technology Data Exchange (ETDEWEB)

    Dogan, N.M.; Dodge, C.; Kantar, C.; Gulcan, S.; Yilmaz, B.C.; Mazmanci, M.A.

    2011-02-14

    Laboratory batch and column experiments were conducted to investigate the role of microbial exudates, e.g., exopolymeric substance (EPS) and alginic acid, on microbial Cr(VI) reduction by two different Pseudomonas strains (P. putida P18 and P. aeuroginosa P16) as a method for treating subsurface environment contaminated with Cr(VI). Our results indicate that microbial exudates significantly enhanced microbial Cr(VI) reduction rates by forming less toxic and highly soluble organo-Cr(III) complexes despite the fact Cr(III) has a very low solubility under the experimental conditions studied (e.g., pH 7). The formation of soluble organo-Cr(III) complexes led to the protection of the cells and chromate reductases from inactivation. In systems with no organic ligands, soluble organo-Cr(III) end products were formed between Cr(III) and the EPS directly released by bacteria due to cell lysis. Our results also provide evidence that cell lysis played an important role in microbial Cr(VI) reduction by Pseudomonas bacteria due to the release of constitutive reductases that intracellularly and/or extracellularly catalyzed the reduction of Cr(VI) to Cr(III). The overall results highlight the need for incorporation of the release and formation of organo-Cr(III) complexes into reactive transport models to more accurately design and monitor in situ microbial remediation techniques for the treatment of subsurface systems contaminated with Cr(VI).

  4. Computational Challenges in Characterization of Bacteria and Bacteria-Host Interactions Based on Genomic Data

    Institute of Scientific and Technical Information of China (English)

    Chao Zhang; Guo-lu Zheng; Shun-Fu Xu; Dong Xu

    2012-01-01

    With the rapid development of next-generation sequencing technologies,bacterial identification becomes a very important and essential step in processing genomic data,especially for metagenomic data.Many computational methods have been developed and some of them are widely used to address the problems in bacterial identification.In this article we review the algorithms of these methods,discuss their drawbacks,and propose future computational methods that use genomic data to characterize bacteria.In addition,we tackle two specific computational problems in bacterial identification,namely,the detection of host-specific bacteria and the detection of disease-associated bacteria,by offering potential solutions as a starting point for those who are interested in the area.

  5. Motility of electric cable bacteria

    DEFF Research Database (Denmark)

    Bjerg, Jesper Tataru; Damgaard, Lars Riis; Holm, Simon Agner

    2016-01-01

    Cable bacteria are filamentous bacteria that electrically couple sulfide oxidation and oxygen reduction at centimeter distances, and observations in sediment environments have suggested that they are motile. By time-lapse microscopy, we found that cable bacteria used gliding motility on surfaces...... with a highly variable speed of 0.50.3 ms1 (meanstandard deviation) and time between reversals of 155108 s. They frequently moved forward in loops, and formation of twisted loops revealed helical rotation of the filaments. Cable bacteria responded to chemical gradients in their environment, and around the oxic......-anoxic interface, they curled and piled up, with straight parts connecting back to the source of sulfide. Thus, it appears that motility serves the cable bacteria in establishing and keeping optimal connections between their distant electron donor and acceptors in a dynamic sediment environment....

  6. Psychrotrophic ~Iydrolytic Bacteria from Antarctica &,Other I. Low Temperature Habitats

    Directory of Open Access Journals (Sweden)

    K.V. Ramana

    2000-04-01

    Full Text Available Samples of water, soil,llake sediments and blue-green algal mats from Antarctica were'processed for enumeration, isolation and screening of psychrotrophic hydrolytic bacteria. Amylolytic bacteria were preponderant (75 per cent in the blue-green algal htat samples. Protease, lipase, an1ylase and urerse producing/bacteria were also isolated from the samples. ,Biochemical characteristics indicated that the isolates ;mainly comprised Pseudomonas and Bacillus species. Proteases and lipases of antarctic bacterial strains preferably hydrolysed denatured protein substrate and water soluble monomeric synthetic lipid substrates, respectively.

  7. Assessment of performance of UV sterilizer for room air bacteria.

    Science.gov (United States)

    Joshi, P V

    2002-02-01

    Paper presents a technique for performance of UV sterilizer for room air bacteria. Patterns of decay of room air bacteria concentration during sterilization and build-up there after as a function of time is studied. Decay process seems to follow exponential pattern. Half-lives during decay are estimated. For single sterilizer unit with a dose of 16 W the decay half-life is around 8.6 min. For the dose of 32 W (2 sterilizers), half-life is estimated to be 6.18 min. The removal rates of room air bacteria due to sterilizer are compared with the natural decay of aerosols at steady state. The importance of decay half-life in the assessment has been stated. The bacteria concentration buildup process after putting off the sterilizers seems to be sigmoidal in nature. The buildup half-life is estimated to be around 53 min for present experimental conditions.

  8. Encapsulation of bacteria and viruses in electrospun nanofibres

    Science.gov (United States)

    Salalha, W.; Kuhn, J.; Dror, Y.; Zussman, E.

    2006-09-01

    Bacteria and viruses were encapsulated in electrospun polymer nanofibres. The bacteria and viruses were suspended in a solution of poly(vinyl alcohol) (PVA) in water and subjected to an electrostatic field of the order of 1 kV cm-1. Encapsulated bacteria in this work, (Escherichia coli, Staphylococcus albus) and bacterial viruses (T7, T4, λ) managed to survive the electrospinning process while maintaining their viability at fairly high levels. Subsequently the bacteria and viruses remain viable during three months at -20 and -55 °C without a further decrease in number. The present results demonstrate the potential of the electrospinning process for the encapsulation and immobilization of living biological material.

  9. Encapsulation of bacteria and viruses in electrospun nanofibres

    Energy Technology Data Exchange (ETDEWEB)

    Salalha, W [Faculty of Mechanical Engineering, Technion, Israel Institute of Technology, Haifa 32000 (Israel); Kuhn, J [Faculty of Biology, Technion, Israel Institute of Technology, Haifa 32000 (Israel); Dror, Y [Faculty of Mechanical Engineering, Technion, Israel Institute of Technology, Haifa 32000 (Israel); Zussman, E [Faculty of Mechanical Engineering, Technion, Israel Institute of Technology, Haifa 32000 (Israel)

    2006-09-28

    Bacteria and viruses were encapsulated in electrospun polymer nanofibres. The bacteria and viruses were suspended in a solution of poly(vinyl alcohol) (PVA) in water and subjected to an electrostatic field of the order of 1 kV cm{sup -1}. Encapsulated bacteria in this work (Escherichia coli, Staphylococcus albus) and bacterial viruses (T7, T4, {lambda}) managed to survive the electrospinning process while maintaining their viability at fairly high levels. Subsequently the bacteria and viruses remain viable during three months at -20 and -55 deg. C without a further decrease in number. The present results demonstrate the potential of the electrospinning process for the encapsulation and immobilization of living biological material.

  10. Modelling the fate and transport of faecal bacteria in estuarine and coastal waters.

    Science.gov (United States)

    Gao, Guanghai; Falconer, Roger A; Lin, Binliang

    2015-11-15

    This paper details a numerical model developed to predict the fate and transport of faecal bacteria in receiving surface waters. The model was first validated by comparing model predicted faecal bacteria concentrations with available field measurements. The model simulations agreed well with the observation data. After calibration, the model was applied to investigate the effects of different parameters, including: tidal processes, river discharges from the upstream boundaries and bacteria inputs from the upstream boundaries, wastewater treatment works (WwTWs), rivers and combined sewer overflows (CSO), on the concentrations of faecal bacteria in the Ribble Estuary. The results revealed that the tide and upstream boundary bacteria inputs were the primary factors controlling the distribution of faecal bacteria. The bacteria inputs from the WwTWs in the model domain were generally found not to have a significant impact on distribution of faecal bacteria in the estuary.

  11. Antibiotic resistance in probiotic bacteria

    Directory of Open Access Journals (Sweden)

    Miguel eGueimonde

    2013-07-01

    Full Text Available Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. The main probiotic bacteria are strains belonging to the genera Lactobacillus and Bifidobacterium, although other representatives, such as Bacillus or Escherichia coli strains, have also been used. Lactobacillus and Bifidobacterium are two common inhabitants of the human intestinal microbiota. Also, some species are used in food fermentation processes as starters, or as adjunct cultures in the food industry. With some exceptions, antibiotic resistance in these beneficial microbes does not constitute a safety concern in itself, when mutations or intrinsic resistance mechanisms are responsible for the resistance phenotype. In fact, some probiotic strains with intrinsic antibiotic resistance could be useful for restoring the gut microbiota after antibiotic treatment. However, specific antibiotic resistance determinants carried on mobile genetic elements, such as tetracycline resistance genes, are often detected in the typical probiotic genera, and constitute a reservoir of resistance for potential food or gut pathogens, thus representing a serious safety issue.

  12. Process of producing mannitol and homopolysaccharides

    NARCIS (Netherlands)

    Geel-Schutten, Gerritdina Hendrika van; Binnema, Doede J.; van der Maarel, Marc

    2004-01-01

    The present invention relates to a process producing mannitol and one or more homopolysaccharides by bacteria. The bacteria according to the invention express mannitol-2-dehydrogenase activity and one or more sucrase activities. Preferably, the bacteria are lactic acid bacteria and more preferably t

  13. 乳酸菌发酵茶饮料加工工艺优化%Optimization of the Processing Technique of Fermented Lactic Acid Bacteria Beverage Containing the Extract from Tea

    Institute of Scientific and Technical Information of China (English)

    胡燕; 陈忠杰

    2011-01-01

    [ Objective ] The reference for the preparation of the fermented lactic acid bacteria drink containing the extract from tea was provided through the optimaization of the technology. [ Method] The material from tea was extracted in 100 times' water and at 60 ℃ for 15 minutes,and the extract solution, added with sugar and milk powder, was fermented under condition of lactic acid bacteria. The factor such as the adding amount of lactic acid bacteria, sugar and milk powder, the fermentation time affecting the fermentation was optimized with the singlefactor experiments and orthogonal test. [ Result] The optimum technical condition was as follows: the adding amount of sugar was 8%; milk powder, 13%; lactic acid bacteria, 3%; the fermentation temperature, 40 ℃ and the fermentation time, 6 hours. [ Conclusion] The beverage had both the different fragrancematerials from the tea and the nutritional ingredients of milk, and also, was with good taste and quality.The product was a kind of multi-function drinking which was both nutrient and healthy.%[目的]时用乳酸菌发酵茶饮料的加工工艺进行优化,为利用茶叶浸提液发酵制备饮料的生产提供参考.[方法]以茶叶为原料,加入100倍的水,在60℃下浸提15 min.浸提液中加入糖、乳粉等原料进行乳酸菌发酵.通过单因素试验和正交试验对影响乳酸菌发酵的几个因素(乳酸菌接种量、加糖量、加乳粉量、发酵时间)进行优化.[结果]得到乳酸菌发酵的最佳工艺条件为:茶叶浸提液中加蔗糖量为8%,加乳粉量为13%,接种量为3%,发酵温度为40℃,发酵时间为6 h.[结论]在优化后的工艺条件下制作的茶饮料,既有茶的各种香味物质和营养成分,又有酸奶的营养成分,而且风味好、品质佳,是一种集营养和保健作用于一体的多功能饮品.

  14. Have sex or not? Lessons from bacteria.

    Science.gov (United States)

    Lodé, T

    2012-01-01

    Sex is one of the greatest puzzles in evolutionary biology. A true meiotic process occurs only in eukaryotes, while in bacteria, gene transcription is fragmentary, so asexual reproduction in this case really means clonal reproduction. Sex could stem from a signal that leads to increased reproductive output of all interacting individuals and could be understood as a secondary consequence of primitive metabolic reactions. Meiotic sex evolved in proto-eukaryotes to solve a problem that bacteria did not have, namely a large amount of DNA material, occurring in an archaic step of proto-cell formation and genetic exchanges. Rather than providing selective advantages through reproduction, sex could be thought of as a series of separate events which combines step-by-step some very weak benefits of recombination, meiosis, gametogenesis and syngamy.

  15. Characterization of Ice nucleation Bacteria and their Applications

    OpenAIRE

    ZONOURİ, Sheida Sadat; FATEHİNİA, Milad; NURİTABAR, Safoura; MANUCHEHRİ, Sara

    2015-01-01

    Abstract. Supercooled water can remain in liquid state at the temperatures below zero and needs catalysts for changing from liquid to solid state, and accumulation of water molecules causes nucleation and creates ice embryo. Then, other molecules are attached to this core and will make ice crystals “visible”. These catalysts may be inorganic materials, bacteria and/or fungal spores. In this review article, bacteria such as P. Syringae have been studied in ice nucleation process and their indu...

  16. Motility of Electric Cable Bacteria

    OpenAIRE

    Bjerg, Jesper Tataru; Damgaard, Lars Riis; Holm, Simon Agner; Schramm, Andreas; Nielsen, Lars Peter

    2016-01-01

    Cable bacteria are filamentous bacteria that electrically couple sulfide oxidation and oxygen reduction at centimeter distances, and observations in sediment environments have suggested that they are motile. By time-lapse microscopy, we found that cable bacteria used gliding motility on surfaces with a highly variable speed of 0.5 ± 0.3 μm s−1 (mean ± standard deviation) and time between reversals of 155 ± 108 s. They frequently moved forward in loops, and formation of twisted loops revealed ...

  17. Bacteria of the year. Industry fights legionella; Bacterie van het jaar. Industrie zet zich schrap tegen legionella

    Energy Technology Data Exchange (ETDEWEB)

    Dijkgraaf, A. [ed.

    2000-09-07

    More and more businesses discover the feared bacteria legionella in their cooling water and process water systems. A brief overview is given of what businesses and umbrella organizations are doing to prevent the occurrence of that dangerous bacteria.

  18. Beer spoilage bacteria and hop resistance

    NARCIS (Netherlands)

    Sakamoto, K; Konings, WN

    2003-01-01

    For brewing industry, beer spoilage bacteria have been problematic for centuries. They include some lactic acid bacteria such as Lactobacillus brevis, Lactobacillus lindneri and Pediococcus damnosus, and some Gram-negative bacteria such as Pectinatus cerevisiiphilus, Pectinatus frisingensis and Mega

  19. Fuel from Bacteria, CO2, Water, and Solar Energy: Engineering a Bacterial Reverse Fuel Cell

    Energy Technology Data Exchange (ETDEWEB)

    None

    2010-07-01

    Electrofuels Project: Harvard is engineering a self-contained, scalable Electrofuels production system that can directly generate liquid fuels from bacteria, carbon dioxide (CO2), water, and sunlight. Harvard is genetically engineering bacteria called Shewanella, so the bacteria can sit directly on electrical conductors and absorb electrical current. This current, which is powered by solar panels, gives the bacteria the energy they need to process CO2 into liquid fuels. The Harvard team pumps this CO2 into the system, in addition to water and other nutrients needed to grow the bacteria. Harvard is also engineering the bacteria to produce fuel molecules that have properties similar to gasoline or diesel fuel—making them easier to incorporate into the existing fuel infrastructure. These molecules are designed to spontaneously separate from the water-based culture that the bacteria live in and to be used directly as fuel without further chemical processing once they’re pumped out of the tank.

  20. Vitamin D₃ and monomethyl fumarate enhance natural killer cell lysis of dendritic cells and ameliorate the clinical score in mice suffering from experimental autoimmune encephalomyelitis.

    Science.gov (United States)

    Al-Jaderi, Zaidoon; Maghazachi, Azzam A

    2015-11-13

    Experimental autoimmune encephalomyelitis (EAE) is a CD4⁺ T cell mediated inflammatory demyelinating disease that is induced in mice by administration of peptides derived from myelin proteins. We developed EAE in SJL mice by administration of PLP139-151 peptide. The effect of treating these mice with 1α,25-Dihydroxyvitamin D₃ (vitamin D₃), or with monomethyl fumarate (MMF) was then examined. We observed that both vitamin D₃ and MMF inhibited and/or prevented EAE in these mice. These findings were corroborated with isolating natural killer (NK) cells from vitamin D₃-treated or MMF-treated EAE mice that lysed immature or mature dendritic cells. The results support and extend other findings indicating that an important mechanism of action for drugs used to treat multiple sclerosis (MS) is to enhance NK cell lysis of dendritic cells.

  1. Toll recognition signal activates oenocytoid cell lysis via a crosstalk between plasmatocyte-spreading peptide and eicosanoids in response to a fungal infection.

    Science.gov (United States)

    Park, Jung-A; Kim, Yonggyun

    2012-10-01

    Plasmatocyte-spreading peptide (PSP) activates hemocyte-spreading behavior in response to various microbial pathogens. Its homolog, growth-blocking peptide, has several functions that activate immune cells and induce oenocytoid cell lysis (OCL). OCL is required for release of prophenoloxidase from oenocytoids in the beet armyworm, Spodoptera exigua. Injection of PSP to S. exigua larvae significantly induced in vivo OCL and resulted in significant increase of phenoloxidase (PO) activity. A fungal infection induced PSP expression and also significantly increased OCL. RNA interference (RNAi) of PSP expression significantly suppressed OCL induction and subsequently inhibited PO activation. Interestingly, an addition of dexamethasone (a specific phospholipase A₂ inhibitor) inhibited the PSP activity to induce OCL. Toll signal pathway was associated with PSP action on inducing OCL because RNAi of Toll expression suppressed PSP expression and subsequent OCL induction. However, an addition of PSP to the larvae under RNAi of Toll expression rescued the progress of OCL.

  2. Lysis to Kill: Evaluation of the Lytic Abilities, and Genomics of Nine Bacteriophages Infective for Gordonia spp. and Their Potential Use in Activated Sludge Foam Biocontrol.

    Directory of Open Access Journals (Sweden)

    Zoe A Dyson

    Full Text Available Nine bacteriophages (phages infective for members of the genus Gordonia were isolated from wastewater and other natural water environments using standard enrichment techniques. The majority were broad host range phages targeting more than one Gordonia species. When their genomes were sequenced, they all emerged as double stranded DNA Siphoviridae phages, ranging from 17,562 to 103,424 bp in size, and containing between 27 and 127 genes, many of which were detailed for the first time. Many of these phage genomes diverged from the expected modular genome architecture of other characterized Siphoviridae phages and contained unusual lysis gene arrangements. Whole genome sequencing also revealed that infection with lytic phages does not appear to prevent spontaneous prophage induction in Gordonia malaquae lysogen strain BEN700. TEM sample preparation techniques were developed to view both attachment and replication stages of phage infection.

  3. Simple and Efficient Method for Measuring Anti-Toxoplasma Immunoglobulin Antibodies in Human Sera Using Complement-Mediated Lysis of Transgenic Tachyzoites Expressing β-Galactosidase

    Science.gov (United States)

    Dando, Caroline; Gabriel, Katie E.; Remington, Jack S.; Parmley, Stephen F.

    2001-01-01

    A simple and efficient method using transgenic Toxoplasma gondii tachyzoites expressing β-galactosidase was developed for detection of specific antibodies against the parasite in sera of patients. The titers obtained with the new test were similar to those obtained with the Sabin-Feldman dye test run in parallel. Although significant changes in endpoint titers were not observed when sera drawn sequentially at 2- to 3-week intervals were tested with both procedures, apparent differences in antibody affinity were observed with the new test which were not perceptible with the Sabin-Feldman dye test. Like the Sabin-Feldman dye test, the new test is based on complement lysis of tachyzoites, but it is much easier to perform and the reaction is read colorimetrically instead of visually. PMID:11376045

  4. LACTIC ACID BACTERIA: PROBIOTIC APPLICATIONS

    OpenAIRE

    NEENA GARG

    2015-01-01

    Lactic acid bacteria (LAB) is a heterotrophic Gram-positive bacteria which under goes lactic acid fermentations and leads to production of lactic acid as an end product. LAB includes Lactobacillus, Leuconostoc, Pediococcus, Lactococcus and Streptococcus which are grouped together in the family lactobacillaceae. LAB shows numerous antimicrobial activities due to production of antibacterial and antifungal compounds such as organic acids, bacteriocins, diacetyl, hydrogen peroxide and reutrin. LA...

  5. Thymidine kinase diversity in bacteria

    DEFF Research Database (Denmark)

    Sandrini, Michael; Clausen, A.R.; Munch-Petersen, B.

    2006-01-01

    Thymidine kinases (TKs) appear to be almost ubiquitous and are found in nearly all prokaryotes, eukaryotes, and several viruses. They are the key enzymes in thymidine salvage and activation of several anti-cancer and antiviral drugs. We show that bacterial TKs can be subdivided into 2 groups. The....... The TKs from Gram-positive bacteria are more closely related to the eukaryotic TK1 enzymes than are TKs from Gram-negative bacteria....

  6. A comparative effect of 3 disinfectants on heterotrophic bacteria, iron bacteria and sulfate-reducing bacteria

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The disinfection effect of chlorine dioxide, chlorine and their mixture on heterotrophic bacteria, iron bacteria and sulfate-reducing bacteria in circulating cooling water was studied. The results of the test indicated that high purity chlorine dioxide was the most effective biocide in the 3 disinfectants, and with a dosage of 0.5mg/L, chlorine dioxide could obtain perfect effect. High purity chloride dioxide could have the excellent effect with the pH value of 6 to 10, and could keep it within 72 h. Chlorine and their mixture couldn't reach the effect of chlorine dioxide.

  7. Redirected lysis of human melanoma cells by a MCSP/CD3-bispecific BiTE antibody that engages patient-derived T cells.

    Science.gov (United States)

    Torisu-Itakura, Hitoe; Schoellhammer, Hans F; Sim, Myung-Shin; Irie, Reiko F; Hausmann, Susanne; Raum, Tobias; Baeuerle, Patrick A; Morton, Donald L

    2011-10-01

    Melanoma-associated chondroitin sulfate proteoglycan (MCSP; also called HMW-MAA, CSPG4, NG2, MSK16, MCSPG, MEL-CSPG, or gp240) is a well characterized melanoma cell-surface antigen. In this study, a new bispecific T-cell engaging (BiTE) antibody that binds to MCSP and human CD3 (MCSP-BiTE) was tested for its cytotoxic activity against human melanoma cell lines. When unstimulated peripheral mononuclear blood cells (PBMCs) derived from healthy donors were cocultured with melanoma cells at effector:target ratios of 1:1, 1:5, or 1:10, and treated with MCSP-BiTE antibody at doses of 10, 100, or 1000 ng/mL, all MCSP-expressing melanoma cell lines (n=23) were lysed in a dose-dependent and effector:target ratio-dependent manner, whereas there was no cytotoxic activity against MCSP-negative melanoma cell lines (n=2). To investigate whether T cells from melanoma patients could act as effector cells, we cocultured unstimulated PBMCs with allogeneic melanoma cells from 13 patients (4 stage I/II, 3 stage III, and 6 stage IV) or with autologous melanoma cells from 2 patients (stage IV). Although cytotoxic activity varied, all 15 PBMC samples mediated significant redirected lysis by the BiTE antibody. When PBMC or CD8 T cells were prestimulated by anti-CD3 antibody OKT-3 and interleukin-2, the MCSP-BiTE concentrations needed for melanoma cell lysis decreased up to 1000-fold. As MCSP is expressed on most human melanomas, immunotherapy with MCSP/CD3-bispecific antibodies merits clinical investigation.

  8. In black South Africans from rural and urban communities, the 4G/5G PAI-1 polymorphism influences PAI-1 activity, but not plasma clot lysis time.

    Science.gov (United States)

    de Lange, Zelda; Rijken, Dingeman C; Hoekstra, Tiny; Conradie, Karin R; Jerling, Johann C; Pieters, Marlien

    2013-01-01

    Data on genetic and environmental factors influencing PAI-1 levels and their consequent effect on clot lysis in black African populations are limited. We identified polymorphisms in the promoter area of the PAI-1 gene and determined their influence on PAI-1act levels and plasma clot lysis time (CLT). We also describe gene-environment interactions and the effect of urbanisation. Data from 2010 apparently healthy urban and rural black participants from the South African arm of the PURE study were cross-sectionally analysed. The 5G allele frequency of the 4G/5G polymorphism was 0.85. PAI-1act increased across genotypes in the urban subgroup (p = 0.009) but not significantly in the rural subgroup, while CLT did not differ across genotypes. Significant interaction terms were found between the 4G/5G polymorphism and BMI, waist circumference and triglycerides in determining PAI-1act, and between the 4G/5G polymorphism and fibrinogen and fibrinogen gamma prime in determining CLT. The C428T and G429A polymorphisms did not show direct relationships with PAI-1act or CLT but they did influence the association of other environmental factors with PAI-1act and CLT. Several of these interactions differed significantly between rural and urban subgroups, particularly in individuals harbouring the mutant alleles. In conclusion, although the 4G/5G polymorphism significantly affected PAI-1act, it contributed less than 1% to the PAI-1act variance. (Central) obesity was the biggest contributor to PAI-1act variance (12.5%). Urbanisation significantly influenced the effect of the 4G/5G polymorphism on PAI-1act as well as gene-environment interactions for the C428T and G429A genotypes in determining PAI-1act and CLT.

  9. In black South Africans from rural and urban communities, the 4G/5G PAI-1 polymorphism influences PAI-1 activity, but not plasma clot lysis time.

    Directory of Open Access Journals (Sweden)

    Zelda de Lange

    Full Text Available Data on genetic and environmental factors influencing PAI-1 levels and their consequent effect on clot lysis in black African populations are limited. We identified polymorphisms in the promoter area of the PAI-1 gene and determined their influence on PAI-1act levels and plasma clot lysis time (CLT. We also describe gene-environment interactions and the effect of urbanisation. Data from 2010 apparently healthy urban and rural black participants from the South African arm of the PURE study were cross-sectionally analysed. The 5G allele frequency of the 4G/5G polymorphism was 0.85. PAI-1act increased across genotypes in the urban subgroup (p = 0.009 but not significantly in the rural subgroup, while CLT did not differ across genotypes. Significant interaction terms were found between the 4G/5G polymorphism and BMI, waist circumference and triglycerides in determining PAI-1act, and between the 4G/5G polymorphism and fibrinogen and fibrinogen gamma prime in determining CLT. The C428T and G429A polymorphisms did not show direct relationships with PAI-1act or CLT but they did influence the association of other environmental factors with PAI-1act and CLT. Several of these interactions differed significantly between rural and urban subgroups, particularly in individuals harbouring the mutant alleles. In conclusion, although the 4G/5G polymorphism significantly affected PAI-1act, it contributed less than 1% to the PAI-1act variance. (Central obesity was the biggest contributor to PAI-1act variance (12.5%. Urbanisation significantly influenced the effect of the 4G/5G polymorphism on PAI-1act as well as gene-environment interactions for the C428T and G429A genotypes in determining PAI-1act and CLT.

  10. T4噬菌体的组装、感染及细胞裂解%Assembly, Infection and Cell Lysis of Bacteriophage T4

    Institute of Scientific and Technical Information of China (English)

    颜晨; 逯凯; 任慧英

    2011-01-01

    To further develop and utilize T4 phage display and phage therapy, and open the broader prospect for the progress in study of T4 phage, the important components of T4 phage particles was analyzed by molecular biology techniques, such as the head, the tail and the basement. The protein composition of these structure were confirmed. DNA packing which was an important part in the assembly of T4 phage particle was described in detail. Not only the phenomenon of infection and lysis, but also the superinfection exclusion and lysis inhibition in T4 life cycles introduced with molecular mechanism and impact factor. The assembly and infection mechanism of T4 phage would be known much better by these research.%为深入开发利用T4噬菌体展示系统及T4噬菌体的临床细菌病治疗,应用现代各种分子生物学手段,在蛋白分子和组成结构上对T4噬菌体颗粒的各个重要组成部分进行分析,例如头部、尾部和尾板,描述了其蛋白构成和各种影响因子,并着重介绍了在噬菌体装配中最为重要的DNA的包装.同时针对噬菌体T4的感染与超感染免疫性和感染所导致的细胞裂解和裂解抑制等方面,介绍了这些现象发生的分子机制和影响因素.以期对T4噬菌体的分子装配和侵染机制有更为深刻的了解.

  11. Role of serum sodium in assessing hospital mortality in cancer patients with spontaneous tumour lysis syndrome inducing acute uric acid nephropathy.

    Science.gov (United States)

    Hsu, H-H; Chen, Y-C; Tian, Y-C; Chan, Y-L; Kuo, M-C; Tang, C-C; Fang, J-T; Lee, S-Y; Yang, C-W

    2009-05-01

    Spontaneous tumour lysis syndrome (STLS) inducing acute uric acid nephropathy, a rare and neglected disease, presents more insidiously than conventional post-treatment tumour lysis syndrome. Although STLS is a serious and potentially fatal complication in patients with neoplastic disorders, few investigations have addressed the relevance of clinical and laboratory features in assessing prognosis. A retrospective study was conducted, reviewing the records of all patients who developed acute renal failure (ARF) at Chang Gung memorial hospital between 1 July 1999 and 30 June 2003. STLS-induced acute uric acid nephropathy was identified in 12 of 1072 ARF patients (1.1%) during the study period. All patients had advanced stage tumours with large tumour burden, and 66.7% of cases had abdominal organ involvement. All 12 hyperuricemic patients became oliguric despite conservative therapy, and remained hyperuricemic (21.6 +/- 5.2 mg/dl) before dialysis therapy. Diuresis developed in eight patients (66.7%), with associated resolution of hyperuricemia, azotemia and metabolic derangements following dialysis initiation. Overall hospital mortality was 58.3%. Death in most patients was related to hyponatremia and hypoalbuminemia on admission. The serum sodium was found to have the best Youden index (0.86) and highest overall prediction accuracy (93%). Moreover, serum sodium and serum albumin for individual patients were significantly and positively correlated (r = 0.617, p = 0.032). This investigation confirms a grave prognosis for cancer patients with STLS inducing acute uric acid nephropathy. Hyponatremia and hypoalbuminemia on the first day of admission indicate poor prognosis in such patients.

  12. Vanadium removal from LD converter slag using bacteria and fungi.

    Science.gov (United States)

    Mirazimi, S M J; Abbasalipour, Z; Rashchi, F

    2015-04-15

    Removal of vanadium from Linz-Donawits (LD) converter slag was investigated by means of three different species of microbial systems: Acidithiobacillus thiooxidans (autotrophic bacteria), Pseudomonas putida (heterotrophic bacteria) and Aspergillus niger (fungi). The bioleaching process was carried out in both one-step and two-step process and the leaching efficiencies in both cases were compared. Formation of inorganic and organic acids during the leaching process caused mobilization of vanadium. In order to reduce toxic effects of the metal species on the above mentioned microorganisms, a prolonged adaptation process was performed. Both bacteria, A. thiooxidans and P. putida were able to remove more than 90% of vanadium at slag concentrations of 1-5 g L(-1) after 15 days. Also, the maximum achievable vanadium removal in the fungal system was approximately 92% at a slag concentration of 1 g L(-1) after 22 days.

  13. Survivability of bacteria ejected from icy surfaces after hypervelocity impact.

    Science.gov (United States)

    Burchell, Mark J; Galloway, James A; Bunch, Alan W; Brandão, Pedro F B

    2003-02-01

    Both the Saturnian and Jovian systems contain satellites with icy surfaces. If life exists on any of these icy bodies (in putative subsurface oceans for example) then the possibility exists for transfer of life from icy body to icy body. This is an application of the idea of Panspermia, wherein life migrates naturally through space. A possible mechanism would be that life, here taken as bacteria, could become frozen in the icy surface of one body. If a high-speed impact occurred on that surface, ejecta containing the bacteria could be thrown into space. It could then migrate around the local region of space until it arrived at a second icy body in another high-speed impact. In this paper we consider some of the necessary steps for such a process to occur, concentrating on the ejection of ice bearing bacteria in the initial impact, and on what happens when bacteria laden projectiles hit an icy surface. Laboratory experiments using high-speed impacts with a light gas gun show that obtaining icy ejecta with viable bacterial loads is straightforward. In addition to demonstrating the viability of the bacteria carried on the ejecta, we have also measured the angular and size distribution of the ejecta produced in hypervelocity impacts on ice. We have however been unsuccessful at transferring viable bacteria to icy surfaces from bacteria laden projectiles impacting at hypervelocities.

  14. Chemotaxis signaling systems in model beneficial plant-bacteria associations.

    Science.gov (United States)

    Scharf, Birgit E; Hynes, Michael F; Alexandre, Gladys M

    2016-04-01

    Beneficial plant-microbe associations play critical roles in plant health. Bacterial chemotaxis provides a competitive advantage to motile flagellated bacteria in colonization of plant root surfaces, which is a prerequisite for the establishment of beneficial associations. Chemotaxis signaling enables motile soil bacteria to sense and respond to gradients of chemical compounds released by plant roots. This process allows bacteria to actively swim towards plant roots and is thus critical for competitive root surface colonization. The complete genome sequences of several plant-associated bacterial species indicate the presence of multiple chemotaxis systems and a large number of chemoreceptors. Further, most soil bacteria are motile and capable of chemotaxis, and chemotaxis-encoding genes are enriched in the bacteria found in the rhizosphere compared to the bulk soil. This review compares the architecture and diversity of chemotaxis signaling systems in model beneficial plant-associated bacteria and discusses their relevance to the rhizosphere lifestyle. While it is unclear how controlling chemotaxis via multiple parallel chemotaxis systems provides a competitive advantage to certain bacterial species, the presence of a larger number of chemoreceptors is likely to contribute to the ability of motile bacteria to survive in the soil and to compete for root surface colonization.

  15. Filtrating forms of soil bacteria

    Science.gov (United States)

    Van'kova, A. A.; Ivanov, P. I.; Emtsev, V. T.

    2013-03-01

    Filtrating (ultramicroscopic) forms (FF) of bacteria were studied in a soddy-podzolic soil and the root zone of alfalfa plants as part of populations of the most widespread physiological groups of soil bacteria. FF were obtained by filtering soil solutions through membrane filters with a pore diameter of 0.22 μm. It was established that the greater part of the bacteria in the soil and in the root zone of the plants has an ultramicroscopic size: the average diameter of the cells is 0.3 μm, and their length is 0.6 μm, which is significantly less than the cell size of banal bacteria. The number of FF varies within a wide range depending on the physicochemical conditions of the habitat. The FF number's dynamics in the soil is of a seasonal nature; i.e., the number of bacteria found increases in the summer and fall and decreases in the winter-spring period. In the rhizosphere of the alfalfa, over the vegetation period, the number of FF and their fraction in the total mass of the bacteria increase. A reverse tendency is observed in the rhizoplane. The morphological particularities (identified by an electron microscopy) and the nature of the FF indicate their physiological activity.

  16. Bioreporter bacteria for landmine detection

    Energy Technology Data Exchange (ETDEWEB)

    Burlage, R.S. [Oak Ridge National Lab., TN (United States); Youngblood, T. [Frisby Technologies, Aiken, SC (United States); Lamothe, D. [American Technologies, Inc., Huntsville, AL (United States). Ordnance/Explosives Environmental Services Div.

    1998-04-01

    Landmines (and other UXO) gradually leak explosive chemicals into the soil at significant concentrations. Bacteria, which have adapted to scavenge low concentrations of nutrients, can detect these explosive chemicals. Uptake of these chemicals results in the triggering of specific bacterial genes. The authors have created genetically recombinant bioreporter bacteria that detect small concentrations of energetic chemicals. These bacteria are genetically engineered to produce a bioluminescent signal when they contact specific explosives. A gene for a brightly fluorescent compound can be substituted for increased sensitivity. By finding the fluorescent bacteria, you find the landmine. Detection might be accomplished using stand-off illumination of the minefield and GPS technology, which would result in greatly reduced risk to the deminers. Bioreporter technology has been proven at the laboratory scale, and will be tested under field conditions in the near future. They have created a bacterial strain that detects sub-micromolar concentrations of o- and p-nitrotoluene. Related bacterial strains were produced using standard laboratory protocols, and bioreporters of dinitrotoluene and trinitrotoluene were produced, screening for activity with the explosive compounds. Response time is dependent on the growth rate of the bacteria. Although frill signal production may require several hours, the bacteria can be applied over vast areas and scanned quickly, producing an equivalent detection speed that is very fast. This technology may be applicable to other needs, such as locating buried explosives at military and ordnance/explosive manufacturing facilities.

  17. Cable Bacteria in Freshwater Sediments.

    Science.gov (United States)

    Risgaard-Petersen, Nils; Kristiansen, Michael; Frederiksen, Rasmus B; Dittmer, Anders Lindequist; Bjerg, Jesper Tataru; Trojan, Daniela; Schreiber, Lars; Damgaard, Lars Riis; Schramm, Andreas; Nielsen, Lars Peter

    2015-09-01

    In marine sediments cathodic oxygen reduction at the sediment surface can be coupled to anodic sulfide oxidation in deeper anoxic layers through electrical currents mediated by filamentous, multicellular bacteria of the Desulfobulbaceae family, the so-called cable bacteria. Until now, cable bacteria have only been reported from marine environments. In this study, we demonstrate that cable bacteria also occur in freshwater sediments. In a first step, homogenized sediment collected from the freshwater stream Giber Å, Denmark, was incubated in the laboratory. After 2 weeks, pH signatures and electric fields indicated electron transfer between vertically separated anodic and cathodic half-reactions. Fluorescence in situ hybridization revealed the presence of Desulfobulbaceae filaments. In addition, in situ measurements of oxygen, pH, and electric potential distributions in the waterlogged banks of Giber Å demonstrated the presence of distant electric redox coupling in naturally occurring freshwater sediment. At the same site, filamentous Desulfobulbaceae with cable bacterium morphology were found to be present. Their 16S rRNA gene sequence placed them as a distinct sister group to the known marine cable bacteria, with the genus Desulfobulbus as the closest cultured lineage. The results of the present study indicate that electric currents mediated by cable bacteria could be important for the biogeochemistry in many more environments than anticipated thus far and suggest a common evolutionary origin of the cable phenotype within Desulfobulbaceae with subsequent diversification into a freshwater and a marine lineage.

  18. Relationship between repair processes and mutation induction in bacteria. [UV radiation; methyl methanesulfonate; N-methyl-N/sup 1/-nitro-N-nitrosoguanidine; N-methyl-N-nitrosourea; N-ethyl-N-nitrosourea; ethyl methanesulfanate; N-ethyl-nitrosoguanidine; 4-nitroquinoline 1-oxide

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R F

    1979-01-01

    The main repair and replication-associated processes that can influence the induction of mutations by various mutagens in bacteria are reviewed. These include both constitutive and induced, error-free and error-prone systems. The mutation yield from a treatment with a mutagen can be markedly affected by which of these systems is operating in a given bacterial species or strain. The effect of these systems on mutation induction by ultraviolet light, monofunctional alkylating agents, base analogues, and frameshift mutagens is discussed in some detail. The bearing of these studies on the practical problems of estimating hazards is briefly considered. 79 references.

  19. Glutathione transferases in bacteria.

    Science.gov (United States)

    Allocati, Nerino; Federici, Luca; Masulli, Michele; Di Ilio, Carmine

    2009-01-01

    Bacterial glutathione transferases (GSTs) are part of a superfamily of enzymes that play a key role in cellular detoxification. GSTs are widely distributed in prokaryotes and are grouped into several classes. Bacterial GSTs are implicated in a variety of distinct processes such as the biodegradation of xenobiotics, protection against chemical and oxidative stresses and antimicrobial drug resistance. In addition to their role in detoxification, bacterial GSTs are also involved in a variety of distinct metabolic processes such as the biotransformation of dichloromethane, the degradation of lignin and atrazine, and the reductive dechlorination of pentachlorophenol. This review article summarizes the current status of knowledge regarding the functional and structural properties of bacterial GSTs.

  20. A synthetic peptide corresponding to the C-terminal 25 residues of phage MS2 coded lysis protein dissipates the protonmotive force in Escherichia coli membrane vesicles by generating hydrophilic pores

    NARCIS (Netherlands)

    Goessens, Wil H.F.; Driessen, Arnold J.M.; Wilschut, Jan; Duin, Jan van

    1988-01-01

    The RNA phage MS2 encodes a protein, 75 amino acids long, that is necessary and sufficient for lysis of the host cell. DNA deletion analysis has shown that the lytic activity is confined to the C-terminal half of the protein. We have examined the effects of a synthetic peptide, covering the C-termin

  1. Bioleaching of marmatite using moderately thermophilic bacteria

    Institute of Scientific and Technical Information of China (English)

    ZHOU Hong-bo; LIU Fei-fei; ZOU Ying-qin; ZENG Xiao-xi; QIU Guan-zhou

    2008-01-01

    The process of bioleaching marmatite using moderately thermophilic bacteria was studied by comparing marmatite leaching performance of mesophiles and moderate thermophiles and valuating the effect of venting capacity as well as pulp density on marmatite leaching performance of moderate thermophiles. The results show that moderate thermophiles have more advantages over mesophilies in bioleaching marmatite at 45℃ and the pulp density of 50g/L, and the zinc extraction efficiency reaches 93.1% in 20d. Aeration agitation can improve the transfer of O2 and CO2 in solution and promote the growth of bacteria and therefore, enhance the leaching efficiency. Under the venting levels of 50, 200 and 800mL/min, the zinc extraction efficiencies by moderate thermophiles are 57.8%, 92.5% and 96.0%, respectively. With the increase of pulp density, the total leaching amount of valuable metals increases, however, the extraction efficiency decreases due to many reasons, such as increasing shear force leading to poorly growth condition for bacteria, etc. The zinc extraction decreases remarkably to 58.9% while the pulp density mounts up 20%.

  2. Thermophilic, lignocellulolytic bacteria for ethanol production: current state and perspectives

    DEFF Research Database (Denmark)

    Chang, Tinghong; Yao, Shuo

    2011-01-01

    , in particular with emphasis on improving ethanol yield, and this facilitates their employment for ethanol production. Finally, different processes for second-generation ethanol production based on thermophilic bacteria have been proposed with the aim to achieve cost-competitive processes. However, thermophilic...

  3. μHall chip for sensitive detection of bacteria.

    Science.gov (United States)

    Issadore, David; Chung, Hyun Jung; Chung, Jaehoon; Budin, Ghyslain; Weissleder, Ralph; Lee, Hakho

    2013-09-01

    Sensitive, rapid and phenotype-specific enumeration of pathogens is essential for the diagnosis of infectious disease, monitoring of food chains, and for defense against bioterrorism. Microbiological culture and genotyping, techniques that sensitively and selectively detect bacteria in laboratory settings, have limited application in clinical environments due to high cost, slow response times, and the need for specially trained staff and laboratory infrastructure. To address these challenges, we developed a microfluidic chip-based micro-Hall (μHall) platform capable of measuring single, magnetically tagged bacteria directly in clinical specimens with minimal sample processing. We demonstrated the clinical utility of the μHall chip by enumerating Gram-positive bacteria. The overall detection limit of the system was similar to that of culture tests (~10 bacteria), but the assay time was 50-times faster. This low-cost, single-cell analytical technique is especially well-suited to diagnose infectious diseases in resource-limited clinical settings.

  4. Swimming and transport of bacteria in time-periodic flows

    Science.gov (United States)

    Winter, Rebecca; Patteson, Alison; Gagnon, David; Arratia, Paulo

    The transport of bacteria can be highly influenced by external flows in oceans, rivers, and intestinal tracts. This has implications in biological systems for the performance of major biological processes, such as biofilm formation. In this study, we experimentally investigate the aggregation and transport of swimming Vibrio cholerae bacteria in time-periodic flows. Bacteria are placed in a well-characterized flow, and bacterial concentrations are recorded for a range of Reynolds numbers (Re) that spans two orders of magnitude, from 0.1 to 10. It is generally found that bacteria deplete from regions of high deformation rate and accumulate near vortices. This phenomenon seems to be dictated by a combination of bacterial activity and background flow vorticity. R.W. supported by NSF-GRFP.

  5. Airborne bacteria in the atmosphere: Presence, purpose, and potential

    Science.gov (United States)

    Smets, Wenke; Moretti, Serena; Denys, Siegfried; Lebeer, Sarah

    2016-08-01

    Numerous recent studies have highlighted that the types of bacteria present in the atmosphere often show predictable patterns across space and time. These patterns can be driven by differences in bacterial sources of the atmosphere and a wide range of environmental factors, including UV intensity, precipitation events, and humidity. The abundance of certain bacterial taxa is of interest, not only for their ability to mediate a range of chemical and physical processes in the atmosphere, such as cloud formation and ice nucleation, but also for their implications -both beneficial and detrimental-for human health. Consequently, the widespread importance of airborne bacteria has stimulated the search for their applicability. Improving air quality, modelling the dispersal of airborne bacteria (e.g. pathogens) and biotechnological purposes are already being explored. Nevertheless, many technological challenges still need to be overcome to fully understand the roles of airborne bacteria in our health and global ecosystems.

  6. Anaerobic degradation of benzoate by sulfate-reducing bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Silva, S.P.; Adorno, M.A.T.; Moraes, E.M.; Varesche, M.B.A. [Sao Paulo Univ., Sao Carlos, SP (Brazil). Biological Processes Laboratory

    2004-07-01

    Anaerobic processes are an efficient way to degrade aromatic compounds in industrial wastewater, such as phenol, cresol and benzoate. This study characterized the bacteria that degrades benzoate, an anaerobic degradation intermediate of several complex aromatic compounds. In particular, the study assessed the capacity to use benzoate with sulfate reducing bacteria in mesophilic conditions. Biofilm from polyurethane foam matrices of a fixed bed reactor was used as the cellular inoculum to treat industrial wastewater containing organic peroxide. Dilution techniques were used to purify the material and obtain cultures of cocci. The benzoate consumption capacity in sulfidogenic conditions was observed when the purified inoculum was applied to batch reactors with different benzoate/sulfate relations. Results indicate that purification was positive to bacteria that can degrade aromatic compounds. Desulfococcus multivorans bacteria was identified following the physiologic and kinetic experiments. The 0.6 benzoate/sulfate relation was considered ideal for complete consumption of carbon and total use of sulfur. 10 refs., 3 figs.

  7. Identification of Lactic Acid Bacteria and Propionic Acid Bacteria using FTIR Spectroscopy and Artificial Neural Networks

    Directory of Open Access Journals (Sweden)

    Beata Nalepa

    2012-01-01

    Full Text Available In the present study, lactic acid bacteria and propionic acid bacteria have been identified at the genus level with the use of artificial neural networks (ANNs and Fourier transform infrared spectroscopy (FTIR. Bacterial strains of the genera Lactobacillus, Lactococcus, Leuconostoc, Streptococcus and Propionibacterium were analyzed since they deliver health benefits and are routinely used in the food processing industry. The correctness of bacterial identification by ANNs and FTIR was evaluated at two stages. At first stage, ANNs were tested based on the spectra of 66 reference bacterial strains. At second stage, the evaluation involved 286 spectra of bacterial strains isolated from food products, deposited in our laboratory collection, and identified by genus-specific PCR. ANNs were developed based on the spectra and their first derivatives. The most satisfactory results were reported for the probabilistic neural network, which was built using a combination of W5W4W3 spectral ranges. This network correctly identified the genus of 95 % of the lactic acid bacteria and propionic acid bacteria strains analyzed.

  8. Effects of viruses on bacterial functions under contrasting nutritional conditions for four species of bacteria isolated from Hong Kong waters

    Science.gov (United States)

    Liu, Hao; Yuan, Xiangcheng; Xu, Jie; Harrison, Paul J.; He, Lei; Yin, Kedong

    2015-09-01

    Free living viruses are ubiquitous in marine waters and concentrations are usually several times higher than the bacterial abundance. These viruses are capable of lysing host bacteria and therefore, play an important role in the microbial loop in oligotrophic waters. However, few studies have been conducted to compare the role of viruses in regulating bacterial abundance and heterotrophic activities between natural oligotrophic waters and anthropogenic influenced eutrophic waters. In this study, we examined viral effects on bacterial functions of four single bacterial species incubated with natural viral assemblages in seawater samples from eutrophic and oligotrophic waters. The viral-lysis of bacteria was significantly higher in eutrophic than oligotrophic waters. This suggests that viruses were capable of controlling bacterial abundance, respiration and production in the eutrophic waters. Cellular bacterial respiration and production was higher with viruses than without viruses, which was more evident in the oligotrophic waters. These results indicate that viruses can slow down bacterial consumption of oxygen and reduce bacteria-induced eutrophication effects in anthropogenic eutrophic waters, but switch to the role of sustaining the bacterial population when nutrients are limiting. There were bacterial species differences in resisting viral attack, which can influence the dominance and biodiversity of bacterial species in coastal waters.

  9. Permissivity of fish cell lines to three Chlamydia-related bacteria: Waddlia chondrophila, Estrella lausannensis and Parachlamydia acanthamoebae.

    Science.gov (United States)

    Kebbi-Beghdadi, Carole; Batista, Claudia; Greub, Gilbert

    2011-12-01

    Epitheliocystis is an infectious disease affecting gills and skin of various freshwater and marine fishes, associated with high mortality and reduced growth of survivors. Candidatus Piscichlamydia salmonis and Clavochlamydia salmonicola have recently been identified as aetiological agents of epitheliocystis in Atlantic Salmon. In addition, several other members of the Chlamydiales order have been identified in other fish species. To clarify the pathogenicity of Chlamydia-like organisms towards fishes, we investigated the permissivity of two fish cell lines, EPC-175 (Fathead Minnow) and RTG-2 (rainbow trout) to three Chlamydia-related bacteria: Waddlia chondrophila, Parachlamydia acanthamoebae and Estrella lausannensis. Quantitative PCR and immunofluorescence demonstrated that W. chondrophila and, to a lesser extent, E. lausannensis were able to replicate in the two cell lines tested. Waddlia chondrophila multiplied rapidly in its host cell and a strong cytopathic effect was observed. During E. lausannensis infection, we observed a limited replication of the bacteria not followed by host cell lysis. Very limited replication of P. acanthamoebae was observed in both cell lines tested. Given its high infectivity and cytopathic effect towards fish cell lines, W. chondrophila represents the most interesting Chlamydia-related bacteria to be used to develop an in vivo model of epitheliocystis disease in fishes.

  10. Bacterial lysis liberates the neutrophil migration suppressor YbcL from the periplasm of uropathogenic Escherichia coli.

    Science.gov (United States)

    Lau, Megan E; Danka, Elizabeth S; Tiemann, Kristin M; Hunstad, David A

    2014-12-01

    Uropathogenic Escherichia coli (UPEC) modulates aspects of the innate immune response during urinary tract infection to facilitate bacterial invasion of the bladder epithelium, a requirement for the propagation of infection. For example, UPEC-encoded YbcL suppresses the traversal of bladder epithelia by neutrophils in both an in vitro model and an in vivo murine cystitis model. The suppressive activity of YbcL requires liberation from the bacterial periplasm, though the mechanism of release is undefined. Here we present findings on the site of action of YbcL and demonstrate a novel mode of secretion for a UPEC exoprotein. Suppression of neutrophil migration by purified YbcL(UTI), encoded by cystitis isolate UTI89, required the presence of a uroepithelial layer; YbcL(UTI) did not inhibit neutrophil chemotaxis directly. YbcL(UTI) was released to a greater extent during UPEC infection of uroepithelial cells than during that of neutrophils. Release of YbcL(UTI) was maximal when UPEC and bladder epithelial cells were in close proximity. Established modes of secretion, including outer membrane vesicles, the type II secretion system, and the type IV pilus, were dispensable for YbcL(UTI) release from UPEC. Instead, YbcL(UTI) was liberated during bacterial death, which was augmented upon exposure to bladder epithelial cells, as confirmed by detection of bacterial cytoplasmic proteins and DNA in the supernatant and enumeration of bacteria with compromised membranes. As YbcL(UTI) acts on the uroepithelium to attenuate neutrophil migration, this mode of release may represent a type of altruistic cooperation within a UPEC population during colonization of the urinary tract.

  11. [Genetic resources of nodule bacteria].

    Science.gov (United States)

    Rumiantseva, M L

    2009-09-01

    Nodule bacteria (rhizobia) form highly specific symbiosis with leguminous plants. The efficiency of accumulation of biological nitrogen depends on molecular-genetic interaction between the host plant and rhizobia. Genetic characteristics of microsymbiotic strains are crucial in developing highly productive and stress-resistant symbiotic pairs: rhizobium strain-host plant cultivar (species). The present review considers the issue of studying genetic resources of nodule bacteria to identify genes and their blocks, responsible for the ability of rhizobia to form highly effective symbiosis in various agroecological conditions. The main approaches to investigation of intraspecific and interspecific genetic and genomic diversity of nodule bacteria are considered, from MLEE analysis to the recent methods of genomic DNA analysis using biochips. The data are presented showing that gene centers of host plants are centers of genetic diversification of nodule bacteria, because the intraspecific polymorphism of genetic markers of the core and the accessory rhizobial genomes is extremely high in them. Genotypic features of trapped and nodule subpopulations of alfalfa nodule bacteria are discussed. A survey of literature showed that the genomes of natural strains in alfalfa gene centers exhibit significant differences in genes involved in control of metabolism, replication, recombination, and the formation of defense response (hsd genes). Natural populations of rhizobia are regarded as a huge gene pool serving as a source of evolutionary innovations.

  12. Process optimization of brewers' grains fermentation using photosynthetic bacteria to make fish feed%光合菌发酵啤酒糟制鱼饲料工艺优化

    Institute of Scientific and Technical Information of China (English)

    张健; 冯学愚; 刘小彬; 谢刚; 方勤; 罗辉

    2011-01-01

    In order to get high-quality fish feed from brewers' spent grains, an anaerobic fermentation installation was set up, and the brewers' grains as raw material were pretreated by anaerobic photosynthetic bacteria separated from drainage ditch of brewers' grains in a beer factory. The orthogonal experiments were conducted based on single factors to study the semisolid fermentation technique for anaerobic photosynthetic bacteria culture. The results showed that the satisfied fermentation effect was attained under the technical conditions of dry brewers' grains/water 1:9 (mass fraction), inoculation amount 9%, illumination intensity 1100 Lx, fermentation time 5 d, fermentation temperature 30 ℃, and material thickness 3 cm. After fermentation, the real protein concentration of brewers' grains increased from 17.0% to 41.6%, the cellulose concentration decreased from 15.3% to 7.0%, the crude fat concentration increased from 5.9% to 6.6%, the crude ash concentration increased from 3.8% to 4.1%, the total phosphorus concentration increased from 0.6% to 1.3%, and the water concentration of dried brewers' grains was 9.8%. All the six indices satisfied the China standard of Grass Carp's fingerling (SC/T 1024-2002). The results can provide references for making active photosynthetic bacteria fish feed from brewers' grains.%为把啤酒糟加工成优质鱼饲料,从啤酒厂啤酒糟排水沟中以厌氧法分离出一株光合菌为菌种,对原料啤酒糟进行预处理,自组装厌氧发酵装置,采用单因素与正交试验法,对啤酒糟半固态培养光合菌的条件进行了研究.结果表明,当料水比1∶9(干糟:水,g/g)、接种量9%、光照强度1 100Lx、发酵时间5d、发酵温度30℃、料层厚度3 cm时,发酵效果较好.发酵后啤酒干糟真蛋白质量分数可从17.0%上升至41.6%、粗纤维素从15.3%下降至7.0%、粗脂肪从5.9%上升至6.6%、粗灰分从3.8%上升至4.1%、总磷从0.6%上升至1.3

  13. Co-electrospinning of bacteria and viruses

    Science.gov (United States)

    Salalha, Wael; Kuhn, Jonathan; Chervinsky, Shmuel; Zussman, Eyal

    2006-03-01

    Co-electrospinning provides a novel and highly versatile approach towards composite fibers with diameters ranging from a few hundred nm down to 30 nm with embedded elements. In the present work, co-electrospinning of poly(vinyl alcohol) (PVA) and viruses (T7, T4, λ) or bacteria (Escherichia coli, Staphylococcus albus) was carried out. These preparations should have applications for tissue engineering, gene therapy, phage therapy and biosensing. The average diameter of the co-spun nanofibers was about 300 nm. We found that the encapsulated viruses and bacteria manage to survive the electrospinning process, its pressure buildup in the core of the fiber and the electrostatic field in the co-electrospinning process. Approximately 10% of the Escherichia coli and 20% of Staphylococcus albus cells are viable after spinning. Approximately 5% of the bacterial viruses were also viable after the electrospinning. It should be noted that the encapsulated cells and viruses remain stable for two months without a further decrease in number. These results demonstrate the potential of the co-electrospinning process for the encapsulation and immobilization of bio-objects and the possibility of adapting them to technical applications (e.g., bio-chips).

  14. A Study on the Change of Bacteria Structure of Roast Duck in the Process of Storage%烤鸭肉在储存过程中菌群结构变化研究

    Institute of Scientific and Technical Information of China (English)

    张红琳; 周红霞; 周东蕊; 吴文婕

    2011-01-01

    The bacterial community change of roast duck was studied to find the theoretical bases for extending the storage period. Total bacteria genomic DNA was extracted from the duck samples by the method of phenol-chloroform after pre-treatment for 16S r DNA V3 region PCR amplification, and PCR products were detected and analyzed by denaturing gradient gel electrophoresis (DGGE). Then the similarity of DGGE fingerprints of the bacterial community was analysed by the Bionumerics software. The results showed that the DNA of the bacteria was extracted and amplified successfully; DGGE results showed that there were significant differences between bands in DGGE fingerprints, and the similarities were always changing, indicating that the bacterial community of roast duck had changed in the stored procedure. The study is to provide a theoretical reference for extending the storage period of duck meat.%探究烤鸭储存过程中菌群的变化,为延长其贮存期奠定理论基础.用酚、氯仿、异戊醇法提取烤鸭肉中细菌的基因组DNA,PCR扩增16SRNAV3区,用DGGE电泳技术分析菌群结构的变化情况,然后用Bionumerics软件对DGGE分子指纹图谱进行烤鸭肉菌群结构相似性分析.PCR结果表明,成功扩增了细菌基因组DNA,为230bp;DGGE实验结果表明,DGGE分子指纹图谱上条带存在显著差异;用bionumberics软件分析,发现样本相似度不断变化,表明在储存过程中,烤鸭肉菌群结构发生了变化.该研究为延长烤鸭肉保质期提供理论参考依据.

  15. 基因组提取过程中菌液浓度测定及提取方法优化%The Determination of Bacteria Concentration and Optimization of Extraction Method in the Process of Extracting Genome

    Institute of Scientific and Technical Information of China (English)

    孟静; 霍胜楠; 郑世超; 吴裕健

    2016-01-01

    Bacterial genomic DNA extraction quality directly affect the rapid test results based on PCR , including the detection speed, sensitivity, detection limit and other indicators. The concentration of cell culture is very important, too thick or too low concentrations will bring interference to the subsequent operation of PCR. In order to get the concentration of cell culture simply and conveniently, the Maxwell nephelometer were used to determine the cell concentration in the supernatant, at the same time validated the data using the plate count method. It showed that Maxwell nephelometer could match the determination purpose and measure the concentration of bacteria quickly and accurately. The standard curve of bacteria concentration were made , compared with the genome content, they have relevance. Different methods of extracting genome were compared to determine the extraction efficiency, at last the third:modified Universal Genomic DNA Extraction Lit Ver.3.0 method was selected.%细菌基因组DNA的提取质量直接影响以PCR为基础的快速检测结果,影响检测速度、灵敏度、检测限度等指标。其中菌体的培养浓度是重要的一个环节,过浓或浓度太低都会对后续的PCR操作带来干扰。为简单方便的测定菌体培养液的浓度,用麦氏比浊仪测定培养液中的菌体浓度,同时用平板计数法进行验证,结果表明麦氏比浊仪可达到测定目的,制定了相关菌种的标准曲线,可快速准确的测定菌液浓度。同时通过测定提取到的基因组含量,验证了与菌体浓度的相关性。另外也对比了不同的基因组提取方法的提取效率并评价,选择了最适合本试验的方法三:改良Universal Genomic DNA Extraction Lit Ver.3.0方法。

  16. Inhibiting function of denitrification in reducing the activities of sulfate bacteria in the crude oil refilling and transportation process%反硝化细菌抑制石油集输系统中硫酸盐还原菌试验研究

    Institute of Scientific and Technical Information of China (English)

    王堂彪; 金鹏康; 周立辉; 李岩

    2012-01-01

    The present paper intends to report our research on the inhibiting function of denitrification in reducing the activities of sulfate bacteria in the crude oil refilling and transportation process. In our research, we have prepared two sets of UASB (up-flow anaerobic sludge blanket) reactors ( UASB - 1 and UASB - 2) . The first reactor is to simulate the current environment of sulfate-reducing bacteria in the crude oil refilling and transportation process in order to promote the breeding of such bacteria for producing hydrogen sulfide, whereas the second reactor, the subsequent reactor, is mainly used to form the denitrification condition to let the oil liquid in so as to control the sulfate-reducing bacteria in producing hydrogen sulfide. Through adjusting the ratio of w(SO42-) : w( NO2-), DNB could be made to be the superior population in UASB - 2, which helps to control the hydrogen sulfide. The results of our experiments indicate that the optimal ORP (Oxidation-Reduction Potential) value with sulfate-reducing bacteria can be made to increase from - 370 mV to - 300 mV when - 150 mV to - 50 mV are just needed for denitrifying bacteria. Adding sodium nitrite to UASB - 2 is not only helpful to improve ORP value but also able to provide sufficient nitrogen sources. In our experiments, we have succeeded in achieving the best inhibitory effect on the decrease of hydrogen sulfide by 10% in the ratio of w(SO42- ):w(N02-) is 8: 1.2. We have also studied the change taking place in the microbial communities by using 16S rDNA gene based method and denaturing gradient gel electrophoresis ( DGGE) analysis. The results prove that the sulfate-reducing bacteria could be successfully inhibited by the denitrifying bacteria, with the three typical SRB (Desulfovibrio, Desulfotomaculum, Desulfomonas) gradually vanishing. At the same time, both the genera and the amount of the denitrifying bacteria were found increased remarkably. The analysis of the microorganism growth reveals that

  17. IDENTIFICATION OF BACTERIA IN LATEX PAINTS

    Directory of Open Access Journals (Sweden)

    Rojas, J.

    2008-01-01

    Full Text Available The bacteria are prokaryote organisms with a high capacity to colonize many types of habits. This research was developed with the object to identify extremophiles bacteria presents in latex paint. The bacteria were cultivated in culture mediums TSA, Blood Agar, Mc Conkey and finally the biochemical proof API-NF® for bacteria's isolation and identification, respectively. Characterization showed bacterial profile of Pasteurella sp. Hypothesis that could be found extremophiles bacteria in latex paint were demonstrated.

  18. Methylotrophic bacteria in sustainable agriculture.

    Science.gov (United States)

    Kumar, Manish; Tomar, Rajesh Singh; Lade, Harshad; Paul, Diby

    2016-07-01

    Excessive use of chemical fertilizers to increase production from available land has resulted in deterioration of soil quality. To prevent further soil deterioration, the use of methylotrophic bacteria that have the ability to colonize different habitats, including soil, sediment, water, and both epiphytes and endophytes as host plants, has been suggested for sustainable agriculture. Methylotrophic bacteria are known to play a significant role in the biogeochemical cycle in soil ecosystems, ultimately fortifying plants and sustaining agriculture. Methylotrophs also improve air quality by using volatile organic compounds such as dichloromethane, formaldehyde, methanol, and formic acid. Additionally, methylotrophs are involved in phosphorous, nitrogen, and carbon cycling and can help reduce global warming. In this review, different aspects of the interaction between methylotrophs and host plants are discussed, including the role of methylotrophs in phosphorus acquisition, nitrogen fixation, phytohormone production, iron chelation, and plant growth promotion, and co-inoculation of these bacteria as biofertilizers for viable agriculture practices.

  19. Chitin Degradation In Marine Bacteria

    DEFF Research Database (Denmark)

    Paulsen, Sara; Machado, Henrique; Gram, Lone

    2015-01-01

    Introduction: Chitin is the most abundant polymer in the marine environment and the second most abundant in nature. Chitin does not accumulate on the ocean floor, because of microbial breakdown. Chitin degrading bacteria could have potential in the utilization of chitin as a renewable carbon...... and nitrogen source in the fermentation industry.Methods: Here, whole genome sequenced marine bacteria were screened for chitin degradation using phenotypic and in silico analyses.Results: The in silico analyses revealed the presence of three to nine chitinases in each strain, however the number of chitinases...... chitin regulatory system.Conclusions: This study has provided insight into the ecology of chitin degradation in marine bacteria. It also served as a basis for choosing a more efficient chitin degrading production strain e.g. for the use of chitin waste for large-scale fermentations....

  20. [Systematic analysis and metabolic regulation of physiological functions for lactic acid bacteria--a review].

    Science.gov (United States)

    Wu, Chongde; Zhang, Juan; Liu, Liming

    2012-01-01

    As cell factories, lactic acid bacteria are widely used in food, agriculture, medicine and other industries, and play a great role in industrial processes. However, lactic acid bacteria encounter various environmental stresses both in industrial processes and in the gastrointestinal tract, which impair their physiological functions and food manufacture efficiency. Recently, the development of metabolic engineering and system biology brings unprecedented opportunity for the physiological modification of lactic acid bacteria. In this review, we addresses the progress of lactic acid bacterium system biology, and based on this, the metabolic engineering strategies for manipulating and optimizing lactic acid bacteria physiological function were summarized.

  1. Bacteria transport through porous media. Annual report, December 31, 1984

    Energy Technology Data Exchange (ETDEWEB)

    Yen, T.F.

    1986-09-01

    The following five chapters in this report have been processed separately for inclusion in the Energy Data Base: (1) theoretical model of convective diffusion of motile and non-motile bacteria toward solid surfaces; (2) interfacial electrochemistry of oxide surfaces in oil-bearing sands and sandstones; (3) effects of sodium pyrophosphate additive on the ''huff and puff''/nutrient flooding MEOR process; (4) interaction of Escherichia coli B, B/4, and bacteriophage T4D with Berea sandstone rock in relation to enhanced oil recovery; and (5) transport of bacteria in porous media and its significance in microbial enhanced oil recovery.

  2. Adaptation, Bacteria and Maxwell's Demons

    Science.gov (United States)

    Galajda, Peter; Keymer, Juan E.; Austin, Robert H.

    2007-03-01

    We propose a method to study the adaptation of bacterial populations with an asymmetric wall of Maxwell Demon openings. A Maxwell Demon opening is a funnel which is easier to enter than to leave. The interaction of swimming cells with such a Maxwell Demon Wall results in a population density separation, in apparent (but not real) violation of the Second Law of Thermodynamics, as we will show. Bacteria can be exposed to spatial challenges in order to move to e. g. higher food levels. The question we address in these experiments is: do the bacteria adapt and overcome the Maxwell Demon Wall?

  3. Close Encounters of Lymphoid Cells and Bacteria

    Science.gov (United States)

    Cruz-Adalia, Aranzazu; Veiga, Esteban

    2016-01-01

    During infections, the first reaction of the host against microbial pathogens is carried out by innate immune cells, which recognize conserved structures on pathogens, called pathogen-associated molecular patterns. Afterward, some of these innate cells can phagocytose and destroy the pathogens, secreting cytokines that would modulate the immune response to the challenge. This rapid response is normally followed by the adaptive immunity, more specific and essential for a complete pathogen clearance in many cases. Some innate immune cells, usually named antigen-presenting cells, such as macrophages or dendritic cells, are able to process internalized invaders and present their antigens to lymphocytes, triggering the adaptive immune response. Nevertheless, the traditional boundary of separated roles between innate and adaptive immunity has been blurred by several studies, showing that very specialized populations of lymphocytes (cells of the adaptive immunity) behave similarly to cells of the innate immunity. These “innate-like” lymphocytes include γδ T cells, invariant NKT cells, B-1 cells, mucosal-associated invariant T cells, marginal zone B cells, and innate response activator cells, and together with the newly described innate lymphoid cells are able to rapidly respond to bacterial infections. Strikingly, our recent data suggest that conventional CD4+ T cells, the paradigm of cells of the adaptive immunity, also present innate-like behavior, capturing bacteria in a process called transinfection. Transinfected CD4+ T cells digest internalized bacteria like professional phagocytes and secrete large amounts of proinflammatory cytokines, protecting for further bacterial challenges. In the present review, we will focus on the data showing such innate-like behavior of lymphocytes following bacteria encounter.

  4. Preliminary Clinical Application of Massage Lysis on Posterior Atlantooccipital Membrane%寰枕后膜推拿松解术的初步临床应用

    Institute of Scientific and Technical Information of China (English)

    于高潞; 英健民; 李茵; 史华敬; 王琳

    2012-01-01

    Objective: To observe the curative effect of massage lysis on posterior atlantooccipital membrane (PAM) for treating contracture syndrome of PAM. Methods: All 60 patients in our division from October 2008 to May 2010 were randomly divided into treatment group and control group, with 30 cases in each group. The patients of treatment group were treated with massage lysis on PAM, and the patients of control group were treated with dexamethasone and chlorzoxazone tablets. Ten days was a course of treatment, and observed for two courses. All the patients were followed up. Results: The atlantooccipital space in the treatment group was significantly improved than control group (P<0. 01). All the CDFI index of vertebral artery in the treatment group were improved obviously than the control group (P<0. 05). The total effective rate of the treatment group and control group was 93. 33% and 56. 70% respectively (P<0. 05). Conclusion: The method of massage lysis of PAM, as a new way to treat contracture syndrome of PAM, is used feasibly and practically in clinic for its advantages of safe, convenient, and cheap.%目的:观察寰枕后膜推拿松解术治疗寰枕后膜挛缩症的临床疗效.方法:对2008年10月~2010年5月我科收治寰枕后膜挛缩症患者60例,运用SPSS16.0随机分为治疗组和对照组,每组30例.治疗组采用寰枕后膜松解术;对照组给予地塞米松和氯唑沙宗口服,10d为1个疗程,观察2个疗程.所有患者均进行随访.结果:治疗组寰枕后间隙间距改善情况明显高于对照组(P<0.01);治疗组椎动脉各项指标(CDFI)改善情况明显高于对照组(P<0.05);治疗组总有效率为93.33%,对照组总有效率56.70%,2组比较差异有统计学意义(P<0.05).结论:寰枕后膜推拿松解术作为治疗寰枕后膜挛缩症的新方法,具有安全、简便、廉验的特点,具有一定临床实用及推广价值.

  5. A review on regulation methods of nitrite oxidizing bacteria in one-stage anaerobic ammonia oxidation process%一段式厌氧氨氧化工艺亚硝酸盐氧化菌抑制方法研究进展

    Institute of Scientific and Technical Information of China (English)

    谢丽; 殷紫; 尹志轩; 王悦超; 周琪

    2016-01-01

    近年来,厌氧氨氧化工艺(anaerobic ammonium oxidation, Anammox)作为一种新型的脱氮技术,由于其耗能少、效率高而被应用于高氨氮废水的处理中。然而,实际运行的厌氧氨氧化工程中有时会出现亚硝酸盐氧化菌(nitrite oxidizing bacteria, NOB)大量繁殖的情况,导致硝酸盐积累,脱氮效率下降。在一段式 Anammox 反应器中,通过控制某些影响因素,如调节体系中的溶解氧,控制游离氨和游离亚硝酸的浓度,调控碳源浓度以及外加中间产物(N2H4、NO 和 NH2OH)等方式,能够在维持 Anammox 工艺脱氮效率的同时有效抑制 NOB。除了系统地综述一段式 Anammox 工艺中 NOB 抑制手段以外,将进一步讨论实际 Anammox 工程应用中抑制 NOB 大量繁殖行之有效的手段。%In recent years,anaerobic ammonium oxidation (Anammox), a new technology for nitrogen removal, has been used in the treatment of high-strength ammonia wastewater due to its low energy consumption and high treatment efficiency. Whereas, the accumulation of nitrite oxidizing bacteria (NOB) often occurs in full-scale Anammox process, leading to the accumulation of nitrate and deterioration of nitrogen removal effectiveness. In two-stage Anammox processes, NOB accumulation often occurs in partial nitritation stage, the inhibition of which has been discussed in details. While in one-stage Anammox process, NOB accumulation is more common and fatal due to the complexity brought by the coexistence of functional bacteria like ammonium oxidizing bacteria (AOB), NOB, anaerobic ammonia oxidizing bacteria (AnAOB) and denitrifiers. It has been reported that NOB could be effectively suppressed in the one-stage Anammox process by some methods, e.g. regulating dissolved oxygen, altering the free ammonia and free nitrous acid concentration, adjusting carbon source and adding externally intermediate products (N2H4, NO, NH2OH), etc. The regulation methods

  6. The impact of lactic acid bacteria on sourdough fermentation

    Directory of Open Access Journals (Sweden)

    Savić Dragiša S.

    2005-01-01

    Full Text Available The baking of sourdough breads represents one of the oldest biotechnological processes. Despite traditionality, sourdough bread has great potential because of its benefits. Sourdough is a mixture of flour and water that is dominated by a complex microflora composed of yeasts and lactic acid bacteria that are crucial in the preparation of bread dough. Lactic acid bacteria cause acidification by producing lactic acid that increases the shelf life of bread by preventing the growth of undesirable microorganisms and affects the nutritional value of bread by increasing the availability of minerals. In addition to these advantages, the use of sourdough fermentation also improves dough machinability, breadcrumb structure and the characteristic flavour of bread. Lactic acid bacteria in sourdough fermentation are well known representing both homofermentative and heterofermentative bacteria. They may originate from selected natural contaminants in the flour or from a starter culture containing one or more known species of lactic acid bacteria. Sourdough can be cultivated in bakeries or obtained from commercial suppliers. However, many bakeries in Europe still use spontaneously fermented sourdoughs, which have been kept metabolically active for decades by the addition of flour and water at regular intervals. The impact of lactic acid bacteria on sourdough fermentation and their influence on dough and bread quality was discussed on the basis of research and literature data.

  7. The role of bacteria and mycorrhiza in plant sulfur supply

    Science.gov (United States)

    Gahan, Jacinta; Schmalenberger, Achim

    2014-01-01

    Plant growth is highly dependent on bacteria, saprophytic, and mycorrhizal fungi which facilitate the cycling and mobilization of nutrients. Over 95% of the sulfur (S) in soil is present in an organic form. Sulfate-esters and sulfonates, the major forms of organo-S in soils, arise through deposition of biological material and are transformed through subsequent humification. Fungi and bacteria release S from sulfate-esters using sulfatases, however, release of S from sulfonates is catalyzed by a bacterial multi-component mono-oxygenase system. The asfA gene is used as a key marker in this desulfonation process to study sulfonatase activity in soil bacteria identified as Variovorax, Polaromonas, Acidovorax, and Rhodococcus. The rhizosphere is regarded as a hot spot for microbial activity and recent studies indicate that this is also the case for the mycorrhizosphere where bacteria may attach to the fungal hyphae capable of mobilizing organo-S. While current evidence is not showing sulfatase and sulfonatase activity in arbuscular mycorrhiza, their effect on the expression of plant host sulfate transporters is documented. A revision of the role of bacteria, fungi and the interactions between soil bacteria and mycorrhiza in plant S supply was conducted. PMID:25566295

  8. Mechanisms of polymyxin resistance: acquired and intrinsic resistance in bacteria

    Directory of Open Access Journals (Sweden)

    Abiola Olumuyiwa Olaitan

    2014-11-01

    Full Text Available Polymyxins are polycationic antimicrobial peptides that are currently the last-resort antibiotics for the treatment of multidrug-resistant, Gram-negative bacterial infections. The reintroduction of polymyxins for antimicrobial therapy has been followed by an increase in reports of resistance among Gram-negative bacteria. Some bacteria, such as Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii, develop resistance to polymyxins in a process referred to as acquired resistance, whereas other bacteria, such as Proteus spp., Serratia spp. and Burkholderia spp., are naturally resistant to these drugs. Reports of polymyxin resistance in clinical isolates have recently increased, including acquired and intrinsically resistant pathogens. This increase is considered a serious issue, prompting concern due to the low number of currently available effective antibiotics. This review summarizes current knowledge concerning the different strategies bacteria employ to resist the activities of polymyxins.Gram-negative bacteria employ several strategies to protect themselves from polymyxin antibiotics (polymyxin B and colistin, including a variety of lipopolysaccharide (LPS modifications, such as modifications of lipid A with phosphoethanolamine and 4-amino-4-deoxy-L-arabinose, in addition to the use of efflux pumps, the formation of capsules and overexpression of the outer membrane protein OprH, which are all effectively regulated at the molecular level. The increased understanding of these mechanisms is extremely vital and timely to facilitate studies of antimicrobial peptides and find new potential drugs targeting clinically relevant Gram-negative bacteria.

  9. The role of bacteria and mycorrhiza in plant sulfur supply

    Directory of Open Access Journals (Sweden)

    Jacinta Mariea Gahan

    2014-12-01

    Full Text Available Plant growth is highly dependent on bacteria, saprophytic and mycorrhizal fungi which facilitate the cycling and mobilization of nutrients. Over 95% of the sulfur (S in soil is present in an organic form. Sulfate-esters and sulfonates, the major forms of organo-S in soils, arise through deposition of biological material and are transformed through subsequent humification. Fungi and bacteria release S from sulfate-esters using sulfatases, however, release of S from sulfonates is catalyzed by a bacterial multi-component mono-oxygenase system. The asfA gene is used as a key marker in this desulfonation process to study sulfonatase activity in soil bacteria identified as Variovorax, Polaromonas, Acidovorax and Rhodococcus. The rhizosphere is regarded as a hot spot for microbial activity and recent studies indicate that this is also the case for the mycorrhizosphere where bacteria may attach to the fungal hyphae capable of mobilizing organo-S. While current evidence is not showing sulfatase and sulfonatase activity in arbuscular mycorrhiza, their effect on the expression of plant host sulfate transporters is documented. A revision of the role of bacteria, fungi and the interactions between soil bacteria and mycorrhiza in plant S supply was conducted.

  10. Isolation and Identification of Epiphytic Lactic Acid Bacteria from Guinea Grass (Panicum maximum

    Directory of Open Access Journals (Sweden)

    M. Pasebani

    2010-01-01

    Full Text Available Problem statement: Bacteria can perform a variety of beneficial functions, for example many lactic acid bacteria are responsible for fermentation of silage in the process of forage conservation. In the making of silage, epiphytic lactic acid bacteria are usually insufficient in numbers to promote efficient lactate fermentation. This study was conducted to identify the predominant indigenous bacteria, with emphasis on lactic acid bacteria, from Guinea grass (Panicum maximum. Approach: Two different condition of growth using nutrient and MRS agar were prepared for isolation of the bacteria. In total, 18 purified isolates were identified by BIOLOG identification system which comprised of 9 bacterial species. Standard plate count in the both conditions was considered. Results: Three bacterial species based on the first condition of growth were identified which were belonging to Flavimonas oryzihabitans, Enerobacter cloacae, Sphingomonas paucimobilis B. Lactic acid bacteria based on the second condition of growth were belonging to Weissella confusa, Weissella paramesenteroides, Leuconostoc mesenteroides ssp. dextranicum, Lactococcus lactis ssp. hordniae. Result of plate count showed that 8.3×103 CFU lactic acid bacteria are available per gram of fresh guinea grass. Conclusion: Three hetero-fermentative and one homo-fermentative lactic acid bacteria were identified which would be suggested to use as bacterial inoculants because of the insufficient amount of epiphytic lactic acid bacteria and the availability of pathogenic bacteria in the grass.

  11. Sensitive detection of maltose and glucose based on dual enzyme-displayed bacteria electrochemical biosensor.

    Science.gov (United States)

    Liu, Aihua; Lang, Qiaolin; Liang, Bo; Shi, Jianguo

    2017-01-15

    Glucoamylase-displayed bacteria (GA-bacteria) and glucose dehydrogenase-displayed bacteria (GDH-bacteria) were co-immobilized on multi-walled carbon nanotubes (MWNTs) modified glassy carbon electrode (GCE) to construct GA-bacteria/GDH-bacteria/MWNTs/GCE biosensor. The biosensor was developed by optimizing the loading amount and the ratio of GA-bacteria to GDH-bacteria. The as-prepared biosensor exhibited a wide dynamic range of 0.2-10mM and a low detection limit of 0.1mM maltose (S/N=3). The biosensor also had a linear response to glucose in the range of 0.1-2.0mM and a low detection limit of 0.04mM glucose (S/N=3). Interestingly, at the same concentration, glucose was 3.75-fold sensitive than that of maltose at the proposed biosensor. No interferences were observed for other possible mono- and disaccharides. The biosensor also demonstrated good long-term storage stability and repeatability. Further, using both GDH-bacteria/MWNTs/GCE biosensor and GA-bacteria/GDH-bacteria/MWNTs/GCE biosensor, glucose and maltose in real samples can be detected. Therefore, the proposed biosensor is capable of monitoring the food manufacturing and fermentation process.

  12. T cells kill bacteria captured by transinfection from dendritic cells and confer protection in mice.

    Science.gov (United States)

    Cruz-Adalia, Aránzazu; Ramirez-Santiago, Guillermo; Calabia-Linares, Carmen; Torres-Torresano, Mónica; Feo, Lidia; Galán-Díez, Marta; Fernández-Ruiz, Elena; Pereiro, Eva; Guttmann, Peter; Chiappi, Michele; Schneider, Gerd; Carrascosa, José López; Chichón, Francisco Javier; Martínez Del Hoyo, Gloria; Sánchez-Madrid, Francisco; Veiga, Esteban

    2014-05-14

    Dendritic cells (DCs) phagocytose, process, and present bacterial antigens to T lymphocytes to trigger adaptive immunity. In vivo, bacteria can also be found inside T lymphocytes. However, T cells are refractory to direct bacterial infection, leaving the mechanisms by which bacteria invade T cells unclear. We show that T cells take up bacteria from infected DCs by the process of transinfection, which requires direct contact between the two cells and is enhanced by antigen recognition. Prior to transfer, bacteria localize to the immunological synapse, an intimate DC/T cell contact structure that activates T cells. Strikingly, T cells efficiently eliminate the transinfecting bacteria within the first hours after infection. Transinfected T cells produced high levels of proinflammatory cytokines and were able to protect mice from bacterial challenge following adoptive transfer. Thus, T lymphocytes can capture and kill bacteria in a manner reminiscent of innate immunity.

  13. Attachment of an anti-receptor antibody to non-target cells renders them susceptible to lysis by a clone of cytotoxic T lymphocytes.

    Science.gov (United States)

    Kranz, D M; Tonegawa, S; Eisen, H N

    1984-12-01

    The molecular basis for the dependence of antigen recognition by T cells on products of the major histocompatibility complex (MHC) is unknown, and the antigenic structures that are actually bound by T-cell receptors are ill-defined. In this study, we asked whether a monoclonal antibody (mAb) that reacts with the T-cell receptor of a clone of murine cytotoxic T lymphocytes (CTL) and not with the receptors of other CTL clones can substitute for that clone's natural ligand in specific cytolytic reactions. To answer the question, a mAb (1B2) to the receptor of a CTL clone (2C) was attached covalently to 51Cr-labeled cells that were not otherwise susceptible to lysis by clone 2C, and the cells thus modified were then tested as targets for clone 2C and other CTL clones of similar specificity. All labeled cells modified in this way, including a murine cell line that expresses no cell-surface MHC class I molecules and a human cell line, were lysed by clone 2C but not by other CTL clones. If, however, instead of attaching the mAb to the receptor of clone 2C, the cells were modified by attaching to them mAbs to other surface antigens on CTL [lymphocyte function-associated antigen (LFA-1), Thy-1.2], they were not lysed. In cytolytic titrations, the cells that had been converted by attachment of mAb 1B2 into specific targets for clone 2C were just as susceptible to lysis by that clone as the clone's natural H-2d targets (e.g., P815 cells). However, some accessory surface molecules (LFA-1, Lyt-2) that are required for clone 2C to lyse its natural H-2d targets seemed not to be required for this clone to lyse the mAb-converted target cells. By demonstrating that a variety of different cell types can be thus converted into target cells for CTL, the approach described in this study may provide opportunities to analyze further the mechanisms by which CTL destroy target cells.

  14. Associations of Europium(III) with gram-negative bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Ozaki, T.; Ohnuki, T. [Advanced Science Research Center, Japan Atomic Energy Research Institute, Tokai, Ibaraki 319-1195 (Japan); Kimura, T. [Department of Materials Science, Japan Atomic Energy Research Institute, Tokai, Ibaraki 319-1195 (Japan); Takahashi, Y. [Department of Earth and Planetary Systems Science, Hiroshima University, Hiroshima 739- 8526 (Japan); Francis, A.J. [Brookhaven National Laboratory, Upton, New York 11973 (United States)

    2005-07-01

    Full text of publication follows: Migration of radionuclides in the environment is greatly affected by bacteria. Gram-negative bacteria are ubiquitous in the environment and can preferentially bind radionuclides because of the presence of the cell envelop consisting of two membrane bilayers with an intervening thin peptidoglycan layer, where carboxyl and phosphate functional groups are mainly involved in metal cation adsorption. In this study, we investigated the association of Eu(III) with four Gram-negative bacteria Pseudomonas fluorescens, Alcaligenes faecalis, Shewanella putrefaciens, and Paracoccus denitrificans. Europium(III) is a good analogue of Am(III) and Cm(III). The association of Eu(III) with the bacteria were determined by time-resolved laser-induced fluorescence spectroscopy (TRLFS). The kinetics study showed that the Eu(III) adsorption on the bacteria proceeded rapidly. The Eu(III) adsorption on P. fluorescens at pH 3, A. faecalis and P. denitrificans at pHs 3, 4, and 5, and S. putrefaciens at pHs 4 and 5 reached a maximum within 5 minutes after contact. For P. denitrificans, the percent adsorption of Eu(III) decreased after the maximum percent adsorption was attained, which suggests the existence of exudates with an affinity with Eu(III). TRLFS showed that the coordination of Eu(III) on these bacteria is multi-dentate through an inner-spherical process. The ligand field of Eu(III) on P. denitrificans was as strong as the ones observed for halo-philic microorganisms, while that of P. fluorescens, A. faecalis, and S. putrefaciens was the typical one observed for non-halo-philic microorganisms. The coordination environment of Eu(III) on the bacteria differed from each other, though they are categorized as Gram-negative bacteria with the similar cell wall components. (authors)

  15. Deodorant bacteria; Des bacteries desodorisantes

    Energy Technology Data Exchange (ETDEWEB)

    Fanlo, J.L. [Ecole Nationale Superieure des Mines, 30 - Ales (France)

    1998-02-01

    Purifying bacteria: if this concept is not new, its application to gases cleansing has only been developed recently. This method allows to eliminate the volatile organic compounds and the gaseous effluents odors which come from industrial sites. Three bioreactors types exist at the present time. Their principles are explained. (O.M.) 6 refs.

  16. Functional genomics of intracellular bacteria.

    Science.gov (United States)

    de Barsy, Marie; Greub, Gilbert

    2013-07-01

    During the genomic era, a large amount of whole-genome sequences accumulated, which identified many hypothetical proteins of unknown function. Rapidly, functional genomics, which is the research domain that assign a function to a given gene product, has thus been developed. Functional genomics of intracellular pathogenic bacteria exhibit specific peculiarities due to the fastidious growth of most of these intracellular micro-organisms, due to the close interaction with the host cell, due to the risk of contamination of experiments with host cell proteins and, for some strict intracellular bacteria such as Chlamydia, due to the absence of simple genetic system to manipulate the bacterial genome. To identify virulence factors of intracellular pathogenic bacteria, functional genomics often rely on bioinformatic analyses compared with model organisms such as Escherichia coli and Bacillus subtilis. The use of heterologous expression is another common approach. Given the intracellular lifestyle and the many effectors that are used by the intracellular bacteria to corrupt host cell functions, functional genomics is also often targeting the identification of new effectors such as those of the T4SS of Brucella and Legionella.

  17. Hydrocarbon degradation by antarctic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Cavanagh, J.A.E.; Nichols, P.D.; McMeekin, T.A.; Franzmann, P.D. [Univ. of Tasmania (Australia)] [and others

    1996-12-31

    Bacterial cultures obtained from sediment samples collected during a trial oil spill experiment conducted at Airport beach, Eastern Antarctica were selectively enriched for n-alkane-degrading and phenanthrenedegrading bacteria. Samples were collected from a control site and sites treated with different hydrocarbon mixtures - Special Antarctic blend (SAB), BP-Visco and orange roughy oils. One set of replicate sites was also treated with water from Organic Lake which had previously been shown to contain hydrocarbon-degrading bacteria. No viable bacteria were obtained from samples collected from sites treated with orange roughy oil. Extensive degradation of n-alkanes by enrichment cultures obtained from sites treated with SAB and BP-Visco occurred at both 25{degrees}C and 10{degrees}C. Extensive degradation of phenanthrene also occurred in enrichment cultures from these sites grown at 25{degrees}C. Concurrent increases of polar lipid in these cultures were also observed. The presence of 1,4-naphthaquinone and 1-naphthol during the growth of the cultures on phenanthrene is unusual and warrants further investigation of the mechanism of phenanthrene-degradation by these Antarctic bacteria.

  18. Synthetic Biology in Streptomyces Bacteria

    NARCIS (Netherlands)

    Medema, Marnix H.; Breitling, Rainer; Takano, Eriko

    2011-01-01

    Actinomycete bacteria of the genus Streptomyces are major producers of bioactive compounds for the biotechnology industry. They are the source of most clinically used antibiotics, as well as of several widely used drugs against common diseases, including cancer . Genome sequencing has revealed that

  19. SYNTHETIC BIOLOGY IN STREPTOMYCES BACTERIA

    NARCIS (Netherlands)

    Medema, Marnix H.; Breitling, Rainer; Takano, Eriko; Voigt, C

    2011-01-01

    Actinomycete bacteria of the genus Streptomyces are major producers of bioactive compounds for the biotechnology industry. They are the source of most clinically used antibiotics, as well as of several widely used drugs against common diseases, including cancer. Genome sequencing has revealed that t

  20. Manipulating Genetic Material in Bacteria

    Science.gov (United States)

    1998-01-01

    Lisa Crawford, a graduate research assistant from the University of Toledo, works with Laurel Karr of Marshall Space Flight Center (MSFC) in the molecular biology laboratory. They are donducting genetic manipulation of bacteria and yeast for the production of large amount of desired protein. Photo credit: NASA/Marshall Space Flight Center (MSFC)