Characterizing aeroallergens by infrared spectroscopy of fungal spores and pollen.

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Boris Zimmermann

Full Text Available Fungal spores and plant pollen cause respiratory diseases in susceptible individuals, such as asthma, allergic rhinitis and hypersensitivity pneumonitis. Aeroallergen monitoring networks are an important part of treatment strategies, but unfortunately traditional analysis is time consuming and expensive. We have explored the use of infrared spectroscopy of pollen and spores for an inexpensive and rapid characterization of aeroallergens.The study is based on measurement of spore and pollen samples by single reflectance attenuated total reflectance Fourier transform infrared spectroscopy (SR-ATR FTIR. The experimental set includes 71 spore (Basidiomycota and 121 pollen (Pinales, Fagales and Poales samples. Along with fresh basidiospores, the study has been conducted on the archived samples collected within the last 50 years.The spectroscopic-based methodology enables clear spectral differentiation between pollen and spores, as well as the separation of confamiliar and congeneric species. In addition, the analysis of the scattering signals inherent in the infrared spectra indicates that the FTIR methodology offers indirect estimation of morphology of pollen and spores. The analysis of fresh and archived spores shows that chemical composition of spores is well preserved even after decades of storage, including the characteristic taxonomy-related signals. Therefore, biochemical analysis of fungal spores by FTIR could provide economical, reliable and timely methodologies for improving fungal taxonomy, as well as for fungal identification and monitoring. This proof of principle study shows the potential for using FTIR as a rapid tool in aeroallergen studies. In addition, the presented method is ready to be immediately implemented in biological and ecological studies for direct measurement of pollen and spores from flowers and sporocarps.

Airspora concentrations in the Vaal-triangle-monitoring and potential health-effects.2, fungal spores

CSIR Research Space (South Africa)

Vismer, HF

1995-08-01

Full Text Available Atmospheric fungal spores were monitored in Vanderbijlpark for the period 1991-92 as part of the Vaal triangle air pollution health study of the medical research council and the CSIR. Cladosporium, Aspergillus/ Penicillium, Alternaria and Epicoccum...

Annual distribution of allergenic fungal spores in atmospheric particulate matter in the Eastern Mediterranean; a comparative study between ergosterol and quantitative PCR analysis

Science.gov (United States)

Lang-Yona, N.; Dannemiller, K.; Yamamoto, N.; Burshtein, N.; Peccia, J.; Yarden, O.; Rudich, Y.

2012-03-01

Airborne fungal spores are an important fraction of atmospheric particulate matter and are major causative agents of allergenic and infectious diseases. Predicting the variability and species of allergy-causing fungal spores requires detailed and reliable methods for identification and quantification. There are diverse methods for their detection in the atmosphere and in the indoor environments; yet, it is important to optimize suitable methods for characterization of fungal spores in atmospheric samples. In this study we sampled and characterized total and specific airborne fungal spores from PM10 samples collected in Rehovot, Israel over an entire year. The total fungal spore concentrations vary throughout the year although the species variability was nearly the same. Seasonal equivalent spore concentrations analyzed by real-time quantitative-PCR-based methods were fall > winter > spring > summer. Reported concentrations based on ergosterol analysis for the same samples were and fall > spring > winter > summer. Correlation between the two analytical methods was found only for the spring season. These poor associations may be due to the per-spore ergosterol variations that arise from both varying production rates, as well as molecular degradation of ergosterol. While conversion of genome copies to spore concentration is not yet straightforward, the potential for improving this conversion and the ability of qPCR to identify groups of fungi or specific species makes this method preferable for environmental spore quantification. Identifying tools for establishing the relation between the presence of species and the actual ability to induce allergies is still needed in order to predict the effect on human health.

Meteorological factors associated with abundance of airborne fungal spores over natural vegetation

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Crandall, Sharifa G.; Gilbert, Gregory S.

2017-08-01

The abundance of airborne fungal spores in agricultural and urban settings increases with greater air temperature, relative humidity, or precipitation. The same meteorological factors that affect temporal patterns in spore abundance in managed environments also vary spatially across natural habitats in association with differences in vegetation structure. Here we investigated how temporal and spatial variation in aerial spore abundance is affected by abiotic (weather) and biotic (vegetation) factors as a foundation for predicting how fungi may respond to changes in weather and land-use patterns. We measured the phenology of airborne fungal spores across a mosaic of naturally occurring vegetation types at different time scales to describe (1) how spore abundance changes over time, (2) which local meteorological variables are good predictors for airborne spore density, and (3) whether spore abundance differs across vegetation types. Using an air volumetric vacuum sampler, we collected spore samples at 3-h intervals over a 120-h period in a mixed-evergreen forest and coastal prairie to measure diurnal, nocturnal, and total airborne spore abundance across vegetation types. Spore samples were also collected at weekly and monthly intervals in mixed-evergreen forest, redwood forest, and maritime chaparral vegetation types from 12 field sites across two years. We found greater airborne spore densities during the wetter winter months compared to the drier summer months. Mean total spore abundance in the mixed-evergreen forest was twice than in the coastal prairie, but there were no significant differences in total airborne spore abundance among mixed-evergreen forest, redwood forest, and maritime chaparral vegetation types. Weekly and monthly peaks in airborne spore abundance corresponded with rain events and peaks in soil moisture. Overall, temporal patterns in meteorological factors were much more important in determining airborne fungal spore abundance than the

Determination of fungal spore release from wet building materials

DEFF Research Database (Denmark)

Kildesø, J.; Wurtz, H.; Nielsen, Kristian Fog

2003-01-01

The release and transport of fungal spores from water-damaged building materials is a key factor for understanding the exposure to particles of fungal origin as a possible cause of adverse health effects associated to growth of fungi indoors. In this study, the release of spores from nine species...... of typical indoor fungi has been measured under controlled conditions. The fungi were cultivated for a period of 4-6 weeks on sterilized wet wallpapered gypsum boards at a relative humidity (RH) of approximately 97%. A specially designed small chamber (P-FLEC) was placed on the gypsum board. The release...

Significance of air humidity and air velocity for fungal spore release into the air

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Pasanen, A.-L.; Pasanen, P.; Jantunen, M. J.; Kalliokoski, P.

Our previous field studies have shown that the presence of molds in buildings does not necessarily mean elevated airborne spore counts. Therefore, we investigated the release of fungal spores from cultures of Aspergillus fumigatus, Penicillium sp. and Cladosporium sp. at different air velocities and air humidities. Spores of A. fumigatus and Penicillium sp. were released from conidiophores already at air velocity of 0.5 ms -1, whereas Cladosporium spores required at least a velocity of 1.0 ms -1. Airborne spore counts of A. fumigatus and Penicillium sp. were usually higher in dry than moist air, being minimal at relative humidities (r.h.) above 70%, while the effect of r.h. on the release of Cladosporium sp. was ambivalent. The geometric mean diameter of released spores increased when the r.h. exceeded a certain level which depends on fungal genus. Thus, spores of all three fungi were hygroscopic but the hygroscopicity of various spores appeared at different r.h.-ranges. This study indicates that spore release is controlled by external factors and depends on fungal genus which can be one reason for considerable variation of airborne spore counts in buildings with mold problems.

A Generic Method for Fungal Spore Detection: The use of a monoclonal antibody and surface plasmon resonance

DEFF Research Database (Denmark)

Skottrup, Peter; Hearty, Stephen; Frøkiær, Hanne

2005-01-01

This study describes a biosensing principle for detection of fungal spores using surface plasmon resonance (SPR). The approach involves the use of a monoclonal antibody (mab) and a SPR sensor for label-free detection of the model organism Puccinia striiformis f.sp. tritici (Pst) a biotrophic fungus...... causing wheat yellow rust. We have developed mabs towards intact whole spores and used a subtractive inhibition format for detection of spores in solution. The antibody was incubated with different spore concentrations and the remaining free antibody was quantified using a BIAcore® 3000 sensor. Decreasing...

Increased levels of airborne fungal spores to Populus tremuloides grown under elevated atmospheric CO{sub 2}

Energy Technology Data Exchange (ETDEWEB)

Klinoromos, J. N. [Guelph Univ., ON (Canada). Dept. of Botany; Rillig, M. C.; Allen, M. F. [San Diego State Univ., CA (United States). Dept. of Biology; Zak, D. R. [Michigan Univ., Ann Arbor, MI (United States). School of Natural Resources and Environment; Pregitzer, K. S.; Kubiske, M. E. [Michigan Technological Univ., Houghton, MI (United States). School of Forestry and Wood Products

1997-10-01

The objective of this study was to test the hypothesis that soil fungi sporulation would be facilitated by increase levels of CO{sub 2} concentration, leading to higher concentrations of fungal population in the atmosphere. Results showed that airborne fungal propagules were increased fourfold under twice-ambient CO{sub 2} concentration, and the decomposing leaf litter, the main source of fungal propagules, produced a fivefold increase of spores under elevated CO{sub 2} conditions. These results confirm the hypothesis that CO{sub 2} concentrations have a direct effect on microbial functions, which in turn will affect decomposition and plant pathogen dynamics. Since there is increasing evidence for causal relationship and exposure to aeroallergens and development of asthma in humans, there is a compelling need to study fungal epidemiology in the context of a globally changing environment. 28 refs., 3 figs.

A novel photosensitization treatment for the inactivation of fungal spores and cells mediated by curcumin.

Science.gov (United States)

Al-Asmari, Fahad; Mereddy, Ram; Sultanbawa, Yasmina

2017-08-01

The global concerns regarding the emergence of fungicide-resistant strains and the impact of the excessive use of fungicidal practises on our health, food, and environment have increased, leading to a demand for alternative clean green technologies as treatments. Photosensitization is a treatment that utilises a photosensitiser, light and oxygen to cause cell damage to microorganisms. The effect of photosensitization mediated by curcumin on Aspergillus niger, Aspergillus flavus, Penicillium griseofulvum, Penicillium chrysogenum, Fusarium oxysporum, Candida albicans and Zygosaccharomyces bailii was investigated using three methods. The viability of spores/cells suspended in aqueous buffer using different concentrations of curcumin solution (100-1000μM) and light dose (0, 24, 48, 72 and 96J/cm 2 ) were determined. Spraying curcumin solution on inoculated surfaces of agar plates followed by irradiation and soaking spores/cells in curcumin solution prior to irradiation was also investigated. In aqueous mixtures, photosensitised spores/cells of F. oxysporum and C. albicans were inhibited at all light doses and curcumin concentrations, while inactivation of A. niger, A. flavus P. griseofulvum, P. chrysogenum and Z. bailii were highly significant (Pcurcumin at 800μM showed complete inhibition for A. niger, F. oxysporum, C. albicans and Z. bailii, while A. flavus P. griseofulvum, and P. chrysogenum reduced by 75%, 80.4% and 88.5% respectively. Soaking spores/cells with curcumin solution prior to irradiation did not have a significant effect on the percentage reduction. These observations suggest that a novel photosensitization mediated curcumin treatment is effective against fungal spores/cells and the variation of percentage reduction was dependent on curcumin concentration, light dosage and fungal species. Copyright © 2017 Elsevier B.V. All rights reserved.

Fungal spores overwhelm biogenic organic aerosols in a midlatitudinal forest

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C. Zhu

2016-06-01

Full Text Available Both primary biological aerosol particles (PBAPs and oxidation products of biogenic volatile organic compounds (BVOCs contribute significantly to organic aerosols (OAs in forested regions. However, little is known about their relative importance in diurnal timescales. Here, we report biomarkers of PBAP and secondary organic aerosols (SOAs for their diurnal variability in a temperate coniferous forest in Wakayama, Japan. Tracers of fungal spores, trehalose, arabitol and mannitol, showed significantly higher levels in nighttime than daytime (p < 0.05, resulting from the nocturnal sporulation under near-saturated relative humidity. On the contrary, BVOC oxidation products showed higher levels in daytime than nighttime, indicating substantial photochemical SOA formation. Using tracer-based methods, we estimated that fungal spores account for 45 % of organic carbon (OC in nighttime and 22 % in daytime, whereas BVOC oxidation products account for 15 and 19 %, respectively. To our knowledge, we present for the first time highly time-resolved results that fungal spores overwhelmed BVOC oxidation products in contributing to OA especially in nighttime. This study emphasizes the importance of both PBAPs and SOAs in forming forest organic aerosols.

Demulsification of crude oil-in-water emulsions by means of fungal spores.

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Alba Adriana Vallejo-Cardona

Full Text Available The present feature describes for the first time the application of spores from Aspergillus sp. IMPMS7 to break out crude oil-in-water emulsions (O/W. The fungal spores were isolated from marine sediments polluted with petroleum hydrocarbons. The spores exhibited the ability to destabilize different O/W emulsions prepared with medium, heavy or extra-heavy Mexican crude oils with specific gravities between 10.1 and 21.2°API. The isolated fungal spores showed a high hydrophobic power of 89.3 ± 1.9% and with 2 g of spores per liter of emulsion, the half-life for emulsion destabilization was roughly 3.5 and 0.7 h for extra-heavy and medium crude oil, respectively. Then, the kinetics of water separation and the breaking of the O/W emulsion prepared with heavy oil through a spectrofluorometric technique were studied. A decrease in the fluorescence ratio at 339 and 326 nm (I339/I326 was observed in emulsions treated with spores, which is similar to previously reported results using chemical demulsifiers.

Monitoring of fungal spores in the indoor air of preschool institution facilities in Novi Sad

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Novaković Milana S.

2013-01-01

Full Text Available Fungal spores can cause a range of health problems in humans such as respiratory diseases and mycotoxicoses. Since children are the most vulnerable, the presence of fungal spores in the facilities of preschool and school institutions should be investigated readily. In order to estimate air contamination by fungal spores, air sampling was conducted in eight facilities of the preschool institution in Novi Sad during February and March, 2007. Sedimentation plate method was used for the detection of viable fungal spores, mostly being members of subdv. Deuteromycota (Fungi imperfecti. In 32 samples a total of 148 colonies were developed, among which five genera were identified: Penicillium, Cladosporium, Aspergillus, Alternaria and Acremonium while non-sporulating fungal colonies were labeled as sterile mycelia. Most frequently recorded genera were Penicillium with 46 colonies and Cladosporium with 44 colonies. The genera Aspergillus and Alternaria were represented with 3 colonies each and Acremonium with only 1 colony. The greatest number of colonies emerged in the samples from the day care facilities “Vendi” (58 colonies and “Panda” (49 colonies. Most diverse samples were obtained from the day care center “Zvončica”, with presence of all identified genera. These results showed notable presence of fungal spores in the indoor air of Preschool institution facilities and indicated the need for further, more complete seasonal research. Obtained information is considered useful for the evaluation of potential mycofactors that endanger health of children. [Projekat Ministarstva nauke Republike Srbije, br. III43002

Rarely reported fungal spores and structures: An overlooked source of probative trace evidence in criminal investigations.

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Hawksworth, David L; Wiltshire, Patricia E J; Webb, Judith A

2016-07-01

The value of pollen and plant spores as trace evidence has long been established, but it is only in the last eight years that fungal spores have been analysed routinely from the same palynological samples. They have greatly enhanced the specificity of links between people, objects, and places. Most fungal species occupy restricted ecological niches and their distributions can be limited both spatially and geographically. Spores may be dispersed over very short distances from the fungal sporophore,(1) and their presence in any palynological assemblage may indicate a restricted area of ground, or the presence of particular plants (even specific dead plant material). Fungal spores can represent primary, secondary, or even tertiary proxy evidence of a location, and can indicate the presence of a plant even though the plant is not obvious at a crime scene. In some cases, spores from fungi which have rarely been reported, and are considered to be rare, have been of particular value in providing intelligence or evidence of contact. Ten examples are given from case work in which rarely reported or unusual fungi have proved to be important in criminal investigations. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

A continental view of pine-associated ectomycorrhizal fungal spore banks: a quiescent functional guild with a strong biogeographic pattern.

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Glassman, Sydney I; Peay, Kabir G; Talbot, Jennifer M; Smith, Dylan P; Chung, Judy A; Taylor, John W; Vilgalys, Rytas; Bruns, Thomas D

2015-03-01

Ecologists have long acknowledged the importance of seed banks; yet, despite the fact that many plants rely on mycorrhizal fungi for survival and growth, the structure of ectomycorrhizal (ECM) fungal spore banks remains poorly understood. The primary goal of this study was to assess the geographic structure in pine-associated ECM fungal spore banks across the North American continent. Soils were collected from 19 plots in forests across North America. Fresh soils were pyrosequenced for fungal internal transcribed spacer (ITS) amplicons. Adjacent soil cores were dried and bioassayed with pine seedlings, and colonized roots were pyrosequenced to detect resistant propagules of ECM fungi. The results showed that ECM spore banks correlated strongly with biogeographic location, but not with the identity of congeneric plant hosts. Minimal community overlap was found between resident ECM fungi vs those in spore banks, and spore bank assemblages were relatively simple and dominated by Rhizopogon, Wilcoxina, Cenococcum, Thelephora, Tuber, Laccaria and Suillus. Similar to plant seed banks, ECM fungal spore banks are, in general, depauperate, and represent a small and rare subset of the mature forest soil fungal community. Yet, they may be extremely important in fungal colonization after large-scale disturbances such as clear cuts and forest fires. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

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

NARCIS (Netherlands)

Farenhorst, M.; Knols, B.G.J.

2010-01-01

Background: Interest in the use of fungal entomopathogens against malaria vectors is growing. Fungal spores infect insects via the cuticle and can be applied directly on the insect to evaluate infectivity. For flying insects such as mosquitoes, however, application of fungal suspensions on resting

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

NARCIS (Netherlands)

Farenhorst, Marit; Knols, Bart G. J.

2010-01-01

Interest in the use of fungal entomopathogens against malaria vectors is growing. Fungal spores infect insects via the cuticle and can be applied directly on the insect to evaluate infectivity. For flying insects such as mosquitoes, however, application of fungal suspensions on resting surfaces is

Forecasting methodologies for Ganoderma spore concentration using combined statistical approaches and model evaluations

Science.gov (United States)

Sadyś, Magdalena; Skjøth, Carsten Ambelas; Kennedy, Roy

2016-04-01

High concentration levels of Ganoderma spp. spores were observed in Worcester, UK, during 2006-2010. These basidiospores are known to cause sensitization due to the allergen content and their small dimensions. This enables them to penetrate the lower part of the respiratory tract in humans. Establishment of a link between occurring symptoms of sensitization to Ganoderma spp. and other basidiospores is challenging due to lack of information regarding spore concentration in the air. Hence, aerobiological monitoring should be conducted, and if possible extended with the construction of forecast models. Daily mean concentration of allergenic Ganoderma spp. spores in the atmosphere of Worcester was measured using 7-day volumetric spore sampler through five consecutive years. The relationships between the presence of spores in the air and the weather parameters were examined. Forecast models were constructed for Ganoderma spp. spores using advanced statistical techniques, i.e. multivariate regression trees and artificial neural networks. Dew point temperature along with maximum temperature was the most important factor influencing the presence of spores in the air of Worcester. Based on these two major factors and several others of lesser importance, thresholds for certain levels of fungal spore concentration, i.e. low (0-49 s m-3), moderate (50-99 s m-3), high (100-149 s m-3) and very high (150 < n s m-3), could be designated. Despite some deviation in results obtained by artificial neural networks, authors have achieved a forecasting model, which was accurate (correlation between observed and predicted values varied from r s = 0.57 to r s = 0.68).

Fungal spore concentrations in indoor and outdoor air in university libraries, and their variations in response to changes in meteorological variables.

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Flores, María Elena Báez; Medina, Pável Gaxiola; Camacho, Sylvia Páz Díaz; de Jesús Uribe Beltrán, Magdalena; De la Cruz Otero, María del Carmen; Ramírez, Ignacio Osuna; Hernández, Martín Ernesto Tiznado

2014-08-01

The fungal spore concentration (FSC) in the air poses a risk for human health. This work studied the FSC in university libraries and how it is affected by environmental factors. A total of 347 samples were obtained using a Microbio MB2(®) Aerosol Sampler. The wind speed (WS), cross wind (CW), temperature (T), relative humidity (HR), barometric pressure (BP) and dew point (DP) were recorded using a Kestrel(®) 4500 weather station. The median indoor/outdoor FSC was 360/1230 CFU m(-3). FSC correlated inversely with BP, HR and DP; and positively with WS and CW; whereas T showed negative or positive correlation with FSC, depending on the region or sampling time. Eleven fungal genera were found and the dominant isolates were identified as Aspergillus niger, Aspergillus tamarii and Aspergillus oryzae. All fungi identified are known to be allergenic. It was concluded that environmental variables can influence the air FSC in different ways.

Evaluation of hirst-type spore trap to monitor environmental fungal load in hospital.

Science.gov (United States)

Dananché, Cédric; Gustin, Marie-Paule; Cassier, Pierre; Loeffert, Sophie Tiphaine; Thibaudon, Michel; Bénet, Thomas; Vanhems, Philippe

2017-01-01

The main purpose was to validate the use of outdoor-indoor volumetric impaction sampler with Hirst-type spore traps (HTSTs) to continuously monitor fungal load in order to prevent invasive fungal infections during major structural work in hospital settings. For 4 weeks, outdoor fungal loads were quantified continuously by 3 HTSTs. Indoor air was sampled by both HTST and viable impaction sampler. Results were expressed as particles/m3 (HTST) or colony-forming units (CFU)/m3 (biocollector). Paired comparisons by day were made with Wilcoxon's paired signed-rank test or paired Student's t-test as appropriate. Paired airborne spore levels were correlated 2 by 2, after log-transformation with Pearson's cross-correlation. Concordance was calculated with kappa coefficient (κ). Median total fungal loads (TFLs) sampled by the 3 outdoor HTSTs were 3,025.0, 3,287.5 and 3,625.0 particles/m3 (P = 0.6, 0.6 and 0.3).-Concordance between Aspergillaceae fungal loads (AFLs, including Aspergillus spp. + Penicillium spp.) was low (κ = 0.2). A low positive correlation was found between TFLs sampled with outdoor HTST and indoor HTST with applying a 4-hour time lag, r = 0.30, 95% CI (0.23-0.43), PHTST-I on only 3.6% of the samples. Concordance between Aspergillus spp. loads and AFLs sampled with the 2 methods was very low (κ = 0.1). This study showed a 4-hour time lag between increase of outdoor and indoor TFLs, possibly due to insulation and aeraulic flow of the building. Outdoor HTSTs may permit to quickly identify (after 48 hours) time periods with high outdoor fungal loads. An identified drawback is that a too low sample area read did not seem to enable detection of Aspergillaceae spores efficiently. Indoor HTSTs may not be recommended at this time, and outdoor HTSTs need further study. Air sampling by viable impaction sampler remains the reference tool for quantifying fungal contamination of indoor air in hospitals.

Ectomycorrhizal fungal spore bank recovery after a severe forest fire: some like it hot.

Science.gov (United States)

Glassman, Sydney I; Levine, Carrie R; DiRocco, Angela M; Battles, John J; Bruns, Thomas D

2016-05-01

After severe wildfires, pine recovery depends on ectomycorrhizal (ECM) fungal spores surviving and serving as partners for regenerating forest trees. We took advantage of a large, severe natural forest fire that burned our long-term study plots to test the response of ECM fungi to fire. We sampled the ECM spore bank using pine seedling bioassays and high-throughput sequencing before and after the California Rim Fire. We found that ECM spore bank fungi survived the fire and dominated the colonization of in situ and bioassay seedlings, but there were specific fire adapted fungi such as Rhizopogon olivaceotinctus that increased in abundance after the fire. The frequency of ECM fungal species colonizing pre-fire bioassay seedlings, post-fire bioassay seedlings and in situ seedlings were strongly positively correlated. However, fire reduced the ECM spore bank richness by eliminating some of the rare species, and the density of the spore bank was reduced as evidenced by a larger number of soil samples that yielded uncolonized seedlings. Our results show that although there is a reduction in ECM inoculum, the ECM spore bank community largely remains intact, even after a high-intensity fire. We used advanced techniques for data quality control with Illumina and found consistent results among varying methods. Furthermore, simple greenhouse bioassays can be used to determine which fungi will colonize after fires. Similar to plant seed banks, a specific suite of ruderal, spore bank fungi take advantage of open niche space after fires.

Chlorine inactivation of fungal spores on cereal grains.

Science.gov (United States)

Andrews, S; Pardoel, D; Harun, A; Treloar, T

1997-04-01

Although 0.4% chlorine for 2 min has been recommended for surface disinfection of food samples before direct plating for fungal enumeration, this procedure may not be adequate for highly contaminated products. The effectiveness of a range of chlorine solutions was investigated using barley samples artificially contaminated with four different concentrations of Aspergillus flavus. A. niger, A. ochraceus, Eurotium repens, Penicillium brevicompactum P. chrysogenum and Cladosporium cladosporioides. At initial contamination levels greater than 10(4)/g, 0.4% chlorine did not inactivate sufficient spores to produce less than 20% contamination. Of the test fungi, ascospores of E. repens were the most resistant to chlorine inactivation, whereas the conidia of C. cladosporioides were the most sensitive. Rinsing the samples with 70% ethanol improved the effectiveness of the recommended surface disinfection procedure. However, some ethanol appears to permeate into the grains and may inactivate sensitive internal fungi, although a minimal effect only was observed on wheat infected with Alternaria.

Soil fungal and bacterial biomass determined by epifluorescence microscopy and mycorrhizal spore density in different sugarcane managements

Directory of Open Access Journals (Sweden)

Adriana Pereira Aleixo

2014-04-01

Full Text Available Crop productivity and sustainability have often been related to soil organic matter and soil microbial biomass, especially because of their role in soil nutrient cycling. This study aimed at measuring fungal and bacterial biomass by epifluorescence microscopy and arbuscular mycorrhizal fungal (AMF spore density in sugarcane (Saccharum officinarum L. fields under different managements. We collected soil samples of sugarcane fields managed with or without burning, with or without mechanized harvest, with or without application of vinasse and from nearby riparian native forest. The soil samples were collected at 10cm depth and storage at 4°C until analysis. Fungal biomass varied from 25 to 37µg C g-1 dry soil and bacterial from 178 to 263µg C g-1 dry soil. The average fungal/bacterial ratio of fields was 0.14. The AMF spore density varied from 9 to 13 spores g-1 dry soil. The different sugarcane managements did not affect AMF spore density. In general, there were no significant changes of microbial biomass with crop management and riparian forest. However, the sum of fungal and bacterial biomass measured by epifluorescence microscopy (i.e. 208-301µg C g-1 dry soil was very close to values of total soil microbial biomass observed in other studies with traditional techniques (e.g. fumigation-extraction. Therefore, determination of fungal/bacterial ratios by epifluorescence microscopy, associated with other parameters, appears to be a promising methodology to understand microbial functionality and nutrient cycling under different soil and crop managements.

Fungal spores in four catholic churches in the metropolitan area of Monterrey, Nuevo León State, Mexico – First study

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Alejandra Rocha Estrada

2015-05-01

Full Text Available Introduction. About 500,000 species of fungi have been described to-date, although an estimated between 1 – 1.5 million species may occur. They have a wide distribution in nature, contributing to the decomposition of organic matter and playing a part in the biogeochemical cycles of major nutrients. A small number are considered pathogens of animals and plants. There is ample historical evidence that certain types of allergies are associated with fungi; exposure to fungal allergens occurs in both outdoor and indoor spaces. Many indoor allergens are the same as those found outside buildings, entering through windows and doors, ventilation systems, or through cracks or other fissures in the walls. Objective. To determine the diversity and abundance of fungal spores inside four churches in the metropolitan area of Monterrey city in Mexico. Materials and methods. The study was carried out from July 2009 – January 2010 using a Hirst type volumetric collector (Burkard Manufacturing Co Ltd. Results. A total of 31,629 spores from 54 taxa were registered in the four churches. The building that showed the highest amount of spores was the Santa Catarina Mártir Church with 12,766 spores, followed by Cristo Rey with 7,155 and Nuestra Señora del Roble with 6,887. Regularly high concentrations of spores were recorded from 14:00 – 20:00 hours. The highest concentration value was observed at the church of Santa Catarina Mártir at 16:00 hours with 1153 spores/m 3 air. Conclusions. The most abundant spores in the four churches studied corresponded to Cladosporium, the [i]Aspergillus/Penicillium complex[/i], [i]Coprinus[/i], [i]Ganoderma[/i], [i]Curvularia and Ustilago[/i].

Evaluation of hirst-type spore trap to monitor environmental fungal load in hospital.

Directory of Open Access Journals (Sweden)

Cédric Dananché

Full Text Available The main purpose was to validate the use of outdoor-indoor volumetric impaction sampler with Hirst-type spore traps (HTSTs to continuously monitor fungal load in order to prevent invasive fungal infections during major structural work in hospital settings. For 4 weeks, outdoor fungal loads were quantified continuously by 3 HTSTs. Indoor air was sampled by both HTST and viable impaction sampler. Results were expressed as particles/m3 (HTST or colony-forming units (CFU/m3 (biocollector. Paired comparisons by day were made with Wilcoxon's paired signed-rank test or paired Student's t-test as appropriate. Paired airborne spore levels were correlated 2 by 2, after log-transformation with Pearson's cross-correlation. Concordance was calculated with kappa coefficient (κ. Median total fungal loads (TFLs sampled by the 3 outdoor HTSTs were 3,025.0, 3,287.5 and 3,625.0 particles/m3 (P = 0.6, 0.6 and 0.3.-Concordance between Aspergillaceae fungal loads (AFLs, including Aspergillus spp. + Penicillium spp. was low (κ = 0.2. A low positive correlation was found between TFLs sampled with outdoor HTST and indoor HTST with applying a 4-hour time lag, r = 0.30, 95% CI (0.23-0.43, P<0.001. In indoor air, Aspergillus spp. were detected by the viable impaction sampler on 63.1% of the samples, whereas AFLs were found by HTST-I on only 3.6% of the samples. Concordance between Aspergillus spp. loads and AFLs sampled with the 2 methods was very low (κ = 0.1. This study showed a 4-hour time lag between increase of outdoor and indoor TFLs, possibly due to insulation and aeraulic flow of the building. Outdoor HTSTs may permit to quickly identify (after 48 hours time periods with high outdoor fungal loads. An identified drawback is that a too low sample area read did not seem to enable detection of Aspergillaceae spores efficiently. Indoor HTSTs may not be recommended at this time, and outdoor HTSTs need further study. Air sampling by viable impaction sampler remains the

Surface Modified Long Period Fiber Grating Sensor for Rapid Detection of Aspergillus Niger Fungal Spores

Science.gov (United States)

Gambhir, Monika; Gupta, Shilpi; John, Priya; Mahakud, Ramakanta; Kumar, Jitendra; Prakash, Om

2018-03-01

We present development of a compact and label-free sensor based on the surface modification of copper vapor laser fabricated long period fiber gratings for detection of airborne Aspergillus niger (A. niger) fungal spores. Surface of sensors were functionalized with monoclonal glucose oxidases IgG1 for target-specific covalent binding. In process of functionalization and binding of 103 cfu/ml of pathogenic A. niger fungal spores, notable shorter wave transition in resonance wavelength from 1562.93 nm to 1555.97 nm, and significant reduction in peak loss from 61.72 dB to 57.48 dB were recorded. The implementation was cost effective and yielded instantaneous results.

Palynological investigation of the sediment cores from the Arabian Sea 1 Fungal spores

Digital Repository Service at National Institute of Oceanography (India)

Chandra, A.; Saxena, R.K.; Setty, M.G.A.P.

are new to the record @iInapertisporites@@ is represented in all the samples and is the most common element of the assemblage The distribution of various fungal spore genera and species in each core has been discussed...

Isolation and Identification of Air Borne Fungal Spores and Fragments in Buildings within Usmanu Danfodiyo University Sokoto, Nigeria

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Shinkafi Sa’adatu Aliyu

2014-07-01

Full Text Available Abstract - Indoor air contains a complex mixture of microorganisms, microorganism fragments, and by products such as molds, bacteria, endotoxins, mycotoxins, and volatile microbial organic compounds. Airborne fungi and bacteria can be toxic, allergenic and/or infectious. A research was conducted to determine the number and types of airborne fungal spores in Buildings of Usmanu Danfodiyo University Sokoto, Nigeria. Five (5 areas were chosen within the University for the Survey, these were student Hostel, Staff Quarters, Botanical garden, Microbiology laboratory and city campus of Usmanu Danfodiyo University. . A total number of fifteen (15 petri dishes containing potato dextrose agar each were vertically placed in each sampler and exposed at end of each height and site for 10 and 20 minutes respectively.  A total of thirteen (13 different fungal specie were identified namely; Aspergillus niger, A. flavus, A fumigates, A. ustus, A. terreus, Fusarium solani, F. oxysporum, Alterneria altenata, Rhizopus oryzae,  R. stolonifer, Helminthosporum sp., Penicillum candidum and Absedia corymbifera. Aspergillus niger had the highest frequency of occurrence of (14.9%, Helminthosporus species had the least frequency of occurrence of (1.5%. Conclusively it was observed that the concentration of fungal spores was high in the upper surface than the ground level at the time of the survey.

Selection of inactivation medium for fungal spores in clinical wastes by supercritical carbon dioxide.

Science.gov (United States)

Noman, Efaq; Norulaini Nik Ab Rahman, Nik; Al-Gheethi, Adel; Nagao, Hideyuki; Talip, Balkis A; Ab Kadir, Omar

2018-05-21

The present study aimed to select the best medium for inactivation of Aspergillus fumigatus, Aspergillus spp. in section Nigri, A. niger, A. terreus var. terreus, A. tubingensis, Penicillium waksmanii, P. simplicissimum, and Aspergillus sp. strain no. 145 spores in clinical wastes by using supercritical carbon dioxide (SC-CO 2 ). There were three types of solutions used including normal saline, seawater, distilled water, and physiological saline with 1% of methanol; each solution was tested at 5, 10, and 20 mL of the water contents. The experiments were conducted at the optimum operating parameters of supercritical carbon dioxide (30 MPa, 75 °C, 90 min). The results showed that the inactivation rate was more effective in distilled water with the presence of 1% methanol (6 log reductions). Meanwhile, the seawater decreases inactivation rate more than normal saline (4.5 vs. 5.1 log reduction). On the other hand, the experiments performed with different volumes of distilled water (5, 10, and 20 mL) indicated that A. niger spores were completely inactivated with 10 mL of distilled water. The inactivation rate of fungal spores decreased from 6 to 4.5 log as the amount of distilled water increased from 10 to 20 mL. The analysis for the spore morphology of A. fumigatus and Aspergillus spp. in section Nigri using scanning electron microscopy (SEM) has revealed the role of temperature and pressure in the SC-CO 2 in the destruction of the cell walls of the spores. It can be concluded that the distilled water represent the best medium for inactivation of fungal spores in the clinical solid wastes by SC-CO 2 .

High viscosity and anisotropy characterize the cytoplasm of fungal dormant stress resistant spores

NARCIS (Netherlands)

Dijksterhuis, J.; Nijsse, J.; Hoekstra, F.A.; Golovina, E.A.

2007-01-01

Ascospores of the fungus Talaromyces macrosporus are dormant and extremely stress resistant, whereas fungal conidia¿the main airborne vehicles of distribution¿are not. Here, physical parameters of the cytoplasm of these types of spores were compared. Cytoplasmic viscosity and level of anisotropy as

The effects of meteorological factors on the occurrence of Ganoderma sp. spores in the air

Science.gov (United States)

Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka

2011-03-01

Ganoderma sp. is an airborne fungal spore type known to trigger respiratory allergy symptoms in sensitive patients. Aiming to reduce the risk for allergic individuals, we analysed fungal spore circulation in Szczecin, Poland, and its dependence on meteorological conditions. Statistical models for the airborne spore concentrations of Ganoderma sp.—one of the most abundant fungal taxa in the area—were developed. Aerobiological sampling was conducted over 2004-2008 using a volumetric Lanzoni trap. Simultaneously, the following meteorological parameters were recorded: daily level of precipitation, maximum and average wind speed, relative humidity and maximum, minimum, average and dew point temperatures. These data were used as the explaining variables. Due to the non-linearity and non-normality of the data set, the applied modelling techniques were artificial neural networks (ANN) and mutlivariate regression trees (MRT). The obtained classification and MRT models predicted threshold conditions above which Ganoderma sp. appeared in the air. It turned out that dew point temperature was the main factor influencing the presence or absence of Ganoderma sp. spores. Further analysis of spore seasons revealed that the airborne fungal spore concentration depended only slightly on meteorological factors.

Resting spore formation of aphid-pathogenic fungus Pandora nouryi depends on the concentration of infective inoculum.

Science.gov (United States)

Huang, Zhi-Hong; Feng, Ming-Guang

2008-07-01

Resting spore formation of some aphid-pathogenic Entomophthorales is important for the seasonal pattern of their prevalence and survival but this process is poorly understood. To explore the possible mechanism involved in the process, Pandora nouryi (obligate aphid pathogen) interacted with green peach aphid Myzus persicae on cabbage leaves under favourable conditions. Host nymphs showered with primary conidia of an isolate (LC(50): 0.9-6.7 conidia mm(-2) 4-7 days post shower) from air captures in the low-latitude plateau of China produced resting spores (azygospores), primary conidia or both spore types. Surprisingly, the proportion of mycosed cadavers forming resting spores (P(CFRS)) increased sharply within the concentrations (C) of 28-240 conidia mm(-2), retained high levels at 240-1760, but was zero or extremely low at 0.3-16. The P(CFRS)-C relationship fit well the logistic equation P(CFRS) = 0.6774/[1 + exp(3.1229-0.0270C)] (r(2) = 0.975). This clarified for the first time the dependence of in vivo resting spore formation of P. nouryi upon the concentration of infective inoculum. A hypothesis is thus proposed that some sort of biochemical signals may exist in the host-pathogen interaction so that the fungal pathogen perceives the signals for prompt response to forthcoming host-density changes by either producing conidia for infecting available hosts or forming resting spores for surviving host absence in situ.

Mixed Production of Filamentous Fungal Spores for Preventing Soil-Transmitted Helminth Zoonoses: A Preliminary Analysis

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M. S. Arias

2013-01-01

Full Text Available Helminth zoonoses are parasitic infections shared by humans and animals, being the soil-transmitted helminths (STHs mainly caused by roundworms (ascarids and hookworms. This study was aimed to assess the individual and/or mixed production of two helminth-antagonistic fungi, one ovicide (Mucor circinelloides and other predator (Duddingtonia flagrans. Fungi were grown both in Petri plates and in a submerged culture (composed by water, NaCl, Na2HPO4 · 12 H2O, and wheat (Triticum aestivum. A Fasciola hepatica recombinant protein (FhrAPS was incorporated to the cultures to improve fungal production. All the cultured plates showed fungal growth, without difference in the development of the fungi when grown alone or mixed. High counts of Mucor spores were produced in liquid media cultures, and no significant differences were achieved regarding single or mixed cultures, or the incorporation of the FhrAPS. A significantly higher production of Duddingtonia spores after the incorporation of the FhrAPS was observed. When analyzing the parasiticide efficacy of the fungal mixture, viability of T. canis eggs reduced to 51%, and the numbers of third stage cyathostomin larvae reduced to 4%. It is concluded, the capability of a fungal mixture containing an ovicide (Mucor and a predator species (Duddingtonia for growing together in a submerged medium containing the FhrAPS offers a very interesting tool for preventing STHs.

Subseafloor basalts as fungal habitats

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M. Ivarsson

2012-09-01

Full Text Available The oceanic crust is believed to host the largest potential habitat for microbial life on Earth, yet, still we lack substantial information about the abundance, diversity, and consequence of its biosphere. The last two decades have involved major research accomplishments within this field and a change in view of the ocean crust and its potential to harbour life. Here fossilised fungal colonies in subseafloor basalts are reported from three different seamounts in the Pacific Ocean. The fungal colonies consist of various characteristic structures interpreted as fungal hyphae, fruit bodies and spores. The fungal hyphae are well preserved with morphological characteristics such as hyphal walls, septa, thallic conidiogenesis, and hyphal tips with hyphal vesicles within. The fruit bodies consist of large (∼50–200 µm in diameter body-like structures with a defined outer membrane and an interior filled with calcite. The fruit bodies have at some stage been emptied of their contents of spores and filled by carbonate-forming fluids. A few fruit bodies not filled by calcite and with spores still within support this interpretation. Spore-like structures (ranging from a few µm to ∼20 µm in diameter are also observed outside of the fruit bodies and in some cases concentrated to openings in the membrane of the fruit bodies. The hyphae, fruit bodies and spores are all closely associated with a crust lining the vein walls that probably represent a mineralized biofilm. The results support a fungal presence in deep subseafloor basalts and indicate that such habitats were vital between ∼81 and 48 Ma.

CONTROL OF POSTHARVEST TOMATO ROT BY SPORE SUSPENSION AND ANTIFUNGAL METABOLITES OF TRICHODERMA HARZIANUM

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Momein H. El-Katatny

2012-06-01

Full Text Available Rot of cherry tomato (Lycopersicon esculentum fruits caused by several fungal pathogens is a detrimental disease leading to substantial yield loses worldwide. Alternaria isolates were the most common fungal species isolated from healthy or rotten fruits. Trichoderma harzianum spore suspension and culture filtrate were tested for their antagonistic activity on controlling tomato fruit rot. T. harzianum isolates suppressed or interfered with the growth of different postharvest tomato fungal pathogens albeit at different degrees. Their culture filtrate inhibited pathogen spore germination possibly due to the released extracellular diffusible metabolite(s. Besides, aberrant morphology of conidia was observed with deformation of hyphal tips. Furthermore, the resulting mycelia appeared desiccated with coagulated protoplasm leading to complete collapse of protoplasm in presence of T. harzianum culture filtrate. Application of T. harzianum spores to tomato fruits decreased disease severity significantly with the most profound effect at higher spore concentrations (108 cells per ml. Similarly, culture filtrate of T. harzianum prevented pathogen spore germination on the surface of tomato fruits leading to decreased incidence of rot symptoms at high culture filtrate concentrations. This work provides strong evidence that T. harzianum is a competent antagonist and its spore suspension and culture filtrate can be used efficiently to control postharvest tomato rot.

Aerobyologic monitoring in urban and extra urban areas : analysis of airborne fungal particle concentration; Indagini aerobiologiche in ambiente urbano ed extraurbano componente fungina aerodiffusa

Energy Technology Data Exchange (ETDEWEB)

Bari, A. [ENEA, Saluggia (Italy). Centro Ricerche Energia; Caramiello, R.; Fossa, V.; Potenza, A. [Turin Univ. (Italy), Dip. di Biologia Vegetale

1995-12-01

In the environmental monitoring studies carried out by ENEA (Italian Agency for New Technologies, Energy and the Environment) a research about air quality of Turin and Saluggia (VC) has started in collaboration with the University of Turin. This research concerns the study of pollen and airborne fungal particle concentrations in the atmosphere. In the last few years the interest in airborne spores and hyphal fragments has increased. Indeed these particles play a very important role in allergic reactions, in several other human diseases, and in plant pathology. The incidence and the risk of infections by airborne fungal spores are correlated to the general climate and to the local micro climatic conditions; a complete aerosporological knowledge can be in many cases a useful indicator for infection risks. In this study the concentrations of airborne fungal spores in two different stations (urban station, Turin; and rural station, Saluggia (VC)) has been evaluated and compared, only for the year 1992, in order to verify the influence of climatic conditions and of floristic and vegetational aspects. In order to establish the correlations between the climate and the airborne fungal data, the results of aerosporogical analysis, relative to the Turin station, are reported. This investigation cover a six year period, three of which under standard climatic conditions and three with considerable peculiarities. A comparative evaluation of the efficiency in the collection of aerobiological particles (pollen and spores) by natural traps (mosses) has been attempted in Saluggia. The results obtained by this palinological analysis have been compared with the composition of the local flora and the data monitored by a spore-trap (volumetric pollen trap) positioned on the roof of a building in the ENEA Research Center of Saluggia.

Biomarkers of Aspergillus spores

Science.gov (United States)

Sulc, Miroslav; Peslova, Katerina; Zabka, Martin; Hajduch, Marian; Havlicek, Vladimir

2009-02-01

We applied both matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometric and 1D sodium dodecylsulfate polyacrylamide gel electrophoretic (1D-PAGE) approaches for direct analysis of intact fungal spores of twenty four Aspergillus species. In parallel, we optimized various protocols for protein extraction from Aspergillus spores using acidic conditions, step organic gradient and variable sonication treatment. The MALDI-TOF mass spectra obtained from optimally prepared samples provided a reproducible fingerprint demonstrating the capability of the MALDI-TOF approach to type and characterize different fungal strains within the Aspergillus genus. Mass spectra of intact fungal spores provided signals mostly below 20 kDa. The minimum material amount represented 0.3 [mu]g (10,000 spores). Proteins with higher molecular weight were detected by 1D-PAGEE Eleven proteins were identified from three selected strains in the range 5-25 kDa by the proteomic approach. Hemolysin and hydrophobin have the highest relevance in host-pathogen interactions.

A laboratory assessment of the Waveband Integrated Bioaerosol Sensor (WIBS-4) using individual samples of pollen and fungal spore material

Science.gov (United States)

Healy, David A.; O'Connor, David J.; Burke, Aoife M.; Sodeau, John R.

2012-12-01

A Bioaerosol sensing instrument referred to as WIBS-4, designed to continuously monitor ambient bioaerosols on-line, has been used to record a multiparameter “signature” from each of a number of Primary Biological Aerosol Particulate (PBAP) samples found in air. These signatures were obtained in a controlled laboratory environment and are based on the size, asymmetry (“shape”) and auto-fluorescence of the particles. Fifteen samples from two separate taxonomic ranks (kingdoms), Plantae (×8) and Fungi (×7) were individually introduced to the WIBS-4 for measurement along with two non-fluorescing chemical solids, common salt and chalk. Over 2000 individual-particle measurements were recorded for each sample type and the ability of the WIBS spectroscopic technique to distinguish between chemicals, pollen and fungal spore material was examined by identifying individual PBAP signatures. The results obtained show that WIBS-4 could potentially be a very useful analytical tool for distinguishing between natural airborne PBAP samples, such as the fungal spores and may potentially play an important role in detecting and discriminating the toxic fungal spore, Aspergillus fumigatus, from others in real-time. If the sizing range of the commercial instrument was customarily increased and permitted to operate simultaneously in its two sizing ranges, pollen and spores could potentially be discriminated between. The data also suggest that the gain setting sensitivity on the detector would also have to be reduced by a factor >5, to routinely detect, in-range fluorescence measurements for pollen samples.

Seasonal dynamics in arbuscular mycorrhizal fungal colonization and spore numbers in the rhizosphere of dactylis glomerata l. and trifolium repens L

International Nuclear Information System (INIS)

Xin, G.; Ye, S.; Wang, Y.; Wu, E.; Sugawara, K.

2012-01-01

The seasonal dynamics in the colonization of the rhizosphere of orchardgrass (Dactylis glomerata L.) and white clover (Trifolium repens L.) pastures by arbuscular mycorrhizal (AM) fungi and the production of spores in an artifical Japanese grassland was investigated over 12 months (between December 2001 and December 2002). The results showed that the AM fungal colonization fluctuated seasonally in the rhizosphere of both pastures. The total AM fungal colonization of the two pastures decreased during winter, then increased from March to June as the pastures grew, but slightly decreased again in July and August, and again followed an increase in September. There was significant difference of the colonization by arbuscules and vesicles between the two pastures ( p <0.05). Besides, the vesicular colonization of orchardgrass was higher than that of white clover, but the opposite trend was observed for arbuscular colonization. Similarly, the numbers of AM fungal spores in the pastures varied throughout the year, decreasing from spring to summer, then slowly increasing in late summer, reaching peak levels in winter. There is significant correlation between the frequency of spores in the rhizosphere soil and both soil temperature and pH. (author)

Atmospheric mold spore counts in relation to meteorological parameters

Science.gov (United States)

Katial, R. K.; Zhang, Yiming; Jones, Richard H.; Dyer, Philip D.

Fungal spore counts of Cladosporium, Alternaria, and Epicoccum were studied during 8 years in Denver, Colorado. Fungal spore counts were obtained daily during the pollinating season by a Rotorod sampler. Weather data were obtained from the National Climatic Data Center. Daily averages of temperature, relative humidity, daily precipitation, barometric pressure, and wind speed were studied. A time series analysis was performed on the data to mathematically model the spore counts in relation to weather parameters. Using SAS PROC ARIMA software, a regression analysis was performed, regressing the spore counts on the weather variables assuming an autoregressive moving average (ARMA) error structure. Cladosporium was found to be positively correlated (Pmodel was derived for Cladosporium spore counts using the annual seasonal cycle and significant weather variables. The model for Alternaria and Epicoccum incorporated the annual seasonal cycle. Fungal spore counts can be modeled by time series analysis and related to meteorological parameters controlling for seasonallity; this modeling can provide estimates of exposure to fungal aeroallergens.

A method for detecting fungal contaminants in wall cavities.

Science.gov (United States)

Spurgeon, Joe C

2003-01-01

This article describes a practical method for detecting the presence of both fungal spores and culturable fungi in wall cavities. Culturable fungi were collected in 25 mm cassettes containing 0.8 microm mixed cellulose ester filters using aggressive sampling conditions. Both culturable fungi and fungal spores were collected in modified slotted-disk cassettes. The sample volume was 4 L. The filters were examined microscopically and dilution plated onto multiple culture media. Collecting airborne samples in filter cassettes was an effective method for assessing wall cavities for fungal contaminants, especially because this method allowed the sample to be analyzed by both microscopy and culture media. Assessment criteria were developed that allowed the sample results to be used to classify wall cavities as either uncontaminated or contaminated. As a criterion, wall cavities with concentrations of culturable fungi below the limit of detection (LOD) were classified as uncontaminated, whereas those cavities with detectable concentrations of culturable fungi were classified as contaminated. A total of 150 wall cavities was sampled as part of a field project. The concentrations of culturable fungi were below the LOD in 34% of the samples, whereas Aspergillus and/or Penicillium were the only fungal genera detected in 69% of the samples in which culturable fungi were detected. Spore counting resulted in the detection of Stachybotrys-like spores in 25% of the samples that were analyzed, whereas Stachybotrys chartarum colonies were only detected on 2% of malt extract agar plates and on 6% of corn meal agar plates.

Airborne fungal and bacterial components in PM1 dust from biofuel plants.

Science.gov (United States)

Madsen, Anne Mette; Schlünssen, Vivi; Olsen, Tina; Sigsgaard, Torben; Avci, Hediye

2009-10-01

Fungi grown in pure cultures produce DNA- or RNA-containing particles smaller than spore size ( 3)-beta-D-glucans. In the 29 PM(1) samples, cultivable fungi were found in six samples and with a median concentration below detection level. Using microscopy, fungal spores were identified in 22 samples. The components NAGase and (1 --> 3)-beta-D-glucans, which are mainly associated with fungi, were present in all PM(1) samples. Thermophilic actinomycetes were present in 23 of the 29 PM(1) samples [average = 739 colony-forming units (CFU) m(-3)]. Cultivable and 'total bacteria' were found in average concentrations of, respectively, 249 CFU m(-3) and 1.8 x 10(5) m(-3). DNA- and RNA-containing particles of different lengths were counted by microscopy and revealed a high concentration of particles with a length of 0.5-1.5 microm and only few particles >1.5 microm. The number of cultivable fungi and beta-glucan in the total dust correlated significantly with the number of DNA/RNA-containing particles with lengths of between 1.0 and 1.5 microm, with DNA/RNA-containing particles >1.5 microm, and with other fungal components in PM(1) dust. Airborne beta-glucan and NAGase were found in PM(1) samples where no cultivable fungi were present, and beta-glucan and NAGase were found in higher concentrations per fungal spore in PM(1) dust than in total dust. This indicates that fungal particles smaller than fungal spore size are present in the air at the plants. Furthermore, many bacteria, including actinomycetes, were present in PM(1) dust. Only 0.2% of the bacteria in PM(1) dust were cultivable.

Parapiptadenia rigida MYCORRHIZATION WITH SPORES OF Scleroderma citrinum

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Gerusa Pauli Kist Steffen

2017-06-01

Full Text Available Ectomycorrhizal fungal inoculation in forestry seedlings aids plant establishment and growth in the field. The objectives of this study were: to determine the mycorrhizal capacity of the ectomycorrhizal fungus Scleroderma citrinum in Parapiptadenia rigida (red angico seedlings and to evaluate the viability of a mycorrhizal inoculation technique for forest seedlings involving the use of spores. Mature spores were inoculated in the substrate (75% soil and 25% carbonized rice husk, totaling 1.5 grams of fungal spores per liter of substrate. P. rigida seeds were sown in substrates inoculated or not inoculated with fungal spores in presence or absence of Pinus echinata and Eucalyptus citriodora essential oil: not inoculated (T1, inoculated (T2, inoculated more pine essential oil (T3, inoculated more eucalyptus essential oil (T4. Seedlings of Pinus elliottii were used for a positive control of mycorrhizal inoculation (T5 and not inoculated (T6 with fungal spores. At 90 days after sowing, the base stem diameter, height, fresh and dry weight of shoots and roots, percentage of root colonization and Dickson Index were determined. The presence of fungal structures in P. rigida and P. elliottii roots inoculated with S. citrinum spores was observed, demonstrating the occurrence of an ectomycorrhizal association. The application of pine and eucalyptus essential oils in the substrate increased the percentage of ectomycorrhizal colonization in P. rigida seedlings. The addition of S. citrinum mature spores in the substrate used for seedling production is a viable practice for ectomycorrhizal inoculation and it can be used in forest nurseries in controlled mycorrhization programs.

Fifth international fungus spore conference

Energy Technology Data Exchange (ETDEWEB)

Timberlake, W.E.

1993-04-01

This folio contains the proceedings of the Fifth International Fungal Spore Conference held August 17-21, 1991 at the Unicoi State Park at Helen, Georgia. The volume contains abstracts of each oral presentation as well as a collection of abstracts describing the poster sessions. Presentations were organized around the themes (1) Induction of Sporulation, (2) Nuclear Division, (3) Spore Formation, (4) Spore Release and Dispersal, and (4) Spore Germination.

Arbuscular mycorrhizal fungi spore propagation using single spore as starter inoculum and a plant host.

Science.gov (United States)

Selvakumar, G; Shagol, C C; Kang, Y; Chung, B N; Han, S G; Sa, T M

2018-06-01

The propagation of pure cultures of arbuscular mycorrhizal fungal (AMF) is an essential requirement for their large-scale agricultural application and commercialization as biofertilizers. The present study aimed to propagate AMF using the single-spore inoculation technique and compare their propagation ability with the known reference spores. Arbuscular mycorrhizal fungal spores were collected from salt-affected Saemangeum reclaimed soil in South Korea. The technique involved inoculation of sorghum-sudangrass (Sorghum bicolor L.) seedlings with single, healthy spores on filter paper followed by the transfer of successfully colonized seedlings to 1-kg capacity pots containing sterilized soil. After the first plant cycle, the contents were transferred to 2·5-kg capacity pots containing sterilized soil. Among the 150 inoculated seedlings, only 27 seedlings were colonized by AMF spores. After 240 days, among the 27 seedlings, five inoculants resulted in the production of over 500 spores. The 18S rDNA sequencing of spores revealed that the spores produced through single-spore inoculation method belonged to Gigaspora margarita, Claroideoglomus lamellosum and Funneliformis mosseae. Furthermore, indigenous spore F. mosseae M-1 reported a higher spore count than the reference spores. The AMF spores produced using the single-spore inoculation technique may serve as potential bio-inoculants with an advantage of being more readily adopted by farmers due to the lack of requirement of a skilled technique in spore propagation. The results of the current study describe the feasible and cost-effective method to mass produce AMF spores for large-scale application. The AMF spores obtained from this method can effectively colonize plant roots and may be easily introduced to the new environment. © 2018 The Society for Applied Microbiology.

Release and characteristics of fungal fragments in various conditions

Energy Technology Data Exchange (ETDEWEB)

Mensah-Attipoe, Jacob [Department of Environmental Science, University of Eastern Finland, Yliopistonranta 1D, P. O. Box 1627, FI-70211 Kuopio (Finland); Saari, Sampo [Department of Physics, Tampere University of Technology, Korkeakoulunkatu 3, 33720 Tampere (Finland); Veijalainen, Anna-Maria; Pasanen, Pertti [Department of Environmental Science, University of Eastern Finland, Yliopistonranta 1D, P. O. Box 1627, FI-70211 Kuopio (Finland); Keskinen, Jorma [Department of Physics, Tampere University of Technology, Korkeakoulunkatu 3, 33720 Tampere (Finland); Leskinen, Jari T.T. [SIB Labs, University of Eastern Finland, Yliopistonranta 1E, P. O. Box 1627, FI-70211, Kuopio (Finland); Reponen, Tiina, E-mail: reponeta@ucmail.uc.edu [Department of Environmental Science, University of Eastern Finland, Yliopistonranta 1D, P. O. Box 1627, FI-70211 Kuopio (Finland); Department of Environmental Health, University of Cincinnati, P.O. Box 670056, Cincinnati, OH 45267-0056 (United States)

2016-03-15

Intact spores and submicrometer size fragments are released from moldy building materials during growth and sporulation. It is unclear whether all fragments originate from fungal growth or if small pieces of building materials are also aerosolized as a result of microbial decomposition. In addition, particles may be formed through nucleation from secondary metabolites of fungi, such as microbial volatile organic compounds (MVOCs). In this study, we used the elemental composition of particles to characterize the origin of submicrometer fragments released from materials contaminated by fungi. Particles from three fungal species (Aspergillus versicolor, Cladosporium cladosporioides and Penicillium brevicompactum), grown on agar, wood and gypsum board were aerosolized using the Fungal Spore Source Strength Tester (FSSST) at three air velocities (5, 16 and 27 m/s). Released spores (optical size, d{sub p} ≥ 0.8 μm) and fragments (d{sub p} ≤ 0.8 μm) were counted using direct-reading optical aerosol instruments. Particles were also collected on filters, and their morphology and elemental composition analyzed using scanning electron microscopes (SEMs) coupled with an Energy-Dispersive X-ray spectroscopy (EDX). Among the studied factors, air velocity resulted in the most consistent trends in the release of fungal particles. Total concentrations of both fragments and spores increased with an increase in air velocity for all species whereas fragment–spore (F/S) ratios decreased. EDX analysis showed common elements, such as C, O, Mg and Ca, for blank material samples and fungal growth. However, N and P were exclusive to the fungal growth, and therefore were used to differentiate biological fragments from non-biological ones. Our results indicated that majority of fragments contained N and P. Because we observed increased release of fragments with increased air velocities, nucleation of MVOCs was likely not a relevant process in the formation of fungal fragments. Based

Plutonium uptake by a soil fungus and transport to its spores

International Nuclear Information System (INIS)

Beckert, W.F.; Au, F.H.F.

1976-01-01

Three concentrations of plutonium-238 nitrate, citrate and dioxide were each added to separate plates of malt agar buffered to pH 2.5 and 5.5 to determine the uptake of plutonium from these chemical forms and concentrations by a common soil fungus, Aspergillus niger. After inoculation and incubation, the aerial spores of Aspergillus niger were collected using a technique that excluded the possibility of cross-contamination of the spores by the culture media or by mycelial fragments. 238 Pu was taken up from all three chemical forms and transported to the aerial spores of Aspergillus niger at each concentration and at both pH levels. The specific activities of the spores grown at pH 5.5 were generally at least twice those of the spores grown at pH 2.5. The uptake of plutonium from the dioxide form was about one-third of that from the nitrate and citrate forms at both pH levels. The term 'transport factor' is used as a means to compare the transport of plutonium from the media to the fungal spores; the concentration-independent transport factor is defined as the specific activity of the spores divided by the specific activity of the dry culture medium. Though the transport factors were less than 1, which indicates discrimination against the transport of 238 Pu from the culture media to the spores, these findings suggest that this common soil fungus may be solubilizing soil-deposited plutonium and rendering it more biologically available for higher plants and animals. (author)

A Generic Method for Fungal Spore Detection: The use of a monoclonal antibody and surface plasmon resonance

DEFF Research Database (Denmark)

Skottrup, Peter; Hearty, Stephen; Frøkiær, Hanne

causing wheat yellow rust. We have developed mabs towards intact whole spores and used a subtractive inhibition format for detection of spores in solution. The antibody was incubated with different spore concentrations and the remaining free antibody was quantified using a BIAcore® 3000 sensor. Decreasing...

Fungal monitoring of the indoor air of the Museo de La Plata Herbarium, Argentina.

Science.gov (United States)

Mallo, Andrea C; Elíades, Lorena A; Nitiu, Daniela S; Saparrat, Mario C N

Biological agents, such as fungal spores in the air in places where scientific collections are stored, can attack and deteriorate them. The aim of this study was to gather information on the indoor air quality of the Herbarium of Vascular Plants of the Museo de Ciencias Naturales de La Plata, Argentina, in relation to fungal propagules and inert particles. This study was made using a volumetric system and two complementary sampling methods: (1) a non-viable method for direct evaluation, and (2) a viable method by culture for viable fungal propagules. The non-viable method led to ten spore morphotypes being found from related fungal sources. A total of 4401.88spores/m 3 and 32135.18 inert suspended particles/m 3 were recorded. The viable method led to the finding of nine fungal taxa as viable spores that mostly belonged to anamorphic forms of Ascomycota, although the pigmented yeast Rhodotorula F.C. Harrison (Basidiomycota) was also found. A total count of 40,500fungal CFU/m 3 air was estimated for all the sites sampled. Both the non-viable and viable sampling methods were necessary to monitor the bio-aerosol load in the La Plata Herbarium. The indoor air of this institution seems to be reasonably adequate for the conservation of vascular plants due to the low indoor/outdoor index, low concentrations of air spores, and/or lack of indicators of moisture problems. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

Spores of most common airborne fungi reveal no ice nucleation activity

Science.gov (United States)

Pummer, B. G.; Atanasova, L.; Bauer, H.; Bernardi, J.; Druzhinina, I. S.; Grothe, H.

2013-06-01

Fungal spores are ubiquitous biological aerosols, which are considered to show ice nucleation (IN) activity. In this study the respective IN activity was tested in oil emulsion in the immersion freezing mode. The focus was laid on species of economical, ecological or sanitary significance. For the first time, not only common moulds, but also edible mushrooms (Basidiomycota, Agaricomycetes) were investigated, as they contribute massively to the total amount of fungal spores in the atmosphere. Only Fusarium avenaceum showed freezing events at low subzero-temperatures, while the other investigated fungal spores showed no significant IN activity. Furthermore, we selected a set of fungal strains from different sites and exposed them to occasional freezing stress during cultivation. Although the total protein expression was altered by this treatment, it had no significant impact on the IN activity.

Selection of Infective Arbuscular Mycorrhizal Fungal Isolates for Field Inoculation

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Elisa Pellegrino

2010-09-01

Full Text Available Arbuscular mycorrhizal (AM fungi play a key role in host plant growth and health, nutrient and water uptake, plant community diversity and dynamics. AM fungi differ in their symbiotic performance, which is the result of the interaction of two fungal characters, infectivity and efficiency. Infectivity is the ability of a fungal isolate to establish rapidly an extensive mycorrhizal symbiosis and is correlated with pre-symbiotic steps of fungal life cycle, such as spore germination and hyphal growth. Here, different AM fungal isolates were tested, with the aim of selecting infective endophytes for field inoculation. Greenhouse and microcosm experiments were performed in order to assess the ability of 12 AM fungal isolates to produce spores, colonize host roots and to perform initial steps of symbiosis establishment, such as spore germination and hyphal growth. AM fungal spore production and root colonization were significantly different among AM fungal isolates. Spore and sporocarp densities ranged from 0.8 to 7.4 and from 0.6 to 2.0 per gram of soil, respectively, whereas root colonization ranged from 2.9 to 72.2%. Percentage of spore or sporocarp germination ranged from 5.8 to 53.3% and hyphal length from 4.7 to 79.8 mm. The ordination analysis (Redundancy Analysis, RDA showed that environmental factors explained about 60% of the whole variance and their effect on fungal infectivity variables was significant (P = 0.002. The biplot clearly showed that variables which might be used to detect infective AM fungal isolates were hyphal length and root colonization. Such analysis may allow the detection of the best parameters to select efficient AM fungal isolates to be used in agriculture.

Fifth international fungus spore conference. [Abstracts]: Final technical report

Energy Technology Data Exchange (ETDEWEB)

Timberlake, W.E.

1993-04-01

This folio contains the proceedings of the Fifth International Fungal Spore Conference held August 17-21, 1991 at the Unicoi State Park at Helen, Georgia. The volume contains abstracts of each oral presentation as well as a collection of abstracts describing the poster sessions. Presentations were organized around the themes (1) Induction of Sporulation, (2) Nuclear Division, (3) Spore Formation, (4) Spore Release and Dispersal, and (4) Spore Germination.

Surface tension propulsion of fungal spores by use of microdroplets

OpenAIRE

Noblin, Xavier; Yang, Sylvia; Dumais, Jacques

2010-01-01

Many edible mushrooms eject their spores (about 10 microns in size) at high speed (about 1 m/s) using surface tension forces in a few microseconds. Basically the coalescence of a droplet with the spore generates the necessary momentum to eject the spore. We have detailed this mechanism in \\cite{noblin2}. In this article, we give some details about the high speed movies (up to 250000 fps) of mushrooms' spores ejection attached to this submission. This video was submitted as part of the Gallery...

Variaciones en la estacionalidad de polen y esporas fúngicas en la atmósfera de la ciudad de La Plata (Argentina Seasonal variation of pollen and fungal spores in the atmosphere of Plata city (Argentina

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Daniela S Nitiu

2011-12-01

Full Text Available Se presentan los primeros datos sobre variaciones estacionales de polen y esporas fúngicas en su conjunto en la atmósfera de la ciudad de La Plata durante julio 2000 / junio 2001. El total de bioaerosoles fue de 201889.15 de los cuales el 15% correspondió a 54 tipos polínicos y el 85% a 79 tipos esporales. El Índice Polínico estuvo dado en un 74% por Polen Arbóreo (PA y en un 26% por Polen Herbáceo (PNA. La máxima concentración de PA tuvo lugar en septiembre con granos provenientes de Platanus y Fraxinus. El PNA presentó dos picos, el primero en diciembre procedente de Poaceae y el segundo en marzo debido a Ambrosia. Con respecto al componente fúngico, se identificaron esporas de Oomycota, Zygomycota, Myxomycota, Ascomycota y Basidiomycota. Estos bioaerosoles estuvieron presentes durante todo el año y en altas concentraciones siendo más abundantes en verano - otoño Las mitosporas de Hongos Imperfectos dominan la nube superando el 44% del total del Índice Esporal. El máximo aporte se registró en febrero dado por las esporas de Cladosporium. Se identificaron dos períodos estacionales en los cuales se hallan simultáneamente altas concentraciones de polen y esporas fúngicas. El primero se desarrolla en primavera y está integrado por Platanus, Fraxinus y Aspergillus / Penicillium y el segundo es estival e involucra a Ambrosia, Alternaria y Cladosporium.We present the first data on representativeness and seasonal variation of pollen and fungal spores as a whole in the atmosphere of the city of La Plata during july 2000 / june 2001. Total was 201889.15 bioaerosols, 15% of which was contributed by 54 pollen types and 85% by 79 spore types. The Pollinic Index came from arboreal pollen (PA in 74% and herbaceous pollen (PNA in 26%. The PA maximum concentration took place in september due to Platanus and Fraxinus. The PNA presented two peaks, one in december due to Poaceae, and the other one in march from Ambrosia. In regard to the

Occurrence of Cladosporium spp. and Alternaria spp. spores in Western, Northern and Central-Eastern Poland in 2004-2006 and relation to some meteorological factors

Science.gov (United States)

Grinn-Gofroń, Agnieszka; Rapiejko, Piotr

2009-08-01

The concentration of airborne spores of Cladosporium spp. and Alternaria spp. has been investigated at three monitoring stations situated along the west-north and central-east transect in Poland (Szczecin, Olsztyn, Warszawa,) i.e. from a height of 100 m to 149 m above sea level. The aerobiological monitoring of fungal spores was performed by means of three Lanzoni volumetric spore traps. Cladosporium spp. spores were dominant at all the stations. The highest Cladosporium spp. and Alternaria spp. numbers of spores were observed at all the cities in July and August. Statistically significant correlations have been found between the Cladosporium spp. and Alternaria spp. concentration in the air and the mean air temperature, amount of precipitation, air pressure and relative air humidity. The spore count of Cladosporium spp. and Alternaria spp. is determined by the diversity of local flora and weather conditions, especially by the air temperature. The identification of factors, which influence and shape spore concentrations, may significantly improve the current methods of allergy prevention.

Seasonal variation of Ganoderma spore concentrations in urban and suburban districts of the city of Szczecin, Poland.

Science.gov (United States)

Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka; Przestrzelska, Katarzyna

2015-01-01

According to recent studies, Ganoderma may be the third genus, after Alternaria and Cladosporium, the spores of which cause symptoms of allergy, and concentration is related to meteorological factors. The aerobiology of Ganoderma spores in Szczecin in urban and suburban districts was examined using Lanzoni Volumetric Spore Traps in 2008-2010. Ganoderma spores were present in the atmosphere on more than 90% of the days from June through September with peak concentrations in June, July and September. The number of days with spores was lower in the suburban district, while the total number of spores collected was higher there than in the urban district. Correlation and multiple regression analyses revealed weak relationships between Ganoderma and meteorological conditions, while testing the significance of differences between the districts showed that urban development did not have a clear impact on the values of meteorological parameters. A significantly higher abundance of spores in the suburbs of Szczecin seemed to be conditioned by the closeness of potential area sources. This study indicates that a single measuring site in the city centre insufficiently reflected the dynamics and level of Ganoderma spore concentration in peripheral districts.

Prophenoloxidase-Mediated Ex Vivo Immunity to Delay Fungal Infection after Insect Ecdysis

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Jie Zhang

2017-11-01

Full Text Available Skin immunity protects animals from airborne pathogen infection. Unlike mammals, arthropods, including insects, undergo periodic ecdysis to grow and develop. Newly molted insects emerge with unsclerotized thin cuticles but successfully escape pathogenic infections during the post-molt period. Here we show that prophenoloxidases (PPOs in molting fluids remain bioactive on the integument and impede fungal infection after ecdysis. We found that the purified plasma PPOs or recombinant PPOs could effectively bind to fungal spores (conidia by targeting the cell wall components chitin and β-1,3-glucan. Pretreatment of the spores of the fungal pathogen Beauveria bassiana with PPOs increased spore hydrophilicity and reduced spore adhesion activity, resulting in a significant decrease in virulence as compared with mock infection. We also identified a spore-secreted protease BPS8, a member of peptidase S8 family of protease that degrade PPOs at high levels to benefit fungal infection, but which at lower doses activate PPOs to inhibit spore germination after melanization. These data indicate that insects have evolved a distinct strategy of ex vivo immunity to survive pathogen infections after ecdysis using PPOs in molting fluids retained on the underdeveloped and tender integument of newly molted insects for protection against airborne fungal infection.

Weed control and cover crop management affect mycorrhizal colonization of grapevine roots and arbuscular mycorrhizal fungal spore populations in a California vineyard.

Science.gov (United States)

Baumgartner, Kendra; Smith, Richard F; Bettiga, Larry

2005-03-01

Arbuscular mycorrhizal (AM) fungi naturally colonize grapevines in California vineyards. Weed control and cover cropping may affect AM fungi directly, through destruction of extraradical hyphae by soil disruption, or indirectly, through effects on populations of mycorrhizal weeds and cover crops. We examined the effects of weed control (cultivation, post-emergence herbicides, pre-emergence herbicides) and cover crops (Secale cereale cv. Merced rye, x Triticosecale cv.Trios 102) on AM fungi in a Central Coast vineyard. Seasonal changes in grapevine mycorrhizal colonization differed among weed control treatments, but did not correspond with seasonal changes in total weed frequency. Differences in grapevine colonization among weed control treatments may be due to differences in mycorrhizal status and/or AM fungal species composition among dominant weed species. Cover crops had no effect on grapevine mycorrhizal colonization, despite higher spring spore populations in cover cropped middles compared to bare middles. Cover crops were mycorrhizal and shared four AM fungal species (Glomus aggregatum, G. etunicatum, G. mosseae, G. scintillans) in common with grapevines. Lack of contact between grapevine roots and cover crop roots may have prevented grapevines from accessing higher spore populations in the middles.

Spore Acquisition and Survival of Ambrosia Beetles Associated with the Laurel Wilt Pathogen in Avocados after Exposure to Entomopathogenic Fungi.

Science.gov (United States)

Avery, Pasco B; Bojorque, Verónica; Gámez, Cecilia; Duncan, Rita E; Carrillo, Daniel; Cave, Ronald D

2018-04-25

Laurel wilt is a disease threatening the avocado industry in Florida. The causative agent of the disease is a fungus vectored by ambrosia beetles that bore into the trees. Until recently, management strategies for the vectors of the laurel wilt fungus relied solely on chemical control and sanitation practices. Beneficial entomopathogenic fungi (EPF) are the most common and prevalent natural enemies of pathogen vectors. Laboratory experiments demonstrated that commercial strains of EPF can increase the mortality of the primary vector, Xyleborus glabratus , and potential alternative vectors, Xylosandrus crassiusculus , Xyleborus volvulus and Xyleborus bispinatus (Coleoptera: Curculionidae: Scolytinae). Our study provides baseline data for three formulated commercially-available entomopathogenic fungi used as potential biocontrol agents against X. crassiusculus , X. volvulus and X. bispinatus. The specific objectives were to determine: (1) the mean number of viable spores acquired per beetle species adult after being exposed to formulated fungal products containing different strains of EPF ( Isaria fumosorosea , Metarhizium brunneum and Beauveria bassiana ); and (2) the median and mean survival times using paper disk bioassays. Prior to being used in experiments, all fungal suspensions were adjusted to 2.4 × 10⁶ viable spores/mL. The number of spores acquired by X. crassiusculus was significantly higher after exposure to B. bassiana , compared to the other fungal treatments. For X. volvulus , the numbers of spores acquired per beetle were significantly different amongst the different fungal treatments, and the sequence of spore acquisition rates on X. volvulus from highest to lowest was I. fumosorosea > M. brunneum > B. bassiana . After X. bispinatus beetles were exposed to the different suspensions, the rates of acquisition of spores per beetle amongst the different fungal treatments were similar. Survival estimates (data pooled across two tests) indicated an

Comparison of methods to evaluate the fungal biomass in heating, ventilation, and air-conditioning (HVAC) dust.

Science.gov (United States)

Biyeyeme Bi Mve, Marie-Jeanne; Cloutier, Yves; Lacombe, Nancy; Lavoie, Jacques; Debia, Maximilien; Marchand, Geneviève

2016-12-01

Heating, ventilation, and air-conditioning (HVAC) systems contain dust that can be contaminated with fungal spores (molds), which may have harmful effects on the respiratory health of the occupants of a building. HVAC cleaning is often based on visual inspection of the quantity of dust, without taking the mold content into account. The purpose of this study is to propose a method to estimate fungal contamination of dust in HVAC systems. Comparisons of different analytical methods were carried out on dust deposited in a controlled-atmosphere exposure chamber. Sixty samples were analyzed using four methods: culture, direct microscopic spore count (DMSC), β-N-acetylhexosaminidase (NAHA) dosing and qPCR. For each method, the limit of detection, replicability, and repeatability were assessed. The Pearson correlation coefficients between the methods were also evaluated. Depending on the analytical method, mean spore concentrations per 100 cm 2 of dust ranged from 10,000 to 682,000. Limits of detection varied from 120 to 217,000 spores/100 cm 2 . Replicability and repeatability were between 1 and 15%. Pearson correlation coefficients varied from -0.217 to 0.83. The 18S qPCR showed the best sensitivity and precision, as well as the best correlation with the culture method. PCR targets only molds, and a total count of fungal DNA is obtained. Among the methods, mold DNA amplification by qPCR is the method suggested for estimating the fungal content found in dust of HVAC systems.

Reduced aboveground tree growth associated with higher arbuscular mycorrhizal fungal diversity in tropical forest restoration.

Science.gov (United States)

Holste, Ellen K; Holl, Karen D; Zahawi, Rakan A; Kobe, Richard K

2016-10-01

Establishing diverse mycorrhizal fungal communities is considered important for forest recovery, yet mycorrhizae may have complex effects on tree growth depending on the composition of fungal species present. In an effort to understand the role of mycorrhizal fungi community in forest restoration in southern Costa Rica, we sampled the arbuscular mycorrhizal fungal (AMF) community across eight sites that were planted with the same species ( Inga edulis, Erythrina poeppigiana, Terminalia amazonia, and Vochysia guatemalensis ) but varied twofold to fourfold in overall tree growth rates. The AMF community was measured in multiple ways: as percent colonization of host tree roots, by DNA isolation of the fungal species associated with the roots, and through spore density, volume, and identity in both the wet and dry seasons. Consistent with prior tropical restoration research, the majority of fungal species belonged to the genus Glomus and genus Acaulospora , accounting for more than half of the species and relative abundance found on trees roots and over 95% of spore density across all sites. Greater AMF diversity correlated with lower soil organic matter, carbon, and nitrogen concentrations and longer durations of prior pasture use across sites. Contrary to previous literature findings, AMF species diversity and spore densities were inversely related to tree growth, which may have arisen from trees facultatively increasing their associations with AMF in lower soil fertility sites. Changes to AMF community composition also may have led to variation in disturbance susceptibility, host tree nutrient acquisition, and tree growth. These results highlight the potential importance of fungal-tree-soil interactions in forest recovery and suggest that fungal community dynamics could have important implications for tree growth in disturbed soils.

Spore Acquisition and Survival of Ambrosia Beetles Associated with the Laurel Wilt Pathogen in Avocados after Exposure to Entomopathogenic Fungi

Directory of Open Access Journals (Sweden)

Pasco B. Avery

2018-04-01

Full Text Available Laurel wilt is a disease threatening the avocado industry in Florida. The causative agent of the disease is a fungus vectored by ambrosia beetles that bore into the trees. Until recently, management strategies for the vectors of the laurel wilt fungus relied solely on chemical control and sanitation practices. Beneficial entomopathogenic fungi (EPF are the most common and prevalent natural enemies of pathogen vectors. Laboratory experiments demonstrated that commercial strains of EPF can increase the mortality of the primary vector, Xyleborus glabratus, and potential alternative vectors, Xylosandrus crassiusculus, Xyleborus volvulus and Xyleborus bispinatus (Coleoptera: Curculionidae: Scolytinae. Our study provides baseline data for three formulated commercially-available entomopathogenic fungi used as potential biocontrol agents against X. crassiusculus, X. volvulus and X. bispinatus. The specific objectives were to determine: (1 the mean number of viable spores acquired per beetle species adult after being exposed to formulated fungal products containing different strains of EPF (Isaria fumosorosea, Metarhizium brunneum and Beauveria bassiana; and (2 the median and mean survival times using paper disk bioassays. Prior to being used in experiments, all fungal suspensions were adjusted to 2.4 × 106 viable spores/mL. The number of spores acquired by X. crassiusculus was significantly higher after exposure to B. bassiana, compared to the other fungal treatments. For X. volvulus, the numbers of spores acquired per beetle were significantly different amongst the different fungal treatments, and the sequence of spore acquisition rates on X. volvulus from highest to lowest was I. fumosorosea > M. brunneum > B. bassiana. After X. bispinatus beetles were exposed to the different suspensions, the rates of acquisition of spores per beetle amongst the different fungal treatments were similar. Survival estimates (data pooled across two tests indicated an

Fungal endophytes characterization from four species of Diplazium Swartz

Science.gov (United States)

Affina-Eliya, A. A.; Noraini, T.; Nazlina, I.; Ruzi, A. R.

2014-09-01

Four species on genus Diplazium namely Diplazium tomentosum, D. sorzogonense, D. asperum and D. accedens of Peninsular Malaysia were studied for presence of fungal endophyte. The objective of this study is to characterize fungal endophytes in the rhizome of four Diplazium species. The rhizome was surface sterilized and incubated to isolate fungal endophytes. Characterization of the colonies was performed by macroscopic morphological, microscopic identification, types of hyphae and mycelium, and spore structure. For isolation that produces spores, the structure of conidiophores and conidia were identified. From this study, four fungal have been isolated and determined as Aspergillus sp. (isolates AE 1), Aspergillus fumigatus (isolates AE 2), Aspergillus versicolor (isolates AE 3) and Verticillium sp. (isolates AE 4). The fungal isolates from this study were classified from the same family Moniliaceae.

Biodiversity and Concentration of Airborne Fungi of Suburban Weekly Market Associated Environment

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Apurva K. Pathak

2012-04-01

Full Text Available It is supposed that the vegetable markets of tropical countries are the significant source of airborne fungal spores, which are generated during transportation, handling and putrefaction. The aim of this work was to monitor the prevalence of the airborne cultivable fungi in the air of weekly market associated environments to evaluate whether the vegetable trading zone of market is a source of airborne cultivable fungal spores of weekly market environment or not. Airborne cultivable fungal spore levels were monitored by using Andersen two-stage viable (microbial particle sampler. The Spearman correlation coefficients and stepwise linear regression analysis test was used to analyze the influence of meteorological factors on spore concentration and paired Student’s t-test was used to compare the bioload of total viable cultivable fungi of vegetable trading area and general item trading area of weekly market extramural environment, the percentage frequency and the percentage contribution of the individual genus was also reported. In both areas, Aspergillus, Cladosporium, Alternaria, and Penicillium, were the most abundant fungal types observed. The spp. of Candida was reported only at the vegetable trading area. The bioload of fungal spore presented maximum values during the Monsoon and lowest in the season of summer. There is no significant difference in quantity between the mean values of the bioload of total viable cultivable fungi of vegetable trading area and general item trading area of weekly market extramural environment were observed. For present environment, activities of animals and humans were supposed to be the key factor governing aerosolization of microorganism.

Seasonal variation of [i]Ganoderma[/i] spore concentrations in urban and suburban districts of the city of Szczecin, Poland

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Agnieszka Grinn-Gofroń

2015-02-01

Full Text Available According to recent studies,[i] Ganoderma[/i] may be the third genus, after [i]Alternaria[/i] and [i]Cladosporium[/i], the spores of which cause symptoms of allergy, and concentration is related to meteorological factors. The aerobiology of [i]Ganoderma[/i] spores in Szczecin in urban and suburban districts was examined using Lanzoni Volumetric Spore Traps in 2008–2010. [i]Ganoderma[/i] spores were present in the atmosphere on more than 90% of the days from June through September with peak concentrations in June, July and September. The number of days with spores was lower in the suburban district, while the total number of spores collected was higher there than in the urban district. Correlation and multiple regression analyses revealed weak relationships between [i]Ganoderma[/i] and meteorological conditions, while testing the significance of differences between the districts showed that urban development did not have a clear impact on the values of meteorological parameters. A significantly higher abundance of spores in the suburbs of Szczecin seemed to be conditioned by the closeness of potential area sources. This study indicates that a single measuring site in the city centre insufficiently reflected the dynamics and level of [i]Ganoderma[/i] spore concentration in peripheral districts.

Flavonoids released naturally from alfalfa promote development of symbiotic glomus spores in vitro.

Science.gov (United States)

Tsai, S M; Phillips, D A

1991-05-01

Because flavonoids from legumes induce transcription of nodulation genes in symbiotic rhizobial bacteria, it is reasonable to test whether these compounds alter the development of vesicular-arbuscular mycorrhizal (VAM) fungi that infect those plants. Quercetin-3-O-galactoside, the dominant flavonoid released naturally from alfalfa (Medicago sativa L.) seeds, promoted spore germination of Glomus etunicatum and Glomus macrocarpum in vitro. Quercetin produced the maximum increases in spore germination, hyphal elongation, and hyphal branching in G. etunicatum at 1 to 2.5 muM concentrations. Two flavonoids exuded from alfalfa roots, 4',7-dihydroxyflavone and 4',7-dihydroxyflavanone, also enhanced spore germination of this fungal species. Formononetin, an isoflavone that is released from stressed alfalfa roots, inhibited germination of both Glomus species. These in vitro results suggest that plant flavonoids may facilitate or regulate the development of VAM symbioses and offer new hope for developing pure, plant-free cultures of VAM fungi.

Protein Composition of Infectious Spores Reveals Novel Sexual Development and Germination Factors in Cryptococcus.

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Mingwei Huang

2015-08-01

Full Text Available Spores are an essential cell type required for long-term survival across diverse organisms in the tree of life and are a hallmark of fungal reproduction, persistence, and dispersal. Among human fungal pathogens, spores are presumed infectious particles, but relatively little is known about this robust cell type. Here we used the meningitis-causing fungus Cryptococcus neoformans to determine the roles of spore-resident proteins in spore biology. Using highly sensitive nanoscale liquid chromatography/mass spectrometry, we compared the proteomes of spores and vegetative cells (yeast and identified eighteen proteins specifically enriched in spores. The genes encoding these proteins were deleted, and the resulting strains were evaluated for discernable phenotypes. We hypothesized that spore-enriched proteins would be preferentially involved in spore-specific processes such as dormancy, stress resistance, and germination. Surprisingly, however, the majority of the mutants harbored defects in sexual development, the process by which spores are formed. One mutant in the cohort was defective in the spore-specific process of germination, showing a delay specifically in the initiation of vegetative growth. Thus, by using this in-depth proteomics approach as a screening tool for cell type-specific proteins and combining it with molecular genetics, we successfully identified the first germination factor in C. neoformans. We also identified numerous proteins with previously unknown functions in both sexual development and spore composition. Our findings provide the first insights into the basic protein components of infectious spores and reveal unexpected molecular connections between infectious particle production and spore composition in a pathogenic eukaryote.

Radiation sensitivity of fungal microflora isolated from some pharmaceutical ingredients

Energy Technology Data Exchange (ETDEWEB)

Mostafa, S.A. (Ain Shams Univ., Cairo (Egypt). Botany Dept.); El-Zawahry, Y.A.; Abdel All, S.S.

1983-01-01

The total number of fungal microflora of D-glucose, NaCl, KCl and their solutions was determined. The fungal isolates were identified as Aspergillus fumigatus. Aspergillus niger; Spicaria divaricate and Spicaria silvatica and their response to ..gamma..-radiation was determined, the most predominant isolate Asp. fumigatus was also the most irradiation resistant. The Dio and the lethal dose were determined for each isolate in a pure spore suspension as well as in the presence of the other isolates. The higher lethal dose values obtained for pure spore suspension as compared to that obtained for the natural fungal flora a D-glucose are discussed in terms of spore clumping. The activity of amylase, protease and L-asparaginase of Asp. fumigatus was examined prior to and after exposure to different doses of ..gamma..-radiation. Though all were inhibited at high doses, the effect was not as drastic as it was on cell viability.

Pollen and spore monitoring in the world.

Science.gov (United States)

Buters, J T M; Antunes, C; Galveias, A; Bergmann, K C; Thibaudon, M; Galán, C; Schmidt-Weber, C; Oteros, J

2018-01-01

Ambient air quality monitoring is a governmental duty that is widely carried out in order to detect non-biological ("chemical") components in ambient air, such as particles of monitoring networks are publicly funded and air quality data are open to the public. The situation for biological particles that have detrimental effects on health, as is the case of pollen and fungal spores, is however very different. Most pollen and spore monitoring networks are not publicly funded and data are not freely available. The information regarding which biological particle is being monitored, where and by whom, is consequently often not known, even by aerobiologists themselves. This is a considerable problem, as local pollen data are an important tool for the prevention of allergic symptoms. The aim of this study was to review pollen monitoring stations throughout the world and to create an interactive visualization of their distribution. The method employed to collect information was based on: (a) a review of the recent and historical bibliography related to pollen and fungal spore monitoring, and (b) personal surveys of the managers of national and regional monitoring networks. The interactive application was developed using the R programming language. We have created an inventory of the active pollen and spore monitoring stations in the world. There are at least 879 active pollen monitoring stations in the world, most of which are in Europe (> 500). The prevalent monitoring method is based on the Hirst principle (> 600 stations). The inventory is visualised as an interactive and on-line map. It can be searched, its appearance can be adjusted to the users' needs and it is updated regularly, as new stations or changes to those that already exist can be submitted online. The map shows the current situation of pollen and spore monitoring and facilitates collaboration among those individuals who are interested in pollen and spore counts. It might also help to improve the

Fungal spore germination into yeast or mycelium: possible implications of dimorphism in evolution and human pathogenesis

Science.gov (United States)

Ghormade, Vandana; Deshpande, M. V.

The ability of dimorphism in fungi is conventionally regarded as a reversible change between the two vegetative forms, yeast and mycelium, in response to environmental change. A zygomycetous isolate, Benjaminiella poitrasii, exhibited yeast-mycelium transition in response to the change in temperature (37-28 °C) and decrease in glucose concentration. For the first time the presence of dimorphic response during asexual and sexual spore germination is reported under the dimorphism-triggering conditions in B. poitrasii. The zygospores germinated into budding yeast when subjected to yeast-form supporting conditions. The mycelium-form favoring conditions gave rise to true mycelium. Similarly, the asexual spores displayed a dimorphic response during germination. Our observations suggest that dimorphism is an intrinsic ability present in the vegetative, asexual, and sexual forms of the fungus. As dimorphic fungi are intermediate to the unicellular yeast and the filamentous forms, understanding of the dimorphic character could be useful to trace the evolutionary relationships among taxonomically different fungi. Moreover, the implications of spore germination during the onset of pathogenesis and in drug development for human health care are discussed.

Biological responses of Raw 264.7 macrophage exposed to two strains of Stachybotrys chartarum spores grown on four different wallboard types.

Science.gov (United States)

Kim, J H; Harvey, L A; Evans, A L; Byfield, G E; Betancourt, D A; Dean, T R

2016-06-01

The many benefits of building "green" have motivated the use of sustainable products in the design and execution of the built environment. However, the use of these natural or recycled materials, some of which have been treated with antimicrobials, provides a growth opportunity for microorganisms with the potential to elicit adverse health effects especially in the presence of an antimicrobial. The focus of this research was to determine the effects of Stachybotrys chartarum (strains Houston and 51-11) grown under different conditions on a macrophage cell line (Raw 264.7) using endpoints, including cytotoxicity, and those associated with immunity specifically inflammation and MHC class II expression. The fungi were grown on four different gypsum products, and macrophages were exposed to whole spores of both strains and fragmented spores of strain 51-11. Whole spores of the Houston strain elicited no cytotoxicity with some level of inflammation, while exposure to whole spores of 51-11 caused variable responses depending on the wallboard type supporting the fungal growth. High concentrations of fragmented 51-11 spores primarily resulted in the apoptosis of macrophage with no inflammation. None of the fungal strains caused elevated levels of major histocompatibility complex (MHC) class II expression on the surface of Raw cells. Mycotoxin levels of 51-11 spores from all of the wallboard types measured  >250 ng/μL of T2 equivalent toxin based on activity. Collectively, the data demonstrated that all of the wallboard types supported growth of fungi with the ability to elicit harmful biological responses with the potential to negatively impact human health.

Heater Choice, Dampness and Mould Growth in 26 New Zealand Homes: A Study of Propensity for Mould Growth Using Encapsulated Fungal Spores

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Mikael Boulic

2015-02-01

Full Text Available The relationship between the use of unflued gas heaters (UGH, N = 14 and heat pump heaters (HP, N = 12 located in the living rooms, and mould growth on the living room and bedroom walls, of 26 New Zealand (NZ occupied homes was investigated during winter. Two methods were employed to evaluate the potential of mould growth on walls: (i measurement of daily hyphal growth rate using a fungal detector (encapsulated fungal spores; and (ii estimation of fungal contamination based on a four level scale visual inspection. The average wall psychrometric conditions were significantly different between the two heater type groups, in both the living rooms and the bedrooms with the UGH user homes being colder and damper than HP user homes. The UGHs were found to be a significant additional source of moisture in the living rooms which dramatically increased the capacity for fungi to grow on wall surfaces. The average daily hyphal growth rates were 4 and 16 times higher in the living rooms and in the bedrooms of the UGH user homes, respectively. Results from both mould detection methods gave good agreement, showing that the use of a fungal detector was an efficient method to predict the potential of mould growth on the inside of the external walls in NZ homes.

Combining real-time PCR and next-generation DNA sequencing to provide quantitative comparisons of fungal aerosol populations

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Dannemiller, Karen C.; Lang-Yona, Naama; Yamamoto, Naomichi; Rudich, Yinon; Peccia, Jordan

2014-02-01

We examined fungal communities associated with the PM10 mass of Rehovot, Israel outdoor air samples collected in the spring and fall seasons. Fungal communities were described by 454 pyrosequencing of the internal transcribed spacer (ITS) region of the fungal ribosomal RNA encoding gene. To allow for a more quantitative comparison of fungal exposure in humans, the relative abundance values of specific taxa were transformed to absolute concentrations through multiplying these values by the sample's total fungal spore concentration (derived from universal fungal qPCR). Next, the sequencing-based absolute concentrations for Alternaria alternata, Cladosporium cladosporioides, Epicoccum nigrum, and Penicillium/Aspergillus spp. were compared to taxon-specific qPCR concentrations for A. alternata, C. cladosporioides, E. nigrum, and Penicillium/Aspergillus spp. derived from the same spring and fall aerosol samples. Results of these comparisons showed that the absolute concentration values generated from pyrosequencing were strongly associated with the concentration values derived from taxon-specific qPCR (for all four species, p 0.70). The correlation coefficients were greater for species present in higher concentrations. Our microbial aerosol population analyses demonstrated that fungal diversity (number of fungal operational taxonomic units) was higher in the spring compared to the fall (p = 0.02), and principal coordinate analysis showed distinct seasonal differences in taxa distribution (ANOSIM p = 0.004). Among genera containing allergenic and/or pathogenic species, the absolute concentrations of Alternaria, Aspergillus, Fusarium, and Cladosporium were greater in the fall, while Cryptococcus, Penicillium, and Ulocladium concentrations were greater in the spring. The transformation of pyrosequencing fungal population relative abundance data to absolute concentrations can improve next-generation DNA sequencing-based quantitative aerosol exposure assessment.

Airway inflammation among compost workers exposed to actinomycetes spores

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Kari Kulvik Heldal

2015-05-01

Full Text Available Objectives. To study the associations between exposure to bioaerosols and work-related symptoms, lung function and biomarkers of airway inflammation in compost workers. Materials and method. Personal full-shift exposure measurements were performed on 47 workers employed at five windrow plants (n=20 and five reactor plants (n=27. Samples were analyzed for endotoxins, bacteria, fungal and actinomycetes spores. Health examinations were performed on workers and 37 controls before and after work on the day exposure was measured. The examinations included symptoms recorded by questionnaire, lung function by spirometry and nasal dimensions by acoustic rhinometry (AR. The pneumoproteins CC16, SP-D and SP-A were measured in a blood sample drawn at the end of the day. Results. The levels of endotoxins (median 3 EU/m[sup]3[/sup] , range 0–730 EU/m[sup]3[/sup] and actinomycetes spores (median 0.2 × 10[sup]6[/sup] spores/m[sup]3[/sup] , range 0–590 × 10[sup]6[/sup] spores/m[sup]3[/sup] were significantly higher in reactor plants compared to windrow plants. However, windrow composting workers reported more symptoms than reactor composting workers, probably due to use of respiratory protection. Exposure-response relationships between actinomycetes spores exposure and respiratory effects, found as cough and nose irritation during a shift, was significantly increased (OR 4.3, 95% CI 1.1–16, OR 6.1, 95% CI 1.5–25, respectively, p<0.05 among workers exposed to 0.02–0.3 × 10[sup]6[/sup] actinomycetes spores/m 3 , and FEV1/FVC% decreased cross shift (b=–3.2, SE=1.5%, p<0.01. Effects were weaker in the highest exposed group, but these workers used respiratory protection, frequently limiting their actual exposure. No relationships were found between exposure and pneumoprotein concentrations. Conclusions. The major agent in the aerosol generated at compost plants was actinomycetes spores which was associated with work related cough symptoms and work

Spores of many common airborne fungi reveal no ice nucleation activity in oil immersion freezing experiments

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Pummer, B. G.; Atanasova, L.; Bauer, H.; Bernardi, J.; Druzhinina, I. S.; Fröhlich-Nowoisky, J.; Grothe, H.

2013-12-01

Fungal spores are ubiquitous biological aerosols, which are considered to act as ice nuclei. In this study the ice nucleation (IN) activity of spores harvested from 29 fungal strains belonging to 21 different species was tested in the immersion freezing mode by microscopic observation of water-in-oil emulsions. Spores of 8 of these strains were also investigated in a microdroplet freezing array instrument. The focus was laid on species of economical, ecological or sanitary significance. Besides common molds (Ascomycota), some representatives of the widespread group of mushrooms (Basidiomycota) were also investigated. Fusarium avenaceum was the only sample showing IN activity at relatively high temperatures (about 264 K), while the other investigated fungal spores showed no freezing above 248 K. Many of the samples indeed froze at homogeneous ice nucleation temperatures (about 237 K). In combination with other studies, this suggests that only a limited number of species may act as atmospheric ice nuclei. This would be analogous to what is already known for the bacterial ice nuclei. Apart from that, we selected a set of fungal strains from different sites and exposed them to occasional freezing stress during their cultivation. This was in order to test if the exposure to a cold environment encourages the expression of ice nuclei during growth as a way of adaptation. Although the total protein expression was altered by this treatment, it had no significant impact on the IN activity.

Arthropods associated with fungal galls: do large galls support more abundant and diverse inhabitants?

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Funamoto, Daichi; Sugiura, Shinji

2017-02-01

Fungus-induced galls can attract spore-feeding arthropods as well as gall-feeding ones, resulting in diverse communities. Do large fungal galls support more abundant and diverse arthropod communities than small fungal galls? To address this question, we investigated the structure of the arthropod community associated with bud galls induced by the fungus Melanopsichium onumae on the tree species Cinnamomum yabunikkei (Lauraceae) in central Japan. Thirteen species of arthropods were associated with M. onumae galls. Dominant arthropod species were represented by the larvae of a salpingid beetle (a spore feeder), a nitidulid beetle (a spore feeder), a cosmopterigid moth (a spore feeder), an unidentified moth (a gall tissue feeder), and a drosophilid species (a gall tissue feeder). Arthropod abundance and species richness were positively correlated with gall diameter. The majority of the most abundant species were more frequently found in large galls than in small ones, indicating that large fungal galls, which have more food and/or space for arthropods, could support a more abundant and diverse arthropod community.

VeA of Aspergillus niger increases spore dispersing capacity by impacting conidiophore architecture.

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Wang, Fengfeng; Dijksterhuis, Jan; Wyatt, Timon; Wösten, Han A B; Bleichrodt, Robert-Jan

2015-01-01

Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger maximizes the diameter of the vesicle and the spore chain length. The length and width of the conidiophore stalk and vesicle were reduced nearly twofold in a ΔveA strain. The latter implies a fourfold reduced surface area to develop chains of spores. Over and above this, the conidial chain length was approximately fivefold reduced. The calculated 20-fold reduction in formation of conidia by ΔveA fits the 8- to 17-fold decrease in counted spore numbers. Notably, morphology of the ΔveA conidiophores of A. niger was very similar to that of wild-type Aspergillus sydowii. This suggests that VeA is key in conidiophore architecture diversity in the fungal kingdom. The finding that biomass formation of the A. niger ΔveA strain was reduced twofold shows that VeA not only impacts dispersion capacity but also colonization capacity of A. niger.

Comparative performance of two air samplers for monitoring airborne fungal propagules

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L.G.F. Távora

2003-05-01

Full Text Available Many studies have attempted to evaluate the importance of airborne fungi in the development of invasive fungal infection, especially for immunocompromised hosts. Several kinds of instruments are available to quantitate fungal propagule levels in air. We compared the performance of the most frequently used air sampler, the Andersen sampler with six stages, with a portable one, the Reuter centrifugal sampler (RCS. A total of 84 samples were analyzed, 42 with each sampler. Twenty-eight different fungal genera were identified in samples analyzed with the Andersen instrument. In samples obtained with the RCS only seven different fungal genera were identified. The three most frequently isolated genera in samples analyzed with both devices were Penicillium, Aspergillus and Cladophialophora. In areas supplied with a high efficiency particulate air filter, fungal spore levels were usually lower when compared to areas without these filters. There was a significant correlation between total fungal propagule measurements taken with both devices on each sampling occasion (Pearson coefficient = 0.50. However, the Andersen device recovered a broader spectrum of fungi. We conclude that the RCS can be used for quantitative estimates of airborne microbiological concentrations. For qualitative studies, however, this device cannot be recommended.

Optical and structural properties of plasma-treated Cordyceps bassiana spores as studied by circular dichroism, absorption, and fluorescence spectroscopy

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Lee, Geon Joon, E-mail: gjlee@kw.ac.kr; Sim, Geon Bo; Choi, Eun Ha [Plasma Bioscience Research Center/Department of Electrical and Biological Physics, Kwangwoon University, Seoul 139-701 (Korea, Republic of); Kwon, Young-Wan [KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 136-701 (Korea, Republic of); Kim, Jun Young; Jang, Siun; Kim, Seong Hwan, E-mail: piceae@naver.com [Department of Microbiology and Institute of Basic Sciences, Dankook University, Cheonan 330-714 (Korea, Republic of)

2015-01-14

To understand the killing mechanism of fungal spores by plasma treatment, the optical, structural, and biological properties of the insect pathogenic fungus Cordyceps bassiana spores were studied. A nonthermal atmospheric-pressure plasma jet (APPJ) was used to treat the spores in aqueous solution. Optical emission spectra of the APPJ acquired in air indicated emission peaks corresponding to hydroxyl radicals and atomic oxygen. When the APPJ entered the aqueous solution, additional reactive species were derived from the interaction of plasma radicals with the aqueous solution. Fluorescence and absorption spectroscopy confirmed the generation of hydroxyl radicals and hydrogen peroxide in the plasma-activated water (PAW). Spore counting showed that plasma treatment significantly reduced spore viability. Absorption spectroscopy, circular dichroism (CD) spectroscopy, and agarose gel electrophoresis of the DNA extracted from plasma-treated spores showed a reduction in spore DNA content. The magnitude of the dip in the CD spectrum was lower in the plasma-treated spores than in the control, indicating that plasma treatment causes structural modifications and/or damage to cellular components. Tryptophan fluorescence intensity was lower in the plasma-treated spores than in the control, suggesting that plasma treatment modified cell wall proteins. Changes in spore viability and DNA content were attributed to structural modification of the cell wall by reactive species coming from the APPJ and the PAW. Our results provided evidence that the plasma radicals and the derived reactive species play critical roles in fungal spore inactivation.

Adaptation of the spore discharge mechanism in the basidiomycota.

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Jessica L Stolze-Rybczynski

Full Text Available Spore discharge in the majority of the 30,000 described species of Basidiomycota is powered by the rapid motion of a fluid droplet, called Buller's drop, over the spore surface. In basidiomycete yeasts, and phytopathogenic rusts and smuts, spores are discharged directly into the airflow around the fungal colony. Maximum discharge distances of 1-2 mm have been reported for these fungi. In mushroom-forming species, however, spores are propelled over much shorter ranges. In gilled mushrooms, for example, discharge distances of <0.1 mm ensure that spores do not collide with opposing gill surfaces. The way in which the range of the mechanism is controlled has not been studied previously.In this study, we report high-speed video analysis of spore discharge in selected basidiomycetes ranging from yeasts to wood-decay fungi with poroid fruiting bodies. Analysis of these video data and mathematical modeling show that discharge distance is determined by both spore size and the size of the Buller's drop. Furthermore, because the size of Buller's drop is controlled by spore shape, these experiments suggest that seemingly minor changes in spore morphology exert major effects upon discharge distance.This biomechanical analysis of spore discharge mechanisms in mushroom-forming fungi and their relatives is the first of its kind and provides a novel view of the incredible variety of spore morphology that has been catalogued by traditional taxonomists for more than 200 years. Rather than representing non-selected variations in micromorphology, the new experiments show that changes in spore architecture have adaptive significance because they control the distance that the spores are shot through air. For this reason, evolutionary modifications to fruiting body architecture, including changes in gill separation and tube diameter in mushrooms, must be tightly linked to alterations in spore morphology.

Concentration of petroleum-hydrocarbon contamination shapes fungal endophytic community structure in plant roots

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Guillaume eBourdel

2016-05-01

Full Text Available Plant-root inhabiting fungi are a universal phenomenon found in all ecosystems where plants are able to grow, even in harsh environments. Interactions between fungi and plant roots can vary widely from mutualism to parasitism depending on many parameters. The role of fungal endophytes in phytoremediation of polluted sites, and characterization of the endophytic diversity and community assemblages in contaminated areas remain largely unexplored. In this study, we investigated the composition of endophytic fungal communities in the roots of two plant species growing spontaneously in petroleum-contaminated sedimentation basins of a former petro-chemical plant. The three adjacent basins showed a highly heterogeneous patterns of pollutant concentrations. We combined a culture-based isolation approach with the pyrosequencing of fungal ITS ribosomal DNA. We selected two species, Eleocharis erythropoda Steud. and Populus balsamifera L., and sampled three individuals of each species from each of three adjacent basins, each with a different concentration of petroleum hydrocarbons. We found that contamination level significantly shaped endophytic fungal diversity and community composition in E. erythropoda, with only 9.9% of these fungal Operational Taxonomic Units (OTUs retrieved in all three basins. However, fungal community structure associated with P. balsamifera remained unaffected by the contamination level with 28.2% of fungal OTUs shared among all three basins. This could be explained by the smaller differences of pollutant concentrations in the soil around our set of P. balsamifera sampless compared to that around our set of E. erythropoda samples. Our culture-based approach allowed isolation of 11 and 30 fungal endophytic species from surface-sterilized roots of E. erythropoda and P. balsamifera, respectively. These isolates were ribotyped using ITS, and all were found in pyrosequensing datasets. Our results demonstrate that extreme levels of

[Airborne Fungal Aerosol Concentration and Distribution Characteristics in Air- Conditioned Wards].

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Zhang, Hua-ling; Feng, He-hua; Fang, Zi-liang; Wang, Ben-dong; Li, Dan

2015-04-01

The effects of airborne fungus on human health in the hospital environment are related to not only their genera and concentrations, but also their particle sizes and distribution characteristics. Moreover, the mechanisms of aerosols with different particle sizes on human health are different. Fungal samples were obtained in medicine wards of Chongqing using a six-stage sampler. The airborne fungal concentrations, genera and size distributions of all the sampling wards were investigated and identified in detail. Results showed that airborne fungal concentrations were not correlated to the diseases or personnel density, but were related to seasons, temperature, and relative humidity. The size distribution rule had roughly the same for testing wards in winter and summer. The size distributions were not related with diseases and seasons, the percentage of airborne fungal concentrations increased gradually from stage I to stage III, and then decreased dramatically from stage V to stage VI, in general, the size of airborne fungi was a normal distribution. There was no markedly difference for median diameter of airborne fungi which was less 3.19 μm in these wards. There were similar dominant genera in all wards. They were Aspergillus spp, Penicillium spp and Alternaria spp. Therefore, attention should be paid to improve the filtration efficiency of particle size of 1.1-4.7 μm for air conditioning system of wards. It also should be targeted to choose appropriate antibacterial methods and equipment for daily hygiene and air conditioning system operation management.

Studies on the fungal flora of Garri, processed cassava ( Manihot ...

African Journals Online (AJOL)

In Nsukka area of southeastern Nigeria, garri is usually displayed in the open market for sale which no doubt exposes the food stuff to dust particles from moving vehicles, wind and other sources. Thus, fungal spores from the air and soil environment could serve as major sources of fungal contamination of this product.

Single Spore Isolation as a Simple and Efficient Technique to obtain fungal pure culture

Science.gov (United States)

Noman, E.; Al-Gheethi, AA; Rahman, N. K.; Talip, B.; Mohamed, R.; H, N.; Kadir, O. A.

2018-04-01

The successful identification of fungi by phenotypic methods or molecular technique depends mainly on the using an advanced technique for purifying the isolates. The most efficient is the single spore technique due to the simple requirements and the efficiency in preventing the contamination by yeast, mites or bacteria. The method described in the present work is depends on the using of a light microscope to transfer one spore into a new culture medium. The present work describes a simple and efficient procedure for single spore isolation to purify of fungi recovered from the clinical wastes.

Genetic analysis reveals efficient sexual spore dispersal at a fine spatial scale in Armillaria ostoyae, the causal agent of root-rot disease in conifers.

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Dutech, Cyril; Labbé, Frédéric; Capdevielle, Xavier; Lung-Escarmant, Brigitte

Armillaria ostoyae (sometimes named Armillaria solidipes) is a fungal species causing root diseases in numerous coniferous forests of the northern hemisphere. The importance of sexual spores for the establishment of new disease centres remains unclear, particularly in the large maritime pine plantations of southwestern France. An analysis of the genetic diversity of a local fungal population distributed over 500 ha in this French forest showed genetic recombination between genotypes to be frequent, consistent with regular sexual reproduction within the population. The estimated spatial genetic structure displayed a significant pattern of isolation by distance, consistent with the dispersal of sexual spores mostly at the spatial scale studied. Using these genetic data, we inferred an effective density of reproductive individuals of 0.1-0.3 individuals/ha, and a second moment of parent-progeny dispersal distance of 130-800 m, compatible with the main models of fungal spore dispersal. These results contrast with those obtained for studies of A. ostoyae over larger spatial scales, suggesting that inferences about mean spore dispersal may be best performed at fine spatial scales (i.e. a few kilometres) for most fungal species. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Fungal biological control agents for integrated management of Culicoides spp. (Diptera: Ceratopogonidae of livestock

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B. W. Narladkar

2015-02-01

Full Text Available Aim: Entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana had wide host range against insects and hence these are being exploited as fungal bio-pesticide on a large scale. Both fungi are proved pesticides against many crop pests and farmers are well acquainted with their use on the field. Thus, research was aimed to explore the potency of these fungal spores against larval and adult Culicoides midges, a pest of livestock. Materials and Methods: In-vitro testing of both fungal biological control agents was undertaken in Petri dishes against field collected Culicoides larvae, while in plastic beakers against field collected blood-engorged female Culicoides midges. In-vivo testing was undertaken by spraying requisite concentration of fungal spores on the drainage channel against larvae and resting sites of adult Culicoides midges in the cattle shed. Lethal concentration 50 (LC50 values and regression equations were drawn by following probit analysis using SPSS statistical computerized program. Results: The results of this study revealed LC50 values of 2692 mg and 3837 mg (108 cfu/g for B. bassiana and M. anisopliae, respectively, against Culicoides spp. larvae. Death of Culicoides larvae due to B. bassiana showed greenish coloration in the middle of the body with head and tail showed intense blackish changes, while infection of M. anisopliae resulted in death of Culicoides larvae with greenish and blackish coloration of body along with total destruction, followed by desquamation of intestinal channel. The death of adult Culicoides midges were caused by both the fungi and after death growth of fungus were very well observed on the dead cadavers proving the efficacy of the fungus. Conclusion: Preliminary trials with both funguses (M. anisopliae, B. bassiana showed encouraging results against larvae and adults of Culicoides spp. Hence, it was ascertained that, these two fungal molecules can form a part of biological control and

1-Octanol, a self-inhibitor of spore germination in Penicillium camemberti.

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Gillot, Guillaume; Decourcelle, Nicolas; Dauer, Gaëlle; Barbier, Georges; Coton, Emmanuel; Delmail, David; Mounier, Jérôme

2016-08-01

Penicillium camemberti is a technologically relevant fungus used to manufacture mold-ripened cheeses. This fungal species produces many volatile organic compounds (VOCs) including ammonia, methyl-ketones, alcohols and esters. Although it is now well known that VOCs can act as signaling molecules, nothing is known about their involvement in P. camemberti lifecycle. In this study, spore germination was shown to be self-regulated by quorum sensing in P. camemberti. This phenomenon, also called "crowding effect", is population-dependent (i.e. observed at high population densities). After determining the volatile nature of the compounds involved in this process, 1-octanol was identified as the main compound produced at high-spore density using GC-MS. Its inhibitory effect was confirmed in vitro and 3 mM 1-octanol totally inhibited spore germination while 100 μM only transiently inhibited spore germination. This is the first time that self-inhibition of spore germination is demonstrated in P. camemberti. The obtained results provide interesting perspectives for better control of mold-ripened cheese processes. Copyright © 2016 Elsevier Ltd. All rights reserved.

Decontamination Options for Drinking Water Contaminated with Bacillus anthracis Spores

Energy Technology Data Exchange (ETDEWEB)

Raber, E; Burklund, A

2010-02-16

Five parameters were evaluated with surrogates of Bacillus anthracis spores to determine effective decontamination options for use in a contaminated drinking water supply. The parameters were: (1) type of Bacillus spore surrogate (B. thuringiensis or B. atrophaeus); (2) spore concentration in suspension (10{sup 2} to 10{sup 6} spores/ml); (3) chemical characteristics of decontaminant [sodium dicholor-s-triazinetrione dihydrate (Dichlor), hydrogen peroxide, potassium peroxymonosulfate (Oxone), sodium hypochlorite, and VirkonS{reg_sign}]; (4) decontaminant concentration (0.01% to 5%); and (5) decontaminant exposure time (10 min to 24 hr). Results from 162 suspension tests with appropriate controls are reported. Hydrogen peroxide at a concentration of 5%, and Dichlor and sodium hypochlorite at a concentration of 2%, were effective at spore inactivation regardless of spore type tested, spore exposure time, or spore concentration evaluated. This is the first reported study of Dichlor as an effective decontaminant for B. anthracis spore surrogates. Dichlor's desirable characteristics of high oxidation potential, high level of free chlorine, and more neutral pH than that of other oxidizers evaluated appear to make it an excellent alternative. All three oxidizers were effective against B. atrophaeus spores in meeting EPA's biocide standard of greater than a 6 log kill after a 10-minute exposure time and at lower concentrations than typically reported for biocide use. Solutions of 5% VirkonS{reg_sign} and Oxone were less effective decontaminants than other options evaluated in this study and did not meet the EPA's efficacy standard for biocides. Differences in methods and procedures reported by other investigators make quantitative comparisons among studies difficult.

Subchronic exposures to fungal bioaerosols promotes allergic pulmonary inflammation in naïve mice.

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Nayak, A P; Green, B J; Lemons, A R; Marshall, N B; Goldsmith, W T; Kashon, M L; Anderson, S E; Germolec, D R; Beezhold, D H

2016-06-01

Epidemiological surveys indicate that occupants of mold contaminated environments are at increased risk of respiratory symptoms. The immunological mechanisms associated with these responses require further characterization. The aim of this study was to characterize the immunotoxicological outcomes following repeated inhalation of dry Aspergillus fumigatus spores aerosolized at concentrations potentially encountered in contaminated indoor environments. Aspergillus fumigatus spores were delivered to the lungs of naïve BALB/cJ mice housed in a multi-animal nose-only chamber twice a week for a period of 13 weeks. Mice were evaluated at 24 and 48 h post-exposure for histopathological changes in lung architecture, recruitment of specific immune cells to the airways, and serum antibody responses. Germinating A. fumigatus spores were observed in lungs along with persistent fungal debris in the perivascular regions of the lungs. Repeated exposures promoted pleocellular infiltration with concomitant epithelial mucus hypersecretion, goblet cell metaplasia, subepithelial fibrosis and enhanced airway hyperreactivity. Cellular infiltration in airways was predominated by CD4(+) T cells expressing the pro-allergic cytokine IL-13. Furthermore, our studies show that antifungal T cell responses (IFN-γ(+) or IL-17A(+) ) co-expressed IL-13, revealing a novel mechanism for the dysregulated immune response to inhaled fungi. Total IgE production was augmented in animals repeatedly exposed to A. fumigatus. Repeated inhalation of fungal aerosols resulted in significant pulmonary pathology mediated by dynamic shifts in specific immune populations and their cytokines. These studies provide novel insights into the immunological mechanisms and targets that govern the health outcomes that result from repeated inhalation of fungal bioaerosols in contaminated environments. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

The dynamics of the fungal aerospores Alternaria sp. and Cladosporium sp. in Parisian atmospheric air, in France

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Brezoczki, V. M.

2016-08-01

The bioallergens occurring naturally in the atmospheric air are microorganisms, pollen grains, plant seeds, leaf and stem scrap, or their protein molecules. The presence of various airborne fungal spores determines a high allergenic potential for public health. This effect is due to the high number of produced spores, which under favourable meteorological conditions (dry weather and wind) reach the surrounding air. This paper traces the dynamics of two types of fungi, Alternaria sp and Cladosporium sp, fungi which can be found outdoors, in the surrounding air, as well as indoors, inside houses (especially the conidia of Cladosporium sp). The effects of these fungal spores on human health are varies, ranging from seasonal allergies (hay fever, rhinitis, sinusitis etc.) to sever afflictions of the respiratory system, onset of asthma, disfunctionalities of the nervous systems, of the immune system, zymoses etc. The monitoring of the dynamics of the aerospores Alternaria sp and Cladosporium sp was carried out between 2010 and 2013, over a period of 42 weeks during one calendar year, from February to the end of September, in the surrounding air in the French capital, Paris. The regional and global climate and meteorological conditions are directly involved in the occurrence and development of fungi colonies, the transportation and dispersion of fungal spores in the atmospheric air, as well as in the creation of the environment required for the interaction of chemical and biological components in the air. Knowledge of the dynamics of the studied fungal aerospores, coupled with climate and meteorological changes, offers a series of information on the magnitude of the allergenic potential these airborne spores can determine. Legal regulations in this domain set the allergen risk threshold for the Alternaria sp aerospores at 3500 ÷ 7000 spores/m3 air/week, and for the Cladosporium sp aerospores at 56,000 spores/m3 air/week. Besides these regulations there exist a series of

Contribution of fungi to primary biogenic aerosols in the atmosphere: wet and dry discharged spores, carbohydrates, and inorganic ions

Directory of Open Access Journals (Sweden)

W. Elbert

2007-09-01

the average abundance of mannitol reported for extratropical continental boundary layer air (~25 ng m⁻³, we have also calculated a value of ~17 Tg yr⁻¹ as a first estimate for the global average emission rate of ABS over land surfaces, which is consistent with the typically observed concentrations of ABS (~10³–10⁴ m⁻³; ~0.1–1 μg m⁻³. The global average atmospheric abundance and emission rate of total fungal spores, including wet and dry discharged species, are estimated to be higher by a factor of about three, i.e. 1 μg m⁻³ and ~50 Tg yr⁻¹. Comparisons with estimated rates of emission and formation of other major types of organic aerosol (~47 Tg yr⁻¹ of anthropogenic primary organic aerosol; 12–70 Tg yr⁻¹ of secondary organic aerosol indicate that emissions from fungi should be taken into account as a significant global source of organic aerosol. The effects of fungal spores and related chemical components might be particularly important in tropical regions, where both physicochemical processes in the atmosphere and biological activity at the Earth's surface are particularly intense, and where the abundance of fungal spores and related chemical compounds are typically higher than in extratropical regions.

Dispersal distances for airborne spores based on deposition rates and stochastic modeling

DEFF Research Database (Denmark)

Stockmarr, Anders; Andreasen, Viggo; Østergård, Hanne

2007-01-01

in terms of time to deposition, and show how this concept is equivalent to the deposition rate for fungal spores. Special cases where parameter values for wind and gravitation lead to exponentially or polynomially decreasing densities are discussed, and formulas for one- and two-dimensional densities...

Non-thermal plasma treatment diminishes fungal viability and up-regulates resistance genes in a plant host.

Science.gov (United States)

Panngom, Kamonporn; Lee, Sang Hark; Park, Dae Hoon; Sim, Geon Bo; Kim, Yong Hee; Uhm, Han Sup; Park, Gyungsoon; Choi, Eun Ha

2014-01-01

Reactive oxygen and nitrogen species can have either harmful or beneficial effects on biological systems depending on the dose administered and the species of organism exposed, suggesting that application of reactive species can possibly produce contradictory effects in disease control, pathogen inactivation and activation of host resistance. A novel technology known as atmospheric-pressure non-thermal plasma represents a means of generating various reactive species that adversely affect pathogens (inactivation) while simultaneously up-regulating host defense genes. The anti-microbial efficacy of this technology was tested on the plant fungal pathogen Fusarium oxysporum f.sp. lycopersici and its susceptible host plant species Solanum lycopercicum. Germination of fungal spores suspended in saline was decreased over time after exposed to argon (Ar) plasma for 10 min. Although the majority of treated spores exhibited necrotic death, apoptosis was also observed along with the up-regulation of apoptosis related genes. Increases in the levels of peroxynitrite and nitrite in saline following plasma treatment may have been responsible for the observed spore death. In addition, increased transcription of pathogenesis related (PR) genes was observed in the roots of the susceptible tomato cultivar (S. lycopercicum) after exposure to the same Ar plasma dose used in fungal inactivation. These data suggest that atmospheric-pressure non-thermal plasma can be efficiently used to control plant fungal diseases by inactivating fungal pathogens and up-regulating mechanisms of host resistance.

Arbuscular Mycorrhizal and Dark Septate Endophyte Fungal Associations in South Indian Aquatic and Wetland Macrophytes

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Kumar Seerangan

2014-01-01

Full Text Available Investigations on the prevalence of arbuscular mycorrhizal (AM and dark septate endophyte (DSE fungal symbioses are limited for plants growing in tropical aquatic and wetland habitats compared to those growing on terrestrial moist or dry habitats. Therefore, we assessed the incidence of AM and DSE symbiosis in 8 hydrophytes and 50 wetland plants from four sites in south India. Of the 58 plant species examined, we found AM and DSE fungal symbiosis in 21 and five species, respectively. We reported for the first time AM and DSE fungal symbiosis in seven and five species, respectively. Intermediate-type AM morphology was common, and AM morphology is reported for the first time in 16 plant species. Both AM and DSE fungal colonization varied significantly across plant species and sites. Intact and identifiable AM fungal spores occurred in root zones of nine plant species, but AM fungal species richness was low. Though no clear relationship between AM and DSE fungal colonization was recognized, a significant negative correlation between AM colonization and spore numbers was established. Our study suggests that the occurrence of AM and DSE fungal symbiosis in plants growing in hydrophytic and wetland habitats is not as common as in terrestrial habitats.

Microfungal spores (Ustilago maydis and U. digitariae) immobilised chitosan microcapsules for heavy metal removal.

Science.gov (United States)

Sargın, İdris; Arslan, Gulsin; Kaya, Murat

2016-03-15

Designing effective chitosan-based biosorbents from unexploited biomass for heavy metal removal has received much attention over the past decade. Ustilago, loose smut, is a ubiquitous fungal plant pathogen infecting over 4000 species including maize and weed. This study aimed to establish whether the spores of the phytopathogenic microfungi Ustilago spores can be immobilised in cross-linked chitosan matrix, and it reports findings on heavy metal sorption performance of chitosan/Ustilago composite microcapsules. Immobilisation of Ustilago maydis and U. digitariae spores (from maize and weed) in chitosan microcapsules was achieved via glutaraldehyde cross-linking. The cross-linked microcapsules were characterised using scanning electron microscopy, FT-IR spectroscopy and thermogravimetric analysis. Sorption capacities of chitosan-U. maydis and chitosan-U. digitariae microcapsules were investigated and compared to cross-linked chitosan beads: Cu(II): 66.72, 69.26, 42.57; Cd(II): 49.46, 53.96, 7.87; Cr(III): 35.88, 49.40, 43.68; Ni(II): 41.67, 33.46, 16.43 and Zn(II): 30.73, 60.81, 15.04mg/g, respectively. Sorption experiments were conducted as a function of initial metal ion concentration (2-10mg/L), contact time (60-480min), temperature (25, 35 and 45°C), amount of the sorbent (0.05-0.25g) and pH of the metal solution. The microcapsules with spores exhibited better performance over the plain chitosan beads, demonstrating their potential use in water treatment. Copyright © 2015 Elsevier Ltd. All rights reserved.

Aspergillosis in the common sea fan Gorgonia ventalina: isolation of waterborne hyphae and spores.

Science.gov (United States)

Troeger, Victoria J; Sammarco, Paul W; Caruso, John H

2014-07-03

The octocoral disease aspergillosis is caused by the terrestrial fungus Aspergillus sydowii. The possibility of secondary (horizontal) transmission of aspergillosis among common sea fans Gorgonia ventalina would require waterborne transmission of hyphae and/or spores. A laboratory filtration experiment confirmed that fungal hyphae and spores were shed into the water by infected fans. This suggests that secondary infection might be possible in this species. It remains to be determined whether healthy fans actually develop aspergillosis after contact with hyphae-laden water.

Fungal transmission of plant viruses.

Science.gov (United States)

Campbell, R N

1996-01-01

Thirty soilborne viruses or virus-like agents are transmitted by five species of fungal vectors. Ten polyhedral viruses, of which nine are in the family Tombusviridae, are acquired in the in vitro manner and do not occur within the resting spores of their vectors, Olpidium brassicae and O. bornovanus. Fungal vectors for other viruses in the family should be sought even though tombusviruses are reputed to be soil transmitted without a vector. Eighteen rod-shaped viruses belonging to the furo- and bymovirus groups and to an unclassified group are acquired in the in vivo manner and survive within the resting spores of their vector, O. brassicae, Polymyxa graminis, P. betae, and Spongospora subterranea. The viral coat protein has an essential role in in vitro transmission. With in vivo transmission a site in the coat protein-read through protein (CP-RT) of beet necrotic yellow vein furovirus determines vector transmissibility as does a site in a similar 98-kDa polyprotein of barley mild mosaic bymovirus. The mechanisms by which virions move (or are moved) into and out of the protoplasm of zoospores or of thalli needs study.

Non-Thermal Plasma Treatment Diminishes Fungal Viability and Up-Regulates Resistance Genes in a Plant Host

Science.gov (United States)

Panngom, Kamonporn; Lee, Sang Hark; Park, Dae Hoon; Sim, Geon Bo; Kim, Yong Hee; Uhm, Han Sup; Park, Gyungsoon; Choi, Eun Ha

2014-01-01

Reactive oxygen and nitrogen species can have either harmful or beneficial effects on biological systems depending on the dose administered and the species of organism exposed, suggesting that application of reactive species can possibly produce contradictory effects in disease control, pathogen inactivation and activation of host resistance. A novel technology known as atmospheric-pressure non-thermal plasma represents a means of generating various reactive species that adversely affect pathogens (inactivation) while simultaneously up-regulating host defense genes. The anti-microbial efficacy of this technology was tested on the plant fungal pathogen Fusarium oxysporum f.sp. lycopersici and its susceptible host plant species Solanum lycopercicum. Germination of fungal spores suspended in saline was decreased over time after exposed to argon (Ar) plasma for 10 min. Although the majority of treated spores exhibited necrotic death, apoptosis was also observed along with the up-regulation of apoptosis related genes. Increases in the levels of peroxynitrite and nitrite in saline following plasma treatment may have been responsible for the observed spore death. In addition, increased transcription of pathogenesis related (PR) genes was observed in the roots of the susceptible tomato cultivar (S. lycopercicum) after exposure to the same Ar plasma dose used in fungal inactivation. These data suggest that atmospheric-pressure non-thermal plasma can be efficiently used to control plant fungal diseases by inactivating fungal pathogens and up-regulating mechanisms of host resistance. PMID:24911947

Non-thermal plasma treatment diminishes fungal viability and up-regulates resistance genes in a plant host.

Directory of Open Access Journals (Sweden)

Kamonporn Panngom

Full Text Available Reactive oxygen and nitrogen species can have either harmful or beneficial effects on biological systems depending on the dose administered and the species of organism exposed, suggesting that application of reactive species can possibly produce contradictory effects in disease control, pathogen inactivation and activation of host resistance. A novel technology known as atmospheric-pressure non-thermal plasma represents a means of generating various reactive species that adversely affect pathogens (inactivation while simultaneously up-regulating host defense genes. The anti-microbial efficacy of this technology was tested on the plant fungal pathogen Fusarium oxysporum f.sp. lycopersici and its susceptible host plant species Solanum lycopercicum. Germination of fungal spores suspended in saline was decreased over time after exposed to argon (Ar plasma for 10 min. Although the majority of treated spores exhibited necrotic death, apoptosis was also observed along with the up-regulation of apoptosis related genes. Increases in the levels of peroxynitrite and nitrite in saline following plasma treatment may have been responsible for the observed spore death. In addition, increased transcription of pathogenesis related (PR genes was observed in the roots of the susceptible tomato cultivar (S. lycopercicum after exposure to the same Ar plasma dose used in fungal inactivation. These data suggest that atmospheric-pressure non-thermal plasma can be efficiently used to control plant fungal diseases by inactivating fungal pathogens and up-regulating mechanisms of host resistance.

A Qualitative and Quantitative Survey on Air-Transmitted Fungal Contamination in Different Wards of Kamkar Hospital in Qom, Iran, in 2007

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M Azizifar

2012-05-01

Full Text Available

Background and Objectives: Fungi spores can be found everywhere. The amount and variety of fungal spores and their vast spread could be a preliminary step to the initiation of different diseases in people with different levels of health.

Methods: In the present study six wards including nephrology, internal ward for women, surgery ward for men, operating theater for E.N.T., ophthalmology, infectious diseases ward and the laboratory were chosen for sampling on the basis of their types of activities and their in-patients. We used Anderson sampling method, collected samples within two minutes with flow rate of 28.3 L/Min in sabouraud medium.

Results: Maximum contamination in the infectious diseases ward was 300 CFU/m³ and minimum contamination in E.N.T. was 94 CFU/m³. The maximum percentage of fungal spores in the hospital air was observed to be as follows: penicillin with 36.36%, Cladosporium 24.74%, A.niger 17.97%, Rhizopus 10.57% and A.flavus 2.74A%.

Conclusion: Fungal contamination concentration in hospital indoor air in this study was higher than the recommended limits and other similar studies from a quantitative point of view, but it was similar to other studies in terms of identified species.

VeA of Aspergillus niger increases spore dispersing capacity by impacting conidiophore architecture

NARCIS (Netherlands)

Wang, Fengfeng; Dijksterhuis, Jan; Wyatt, Timon; Wösten, Han A B; Bleichrodt, Robert-Jan

Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger

Transport of Phanerochaete chrysosporium and Mucor hiemalis f. irnsingii spores through water-saturated quartz sands; Transport von Phanerochaete chrysosporium- und Mucor hiemalis f. irnsingii-Sporen durch wassergesaettigten Quarzsand

Energy Technology Data Exchange (ETDEWEB)

Hoque, E.; Klotz, D.; Teichmann, G.; Lang, H.; Wolf, M. [GSF - Forschungszentrum fuer Umwelt und Gesundheit GmbH, Neuherberg (Germany). Inst. fuer Hydrologie; Beisker, W. [GSF, Inst. fuer Pathologie (Germany)

2001-11-01

Just as bacteria so can aquatic fungi contribute effectively to pollutant elimination, at least in superficial groundwaters. However, very little is known about this. Nor is it known whether fungal spores are capable of being transported through sedimentary cavities. At least fungal spores are for some part of similar size as bacteria. We here report for the first time on a demonstration of the migration through quartz sand of spores of the two fungal species Phanerochaete chrysosporium and Mucor hiemalis f. irnsingii. [German] Neben Bakterien koennen auch aquatische Pilze, zumindest im oberflaechennahen Grundwasser, effiziente Beitraege zur Schadstoffentgiftung leisten. Darueber ist aber sehr wenig bekannt. Es ist auch nicht bekannt, ob ueberhaupt Pilzsporen durch Sedimenthohlraeume transportiert werden koennen, obwohl die Pilzsporen z.T. aehnliche Abmessungen wie Bakterien haben. Hier berichten wir erstmalig ueber den Nachweis der Migration von Sporen der beiden Pilzarten Phanerochaete chrysosporium und Mucor hiemalis f. irnsingii durch Quarzsand. (orig.)

Optical properties of arbuscular mycorrhizal fungal structures

International Nuclear Information System (INIS)

Perez, Adverdi; V-Hernandez, Alejandra; Rudamas, Carlos; Dreyer, Beatriz

2008-01-01

It was already reported by B. Dreyer at al. [1] that all fungal structures, both intra- and extra-radical fluoresced under blue light excitation regardless of their state (dead or alive). The source of the so called autofluorescence appears to be localized in the fungal cell wall. This supports the use of photoluminescence for the evaluation of AM colonization. However, the interpretation of these results is still in discussion [1-4]. In this work, arbuscular mycorrhizal spores were isolated from the rhizosphere of mango (Mangifera indica L.) plants by the method of wet sieving and decanting of Gerdemann and Nicolson [5] and studied by photoluminescence spectroscopy. Our experimental setup consists of an epifluorescence microscope (EM) coupled to a CCD-spectrometer through an arrangement of a home-made-telescope + fiber optic. This experimental setup allows the capture of images of the mycorrhizal structures (as usual in a standard epifluorescence microscope) combined with measurements of their corresponding emission bands. The preliminary results based on images obtained by standard EM do not clearly show that the emission is originated in the fungal cell walls as reported in Ref. 1. On the other hand, a very broad emission band in the visible part of the electromagnetic spectrum was observed in these spores by exciting at 450-490 nm and 300- 380 nm. We obtain a Full Width at Half Maximum (FWHM) of around 200 nm for this emission band whichis centered at 515 nm. This broad band seems to be composed of two narrower bands peaked around 494 and 547 nm and with FWHM of 50 nm and 150 nm, respectively. The profile of the observed emission band is in good agreement with the bands reported in Ref. 1 for vesicles, arbuscules and spores measured using the λ-Scan of a confocal laser scanning microscope. However, our results for spores show that the maxima of the narrower bands are shifted to higher energies in comparison to the corresponding bands observed in Ref. 1

Isolation of fungal homokaryotic lines from heterokaryotic transformants by sonic disruption of mycelia.

Science.gov (United States)

Bashi, Zafer Dallal; Khachatourians, George; Hegedus, Dwayne Daniel

2010-01-01

Fungal hyphae--and in some cases, spores--are multi-nucleate. During genetic transformation of these spores or mycelia, only one nucleus generally receives the transferred T-DNA generating heterokaryotic colonies. Characterization of genetic changes, such as the effects of gene disruption in the transformants, requires purified homokaryotic lines. Hyphal tip transfer has conventionally been used to isolate homokaryons. We developed an alternative method for purification of fungal homokaryons from transformed heterokaryotic lines of Sclerotinia sclerotiorum. Ultrasound pulses were used to generate bi-septate, unicellular hyphal fragments that regenerate under selection to produce homokaryotic lines that can be easily identified using colony PCR. This technique facilitates the purification of transformed lines, which allows for routine genome manipulation, and should be adaptable for other filamentous fungi.

Cell cycle and cell death are not necessary for appressorium formation and plant infection in the fungal plant pathogen Colletotrichum gloeosporioides

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Barhoom Sima

2008-02-01

Full Text Available Abstract Background In order to initiate plant infection, fungal spores must germinate and penetrate into the host plant. Many fungal species differentiate specialized infection structures called appressoria on the host surface, which are essential for successful pathogenic development. In the model plant pathogen Magnaporthe grisea completion of mitosis and autophagy cell death of the spore are necessary for appressoria-mediated plant infection; blocking of mitosis prevents appressoria formation, and prevention of autophagy cell death results in non-functional appressoria. Results We found that in the closely related plant pathogen Colletotrichum gloeosporioides, blocking of the cell cycle did not prevent spore germination and appressoria formation. The cell cycle always lagged behind the morphogenetic changes that follow spore germination, including germ tube and appressorium formation, differentiation of the penetrating hypha, and in planta formation of primary hyphae. Nuclear division was arrested following appressorium formation and was resumed in mature appressoria after plant penetration. Unlike in M. grisea, blocking of mitosis had only a marginal effect on appressoria formation; development in hydroxyurea-treated spores continued only for a limited number of cell divisions, but normal numbers of fully developed mature appressoria were formed under conditions that support appressoria formation. Similar results were also observed in other Colletotrichum species. Spores, germ tubes, and appressoria retained intact nuclei and remained viable for several days post plant infection. Conclusion We showed that in C. gloeosporioides the differentiation of infection structures including appressoria precedes mitosis and can occur without nuclear division. This phenomenon was also found to be common in other Colletotrichum species. Spore cell death did not occur during plant infection and the fungus primary infection structures remained viable

Evaluation of fungal air contamination in selected wards of two tertiary hospitals in Tehran, Iran

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Zahra Kamali Sarwestani

2017-07-01

Conclusion: According to the results of this study, the frequency and diversity of fungal spores in hospital wards were different. In addition, since the fungal contamination in the hospital environment are affected by various environmental factors and the efficiency of ventilation systems, some of these wards require better ventilation system as well as regular monitoring to remove these fungal bioaerosols in order to maintain the health of patients and health care workers.

Assessment of indoor air in Austrian apartments with and without visible mold growth

Science.gov (United States)

Haas, D.; Habib, J.; Galler, H.; Buzina, W.; Schlacher, R.; Marth, E.; Reinthaler, F. F.

Fungal spores are transported across great distances in the outdoor air and are also regularly found indoors. Building conditions and behavior-related problems in apartments may lead to massive growth of mold within a very short period of time. The aim of this study was to evaluate whether the visible growth of mold indoors influences the concentration of fungal spores in the air as well as the variety of their species. Samples were collected from 66 households in Austria. For each sampling, the corresponding outdoor air was measured as reference value. The size of the visible mold growth was categorized in order to correlate the extent of mold growth with the concentration of airborne spores as well as the fungal genera. In order to determine fungal spore concentrations in the air, the one-stage MAS-100 ® air sampler was used. Malt extract agar (MEA) and dichloran glycerol agar (DG18) plates were used as culture media. The total colony forming units (CFU) per m 3 were determined. The fungi were identified from the isolated colonies. The results show that in apartments visibly affected by mold, the median values were significantly higher than those of apartments without visible mold growth. The extent of visible mold growth is significantly correlated with both concentration of fungal spores ( pPenicillium sp. and Aspergillus sp. ( pPenicillium and Aspergillus in the air of apartments is recommended for assessing fungal exposure.

Long-term cryopreservation of non-spore-forming fungi in Microbank™ beads for plant pathological investigations.

Science.gov (United States)

Lakshman, Dilip K; Singh, Vimla; Camacho, Manuel E

2018-05-01

Long-term preservation of experimental fungi without genetic, morphological, and pathogenic changes is of paramount importance in mycological and plant pathological investigations. Several cryogenic and non-cryogenic methods are available for the preservation of fungi, but the methods can be cumbersome, hazardous, expensive, and often not suitable for long-term storage of non-spore-forming (sterile) fungi. A method of preservation of spore-forming fungi in commercially available porous beads (Micrbank™) under cryogenic condition was successfully tested for three non-spore-forming basidiomycetes genera: Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) (n = 19), Ceratobasidium species (n = 1), and Waitea circinata (n = 3), and a non-spore forming ascomycetes, Sclerotinia sclerotiorum (n = 1). For comparison, spore-forming ascomycetous fungi, Alternaria alternata (n = 1), Bauveria basiana (n = 2), Botrytis cinerea (n = 1), Fusarium oxysporum f.sp. gladiolii (n = 1), Trichoderma spp. (n = 3), and Thielaviopsis basicola (n = 2) were also cryopreserved in Microbank beads. Viable fungal isolates of all test species were retrieved after five years of storage at -80 °C, which was longer than the viabilities of the corresponding isolates cryopreserved in agar plugs or colonized wheat seeds. Fungi revived from the Microbank beads maintained identical morphology and cultural characteristics of the parent isolates. Randomly selected Rhizoctonia isolates revived from the Microbank beads maintained respective pathological properties of the parent isolates; also, no mutation was detected in the internal transcribed spacer (ITS) ribosomal DNA when compared with respective cultures maintained at ambient temperature. This finding demonstrated the utility of cryopreservation in Microbank beads as a convenient alternative to conventional long-term preservation of a wide group of fungal cultures for plant pathological investigations

Building and environmental factors that influence bacterial and fungal loading on air conditioning cooling coils.

Science.gov (United States)

Bakker, A; Siegel, J A; Mendell, M J; Peccia, J

2018-05-30

We investigated bacterial and fungal concentrations on cooling coils of commercial AC units and quantified associations between microbial loads and AC unit or building operational parameters. A field campaign was conducted to sample 25 AC units in the humid, subtropical climate of Southern CT, USA and 15 AC units in the hot-summer Mediterranean climate of Sacramento, CA, USA. Median concentrations (with interquartile range) of bacteria and fungi on the cooling coils were 1.2 × 10 7 (5.1 × 10 6 -3.9 × 10 7 ) cells/m 2 and 7.6 × 10 5 (5.6 × 10 4 -4.4 × 10 6 ) spore equivalents (SE)/m 2 , respectively. Concentrations varied among units with median unit concentrations ranging three orders of magnitude for bacteria and seven orders of magnitude for fungi. Controlled comparisons and multivariable regressions indicate that dominant factors associated with AC coil loading include the nominal efficiency of upstream filters (P = .008 for bacteria and P < .001 for fungi) and coil moisture, which was reflected in fungal loading differences between top and bottom halves of the AC coils in Southern CT (P = .05) and the dew points of the two climates considered (P = .04). Environmental and building characteristics explained 42% (P < .001) of bacterial concentration variability and 66% (P < .001) of fungal concentration variability among samples. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Efficacy Of Some Fungal Isolates And Their Applications For Controlling The Immature Stages Of The Medfly, Ceratitis Capitata (Wiedemann) Before And During Sterile Insect Technique (SIT)

International Nuclear Information System (INIS)

OUDA, S.M.; EL-AKHDAR, E.A.

2009-01-01

The application of the sterile insect technique (SIT) needs a suitable and effective cheap method alternative to chemical pesticides for suppressing or controlling the population density of the Mediterranean fruit fly, Ceratitis capitata (Wi ed.). The effectiveness of five different fungal isolates: Trichoderma longibranchiatum, T. harzianum, Aspergillus terreus, A. niger and Penicillium oxalicum was evaluated against the immature stages of med fly. Filtrate and three spore suspension concentrations (104, 106 and 108 spores/ml) of each fungal isolate were applied to both 3 rd larval instar and pupae at two ages (2 and 8 days-old). Percentage pupation, pupal mortality, adult emergence and sex ratio were studied. The percentage of pupation of 3 rd larval instar was not affected. However, there was a significant increase (P<0.05) in the percentage of pupal mortality as well as in adult emergence from pupae 2 and 8 days-old at all fungal treatments. Pupae (2 days-old) were more susceptible than pupae (8 days-old) to all fungal treatments. The microbial biomass of both A. terreus and P. oxalicum was selected to apply with different concentrations (5, 10 and 20 w/w) on the pupae at the two ages (2 and 8 days-old). Best results were recorded with biomass at 20% concentration of both fungal isolates. Percentage of mortality was significantly higher (P<0.05) than 50% at the two ages (2 and 8 days-old) for both fungi. A significant decrease (P<0.05) was obtained in adult females emergence resulted in a reduction in the number of eggs deposited and subsequently the punctures on fruits. Chitinase enzyme (important for degradation of chitin which is a component of insect cuticle) and cellulase enzyme (used for improving soil fertility) were determined. The microbial biomass by A. terreus was appeared to be safe to human and animals and can be used for combating immature stages of medfly.

Extraction of High Molecular Weight DNA from Fungal Rust Spores for Long Read Sequencing.

Science.gov (United States)

Schwessinger, Benjamin; Rathjen, John P

2017-01-01

Wheat rust fungi are complex organisms with a complete life cycle that involves two different host plants and five different spore types. During the asexual infection cycle on wheat, rusts produce massive amounts of dikaryotic urediniospores. These spores are dikaryotic (two nuclei) with each nucleus containing one haploid genome. This dikaryotic state is likely to contribute to their evolutionary success, making them some of the major wheat pathogens globally. Despite this, most published wheat rust genomes are highly fragmented and contain very little haplotype-specific sequence information. Current long-read sequencing technologies hold great promise to provide more contiguous and haplotype-phased genome assemblies. Long reads are able to span repetitive regions and phase structural differences between the haplomes. This increased genome resolution enables the identification of complex loci and the study of genome evolution beyond simple nucleotide polymorphisms. Long-read technologies require pure high molecular weight DNA as an input for sequencing. Here, we describe a DNA extraction protocol for rust spores that yields pure double-stranded DNA molecules with molecular weight of >50 kilo-base pairs (kbp). The isolated DNA is of sufficient purity for PacBio long-read sequencing, but may require additional purification for other sequencing technologies such as Nanopore and 10× Genomics.

Separation of mycotoxin-containing sources in grain dust and determination of their mycotoxin potential.

Science.gov (United States)

Palmgren, M S; Lee, L S

1986-01-01

Two distinct reservoirs of mycotoxins exist in fungal-infected cereal grains--the fungal spores and the spore-free mycelium-substrate matrix. Many fungal spores are of respirable size and the mycelium-substrate matrix can be pulverized to form particles of respirable size during routine handling of grain. In order to determine the contribution of each source to the level of mycotoxin contamination of dust, we developed techniques to harvest and separate mycelium-substrate matrices from spores of fungi. Conventional quantitative chromatographic analyses of separated materials indicated that aflatoxin from Aspergillus parasiticus, norsolorinic acid from a mutant of A. parasiticus, and secalonic acid D from Penicillium oxalicum were concentrated in the mycelium-substrate matrices and not in the spores. In contrast, spores of Aspergillus niger and Aspergillus fumigatus contained significant concentrations of aurasperone C and fumigaclavine C, respectively; only negligible amounts of the toxins were detected in the mycelium-substrate matrices of these two fungi. PMID:3709472

Effect of concentration, exposure time, temperature, and relative humidity on the toxicity of sulfur dioxide to the spores of Botrytis cinerea

Energy Technology Data Exchange (ETDEWEB)

Couey, H.M.; Uota, M.

1961-12-01

When spores of Botrytis cinerea are exposed to SO/sub 2/ gas, the subsequent reduction in spore germination is quantitatively proportional to the SO/sub 2/ concentration and the exposure time. The toxicity of SO/sub 2/ increases with increasing relative humidity. In an atmosphere of 96% RH, SO/sub 2/ is more than 20 times as effective as at 75% RH. The toxicity also increases about 1.5 times for each 10/sup 0/C rise in temperature between 0/sup 0/ and 30/sup 0/C. 8 references, 4 figures, 1 table.

Assessing the anti-fungal efficiency of filters coated with zinc oxide nanoparticles

Science.gov (United States)

Decelis, Stephen; Sardella, Davide; Triganza, Thomas; Brincat, Jean-Pierre; Gatt, Ruben; Valdramidis, Vasilis P.

2017-05-01

Air filters support fungal growth, leading to generation of conidia and volatile organic compounds, causing allergies, infections and food spoilage. Filters that inhibit fungi are therefore necessary. Zinc oxide (ZnO) nanoparticles have anti-fungal properties and therefore are good candidates for inhibiting growth. Two concentrations (0.012 M and 0.12 M) were used to coat two types of filters (melt-blown and needle-punched) for three different periods (0.5, 5 and 50 min). Rhizopus stolonifer and Penicillium expansum isolated from spoiled pears were used as test organisms. Conidial suspensions of 105 to 103 spores ml-1 were prepared in Sabouraud dextrose agar at 50°C, and a modified slide-culture technique was used to test the anti-fungal properties of the filters. Penicillium expansum was the more sensitive organism, with inhibition at 0.012 M at only 0.5 min coating time on the needle-punched filter. The longer the coating time, the more effective inhibition was for both organisms. Furthermore, it was also determined that the coating process had only a slight effect on the Young's Moduli of the needle-punched filters, while the Young's Moduli of the melt-blown filters is more susceptible to the coating method. This work contributes to the assessment of the efficacy of filter coating with ZnO nanopaticles aimed at inhibiting fungal growth.

Bioactivities of Ketones Terpenes: Antifungal Effect on F. verticillioides and Repellents to Control Insect Fungal Vector, S. zeamais

OpenAIRE

Pizzolitto, Romina P.; Herrera, Jimena M.; Zaio, Yesica P.; Dambolena, Jose S.; Zunino, Maria P.; Gallucci, Mauro N.; Zygadlo, Julio A.

2015-01-01

Maize is one the most important staple foods in the world. However, numerous pests, such as fungal pathogens, e.g., Fusarium verticillioides, and insects, such as Sitophlilus zeamais, attack maize grains during storage. Many F. verticillioides strains produce fumonisins, one of the most important mycotoxin that causes toxic effects on human and animal health. This situation is aggravated by the insect fungal vector, Sitophlilus zeamais, which contributes to the dispersal of fungal spores, and...

Spore Coat Architecture of Clostridium novyi-NT spores

Energy Technology Data Exchange (ETDEWEB)

Plomp, M; McCafferey, J; Cheong, I; Huang, X; Bettegowda, C; Kinzler, K; Zhou, S; Vogelstein, B; Malkin, A

2007-05-07

Spores of the anaerobic bacterium Clostridium novyi-NT are able to germinate in and destroy hypoxic regions of tumors in experimental animals. Future progress in this area will benefit from a better understanding of the germination and outgrowth processes that are essential for the tumorilytic properties of these spores. Towards this end, we have used both transmission electron microscopy and atomic force microscopy to determine the structure of dormant as well as germinating spores. We found that the spores are surrounded by an amorphous layer intertwined with honeycomb parasporal layers. Moreover, the spore coat layers had apparently self-assembled and this assembly was likely to be governed by crystal growth principles. During germination and outgrowth, the honeycomb layers as well as the underlying spore coat and undercoat layers sequentially dissolved until the vegetative cell was released. In addition to their implications for understanding the biology of C. novyi-NT, these studies document the presence of proteinaceous growth spirals in a biological organism.

Electrochemical study of AISI 304 stainless steel during the exposure at the disinfectant solutions with fungal suspensions

Energy Technology Data Exchange (ETDEWEB)

Stoica, M.; Bruma, M. [Department of Biochemistry and Technologies, Faculty of Food Science and Engineering, ' ' Dunarea de Jos' ' University of Galati, 111 Domneasca Street, 800201 Galati (Romania); Carac, G. [Department of Chemistry, Faculty of Sciences, ' ' Dunarea de Jos' ' University of Galati, 47 Domneasca Street, 800008 Galati (Romania)

2010-12-15

The effect of fungal suspensions on the disinfectant solution (Actisept with sodium dichloroisocyanurate as active substance, which releases 140 ppm active chlorine) and the corrosion behavior of type AISI 304 stainless steel (SS) were evaluated through linear polarization. The work presents a study of biocide Actisept solution as reference solution and the mixed solutions: reference biocide Actisept solution with three fungal suspensions (10 vol%), Aspergillus niger, Candida mycoderma, and Saccharomyces cerevisiae. The fungal suspension has an important influence on the synergistic effect of the SS corrosion in the following order: Saccharomyces cerevisiae > Candida mycoderma > Aspergillus niger. The E{sub corr} values from the mixed solutions decreased with immersion time of the samples. In the case of Aspergillus niger, the corrosion rate was lower than that of any other type of fungal suspension tested, because the arrangement in solutions, the size and composition of Aspergillus niger spores were more favorable to protect the spores of the chemical and the electrical stressing. The results showed that there is a synergistic effect between the active substances from the disinfectant, fungal suspensions and applied potential, thus this effect can be taken into consideration for the hygienic and safety food industry. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

Quantification of Nonproteolytic Clostridium botulinum Spore Loads in Food Materials.

Science.gov (United States)

Barker, Gary C; Malakar, Pradeep K; Plowman, June; Peck, Michael W

2016-01-04

We have produced data and developed analysis to build representations for the concentration of spores of nonproteolytic Clostridium botulinum in materials that are used during the manufacture of minimally processed chilled foods in the United Kingdom. Food materials are categorized into homogenous groups which include meat, fish, shellfish, cereals, fresh plant material, dairy liquid, dairy nonliquid, mushroom and fungi, and dried herbs and spices. Models are constructed in a Bayesian framework and represent a combination of information from a literature survey of spore loads from positive-control experiments that establish a detection limit and from dedicated microbiological tests for real food materials. The detection of nonproteolytic C. botulinum employed an optimized protocol that combines selective enrichment culture with multiplex PCR, and the majority of tests on food materials were negative. Posterior beliefs about spore loads center on a concentration range of 1 to 10 spores kg(-1). Posterior beliefs for larger spore loads were most significant for dried herbs and spices and were most sensitive to the detailed results from control experiments. Probability distributions for spore loads are represented in a convenient form that can be used for numerical analysis and risk assessments. Copyright © 2016 Barker et al.

Effects of produced water discharges on the colonization potential of Macrocystis pyrifera spores

International Nuclear Information System (INIS)

Lewis, R.J.; Reed, D.C.

1993-01-01

Point sources of pollution (e.g. industrial outfalls) may produce ecological impacts at distant locations if pollutants affect dispersive propagules. The authors used laboratory experiments to determine how exposure to produced water (PW; aqueous fraction of petroleum production that is typically discharged into coastal waters) in the water column influences the colonization potential of giant kelp (Macrocystis pyrifera) spores on the bottom. Spores were maintained in suspension in 18 L containers and exposed to one of five concentrations of PW (0 to 10%) for varying amounts of time. Spore swimming generally decreased with increasing PW concentration and exposure duration, with the specific pattern of decrease differing between experimental trials done at different dates. The effect of exposure duration in the water column on the ability of swimming spores to attach to plastic dishes placed the bottom varied with PW concentration. Spores placed in 1 and 10% PW showed a steady decline in their ability to attach with increased exposure; lower concentrations of PW had no such effects. The proportion of spores that germinated after attachment varied tremendously with exposure duration and date of experimental trial. A low proportion of spores that settled during the first 12 h germinated, indicative of a short period of precompetency. Surprisingly, water column exposure to high concentrations of PW during the first 12 h reduced this precompetent period and greatly improved germination success. The magnitude of this enhancement, however, varied among dates. Delayed expression of PW effects were not observed in developing gametophytes; survival of individuals that successfully germinated and gamete production was not affected by previous exposure to PW as a spore

Enzyme-driven Bacillus spore coat degradation leading to spore killing.

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Mundra, Ruchir V; Mehta, Krunal K; Wu, Xia; Paskaleva, Elena E; Kane, Ravi S; Dordick, Jonathan S

2014-04-01

The bacillus spore coat confers chemical and biological resistance, thereby protecting the core from harsh environments. The primarily protein-based coat consists of recalcitrant protein crosslinks that endow the coat with such functional protection. Proteases are present in the spore coat, which play a putative role in coat degradation in the environment. However these enzymes are poorly characterized. Nonetheless given the potential for proteases to catalyze coat degradation, we screened 10 commercially available proteases for their ability to degrade the spore coats of B. cereus and B. anthracis. Proteinase K and subtilisin Carlsberg, for B. cereus and B. anthracis spore coats, respectively, led to a morphological change in the otherwise impregnable coat structure, increasing coat permeability towards cortex lytic enzymes such as lysozyme and SleB, thereby initiating germination. Specifically in the presence of lysozyme, proteinase K resulted in 14-fold faster enzyme induced germination and exhibited significantly shorter lag times, than spores without protease pretreatment. Furthermore, the germinated spores were shown to be vulnerable to a lytic enzyme (PlyPH) resulting in effective spore killing. The spore surface in response to proteolytic degradation was probed using scanning electron microscopy (SEM), which provided key insights regarding coat degradation. The extent of coat degradation and spore killing using this enzyme-based pretreatment approach is similar to traditional, yet far harsher, chemical decoating methods that employ detergents and strong denaturants. Thus the enzymatic route reduces the environmental burden of chemically mediated spore killing, and demonstrates that a mild and environmentally benign biocatalytic spore killing is achievable. © 2013 Wiley Periodicals, Inc.

Trichoderma asperelloides Spores Downregulate dectin1/2 and TLR2 Receptors of Mice Macrophages and Decrease Candida parapsilosis Phagocytosis Independent of the M1/M2 Polarization

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Andréa G. dos Santos

2017-09-01

Full Text Available The intensive use of pesticides to control pests in agriculture has promoted several issues relating to environment. As chemical pesticides remain controversial, biocontrol agents originating from fungi could be an alternative. Among them, we highlight biocontrol agents derived from the fungi genus Trichoderma, which have been documented in limiting the growth of other phytopathogenic fungus in the roots and leaves of several plant species. An important member of this genus is Trichoderma asperelloides, whose biocontrol agents have been used to promote plant growth while also treating soil diseases caused by microorganisms in both greenhouses and outdoor crops. To evaluate the safety of fungal biological agents for human health, tests to detect potentially adverse effects, such as allergenicity, toxicity, infectivity and pathogenicity, are crucial. In addition, identifying possible immunomodulating properties of fungal biocontrol agents merits further investigation. Thus, the aim of this study was to evaluate the effects of T. asperelloides spores in the internalization of Candida parapsilosis yeast by mice phagocytes, in order to elucidate the cellular and molecular mechanism of this interaction, as a model to understand possible in vivo effects of this fungus. For this, mice were exposed to a fungal spore suspension through-intraperitoneal injection, euthanized and cells from the peripheral blood and peritoneal cavity were collected for functional, quantitative and phenotypic analysis, throughout analysis of membrane receptors gene expression, phagocytosis ability and cells immunophenotyping M1 (CCR7 and CD86 and M2 (CCR2 and CD206. Our analyses showed that phagocytes exposed to fungal spores had reduced phagocytic capacity, as well as a decrease in the quantity of neutrophils and monocytes in the peripheral blood and peritoneal cavity. Moreover, macrophages exposed to T. asperelloides spores did not display the phenotypic profile M1/M2, and

Influence of selected variables on transport of plutonium to spores of Aspergillus niger

International Nuclear Information System (INIS)

Au, F.H.F.; Beckert, W.F.

1975-01-01

Studies were carried out on the influences of different chemical forms and concentrations of Pu at two hydrogen ion concentrations of the culture medium on uptake and transport of 238 Pu to the spores of Aspergillus niger. Results indicated that Pu, when added to the culture medium as dioxide microspheres, nitrate, or citrate complex, was transported to the spores, and that an almost linear relationship existed between transport and concentration. Raising the pH of the culture medium from 2.5 to 5.5 generally increased transport of Pu to spores for all three chemical forms. At Pu concentrations of 224 pCi/g in the culture media, and for both pH 2.5 and 5.5, transport of Pu to spores was approximately three times as high from the nitrate or citrate form as from the dioxide microspheres. (auth)

Human Fungal Pathogens of Mucorales and Entomophthorales.

Science.gov (United States)

Mendoza, Leonel; Vilela, Raquel; Voelz, Kerstin; Ibrahim, Ashraf S; Voigt, Kerstin; Lee, Soo Chan

2014-11-06

In recent years, we have seen an increase in the number of immunocompromised cohorts as a result of infections and/or medical conditions, which has resulted in an increased incidence of fungal infections. Although rare, the incidence of infections caused by fungi belonging to basal fungal lineages is also continuously increasing. Basal fungal lineages diverged at an early point during the evolution of the fungal lineage, in which, in a simplified four-phylum fungal kingdom, Zygomycota and Chytridiomycota belong to the basal fungi, distinguishing them from Ascomycota and Basidiomycota. Currently there are no known human infections caused by fungi in Chytridiomycota; only Zygomycotan fungi are known to infect humans. Hence, infections caused by zygomycetes have been called zygomycosis, and the term "zygomycosis" is often used as a synonym for "mucormycosis." In the four-phylum fungal kingdom system, Zygomycota is classified mainly based on morphology, including the ability to form coenocytic (aseptated) hyphae and zygospores (sexual spores). In the Zygomycota, there are 10 known orders, two of which, the Mucorales and Entomophthorales, contain species that can infect humans, and the infection has historically been known as zygomycosis. However, recent multilocus sequence typing analyses (the fungal tree of life [AFTOL] project) revealed that the Zygomycota forms not a monophyletic clade but instead a polyphyletic clade, whereas Ascomycota and Basidiomycota are monophyletic. Thus, the term "zygomycosis" needed to be further specified, resulting in the terms "mucormycosis" and "entomophthoramycosis." This review covers these two different types of fungal infections. Copyright © 2015 Cold Spring Harbor Laboratory Press; all rights reserved.

Root colonization and spore abundance of arbuscular mycorrhizal fungi in distinct successional stages from an Atlantic rainforest biome in southern Brazil.

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Zangaro, Waldemar; Rostirola, Leila Vergal; de Souza, Priscila Bochi; de Almeida Alves, Ricardo; Lescano, Luiz Eduardo Azevedo Marques; Rondina, Artur Berbel Lírio; Nogueira, Marco Antonio; Carrenho, Rosilaine

2013-04-01

The influence of plant functional groups and moderate seasonality on arbuscular mycorrhizal (AM) fungal status (root colonization and spore density) was investigated during 13 consecutive months in a chronosequence of succession in southern Brazil, consisting of grassland field, scrub vegetation, secondary forest and mature forest, in a region of transition from tropical to subtropical zones. AM root colonization and spore density decreased with advancing succession and were highest in early successional sites with grassland and scrub vegetation, intermediary in the secondary forest and lowest in the mature forest. They were little influenced by soil properties, but were sufficiently influenced by the fine root nutrient status and fine root traits among different functional plant groups. AM root colonization and spore density were higher during the favourable plant growth season (spring and summer) than during the less favourable plant growth season (autumn and winter). Spore density displayed significant seasonal variation at all sites, whilst root colonization displayed significant seasonal variation in grassland, scrub and secondary forest, but not in mature forest. The data suggest that (1) different plant functional groups display different relationships with AM fungi, influencing their abundance differentially; (2) plant species from early successional phases are more susceptible to AM root colonization and maintain higher AM sporulation than late successional species; (3) fine root traits and nutrient status influence these AM fungal attributes; and (4) higher AM spore production and root colonization is associated with the season of higher light incidence and temperature, abundant water in soil and higher plant metabolic activity.

Clonal diversity and population genetic structure of arbuscular mycorrhizal fungi (Glomus spp.) studied by multilocus genotyping of single spores

DEFF Research Database (Denmark)

Holtgrewe-Stukenbrock, Eva; Rosendahl, Søren

2005-01-01

A nested multiplex PCR (polymerase chain reaction) approach was used for multilocus genotyping of arbuscular mycorrhizal fungal populations. This method allowed us to amplify multiple loci from Glomus single spores in a single PCR amplification. Variable introns in the two protein coding genes Gm......FOX2 and GmTOR2 were applied as codominant genetic markers together with the LSU rDNA.   Genetic structure of Glomus spp. populations from an organically and a conventionally cultured field were compared by hierarchical sampling of spores from four plots in each field. Multilocus genotypes were...

Soil spore bank communities of ectomycorrhizal fungi in endangered Chinese Douglas-fir forests.

Science.gov (United States)

Wen, Zhugui; Shi, Liang; Tang, Yangze; Hong, Lizhou; Xue, Jiawang; Xing, Jincheng; Chen, Yahua; Nara, Kazuhide

2018-01-01

Chinese Douglas-fir (Pseudotsuga sinensis) is an endangered Pinaceae species found in several isolated regions of China. Although soil spore banks of ectomycorrhizal (ECM) fungi can play an important role in seedling establishment after disturbance, such as in the well-known North American relative (Pseudotsuga menziesii), we have no information about soil spore bank communities in relict forests of Chinese Douglas-fir. We conducted bioassays of 73 soil samples collected from three Chinese Douglas-fir forests, using North American Douglas-fir as bait seedlings, and identified 19 species of ECM fungi. The observed spore bank communities were significantly different from those found in ECM fungi on the roots of resident trees at the same sites (p = 0.02). The levels of potassium (K), nitrogen (N), organic matter, and the pH of soil were the dominant factors shaping spore bank community structure. A new Rhizopogon species was the most dominant species in the spore banks. Specifically, at a site on Sanqing Mountain, 22 of the 57 surviving bioassay seedlings (representing 21 of the 23 soil samples) were colonized by this species. ECM fungal richness significantly affected the growth of bioassay seedlings (R 2  = 0.20, p = 0.007). Growth was significantly improved in seedlings colonized by Rhizopogon or Meliniomyces species compared with uncolonized seedlings. Considering its specificity to Chinese Douglas-fir, predominance in the soil spore banks, and positive effect on host growth, this new Rhizopogon species could play critical roles in seedling establishment and forest regeneration of endangered Chinese Douglas-fir.

Spore germination of fungi belonging to Aspergillus species under deep-sea conditions

Digital Repository Service at National Institute of Oceanography (India)

Damare, S.R.; Nagarajan, M.; Raghukumar, C.

Culture Collection (Chandigarh, India) were used. 2.2. Media Sediment extract (SE) was prepared using sediments collected from the Central Indian Basin. Sediment (30% (w/v)) was suspended in sterile seawater containing antibiotics (penicillin— 40,000U... extract was used as one of the growth media. Malt extract broth (MEB, HiMedia Laboratories, India) prepared in seawater was used for the comparative studies. 2.3. Fungal spore suspension The cultures were inoculated onto malt extract agar (MEA) plates...

Electron Beam Irradiation Dose Dependently Damages the Bacillus Spore Coat and Spore Membrane

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S. E. Fiester

2012-01-01

Full Text Available Effective control of spore-forming bacilli begs suitable physical or chemical methods. While many spore inactivation techniques have been proven effective, electron beam (EB irradiation has been frequently chosen to eradicate Bacillus spores. Despite its widespread use, there are limited data evaluating the effects of EB irradiation on Bacillus spores. To study this, B. atrophaeus spores were purified, suspended in sterile, distilled water, and irradiated with EB (up to 20 kGy. Irradiated spores were found (1 to contain structural damage as observed by electron microscopy, (2 to have spilled cytoplasmic contents as measured by spectroscopy, (3 to have reduced membrane integrity as determined by fluorescence cytometry, and (4 to have fragmented genomic DNA as measured by gel electrophoresis, all in a dose-dependent manner. Additionally, cytometry data reveal decreased spore size, increased surface alterations, and increased uptake of propidium iodide, with increasing EB dose, suggesting spore coat alterations with membrane damage, prior to loss of spore viability. The present study suggests that EB irradiation of spores in water results in substantial structural damage of the spore coat and inner membrane, and that, along with DNA fragmentation, results in dose-dependent spore inactivation.

Assessment of relevant fungal species in clinical solid wastes.

Science.gov (United States)

Noman, Efaq Ali; Al-Gheethi, A A; Rahman, Nik Norulaini Nik Ab; Nagao, H; Ab Kadir, M O

2016-10-01

The study aimed to determine the fungal diversity in clinical waste samples from a healthcare facility in Penang Malaysia. Different fungi species were detected in 83.75 % of the 92 clinical waste samples that were screened from different sections of the healthcare facility. One hundred fifty fungal isolates comprising of 8 genera and 36 species were obtained. They were purified by using single spore isolation technique. Subsequently, the isolates were identified by phenotypic method based on morphological and culture characteristics on different culture media. Among all fungal isolates, Aspergillus spp. in section Nigri 10.2 %, Aspergillus niger 9.5 %, Aspergillus fumigatus 8.8 %, Penicillium. simplicissium 8 %, Aspergillus tubingensis 7.3 %, Aspergillus terreus var. terreus 6.6 %, Penicillium waksmanii 5.9 % and Curvularia lunata 6.5 % were the most frequent. Among five sections of the Wellness Centre, the clinical wastes collected from the diagnostic labs of haematology section had the highest numbers of fungal species (29 species). Glove wastes had the highest numbers of fungal species (19 species) among 17 types of clinical wastes screened. Among all fungal species, Aspergillus spp. exhibited higher growth at 37 °C than at 28 °C, indicating the potential of these opportunistic fungi to cause diseases in human. These results indicated the potential of hospital wastes as reservoirs for fungal species.

Can fungal zoospores be the source of energy for the rumen protozoa Eudiplodinium maggii?

Science.gov (United States)

Miltko, Renata; Bełżecki, Grzegorz; Kowalik, Barbara; Michałowski, Tadeusz

2014-10-01

Results of our earlier studies showed the ability of ciliates Eudiplodinium maggii to digest and metabolize commercial chitin. The natural source of this polysaccharide in the rumen are fungi. The objectives of present research were to determine the effect of fungal zoospores on the survival and population density of E. maggii to quantify the concentration of chitin in the cells of protozoa and to examine the ability of E. maggii, to ferment chitin of fungal zoospores. The cultivation experiment showed that the survival of protozoa was shorter than 4 days when the culture medium was composed of buffer solution and lyophilized fungal spores. An enrichment of this medium with wheat gluten prolonged the survival of ciliates up to 8 days. The supplementation of the last medium with meadow hay enabled the protozoa to survive for 28 days but a positive effect was observed only during the last 8 days of experiment. The chitin content was 0.27 ng and 0.21-0.35 ng per single zoospore and ciliate, respectively. An increase in the concentration of volatile fatty acids (VFA) was found when protozoa were incubated with zoospores. The production rate of VFA was 46.3 pM/protozoan per h whereas the endogenous production did not exceed 31 pM/protozoan per h. The molar proportion of acetic acid was 77.7% and these of butyric and propionic acids-12.2 and 11.0%, respectively. The obtained results make it evident that carbohydrates present in fungal zoospores were utilized by protozoa in energy yielding processes. Copyright © 2013 Elsevier Ltd. All rights reserved.

Heat activation of Phycomyces blakesleeanus spores: theromdynamics and effect of alcohols, furfural, and high pressure.

Science.gov (United States)

Thevelein, J M; Van Assche, J A; Carlier, A R; Heremans, K

1979-08-01

The thermodynamic parameters for the heat activation of the sporangiospores of Phycomyces blakesleeanus were determined. For the apparent activation enthalpy (DeltaH(#)) a value of 1,151 kJ/mol was found, whereas a value of 3,644 J./ degrees K.mol was calculated for the apparent activation entropy (DeltaS(#)). n-Alcohols (from methanol to octanol), phenethyl alcohol, and furfural lowered the activation temperature of P. blakesleeanus spores. The heat resistance of the spores was lowered concomitantly. The effect of the alcohols was a linear function of the concentration in the range that could be applied. When the log of the concentration needed to produce an equal shift of the activation temperature was plotted for each alochol against the log of the octanol/water partition coefficient, a straight line was obtained. The free energy of adsorption of the n-alcohols to their active sites was calculated to be -2,487 J/mol of CH(2) groups. Although still inconclusive, this points toward an involvement of protein in the activation process. The effect of phenethyl alcohol was similar to the effect of n-alcohols, but furfural produced a greater shift than would be expected from the value of its partition coefficient. When the heat activation of the spores was performed under high pressure, the activation temperature was raised by 2 to 4 degrees K/1,000 atm. However, with pressures higher than 1,000 atm (1.013 x 10(5) kPa) the activation temperature was lowered until the pressure became lethal (more than 2,500 atm). It is known that membrane phase transition temperatures are shifted upward by about 20 degrees K/1,000 atm and that protein conformational changes are shifted upward by 2 to 6 degrees K/1,000 atm. Consequently, heat activation of fungal spores seems to be triggered by a protein conformational change and not by a membrane phase transition. Activation volumes of -54.1 cm(3)/mol at 38 degrees C and -79.3 cm(2)/mol at 40 degrees C were found for the lowering effect

Heat Activation of Phycomyces blakesleeanus Spores: Thermodynamics and Effect of Alcohols, Furfural, and High Pressure

Science.gov (United States)

Thevelein, Johan M.; Van Assche, Jozef A.; Carlier, Albert R.; Heremans, Karel

1979-01-01

The thermodynamic parameters for the heat activation of the sporangiospores of Phycomyces blakesleeanus were determined. For the apparent activation enthalpy (ΔH#) a value of 1,151 kJ/mol was found, whereas a value of 3,644 J./°K·mol was calculated for the apparent activation entropy (ΔS#). n-Alcohols (from methanol to octanol), phenethyl alcohol, and furfural lowered the activation temperature of P. blakesleeanus spores. The heat resistance of the spores was lowered concomitantly. The effect of the alcohols was a linear function of the concentration in the range that could be applied. When the log of the concentration needed to produce an equal shift of the activation temperature was plotted for each alochol against the log of the octanol/water partition coefficient, a straight line was obtained. The free energy of adsorption of the n-alcohols to their active sites was calculated to be −2,487 J/mol of CH2 groups. Although still inconclusive, this points toward an involvement of protein in the activation process. The effect of phenethyl alcohol was similar to the effect of n-alcohols, but furfural produced a greater shift than would be expected from the value of its partition coefficient. When the heat activation of the spores was performed under high pressure, the activation temperature was raised by 2 to 4°K/1,000 atm. However, with pressures higher than 1,000 atm (1.013 × 105 kPa) the activation temperature was lowered until the pressure became lethal (more than 2,500 atm). It is known that membrane phase transition temperatures are shifted upward by about 20°K/1,000 atm and that protein conformational changes are shifted upward by 2 to 6°K/1,000 atm. Consequently, heat activation of fungal spores seems to be triggered by a protein conformational change and not by a membrane phase transition. Activation volumes of −54.1 cm3/mol at 38°C and −79.3 cm2/mol at 40°C were found for the lowering effect of high pressure on the heat activation temperature

Spore communities of arbuscular mycorrhizal fungi and mycorrhizal associations in different ecosystems, south Australia

Directory of Open Access Journals (Sweden)

Z. I. Antoniolli

2002-09-01

Full Text Available Communities of arbuscular mycorrhizal fungi (AMF were surveyed in different South Australian ecosystems. The soil was wet-sieved for spore extraction, followed by the determination of presence and abundance of AMF species as well as the percentage of root colonization. Mycorrhizal associations were common and there was substantial fungal diversity in different ecosystems. Spores were most abundant in the permanent pasture system and less abundant under continuous wheat. The incidence of mycorrhizal associations in different plant species and the occurrence of Arum and Paris type colonization generally conformed with previous information. Spores of seventeen AMF were verified throughout seasonal changes in 1996 and 1997 in the permanent pasture and on four host species (Lolium perenne, Plantago lanceolata, Sorghum sp. and Trifolium subterraneum , set up with the same soils under greenhouse conditions. Glomus mosseae was the dominant spore type at all sampling times and in all trap cultures. Mycorrhizal diversity was significantly affected by different sampling times in trap cultures but not in field-collected soil. P. lanceolata, Sorghum sp. and T. subterraneum as hosts for trap cultures showed no differences in richness and diversity of AMF spores that developed in association with their roots. Abundance and diversity were lowest, however, in association with L. perenne , particularly in December 1996. Results show that the combination of spore identification from field-collected soil and trap cultures is essential to study population and diversity of AMF. The study provides baseline data for ongoing monitoring of mycorrhizal populations using conventional methods and material for the determination of the symbiotic effectiveness of AMF key members.

Handling technique of spore-forming bacteria in radiation sterilization. 1. Preparation of spores

International Nuclear Information System (INIS)

Koshikawa, Tomihiko

1994-01-01

This paper deals with a handling technique of spore-forming bacteria in radiation sterilization. An explanation is given under three sections: (1) life cycle of spore-forming bacteria, medium to form bacterial spores, and colony and purification methods of bacterial spores; (2) methods for measuring the number of bacterial spores and resistance against gamma radiation (D values); and (3) a test method for identifying spore-forming bacteria and a simple identification method. (N.K.)

Ectomycorrhizal fungal diversity in orchards of cultivated pecan (Carya illinoinensis; Juglandaceae).

Science.gov (United States)

Bonito, Gregory; Brenneman, Timothy; Vilgalys, Rytas

2011-10-01

Carya illinoinensis (pecan) belongs to the Juglandaceae (walnut family) and is a major economic nut crop in the southern USA. Although evidence suggests that some species in the Juglandaceae are ectomycorrhizal, investigations on their ectomycorrhizal fungal symbionts are quite limited. Here we assessed the ectomycorrhizal fungal diversity in cultivated orchards of C. illinoinensis. Five pecan orchards in southern Georgia, USA, were studied, three of which were known to fruit the native edible truffle species Tuber lyonii. We sequenced rDNA from single ectomycorrhizal root tips sampled from a total of 50 individual trees. Mycorrhizae were identified by ITS and LSU rDNA sequence-based methods. Forty-four distinct ectomycorrhizal taxa were detected. Sequestrate taxa including Tuber and Scleroderma were particularly abundant. The two most abundant sequence types belonged to T. lyonii (17%) and an undescribed Tuber species (~20%). Because of our interest in the ecology of T. lyonii, we also conducted greenhouse studies to determine whether this species would colonize and form ectomycorrhizae on roots of pecan, oak, or pine species endemic to the region. T. lyonii ectomycorrhizae were formed on pecan and oak seedlings, but not pine, when these were inoculated with spores. That oak and pecan seedling roots were receptive to truffle spores indicates that spore slurry inoculation could be a suitable method for commercial use and that, ecologically, T. lyonii may function as a pioneer ectomycorrhizal species for these hosts. © Springer-Verlag 2011

High-Resolution Spore Coat Architecture and Assembly of Bacillus Spores

Energy Technology Data Exchange (ETDEWEB)

Malkin, A J; Elhadj, S; Plomp, M

2011-03-14

Elucidating the molecular architecture of bacterial and cellular surfaces and its structural dynamics is essential to understanding mechanisms of pathogenesis, immune response, physicochemical interactions, environmental resistance, and provide the means for identifying spore formulation and processing attributes. I will discuss the application of in vitro atomic force microscopy (AFM) for studies of high-resolution coat architecture and assembly of several Bacillus spore species. We have demonstrated that bacterial spore coat structures are phylogenetically and growth medium determined. We have proposed that strikingly different species-dependent coat structures of bacterial spore species are a consequence of sporulation media-dependent nucleation and crystallization mechanisms that regulate the assembly of the outer spore coat. Spore coat layers were found to exhibit screw dislocations and two-dimensional nuclei typically observed on inorganic and macromolecular crystals. This presents the first case of non-mineral crystal growth patterns being revealed for a biological organism, which provides an unexpected example of nature exploiting fundamental materials science mechanisms for the morphogenetic control of biological ultrastructures. We have discovered and validated, distinctive formulation-specific high-resolution structural spore coat and dimensional signatures of B. anthracis spores (Sterne strain) grown in different formulation condition. We further demonstrated that measurement of the dimensional characteristics of B. anthracis spores provides formulation classification and sample matching with high sensitivity and specificity. I will present data on the development of an AFM-based immunolabeling technique for the proteomic mapping of macromolecular structures on the B. anthracis surfaces. These studies demonstrate that AFM can probe microbial surface architecture, environmental dynamics and the life cycle of bacterial and cellular systems at near

The effects of road building on arbuscular mycorrhizal fungal diversity in Huangshan Scenic Area.

Science.gov (United States)

Yang, Anna; Tang, Dongmei; Jin, Xiulong; Lu, Lin; Li, Xiaohong; Liu, Kun

2018-01-22

Arbuscular mycorrhizal (AM) fungi are vital soil microbes that connect many individual plants into a large functional organism via a vast mycelial network under the ground. In this study, the changes of soil AM fungal community in response to road-building disturbance caused by tourism development in Huangshan (Yellow Mountain) Scenic Area are assessed. Road building have brought negative effects on AM fungal community, inducing lower diversity parameters, including species number, spore density and diversity indices. However, the dominant genus and species of AM fungi which play key roles in the AM fungal community composition are quite similar before and after road building. Moreover, there are no significant differences in species richness of AM fungi associated with plants, suggesting the tolerance of AM fungal community to the disturbance of road building.

Two-stage fungal leaching of vanadium from uranium ore residue of the leaching stage using statistical experimental design

International Nuclear Information System (INIS)

Gharehbagheri, Hassan; Safdari, Jaber; Roostaazad, Reza; Rashidi, Abbas

2013-01-01

Highlights: ► In this work, the percent of vanadium recovery from uranium mine waste was 44.8. ► Unlike autotrophs, Aspergillus niger is a suitable microorganism to deal with such a resource. ► In the first step of the present work, citric acid was produced more than the other acids. ► When sterilization of uranium ore waste is not economic, two-step bioleaching is an appropriate method. - Abstract: In this investigation, bioleaching of vanadium from uranium ore residue of the leaching stage was studied by Aspergillus niger in a two-step process at 30 °C and 150 rpm. The first step was initiated by growth of fungi in the absence of mine waste. Using response surface methodology, three factors were surveyed for fungal growth: initial pH, sucrose concentration and spore population. Also concentrations of oxalic, citric, and gluconic acids were measured as response in this step. During 30 days, maximum productions of these acids were 3265, 11578, and 7988 mg/l, respectively. Initial pH and sucrose concentration were significant factors for oxalic and citric acid production; however, for gluconic acid production sucrose concentration and spore population were significant. Then, the content of each flask was filtered and mine waste was added to liquor with pulp density of 3%. During 3 days, in the second step, vanadium recovered about 44.8% in the liquor

Living fungal hyphae-templated porous gold microwires using nanoparticles as building blocks

International Nuclear Information System (INIS)

Rehman, Asma; Majeed, Muhammad Irfan; Ihsan, Ayesha; Hussain, Syed Zajif; Saif-ur-Rehman; Ghauri, Muhammad Afzal; Khalid, Zafar M.; Hussain, Irshad

2011-01-01

A simple and environmentally benign green method is reported to decorate growing fungal hyphae with high loading of gold nanoparticles, which were initially produced using aqueous tea extract as a sole reducing/stabilizing agent. Inoculation of fungal spores in aqueous suspension of nanoparticles led to the growth of intensely red-coloured fungal hyphae due to the accumulation of gold nanoparticles. Heat treatment of these hybrid materials led to the formation of porous gold microwires. This report is thus an interesting example of using green and sustainable approach to produce nanostructured materials which have potential applications in catalysis, sensing and electronics.Graphical AbstractPorous gold microwires are formed by the heat treatment of fungal hyphae–gold nanoparticle composites. These nanoparticle-loaded composites were formed by growing Aspergillus niger in gold nanoparticles suspension produced using tea extract as the sole chemical source in addition to the gold salt.

Computational fluid dynamics modeling of Bacillus anthracis spore deposition in rabbit and human respiratory airways

Energy Technology Data Exchange (ETDEWEB)

Kabilan, S.; Suffield, S. R.; Recknagle, K. P.; Jacob, R. E.; Einstein, D. R.; Kuprat, A. P.; Carson, J. P.; Colby, S. M.; Saunders, J. H.; Hines, S. A.; Teeguarden, J. G.; Straub, T. M.; Moe, M.; Taft, S. C.; Corley, R. A.

2016-09-01

Three-dimensional computational fluid dynamics and Lagrangian particle deposition models were developed to compare the deposition of aerosolized Bacillus anthracis spores in the respiratory airways of a human with that of the rabbit, a species commonly used in the study of anthrax disease. The respiratory airway geometries for each species were derived respectively from computed tomography (CT) and µCT images. Both models encompassed airways that extended from the external nose to the lung with a total of 272 outlets in the human model and 2878 outlets in the rabbit model. All simulations of spore deposition were conducted under transient, inhalation–exhalation breathing conditions using average species-specific minute volumes. Two different exposure scenarios were modeled in the rabbit based upon experimental inhalation studies. For comparison, human simulations were conducted at the highest exposure concentration used during the rabbit experimental exposures. Results demonstrated that regional spore deposition patterns were sensitive to airway geometry and ventilation profiles. Due to the complex airway geometries in the rabbit nose, higher spore deposition efficiency was predicted in the nasal sinus compared to the human at the same air concentration of anthrax spores. In contrast, higher spore deposition was predicted in the lower conducting airways of the human compared to the rabbit lung due to differences in airway branching pattern. This information can be used to refine published and ongoing biokinetic models of inhalation anthrax spore exposures, which currently estimate deposited spore concentrations based solely upon exposure concentrations and inhaled doses that do not factor in species-specific anatomy and physiology for deposition.

Spore-to-spore agar culture of the myxomycete Physarum globuliferum.

Science.gov (United States)

Liu, Pu; Wang, Qi; Li, Yu

2010-02-01

The ontogeny of the myxomycete Physarum globuliferum was observed on corn meal agar and hanging drop cultures without adding sterile oat flakes, bacteria or other microorganisms. Its complete life cycle including spore germination, myxamoebae, swarm cells, plasmodial development, and maturity of fructifications was demonstrated. Details of spore-to-spore development are described and illustrated.

Inactivation kinetics of spores of Bacillus cereus strains treated by a peracetic acid-based disinfectant at different concentrations and temperatures.

Science.gov (United States)

Sudhaus, Nadine; Pina-Pérez, Maria Consuelo; Martínez, Antonio; Klein, Günter

2012-05-01

The purpose of this study was to assess the effect of a commercial peracetic acid-based disinfectant against spores of Bacillus cereus, to identify the most influential factor for the final number of microorganisms after different disinfection procedures, and to evaluate the nature of the inactivation kinetics. The spores of four different strains of B. cereus (DSM 318, 4312, 4313, and 4384) were treated with five different disinfectant concentrations (0.25%, 0.5%, 1.0%, 1.5%, and 2.0% [w/v]) at three different temperatures (10°C, 15°C, and 20°C) with or without protein load. A higher temperature and PES 15/23 concentration resulted in a higher inactivation. Inactivation of B. cereus strain 4312 was around 2 log₁₀ cycles at 10°C and around 7 log₁₀ at 20°C (conc=1% [w/v] PAA; t=60 min; without protein). The protein load at higher concentrations did not significantly reduce the efficacy of the disinfectant (p>0.05). This article indicates the applicability of the Weibull model to fit the B. cereus disinfectant survival curves. A Monte Carlo simulation was used to carry out a sensitivity analysis, which revealed the most influential factors affecting the final number of microorganisms after the disinfection process.

Computational Fluid Dynamics Modeling of Bacillus anthracis Spore Deposition in Rabbit and Human Respiratory Airways

Energy Technology Data Exchange (ETDEWEB)

Kabilan, Senthil; Suffield, Sarah R.; Recknagle, Kurtis P.; Jacob, Rick E.; Einstein, Daniel R.; Kuprat, Andrew P.; Carson, James P.; Colby, Sean M.; Saunders, James H.; Hines, Stephanie; Teeguarden, Justin G.; Straub, Tim M.; Moe, M.; Taft, Sarah; Corley, Richard A.

2016-09-30

Three-dimensional computational fluid dynamics and Lagrangian particle deposition models were developed to compare the deposition of aerosolized Bacillus anthracis spores in the respiratory airways of a human with that of the rabbit, a species commonly used in the study of anthrax disease. The respiratory airway geometries for each species were derived from computed tomography (CT) or µCT images. Both models encompassed airways that extended from the external nose to the lung with a total of 272 outlets in the human model and 2878 outlets in the rabbit model. All simulations of spore deposition were conducted under transient, inhalation-exhalation breathing conditions using average species-specific minute volumes. The highest exposure concentration was modeled in the rabbit based upon prior acute inhalation studies. For comparison, human simulation was also conducted at the same concentration. Results demonstrated that regional spore deposition patterns were sensitive to airway geometry and ventilation profiles. Due to the complex airway geometries in the rabbit nose, higher spore deposition efficiency was predicted in the upper conducting airways compared to the human at the same air concentration of anthrax spores. As a result, higher particle deposition was predicted in the conducting airways and deep lung of the human compared to the rabbit lung due to differences in airway branching pattern. This information can be used to refine published and ongoing biokinetic models of inhalation anthrax spore exposures, which currently estimate deposited spore concentrations based solely upon exposure concentrations and inhaled doses that do not factor in species-specific anatomy and physiology.

Petroleum Contamination and Plant Identity Influence Soil and Root Microbial Communities While AMF Spores Retrieved from the Same Plants Possess Markedly Different Communities.

Science.gov (United States)

Iffis, Bachir; St-Arnaud, Marc; Hijri, Mohamed

2017-01-01

Phytoremediation is a promising in situ green technology based on the use of plants to cleanup soils from organic and inorganic pollutants. Microbes, particularly bacteria and fungi, that closely interact with plant roots play key roles in phytoremediation processes. In polluted soils, the root-associated microbes contribute to alleviation of plant stress, improve nutrient uptake and may either degrade or sequester a large range of soil pollutants. Therefore, improving the efficiency of phytoremediation requires a thorough knowledge of the microbial diversity living in the rhizosphere and in close association with plant roots in both the surface and the endosphere. This study aims to assess fungal ITS and bacterial 16S rRNA gene diversity using high-throughput sequencing in rhizospheric soils and roots of three plant species ( Solidago canadensis, Populus balsamifera , and Lycopus europaeus ) growing spontaneously in three petroleum hydrocarbon polluted sedimentation basins. Microbial community structures of rhizospheric soils and roots were compared with those of microbes associated with arbuscular mycorrhizal fungal (AMF) spores to determine the links between the root and rhizosphere communities and those associated with AMF. Our results showed a difference in OTU richness and community structure composition between soils and roots for both bacteria and fungi. We found that petroleum hydrocarbon pollutant (PHP) concentrations have a significant effect on fungal and bacterial community structures in both soils and roots, whereas plant species identity showed a significant effect only on the roots for bacteria and fungi. Our results also showed that the community composition of bacteria and fungi in soil and roots varied from those associated with AMF spores harvested from the same plants. This let us to speculate that in petroleum hydrocarbon contaminated soils, AMF may release chemical compounds by which they recruit beneficial microbes to tolerate or degrade the

Petroleum Contamination and Plant Identity Influence Soil and Root Microbial Communities While AMF Spores Retrieved from the Same Plants Possess Markedly Different Communities

Directory of Open Access Journals (Sweden)

Bachir Iffis

2017-08-01

Full Text Available Phytoremediation is a promising in situ green technology based on the use of plants to cleanup soils from organic and inorganic pollutants. Microbes, particularly bacteria and fungi, that closely interact with plant roots play key roles in phytoremediation processes. In polluted soils, the root-associated microbes contribute to alleviation of plant stress, improve nutrient uptake and may either degrade or sequester a large range of soil pollutants. Therefore, improving the efficiency of phytoremediation requires a thorough knowledge of the microbial diversity living in the rhizosphere and in close association with plant roots in both the surface and the endosphere. This study aims to assess fungal ITS and bacterial 16S rRNA gene diversity using high-throughput sequencing in rhizospheric soils and roots of three plant species (Solidago canadensis, Populus balsamifera, and Lycopus europaeus growing spontaneously in three petroleum hydrocarbon polluted sedimentation basins. Microbial community structures of rhizospheric soils and roots were compared with those of microbes associated with arbuscular mycorrhizal fungal (AMF spores to determine the links between the root and rhizosphere communities and those associated with AMF. Our results showed a difference in OTU richness and community structure composition between soils and roots for both bacteria and fungi. We found that petroleum hydrocarbon pollutant (PHP concentrations have a significant effect on fungal and bacterial community structures in both soils and roots, whereas plant species identity showed a significant effect only on the roots for bacteria and fungi. Our results also showed that the community composition of bacteria and fungi in soil and roots varied from those associated with AMF spores harvested from the same plants. This let us to speculate that in petroleum hydrocarbon contaminated soils, AMF may release chemical compounds by which they recruit beneficial microbes to tolerate

Petroleum Contamination and Plant Identity Influence Soil and Root Microbial Communities While AMF Spores Retrieved from the Same Plants Possess Markedly Different Communities

Science.gov (United States)

Iffis, Bachir; St-Arnaud, Marc; Hijri, Mohamed

2017-01-01

Phytoremediation is a promising in situ green technology based on the use of plants to cleanup soils from organic and inorganic pollutants. Microbes, particularly bacteria and fungi, that closely interact with plant roots play key roles in phytoremediation processes. In polluted soils, the root-associated microbes contribute to alleviation of plant stress, improve nutrient uptake and may either degrade or sequester a large range of soil pollutants. Therefore, improving the efficiency of phytoremediation requires a thorough knowledge of the microbial diversity living in the rhizosphere and in close association with plant roots in both the surface and the endosphere. This study aims to assess fungal ITS and bacterial 16S rRNA gene diversity using high-throughput sequencing in rhizospheric soils and roots of three plant species (Solidago canadensis, Populus balsamifera, and Lycopus europaeus) growing spontaneously in three petroleum hydrocarbon polluted sedimentation basins. Microbial community structures of rhizospheric soils and roots were compared with those of microbes associated with arbuscular mycorrhizal fungal (AMF) spores to determine the links between the root and rhizosphere communities and those associated with AMF. Our results showed a difference in OTU richness and community structure composition between soils and roots for both bacteria and fungi. We found that petroleum hydrocarbon pollutant (PHP) concentrations have a significant effect on fungal and bacterial community structures in both soils and roots, whereas plant species identity showed a significant effect only on the roots for bacteria and fungi. Our results also showed that the community composition of bacteria and fungi in soil and roots varied from those associated with AMF spores harvested from the same plants. This let us to speculate that in petroleum hydrocarbon contaminated soils, AMF may release chemical compounds by which they recruit beneficial microbes to tolerate or degrade the

Distinction of broken cellular wall Ganoderma lucidum spores and G. lucidum spores using FTIR microspectroscopy

Science.gov (United States)

Chen, Xianliang; Liu, Xingcun; Sheng, Daping; Huang, Dake; Li, Weizu; Wang, Xin

2012-11-01

In this paper, FTIR microspectroscopy was used to identify broken cellular wall Ganoderma lucidum spores and G. lucidum spores. For IR spectra, broken cellular wall G. lucidum spores and G. lucidum spores were mainly different in the regions of 3000-2800, 1660-1600, 1400-1200 and 1100-1000 cm-1. For curve fitting, the results showed the differences in the protein secondary structures and the polysaccharide structures/content between broken cellular wall G. lucidum spores and G. lucidum spores. Moreover, the value of A1078/A1741 might be a potentially useful factor to distinguish broken cellular wall G. lucidum spores from G. lucidum spores. Additionally, FTIR microspectroscopy could identify broken cellular wall G. lucidum spores and G. lucidum spores accurately when it was combined with hierarchical cluster analysis. The result suggests FTIR microspectroscopy is very simple and efficient for distinction of broken cellular wall G. lucidum spores and G. lucidum spores. The result also indicates FTIR microspectroscopy may be useful for TCM identification.

Genetic analysis reveals efficient sexual spore dispersal at a fine spatial scale in Armillaria ostoyae, the causal agent of root-rot disease in conifers

OpenAIRE

Capedevielle, Xavier; Lung, Brigitte; Labbé, Frédéric; Dutech, Cyril; Lung-Escarmant, Brigitte

2017-01-01

Armillaria ostoyae (sometimes named A. solidipes) is a fungal species causing root diseases in numerous coniferous forests of the northern hemisphere. The importance of sexual spores for the establishment of new disease centers remains unclear, particularly in the large maritime pine plantations of southwestern France. An analysis of the genetic diversity of a local fungal population distributed over 500 ha in this French forest showed genetic recombination between genotypes to be frequent, c...

Distinction of broken cellular wall Ganoderma lucidum spores and G. lucidum spores using FTIR microspectroscopy.

Science.gov (United States)

Chen, Xianliang; Liu, Xingcun; Sheng, Daping; Huang, Dake; Li, Weizu; Wang, Xin

2012-11-01

In this paper, FTIR microspectroscopy was used to identify broken cellular wall Ganoderma lucidum spores and G. lucidum spores. For IR spectra, broken cellular wall G. lucidum spores and G. lucidum spores were mainly different in the regions of 3000-2800, 1660-1600, 1400-1200 and 1100-1000 cm(-1). For curve fitting, the results showed the differences in the protein secondary structures and the polysaccharide structures/content between broken cellular wall G. lucidum spores and G. lucidum spores. Moreover, the value of A1078/A1741 might be a potentially useful factor to distinguish broken cellular wall G. lucidum spores from G. lucidum spores. Additionally, FTIR microspectroscopy could identify broken cellular wall G. lucidum spores and G. lucidum spores accurately when it was combined with hierarchical cluster analysis. The result suggests FTIR microspectroscopy is very simple and efficient for distinction of broken cellular wall G. lucidum spores and G. lucidum spores. The result also indicates FTIR microspectroscopy may be useful for TCM identification. Copyright © 2012 Elsevier B.V. All rights reserved.

Pollen and spores of terrestrial plants

Science.gov (United States)

Bernhardt, Christopher E.; Willard, Debra A.; Shennan, Ian; Long, Antony J.; Horton, Benjamin P.

2015-01-01

Pollen and spores are valuable tools in reconstructing past sea level and climate because of their ubiquity, abundance, and durability as well as their reciprocity with source vegetation to environmental change (Cronin, 1999; Traverse, 2007; Willard and Bernhardt, 2011). Pollan is found in many sedimentary environments, from freshwater to saltwater, terrestrial to marine. It can be abundant in a minimal amount of sample material, for example half a gram, as concentrations can be as high as four million grains per gram (Traverse, 2007). The abundance of pollen in a sample lends it to robust statistical analysis for the quantitative reconstruction of environments. The outer cell wall is resistant to decay in sediments and allows palynomorphs (pollen and spores) to record changes in plant communities and sea level over millions of years. These characteristics make pollen and spores a powerful tool to use in sea-level research.This chapter describes the biology of pollen and spores and how they are transported and preserved in sediments. We present a methodology for isolating pollen from sediments and a general language and framework to identify pollen as well as light micrographs of a selection of common pollen grains, We then discuss their utility in sea-level research.

Arbuscular mycorrhizal fungal community composition associated with Juniperus brevifolia in native Azorean forest

Science.gov (United States)

Melo, Catarina Drumonde; Luna, Sara; Krüger, Claudia; Walker, Christopher; Mendonça, Duarte; Fonseca, Henrique M. A. C.; Jaizme-Vega, Maria; da Câmara Machado, Artur

2017-02-01

The communities of glomeromycotan fungi (arbuscular mycorrhizal fungi, AMF) under native Juniperus brevifolia forest from two Azorean islands, Terceira and São Miguel, were compared, mainly by spore morphology, and when possible, by molecular analysis. Thirty-nine morphotypes were detected from 12 genera. Glomeromycotan fungal richness was similar in Terceira and São Miguel, but significantly different among the four fragments of native forest. Spore diversity and community composition differed significantly between the two islands. The less degraded island, Terceira, showed 10 exclusive morphotypes including more rare types, whereas the more disturbed forest on São Miguel showed 13 morphs, mostly of common types. Forests from Terceira were dominated by Acaulosporaceae and Glomeraceae. Whereas members of Acaulosporaceae, Glomeraceae and Ambisporaceae were most frequent and abundant in those from São Miguel. Spore abundance was greatest on Terceira, and correlated with soil chemical properties (pH), average monthly temperature and relative humidity.

Microalgae Harvest through Fungal Pelletization—Co-Culture of Chlorella vulgaris and Aspergillus niger

Directory of Open Access Journals (Sweden)

Sarman Oktovianus Gultom

2014-07-01

Full Text Available Microalgae harvesting is a labor- and energy-intensive process and new approaches to harvesting microalgae need to be developed in order to decrease the costs. In this study; co-cultivatation of filamentous fungus (Aspergillus niger and microalgae (Chlorella vulgaris to form cell pellets was evaluated under different conditions, including organic carbon source (glucose; glycerol; and sodium acetate concentration; initial concentration of fungal spores and microalgal cells and light. Results showed that 2 g/L of glucose with a 1:300 ratio of fungi to microalgae provided the best culturing conditions for the process to reach >90% of cell harvest efficiency. The results also showed that an organic carbon source was required to sustain the growth of fungi and form the cell pellets. The microalgae/fungi co-cultures at mixotrophic conditions obtained much higher total biomass than pure cultures of each individual strains; indicating the symbiotic relationship between two strains. This can benefit the microbial biofuel production in terms of cell harvest and biomass production.

Fungal Fluorescence in Hematoxylin-Eosin Stained Sections

Directory of Open Access Journals (Sweden)

Murat Durdu

2017-06-01

Full Text Available A forty-six-year-old male presented to our dermatology clinic with two-year history of itching on his groin. His medical history revealed various topical corticosteroid creams without improvement of the skin lesion. Dermatological examination revealed erythematous nodules and follicular pustules on erythematous background on the inguinal area (Figure 1a. Potassium hydroxide (KOH examination was negative. Tzanck smear revealed abundant neutrophils without bacteria, fungi, or parasite. The histopathological examination showed granuloma formation with multinuclear giant cells and Periodic acid-Schiff (PAS-positive hyphae and spores around the hair follicles (Figure 1b, 1c. Hematoxylin-eosin (H&E-stained slides were examined under an immunofluorescence microscope, and these hyphae and spores showed autofluorescence (Figure 1d. Based on the clinical and histopathological findings, a Majocchi’s granuloma was considered. All lesions disappeared with topical and systemic terbinafine (250 mg/day treatment for six weeks.\tPearls;\tClinical: Not only bacteria, but also fungi, parasites, and viruses may cause folliculitis. Cytology should be initially done to identify the causes of infectious folliculitis. In case of negative cytology, histopathological examination and molecular methods can be used.\tCytological: To cytologically identify all of the causes of folliculitis, four separate samples should be taken: (i the first sample is stained with May-Grünwald-Giemsa for routine cytological examination; (ii the second sample is used for KOH testing; (iii the third sample is stained with an acid-fast stain to detect mycobacteria; and (iv the last specimen is Gram-stained to identify whether it is Gram-positive or Gram-negative (1. Histopathological: In infectious diseases, a definitive diagnosis should be done to identify the etiologic agent. The detection of fungal elements is challenging, when histopathological examination is performed with the H

Inactivation of Ichthyophonus spores using sodium hypochlorite and polyvinyl pyrrolidone iodine.

Science.gov (United States)

Hershberger, P K; Pacheco, C A; Gregg, J L

2008-11-01

Chlorine and iodine solutions were effective at inactivating Ichthyophonus spores in vitro. Inactivation in sea water increased directly with halogen concentration and exposure duration, with significant differences (P < 0.05) from controls occurring at all chlorine concentrations and exposure durations tested (1.5-13.3 ppm for 1-60 min) and at most iodine concentrations and exposure durations tested (1.2 ppm for 60 min and 5.9-10.7 ppm for 1-60 min). However, 10-fold reductions in spore viability occurred only after exposure to halogen solutions at higher concentrations and/or longer durations (13 ppm total chlorine for 1-60 min, 5.9 ppm total iodine for 60 min, and 10.7 ppm total iodine for 1-60 min). Inactivation efficacy was greater when halogen solutions were prepared in fresh water, presumably because of combined effects of halogen-induced inactivation and general spore instability in fresh water. The results have practical implications for disinfection and biocontainment in research laboratories and other facilities that handle live Ichthyophonus cultures and/or infected fish.

Fungal contamination in hospital environments.

Science.gov (United States)

Perdelli, F; Cristina, M L; Sartini, M; Spagnolo, A M; Dallera, M; Ottria, G; Lombardi, R; Grimaldi, M; Orlando, P

2006-01-01

To assess the degree of fungal contamination in hospital environments and to evaluate the ability of air conditioning systems to reduce such contamination. We monitored airborne microbial concentrations in various environments in 10 hospitals equipped with air conditioning. Sampling was performed with a portable Surface Air System impactor with replicate organism detection and counting plates containing a fungus-selective medium. The total fungal concentration was determined 72-120 hours after sampling. The genera most involved in infection were identified by macroscopic and microscopic observation. The mean concentration of airborne fungi in the set of environments examined was 19 +/- 19 colony-forming units (cfu) per cubic meter. Analysis of the fungal concentration in the different types of environments revealed different levels of contamination: the lowest mean values (12 +/- 14 cfu/m(3)) were recorded in operating theaters, and the highest (45 +/- 37 cfu/m(3)) were recorded in kitchens. Analyses revealed statistically significant differences between median values for the various environments. The fungal genus most commonly encountered was Penicillium, which, in kitchens, displayed the highest mean airborne concentration (8 +/- 2.4 cfu/m(3)). The percentage (35%) of Aspergillus documented in the wards was higher than that in any of the other environments monitored. The fungal concentrations recorded in the present study are comparable to those recorded in other studies conducted in hospital environments and are considerably lower than those seen in other indoor environments that are not air conditioned. These findings demonstrate the effectiveness of air-handling systems in reducing fungal contamination.

New pressure and temperature effects on bacterial spores

Science.gov (United States)

Mathys, A.; Heinz, V.; Knorr, D.

2008-07-01

The mechanism of inactivation of bacterial spores by heat and pressure is still a matter of discussion. Obviously, the change of the dissociation equilibrium under pressure and temperature plays a dominant role in inactivation of microorganisms. Heat and pressure inactivation of Geobacillus. stearothermophilus spores at different initial pH-values in ACES and phosphate buffer confirmed this view. Thermal inactivation in ACES buffer at 122°C resulted in higher logarithmic reductions. Contrary, after pressure treatment at 900 MPa with 80°C phosphate buffer showed higher inactivation. These results indicated the different dissociation equilibrium shifts in buffer systems by heat and pressure. Due to preparation, storage and handling of highly concentrated spore suspensions the clumping and the formation of aggregates can hardly be avoided. Consequently, the impact of the agglomeration size distribution on the quantitative assessment of G. stearothermophilus spore inactivation was determined by using a three-fold dynamic optical backreflexion measurement. Two limiting cases have been discriminated in mathematical modelling: three dimensional, spherical packing for maximum spore count and two dimensional, circular packing for minimum spore count of a particular agglomerate. Thermal inactivation studies have been carried out in thin glass capillaries, where by using numerical simulations the non isothermal conditions were modelled and taken into account. It is shown that the shoulder formation often found in thermal spore inactivation can sufficiently be described by first-order inactivation kinetics when the agglomeration size is considered. In case of high pressure inactivation agglomerations could be strongly changed by high forces at compression and especially decompression phase. The physiological response of Bacillus licheniformis spores to high pressure was investigated using multiparameter flow cytometry. Spores were treated by high pressure at 150 MPa with 37�

New pressure and temperature effects on bacterial spores

Energy Technology Data Exchange (ETDEWEB)

Mathys, A; Knorr, D [Berlin University of Technology, Department of Food Biotechnology and Food Process Engineering, Koenigin-Luise-Str. 22, D-14195 Berlin (Germany); Heinz, V [German Institute of Food Technology, p. o. box 1165, D-49601, Quackenbrueck (Germany)], E-mail: alexander.mathys@tu-berlin.de

2008-07-15

The mechanism of inactivation of bacterial spores by heat and pressure is still a matter of discussion. Obviously, the change of the dissociation equilibrium under pressure and temperature plays a dominant role in inactivation of microorganisms. Heat and pressure inactivation of Geobacillus. stearothermophilus spores at different initial pH-values in ACES and phosphate buffer confirmed this view. Thermal inactivation in ACES buffer at 122 deg. C resulted in higher logarithmic reductions. Contrary, after pressure treatment at 900 MPa with 80 deg. C phosphate buffer showed higher inactivation. These results indicated the different dissociation equilibrium shifts in buffer systems by heat and pressure. Due to preparation, storage and handling of highly concentrated spore suspensions the clumping and the formation of aggregates can hardly be avoided. Consequently, the impact of the agglomeration size distribution on the quantitative assessment of G. stearothermophilus spore inactivation was determined by using a three-fold dynamic optical backreflexion measurement. Two limiting cases have been discriminated in mathematical modelling: three dimensional, spherical packing for maximum spore count and two dimensional, circular packing for minimum spore count of a particular agglomerate. Thermal inactivation studies have been carried out in thin glass capillaries, where by using numerical simulations the non isothermal conditions were modelled and taken into account. It is shown that the shoulder formation often found in thermal spore inactivation can sufficiently be described by first-order inactivation kinetics when the agglomeration size is considered. In case of high pressure inactivation agglomerations could be strongly changed by high forces at compression and especially decompression phase. The physiological response of Bacillus licheniformis spores to high pressure was investigated using multiparameter flow cytometry. Spores were treated by high pressure at 150 MPa

New pressure and temperature effects on bacterial spores

International Nuclear Information System (INIS)

Mathys, A; Knorr, D; Heinz, V

2008-01-01

The mechanism of inactivation of bacterial spores by heat and pressure is still a matter of discussion. Obviously, the change of the dissociation equilibrium under pressure and temperature plays a dominant role in inactivation of microorganisms. Heat and pressure inactivation of Geobacillus. stearothermophilus spores at different initial pH-values in ACES and phosphate buffer confirmed this view. Thermal inactivation in ACES buffer at 122 deg. C resulted in higher logarithmic reductions. Contrary, after pressure treatment at 900 MPa with 80 deg. C phosphate buffer showed higher inactivation. These results indicated the different dissociation equilibrium shifts in buffer systems by heat and pressure. Due to preparation, storage and handling of highly concentrated spore suspensions the clumping and the formation of aggregates can hardly be avoided. Consequently, the impact of the agglomeration size distribution on the quantitative assessment of G. stearothermophilus spore inactivation was determined by using a three-fold dynamic optical backreflexion measurement. Two limiting cases have been discriminated in mathematical modelling: three dimensional, spherical packing for maximum spore count and two dimensional, circular packing for minimum spore count of a particular agglomerate. Thermal inactivation studies have been carried out in thin glass capillaries, where by using numerical simulations the non isothermal conditions were modelled and taken into account. It is shown that the shoulder formation often found in thermal spore inactivation can sufficiently be described by first-order inactivation kinetics when the agglomeration size is considered. In case of high pressure inactivation agglomerations could be strongly changed by high forces at compression and especially decompression phase. The physiological response of Bacillus licheniformis spores to high pressure was investigated using multiparameter flow cytometry. Spores were treated by high pressure at 150 MPa

Water condensation promotes fungal growth in ventilation ducts

Energy Technology Data Exchange (ETDEWEB)

Pasanen, P.; Pasanen, A.-L. (University of Kupopio, Department of Environmental Sciences, Kuopio (Finland)); Jantunen, M. (National Public Health Institute, Kuopio (Finland))

1993-01-01

In a subarctic climate the diurnal variation in temperature may cause water condensation in ducts placed in the unheated spaces of a building. In this study, germination time and sporulation of a fungus, Penicillium verrucosum, were studied on dusty, galvanized steel sheet under different moisture conditions at room temperature. The effect of condensed water in a supply air duct on spore amplification was studied in an experimental ventilation set-up. In the field, air temperatures and the dew point temperature of air in the duct were monitored continuously for a week. P. verrucosum germinated on steel surfaces during five-hour incubation of the surface under humid conditions, when the surface has been moist for half an hour, germ tubes appeared within 17 hours. During 24-hour incubation under moist conditions, P. verrucosum produced hyphae and spores. In the experimental set-up the airborne spore counts increased when the air passed through a water-condensing section of the duct. Penicillium was the most abundant fungus sporulated on the moist duct surface. In the field, during humid weather, the surface temperature on the air stream surface decreased to the dew point temperature of the air in the duct. thus water condensation in air ducts may promote fungal growth. (au)

ANALYSIS OF FUNGAL SPORE MYCOTOXIN AND THE RELATIONSHIP BETWEEN SPORE SURFACE AREA AND MYCOTOXIN CONTENT UTILIZING A PROTEIN TRANSLATION INHIBITION ASSAY

Science.gov (United States)

Due to mounting evidence suggesting that biological contamination in the built environment may cause a myriad of adverse health effects, research aimed at understanding the potential exposure to fungal organisms and their metabolites is of utmost importance. To this end we utiliz...

Assessment and determinants of airborne bacterial and fungal concentrations in different indoor environments: Homes, child day-care centres, primary schools and elderly care centres

Science.gov (United States)

Madureira, Joana; Paciência, Inês; Rufo, João Cavaleiro; Pereira, Cristiana; Teixeira, João Paulo; de Oliveira Fernandes, Eduardo

2015-05-01

Until now the influence of risk factors resulting from exposure to biological agents in indoor air has been far less studied than outdoor pollution; therefore the uncertainty of health risks, and how to effectively prevent these, remains. This study aimed (i) to quantify airborne cultivable bacterial and fungal concentrations in four different types of indoor environment as well as to identify the recovered fungi; (ii) to assess the impact of outdoor bacterial and fungal concentrations on indoor air; (iii) to investigate the influence of carbon dioxide (CO2), temperature and relative humidity on bacterial and fungal concentrations; and (iv) to estimate bacterial and fungal dose rate for children (3-5 years old and 8-10 years old) in comparison with the elderly. Air samples were collected in 68 homes, 9 child day-care centres, 20 primary schools and 22 elderly care centres, in a total of 264 rooms with a microbiological air sampler and using tryptic soy agar and malt extract agar culture media for bacteria and fungi growth, respectively. For each building, one outdoor representative location were identified and simultaneously studied. The results showed that child day-care centres were the indoor microenvironment with the highest median bacterial and fungal concentrations (3870 CFU/m3 and 415 CFU/m3, respectively), whereas the lowest median concentrations were observed in elderly care centres (222 CFU/m3 and 180 CFU/m3, respectively). Indoor bacterial concentrations were significantly higher than outdoor concentrations (p < 0.05); whereas the indoor/outdoor ratios for the obtained fungal concentrations were approximately around the unit. Indoor CO2 levels were associated with the bacterial concentration, probably due to occupancy and insufficient ventilation. Penicillium and Cladosporium were the most frequently occurring fungi. Children's had two times higher dose rate to biological pollutants when compared to adult individuals. Thus, due to children

The fungal colonisation of rock-art caves: experimental evidence.

Science.gov (United States)

Jurado, Valme; Fernandez-Cortes, Angel; Cuezva, Soledad; Laiz, Leonila; Cañaveras, Juan Carlos; Sanchez-Moral, Sergio; Saiz-Jimenez, Cesareo

2009-09-01

The conservation of rock-art paintings in European caves is a matter of increasing interest. This derives from the bacterial colonisation of Altamira Cave, Spain and the recent fungal outbreak of Lascaux Cave, France-both included in the UNESCO World Heritage List. Here, we show direct evidence of a fungal colonisation of rock tablets in a testing system exposed in Altamira Cave. After 2 months, the tablets, previously sterilised, were heavily colonised by fungi and bacteria. Most fungi isolated were labelled as entomopathogens, while the bacteria were those regularly identified in the cave. Rock colonisation was probably promoted by the dissolved organic carbon supplied with the dripping and condensation waters and favoured by the displacement of aerosols towards the interior of the cave, which contributed to the dissemination of microorganisms. The role of arthropods in the dispersal of spores may also help in understanding fungal colonisation. This study evidences the fragility of rock-art caves and demonstrates that microorganisms can easily colonise bare rocks and materials introduced into the cavity.

Bioleaching of heavy metals from soil using fungal-organic acids : bench scale testing

Energy Technology Data Exchange (ETDEWEB)

Cathum, S.J.; Ousmanova, D.; Somers, A.; Punt, M. [SAIC Canada, Ottawa, ON (Canada); Brown, C.E. [Environment Canada, Ottawa, ON (Canada). Emergencies Engineering Division]|[Environment Canada, Ottawa, ON (Canada). Environmental Technology Centre

2006-07-01

The ability of fungi to solubilize metals from solid materials may present new opportunities in environmental remediation. This paper presented details of a bench scale experiment that evaluated the leaching of heavy metals from contaminated soil using in situ fungal-generated organic acids. Rice was used as the growing media for organic acid production by A. foetidus. The cultivated fungus was placed on large pieces of potato-dextrose agar (PDA) plates and suspended in 5 L of sterilized water. The cooked rice was inoculated by pouring the 5 L spore suspension over the rice layer. Soil was obtained from a soil pile impacted with heavy metals at a private industrial site and augmented with Pb-contaminated soil. A polyethylene tub was used with a drain pipe leading to a leachate vessel. Crushed stone was spread over the bottom of the tub to assist leachate drainage. Approximately 45 kg of the contaminated soil was spread evenly over the stone layer to a depth of 10 cm. The concentrated spore suspension was sprinkled over the rice. Each week the leachate collection vessel was removed from the bioleaching system and the fine soil particles were allowed to settle. A control was run using the contaminated soil and solid substrate without fungus. Growth of A. foetidus was observed in both control experiment and test experiment after a period of 35 days. The pH of the leachate was measured as the fungal growth progressed. The process was assessed using ICP Mass Spectroscopy and electron spectroscopy, which showed that approximately 65 g of heavy metals were mobilized from 45 kg of soil, and that the biological leaching process resulted in greater mobilization of heavy metals relative to the control experiment. It was concluded that organic acids generated by A. foetidus were capable of leaching heavy metals from the soil. 30 refs., 4 tabs., 15 figs.

Deposition of Bacillus subtilis spores using an airbrush-spray or spots to study surface decontamination by pulsed light.

Science.gov (United States)

Levy, Caroline; Bornard, Isabelle; Carlin, Frédéric

2011-02-01

Microbial contamination on surfaces of food processing equipment is a major concern in industries. A new method to inoculate a single-cell layer (monolayer) of microorganisms onto polystyrene was developed, using a deposition with an airbrush. A homogeneous dispersion of Bacillus subtilis DSM 402 spores sprayed on the surface was observed using both plate count and scanning electron microscopy. No clusters were found, even with high spore concentrations (10(7) spores/inoculated surface). A monolayer of microorganisms was also obtained after deposition of 10 μL droplets containing 3×10(4) spores/spot on polystyrene disks, but not with a higher spore concentration. Pulsed light (PL) applied to monolayers of B. subtilis spores allowed log reductions higher than 6. As a consequence of clusters formation in spots of 10 μL containing more than 3×10(5) spores, log reductions obtained by PL were significantly lower. The comparative advantages of spot and spray depositions were discussed. Copyright © 2010 Elsevier B.V. All rights reserved.

Long-distance wind-dispersal of spores in a fungal plant pathogen: estimation of anisotropic dispersal kernels from an extensive field experiment.

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Adrien Rieux

Full Text Available Given its biological significance, determining the dispersal kernel (i.e., the distribution of dispersal distances of spore-producing pathogens is essential. Here, we report two field experiments designed to measure disease gradients caused by sexually- and asexually-produced spores of the wind-dispersed banana plant fungus Mycosphaerella fijiensis. Gradients were measured during a single generation and over 272 traps installed up to 1000 m along eight directions radiating from a traceable source of inoculum composed of fungicide-resistant strains. We adjusted several kernels differing in the shape of their tail and tested for two types of anisotropy. Contrasting dispersal kernels were observed between the two types of spores. For sexual spores (ascospores, we characterized both a steep gradient in the first few metres in all directions and rare long-distance dispersal (LDD events up to 1000 m from the source in two directions. A heavy-tailed kernel best fitted the disease gradient. Although ascospores distributed evenly in all directions, average dispersal distance was greater in two different directions without obvious correlation with wind patterns. For asexual spores (conidia, few dispersal events occurred outside of the source plot. A gradient up to 12.5 m from the source was observed in one direction only. Accordingly, a thin-tailed kernel best fitted the disease gradient, and anisotropy in both density and distance was correlated with averaged daily wind gust. We discuss the validity of our results as well as their implications in terms of disease diffusion and management strategy.

A method for the determination of bacterial spore DNA content based on isotopic labelling, spore germination and diphenylamine assay; ploidy of spores of several Bacillus species

International Nuclear Information System (INIS)

Hauser, P.M.; Karamata, D.

1992-01-01

A reliable method for measuring the spore DNA content, based on radioactive DNA labelling, spore germination in absence of DNA replication and diphenylamine assay, was developed. The accuracy of the method, within 10 - 15%, is adequate for determining the number of chromosomes per spore, provided that the genome size is known. B subtilis spores were shown to be invariably monogenomic, while those of larger bacilli Bacillus megaterium, Bacillus cereus and Bacillus thuringiensis, often, if not invariably, contain two genomes. Attempts to modify the spore DNA content of B subtilis by altering the richness of the sporulation medium, the sporulation conditions (liquid or solid medium), or by mutation, were apparently unsuccessful. An increase of spore size with medium richness, not accompanied by an increase in DNA content, was observed. The implication of the apparently species-specific spore ploidy and the influence of the sporulation conditions on spore size and shape are discussed

Modelling fungal sink competitiveness with grains for assimilates in wheat infected by a biotrophic pathogen

Science.gov (United States)

Bancal, Marie-Odile; Hansart, Amandine; Sache, Ivan; Bancal, Pierre

2012-01-01

Background and Aims Experiments have shown that biotrophic fungi divert assimilates for their growth. However, no attempt has been made either to account for this additional sink or to predict to what extent it competes with both grain filling and plant reserve metabolism for carbon. Fungal sink competitiveness with grains was quantified by a mixed experimental–modelling approach based on winter wheat infected by Puccinia triticina. Methods One week after anthesis, plants grown under controlled conditions were inoculated with varying loads. Sporulation was recorded while plants underwent varying degrees of shading, ensuring a range of both fungal sink and host source levels. Inoculation load significantly increased both sporulating area and rate. Shading significantly affected net assimilation, reserve mobilization and sporulating area, but not grain filling or sporulation rates. An existing carbon partitioning (source–sink) model for wheat during the grain filling period was then enhanced, in which two parameters characterize every sink: carriage capacity and substrate affinity. Fungal sink competitiveness with host sources and sinks was modelled by representing spore production as another sink in diseased wheat during grain filling. Key Results Data from the experiment were fitted to the model to provide the fungal sink parameters. Fungal carriage capacity was 0·56 ± 0·01 µg dry matter °Cd−1 per lesion, much less than grain filling capacity, even in highly infected plants; however, fungal sporulation had a competitive priority for assimilates over grain filling. Simulation with virtual crops accounted for the importance of the relative contribution of photosynthesis loss, anticipated reserve depletion and spore production when light level and disease severity vary. The grain filling rate was less reduced than photosynthesis; however, over the long term, yield loss could double because the earlier reserve depletion observed here would shorten the

The Influence of Sporulation Conditions on the Spore Coat Protein Composition of Bacillus subtilis Spores.

Science.gov (United States)

Abhyankar, Wishwas R; Kamphorst, Kiki; Swarge, Bhagyashree N; van Veen, Henk; van der Wel, Nicole N; Brul, Stanley; de Koster, Chris G; de Koning, Leo J

2016-01-01

Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has also been shown that the more mature a spore is, the higher is its heat resistance most likely mediated, at least in part, by the progressive cross-linking of coat proteins. The current study for the first time assesses, at the proteomic level, the effect of two commonly used sporulation conditions on spore protein presence. 14 N spores prepared on solid Schaeffer's-glucose (SG) agar plates and 15 N metabolically labeled spores prepared in shake flasks containing 3-( N -morpholino) propane sulfonic acid (MOPS) buffered defined liquid medium differ in their coat protein composition as revealed by LC-FT-MS/MS analyses. The former condition mimics the industrial settings while the latter conditions mimic the routine laboratory environment wherein spores are developed. As seen previously in many studies, the spores prepared on the solid agar plates show a higher thermal resistance than the spores prepared under liquid culture conditions. The 14 N: 15 N isotopic ratio of the 1:1 mixture of the spore suspensions exposes that most of the identified inner coat and crust proteins are significantly more abundant while most of the outer coat proteins are significantly less abundant for the spores prepared on solid SG agar plates relative to the spores prepared in the liquid MOPS buffered defined medium. Sporulation condition-specific differences and variation in isotopic ratios between the tryptic peptides of expected cross-linked proteins suggest that the coat protein cross-linking may also be condition specific. Since the core dipicolinic acid content is found to be similar in both the spore populations, it appears that the difference in wet heat resistance is connected to the

The Influence of Sporulation Conditions on the Spore Coat Protein Composition of Bacillus subtilis Spores

Science.gov (United States)

Abhyankar, Wishwas R.; Kamphorst, Kiki; Swarge, Bhagyashree N.; van Veen, Henk; van der Wel, Nicole N.; Brul, Stanley; de Koster, Chris G.; de Koning, Leo J.

2016-01-01

Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has also been shown that the more mature a spore is, the higher is its heat resistance most likely mediated, at least in part, by the progressive cross-linking of coat proteins. The current study for the first time assesses, at the proteomic level, the effect of two commonly used sporulation conditions on spore protein presence. 14N spores prepared on solid Schaeffer’s-glucose (SG) agar plates and 15N metabolically labeled spores prepared in shake flasks containing 3-(N-morpholino) propane sulfonic acid (MOPS) buffered defined liquid medium differ in their coat protein composition as revealed by LC-FT-MS/MS analyses. The former condition mimics the industrial settings while the latter conditions mimic the routine laboratory environment wherein spores are developed. As seen previously in many studies, the spores prepared on the solid agar plates show a higher thermal resistance than the spores prepared under liquid culture conditions. The 14N:15N isotopic ratio of the 1:1 mixture of the spore suspensions exposes that most of the identified inner coat and crust proteins are significantly more abundant while most of the outer coat proteins are significantly less abundant for the spores prepared on solid SG agar plates relative to the spores prepared in the liquid MOPS buffered defined medium. Sporulation condition-specific differences and variation in isotopic ratios between the tryptic peptides of expected cross-linked proteins suggest that the coat protein cross-linking may also be condition specific. Since the core dipicolinic acid content is found to be similar in both the spore populations, it appears that the difference in wet heat resistance is connected to the

The influence of sporulation conditions on the spore coat protein composition of Bacillus subtilis spores.

Directory of Open Access Journals (Sweden)

Wishwas R. Abhyankar

2016-10-01

Full Text Available Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has also been shown that the more mature a spore is, the higher is its heat resistance most likely mediated, at least in part, by the progressive cross-linking of coat proteins. The current study for the first time assesses, at the proteomic level, the effect of two commonly used sporulation conditions on spore protein presence. 14N spores prepared on solid SG agar plates and 15N metabolically labelled spores prepared in shake flasks containing MOPS buffered defined liquid medium differ in their coat protein composition as revealed by LC-FT-MS/MS analyses. The former condition mimics the industrial settings while the latter conditions mimic the routine laboratory environment wherein spores are developed. As seen previously in many studies, the spores prepared on the solid agar plates show a higher thermal resistance than the spores prepared under liquid culture conditions. The 14N: 15N isotopic ratio of the 1:1 mixture of the spore suspensions exposes that most of the identified inner coat and crust proteins are significantly more abundant while most of the outer coat proteins are significantly less abundant for the spores prepared on solid SG agar plates relative to the spores prepared in the liquid MOPS buffered defined medium. Sporulation condition-specific differences and variation in isotopic ratios between the tryptic peptides of expected cross-linked proteins suggest that the coat protein cross-linking may also be condition specific. Since the core dipicolinic acid content is found to be similar in both the spore populations, it appears that the difference in wet heat resistance is connected to the differences in the coat protein composition and

Diurnal Variations of Airborne Pollen and Spores in Taipei City, Taiwan

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Yueh-Lin Yang

2003-09-01

Full Text Available The diurnal variation of airborne pollen and spores in Taipei City, Taiwan, was investigated during a two-year survey from 1993 to 1994. The pollen and spores were sampled using a Burkard seven-day volumetric pollen trap. The diurnal trends of the total amount of pollen and spores in 1993 and in 1994 were similar to each other, and peaked at 3 to 10 o’clock. The diurnal patterns of airborne pollen and spores of Broussonetia, Fraxinus, Cyathea and Gramineae in 1993 were similar to those in 1994. High concentrations of Broussonetia and Fraxinus were obtained from midnight to the next morning. Cyathea spores peaked from morning till noon, and Gramineae peaked in the afternoon. The diurnal patterns of airborne pollen of Bischofia, Juniperus, Mallotus, Morus, Trema and Urticaceae in 1993 were different to those in 1994. Regular diurnal patterns also associated with the taxa, which produce large pollen or spores, such as Gramineae and Cyathea. In contrast, Bischofia, Juniperus, Mallotus, Morus, Trema and Urticaceae produce relatively small pollen and the diurnal patterns of their airborne pollen were found irregular. The source plants Broussonetia and Fraxinus were close to the collection site so the diurnal patterns of their airborne pollen were regular, suggesting that the diurnal fluctuations of the pollen or spores in air might be affected by the source of plants and the sizes of pollen or spores. The transportation of the smaller pollen or spores in air is probably more easily affected by instability of air currents; they are therefore more likely to exhibit irregular diurnal patterns.

The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in Clostridium difficile Spore Formation.

Science.gov (United States)

Ribis, John W; Ravichandran, Priyanka; Putnam, Emily E; Pishdadian, Keyan; Shen, Aimee

2017-01-01

The spore-forming bacterial pathogen Clostridium difficile is a leading cause of health care-associated infections in the United States. In order for this obligate anaerobe to transmit infection, it must form metabolically dormant spores prior to exiting the host. A key step during this process is the assembly of a protective, multilayered proteinaceous coat around the spore. Coat assembly depends on coat morphogenetic proteins recruiting distinct subsets of coat proteins to the developing spore. While 10 coat morphogenetic proteins have been identified in Bacillus subtilis , only two of these morphogenetic proteins have homologs in the Clostridia : SpoIVA and SpoVM. C. difficile SpoIVA is critical for proper coat assembly and functional spore formation, but the requirement for SpoVM during this process was unknown. Here, we show that SpoVM is largely dispensable for C. difficile spore formation, in contrast with B. subtilis . Loss of C. difficile SpoVM resulted in modest decreases (~3-fold) in heat- and chloroform-resistant spore formation, while morphological defects such as coat detachment from the forespore and abnormal cortex thickness were observed in ~30% of spoVM mutant cells. Biochemical analyses revealed that C. difficile SpoIVA and SpoVM directly interact, similarly to their B. subtilis counterparts. However, in contrast with B. subtilis , C. difficile SpoVM was not essential for SpoIVA to encase the forespore. Since C. difficile coat morphogenesis requires SpoIVA-interacting protein L (SipL), which is conserved exclusively in the Clostridia , but not the more broadly conserved SpoVM, our results reveal another key difference between C. difficile and B. subtilis spore assembly pathways. IMPORTANCE The spore-forming obligate anaerobe Clostridium difficile is the leading cause of antibiotic-associated diarrheal disease in the United States. When C. difficile spores are ingested by susceptible individuals, they germinate within the gut and

Antitumor effects and mechanisms of Ganoderma extracts and spores oil.

Science.gov (United States)

Chen, Chun; Li, Peng; Li, Ye; Yao, Guan; Xu, Jian-Hua

2016-11-01

Ganoderma lucidum is a popular herbal medicine used in China to promote health. Modern studies have disclosed that the active ingredients of Ganoderma can exhibit several effects, including antitumor effects and immunomodulation. The present study evaluated the antitumor effects of self-prepared Ganoderma extracts and spores oil, and investigated the possible underlying mechanisms by observing the effects of the extracts and oil on topoisomerases and the cell cycle. The results showed that Ganoderma extracts and spores oil presented dose-dependent inhibitory effects on tumor cells. The half maximal inhibitory concentration (IC 50 ) values of Ganoderma extracts on HL60, K562 and SGC-7901 cells for 24 h were 0.44, 0.39 and 0.90 mg/ml, respectively; for Ganoderma spores oil, the IC 50 values were 1.13, 2.27 and 6.29 mg/ml, respectively. In the in vivo study, the inhibitory rates of Ganoderma extracts (4 g/kg/d, intragastrically) on S180 and H22 cells were 39.1 and 44.6%, respectively, and for Ganoderma spores oil (1.2 g/kg/d, intragastrically) the inhibitory rates were 30.9 and 44.9%, respectively. Ganoderma extracts and spores oil inhibited the activities of topoisomerase I and II. Ganoderma spores oil was shown block the cell cycle at the transition between the G1 and S phases and induce a marked decrease in cyclin D1 levels in K562 cells, with no significant change in cyclin E level. These results suggest that the Ganoderma extracts and spores oil possessed antitumor effects in the in vitro and in vivo studies. The antitumor mechanisms of the extracts and spores oil were associated with inhibitory effects on topoisomerase I and II activities, and for Ganoderma spores oil, the antitumor effects may also be associated with decreased cyclin D1 levels, thus inducing G1 arrest in the cell cycle.

The regulated synthesis of a Bacillus anthracis spore coat protein that affects spore surface properties.

Science.gov (United States)

Aronson, A; Goodman, B; Smith, Z

2014-05-01

Examine the regulation of a spore coat protein and the effects on spore properties. A c. 23 kDa band in coat/exosporial extracts of Bacillus anthracis Sterne spores varied in amount depending upon the conditions of sporulation. It was identified by MALDI as a likely orthologue of ExsB of Bacillus cereus. Little if any was present in an exosporial preparation with a location to the inner coat/cortex region established by spore fractionation and immunogold labelling of electron micrograph sections. Because of its predominant location in the inner coat, it has been renamed Cotγ. It was relatively deficient in spores produced at 37°C and when acidic fermentation products were produced a difference attributable to transcriptional regulation. The deficiency or absence of Cotγ resulted in a less robust exosporium positioned more closely to the coat. These spores were less hydrophobic and germinated somewhat more rapidly. Hydrophobicity and appearance were rescued in the deletion strain by introduction of the cotγ gene. The deficiency or lack of a protein largely found in the inner coat altered spore hydrophobicity and surface appearance. The regulated synthesis of Cotγ may be a paradigm for other spore coat proteins with unknown functions that modulate spore properties in response to environmental conditions. © 2014 The Society for Applied Microbiology.

Madeira—a tourist destination for asthma sufferers

Science.gov (United States)

Camacho, Irene; Grinn-Gofroń, Agnieszka; Camacho, Roberto; Berenguer, Pedro; Sadyś, Magdalena

2016-11-01

Madeira Island is a famous tourist destination due to its natural and climatic values. Taking into account optimal weather conditions, flora richness and access to various substrates facilitating fungal growth, we hypothesised a very high risk of elevated fungal spore and pollen grain concentrations in the air of Funchal, the capital of Madeira. Concentration levels of the most allergenic taxa were measured from 2003 to 2009, using a 7-day volumetric air sampler, followed by microscopy analysis. Dependence of bioaerosols on the weather conditions and land use were assessed using spatial and statistical tools. Obtained results were re-visited by a comparison with hospital admission data recorded at the Dr. Nélio Mendonça Hospital in Funchal. Our results showed that despite propitious climatic conditions, overall pollen grain and fungal spore concentrations in the air were very low and did not exceed any clinically established threshold values. Pollen and spore peak concentrations also did not match with asthma outbreaks in the winter. Identification of places that are "free" from biological air pollution over the summer, such as Madeira Island, is very important from the allergic point of view.

Madeira-a tourist destination for asthma sufferers.

Science.gov (United States)

Camacho, Irene; Grinn-Gofroń, Agnieszka; Camacho, Roberto; Berenguer, Pedro; Sadyś, Magdalena

2016-11-01

Madeira Island is a famous tourist destination due to its natural and climatic values. Taking into account optimal weather conditions, flora richness and access to various substrates facilitating fungal growth, we hypothesised a very high risk of elevated fungal spore and pollen grain concentrations in the air of Funchal, the capital of Madeira. Concentration levels of the most allergenic taxa were measured from 2003 to 2009, using a 7-day volumetric air sampler, followed by microscopy analysis. Dependence of bioaerosols on the weather conditions and land use were assessed using spatial and statistical tools. Obtained results were re-visited by a comparison with hospital admission data recorded at the Dr. Nélio Mendonça Hospital in Funchal. Our results showed that despite propitious climatic conditions, overall pollen grain and fungal spore concentrations in the air were very low and did not exceed any clinically established threshold values. Pollen and spore peak concentrations also did not match with asthma outbreaks in the winter. Identification of places that are "free" from biological air pollution over the summer, such as Madeira Island, is very important from the allergic point of view.

Factors influencing the inactivation of Alicyclobacillus acidoterrestris spores exposed to high hydrostatic pressure in apple juice

Science.gov (United States)

Sokołowska, B.; Skąpska, S.; Fonberg-Broczek, M.; Niezgoda, J.; Chotkiewicz, M.; Dekowska, A.; Rzoska, S. J.

2013-03-01

Alicyclobacillus acidoterrestris, a thermoacidophilic and spore-forming bacterium, survives the typical pasteurization process and can cause the spoilage of juices, producing compounds associated with disinfectant-like odour (guaiacol, 2,6 - dibromophenol, 2,6 - dichlorophenol). Therefore, the use of other more effective techniques such as high hydrostatic pressure (HHP) is considered for preserving juices. The aim of this study was to search for factors affecting the resistance of A. acidoterrestris spores to HHP. The baroprotective effect of increased solute concentration in apple juice on A. acidoterrestris spores during high pressure processing was observed. During the 45 min pressurization (200 MPa, 50°C) of the spores in concentrated apple juice (71.1°Bx), no significant changes were observed in their number. However, in the juices with a soluble solids content of 35.7, 23.6 and 11.2°Bx, the reduction in spores was 1.3-2.4 log, 2.6-3.3 log and 2.8-4.0 log, respectively. No clear effect of age of spores on the survival under high pressure conditions was found. Spores surviving pressurization and subjected to subsequent HHP treatment showed increased resistance to pressure, by even as much as 2.0 log.

[Quantitative assessment of fungal risk in the case of construction works in healthcare establishments: Proposed indicators for the determination of the impact of management precautions on the risk of fungal infection].

Science.gov (United States)

Gangneux, J-P; Adjidé, C-C; Bernard, L; Botterel, F; Carel, A; Castel, O; Derouin, F; Hoarau, G; Labussière, H; Lafaurie, M; Millon, L; Pottecher, B; Thiebaut, A; Turco, M; Baron, R

2012-03-01

Construction works in healthcare establishments produce airborne fungal spores and considerably increase the risk of exposure of immunosuppressed patients. It is necessary to reinforce protective measures, or even to implement specific precautions, during this critical phase. The aim of these precautions is to protect both those areas, which are susceptible to dust, and patients at risk of a fungal infection particularly invasive aspergillosis. When construction works are planned in healthcare establishments, the first step consists in the characterisation of the environmental fungal risk and the second one in proposing risk management methods. It is then essential to establish impact indicators in order to evaluate the risk management precautions applied. The working group promoted by the French societies of medical mycology and hospital hygiene (SFMM & SF2H) details here both environmental and epidemiological impact indicators that can be used. Copyright © 2012. Published by Elsevier SAS.

Modeling to control spores in raw milk

NARCIS (Netherlands)

Vissers, M.

2007-01-01

A modeling approach was used to identify measures at the farm that reduce transmission of microorganisms to raw milk. Butyric acid bacteria (BAB) and Bacillus cereus were used as case-studies. Minimizing the concentration of BAB spores in raw milk is important to prevent late-blowing of Gouda-type

Ultrastructure and properties of Paecilomyces lilacinus spores

Energy Technology Data Exchange (ETDEWEB)

Holland, R.J.; Gunasekera, T.S. [Macquarie Univ., Dept. of Biological Sciences, Sydney (Australia); Williams, K.L. [Proteome Systems Ltd., Sydney (Australia); Nevalainen, K.M.H. [Dept. of Biological Sciences, Macquarie University, Sydney (Australia)

2002-10-01

Strains of the filamentous soil fungus Paecilomyces lilacinus are currently being developed for use as biological control agents against root-knot, cyst, and other plant-parasitic nematodes. The inoculum applied in the field consists mainly of spores. This study was undertaken to examine the size, ultrastructure, and rodlet layers of P. lilacinus spores and the effect of the culture method on structural and functional spore properties. A rodlet layer was identified on aerial spores only. Other differences noted between aerial spores and those produced in submerged culture included the size and appearance of spores and thickness of spore coat layers when examined with transmission electron microscopy. The two spore types differed in UV tolerance, with aerial spores being less sensitive to environmentally relevant UV radiation. Also, viability after drying and storage was better with the aerial spores. Both spore types exhibited similar nematophagous ability. (author)

Ultrastructure and properties of Paecilomyces lilacinus spores

International Nuclear Information System (INIS)

Holland, R.J.; Gunasekera, T.S.; Williams, K.L.; Nevalainen, K.M.H.

2002-01-01

Strains of the filamentous soil fungus Paecilomyces lilacinus are currently being developed for use as biological control agents against root-knot, cyst, and other plant-parasitic nematodes. The inoculum applied in the field consists mainly of spores. This study was undertaken to examine the size, ultrastructure, and rodlet layers of P. lilacinus spores and the effect of the culture method on structural and functional spore properties. A rodlet layer was identified on aerial spores only. Other differences noted between aerial spores and those produced in submerged culture included the size and appearance of spores and thickness of spore coat layers when examined with transmission electron microscopy. The two spore types differed in UV tolerance, with aerial spores being less sensitive to environmentally relevant UV radiation. Also, viability after drying and storage was better with the aerial spores. Both spore types exhibited similar nematophagous ability. (author)

Substantiation of the active ingredients rational concentration of ointment for treatment of allergic dermatitis complicated by fungal infection

Directory of Open Access Journals (Sweden)

O. A. Rukhmakova

2015-08-01

Full Text Available Aim. Due to the rapid growth of number of allergic skin diseases complicated by secondary fungal infection, creation of new drugs with an integrated anti-allergic and anti-fungal action for their local treatment is especially important. Methods and results. In the process of developing an ointment under conventional name “Allergolik” microbiological studies have been conducted to determine its rational composition. Concentrations of licorice root extract, terbinafine hydrochloride and lavender essential oil have been substantiated as parts of the studied medication. Study of acute toxicity of the developed dosage form has set that it belongs to the IV class of low-toxic substances due to the standard classification of K.K. Sidorova. Conclusion. This testifies the correctness of the choice of drug active ingredients and their concentrations.

Photometric immersion refractometry of bacterial spores.

Science.gov (United States)

Gerhardt, P; Beaman, T C; Corner, T R; Greenamyre, J T; Tisa, L S

1982-01-01

Photometric immersion refractometry was used to determine the average apparent refractive index (n) of five types of dormant Bacillus spores representing a 600-fold range in moist-heat resistance determined as a D100 value. The n of a spore type increased as the molecular size of various immersion solutes decreased. For comparison of the spore types, the n of the entire spore and of the isolated integument was determined by use of bovine serum albumin, which is excluded from permeating into them. The n of the sporoplast (the structures bounded by the outer pericortex membrane) was determined by use of glucose, which was shown to permeate into the spore only as deeply as the pericortex membrane. Among the various spore types, an exponential increase in the heat resistance correlated with the n of the entire spore and of the sporoplast, but not of the isolated perisporoplast integument. Correlation of the n with the solids content of the entire spore provided a method of experimentally obtaining the refractive index increment (dn/dc), which was constant for the various spore types and enables the calculation of solids and water content from an n. Altogether, the results showed that the total water content is distributed unequally within the dormant spore, with less water in the sporoplast than in the perisporoplast integument, and that the sporoplast becomes more refractile and therefore more dehydrated as the heat resistance becomes greater among the various spore types. PMID:6802796

Analysis of surfaces for characterization of fungal burden - Does it matter?

Science.gov (United States)

Viegas, Carla; Faria, Tiago; Meneses, Márcia; Carolino, Elisabete; Viegas, Susana; Gomes, Anita Quintal; Sabino, Raquel

2016-01-01

Mycological contamination of occupational environments can be a result of fungal spores' dispersion in the air and on surfaces. Therefore, it is very important to assess it in both types of the samples. In the present study we assessed fungal contamination in the air and in the surface samples to show relevance of surfaces sampling in complementing the results obtained in the air samples. In total, 42 settings were assessed by the analysis of air and surfaces samples. The settings were divided into settings with a high fungal load (7 poultry farms and 7 pig farms, 3 cork industries, 3 waste management plants, 2 wastewater treatment plants and 1 horse stable) and a low fungal load (10 hospital canteens, 8 college canteens and 1 maternity hospital). In addition to culture-based methods, molecular tools were also applied to detect fungal burden in the settings with a higher fungal load. From the 218 sampling sites, 140 (64.2%) presented different species in the examined surfaces when compared with the species identified in the air. A positive association in the high fungal load settings was found between the presence of different species in the air and surfaces. Wastewater treatment plants constituted the setting with the highest number of different species between the air and surface. We observed that surfaces sampling and application of molecular tools showed the same efficacy of species detection in high fungal load settings, corroborating the fact that surface sampling is crucial for a correct and complete analysis of occupational scenarios. This work is available in Open Access model and licensed under a CC BY-NC 3.0 PL license.

Aspergillus Sydowii Marine Fungal Bloom in Australian Coastal Waters, Its Metabolites and Potential Impact on Symbiodinium Dinoflagellates

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Aiko Hayashi

2016-03-01

Full Text Available Dust has been widely recognised as an important source of nutrients in the marine environment and as a vector for transporting pathogenic microorganisms. Disturbingly, in the wake of a dust storm event along the eastern Australian coast line in 2009, the Continuous Plankton Recorder collected masses of fungal spores and mycelia (~150,000 spores/m3 forming a floating raft that covered a coastal area equivalent to 25 times the surface of England. Cultured A. sydowii strains exhibited varying metabolite profiles, but all produced sydonic acid, a chemotaxonomic marker for A. sydowii. The Australian marine fungal strains share major metabolites and display comparable metabolic diversity to Australian terrestrial strains and to strains pathogenic to Caribbean coral. Secondary colonisation of the rafts by other fungi, including strains of Cladosporium, Penicillium and other Aspergillus species with distinct secondary metabolite profiles, was also encountered. Our bioassays revealed that the dust-derived marine fungal extracts and known A. sydowii metabolites such as sydowic acid, sydowinol and sydowinin A adversely affect photophysiological performance (Fv/Fm of the coral reef dinoflagellate endosymbiont Symbiodinium. Different Symbiodinium clades exhibited varying sensitivities, mimicking sensitivity to coral bleaching phenomena. The detection of such large amounts of A. sydowii following this dust storm event has potential implications for the health of coral environments such as the Great Barrier Reef.

Spore membrane(s) as the site of damage within heated Clostridium perfringens spores.

Science.gov (United States)

Flowers, R S; Adams, D M

1976-02-01

Clostridium perfringens spores were injured by ultrahigh-temperature treatment at 105 C for 5 min. Injury was manifested as an increased sensitivity to polymyxin and neomycin. Since many of the survivors could not germinate normally the ultrahigh-temperature-treated spores were sensitized to and germinated by lysozyme. Polymyxin reportedly acts upon the cell membrane. Neomycin may inhibit protein synthesis and has surface-active properties. Injured spores were increasingly sensitive to known surface-active agents, sodium lauryl sulfate, sodium deoxycholate, and Roccal, a quaternary ammonium compound. Injured spores sensitive to polymyxin and neomycin also were osmotically fragile and died during outgrowth in a liquid medium unless the medium was supplemented with 20% sucrose, 10% dextran, or 10% polyvinylpyrrolidone. The results suggested that a spore structure destined to become cell membrane or cell wall was the site of injury. Repair of injury during outgrowth in the presence of protein, deoxyribonucleic acid, ribonucleic acid and cell wall synthesis inhibitors was consistent with this hypothesis.

Characteristics of exo- polygalacturonase produced by irradiated Aspergillus niger and Trichoderma viride spores

International Nuclear Information System (INIS)

Youssef, B.M.; Swailam, H.M.; Gomaa, N.M.; EL-Mehallawy, A.A.

2010-01-01

Out of 69 fungal isolates from nine pectin rich fresh fruit wastes-showed pectinolytic activity, two were the powerful and best. They were identified as Aspergillus niger and Trichoderma viride. Gamma irradiation of the spore suspensions of these two isolates (0- 3 kGy) stimulated the exo-polygalacturonase (PG) production. Treatment with 0.25 kGy and 0.25-0.50 kGy was found to be the best doses for inducing PG activity produced from T. viride and Asp. niger respectively . The enzyme characteristics were also studied. The optimum temperature of T. viride enzyme reaction was 5 C compared with 45 degree C for Asp. niger enzyme extract.The optimum incubation time of T. viride enzyme reaction was 70-80 min which greater than that of Asp. niger namely 60 min.The results of enzyme reaction ph revealed that the best PG activity was observed at ph 5.0 for the extract of the two fungal isolates. The stability of the enzyme was affected markedly by each of incubation temp., incubation period and ph value .A. niger and T. viride crude extract enzymes were stimulated with Mn 2+ while Zn 2+ and Ca 2+ were inhibitors. The best volume of crude enzyme extract was 3.00 ml in case of T. viride while in case of A. niger was 2.00 ml. T. viride enzyme extract showed its highest enzyme activity with substrate concentration 1.5 % while that of A. niger was found to be 3.0%.

Endosymbiont-dependent host reproduction maintains bacterial-fungal mutualism.

Science.gov (United States)

Partida-Martinez, Laila P; Monajembashi, Shamci; Greulich, Karl-Otto; Hertweck, Christian

2007-05-01

Bacterial endosymbionts play essential roles for many organisms, and thus specialized mechanisms have evolved during evolution that guarantee the persistence of the symbiosis during or after host reproduction. The rice seedling blight fungus Rhizopus microsporus represents a unique example of a mutualistic life form in which a fungus harbors endobacteria (Burkholderia sp.) for the production of a phytotoxin. Here we report the unexpected observation that in the absence of endosymbionts, the host is not capable of vegetative reproduction. Formation of sporangia and spores is restored only upon reintroduction of endobacteria. To monitor this process, we succeeded in GFP labeling cultured endosymbionts. We also established a laserbeam transformation technique for the first controlled introduction of bacteria into fungi to observe their migration to the tips of the aseptate hyphae. The persistence of this fungal-bacterial mutualism through symbiont-dependent sporulation is intriguing from an evolutionary point of view and implies that the symbiont produces factors that are essential for the fungal life cycle. Reproduction of the host has become totally dependent on endofungal bacteria, which in return provide a highly potent toxin for defending the habitat and accessing nutrients from decaying plants. This scenario clearly highlights the significance for a controlled maintenance of this fungal-bacterial symbiotic relationship.

Antitumor effects and mechanisms of Ganoderma extracts and spores oil

Science.gov (United States)

Chen, Chun; Li, Peng; Li, Ye; Yao, Guan; Xu, Jian-Hua

2016-01-01

Ganoderma lucidum is a popular herbal medicine used in China to promote health. Modern studies have disclosed that the active ingredients of Ganoderma can exhibit several effects, including antitumor effects and immunomodulation. The present study evaluated the antitumor effects of self-prepared Ganoderma extracts and spores oil, and investigated the possible underlying mechanisms by observing the effects of the extracts and oil on topoisomerases and the cell cycle. The results showed that Ganoderma extracts and spores oil presented dose-dependent inhibitory effects on tumor cells. The half maximal inhibitory concentration (IC50) values of Ganoderma extracts on HL60, K562 and SGC-7901 cells for 24 h were 0.44, 0.39 and 0.90 mg/ml, respectively; for Ganoderma spores oil, the IC50 values were 1.13, 2.27 and 6.29 mg/ml, respectively. In the in vivo study, the inhibitory rates of Ganoderma extracts (4 g/kg/d, intragastrically) on S180 and H22 cells were 39.1 and 44.6%, respectively, and for Ganoderma spores oil (1.2 g/kg/d, intragastrically) the inhibitory rates were 30.9 and 44.9%, respectively. Ganoderma extracts and spores oil inhibited the activities of topoisomerase I and II. Ganoderma spores oil was shown block the cell cycle at the transition between the G1 and S phases and induce a marked decrease in cyclin D1 levels in K562 cells, with no significant change in cyclin E level. These results suggest that the Ganoderma extracts and spores oil possessed antitumor effects in the in vitro and in vivo studies. The antitumor mechanisms of the extracts and spores oil were associated with inhibitory effects on topoisomerase I and II activities, and for Ganoderma spores oil, the antitumor effects may also be associated with decreased cyclin D1 levels, thus inducing G1 arrest in the cell cycle. PMID:27900038

Germination and Inactivation of Alicyclobacillus acidoterrestris Spores Induced by Moderate Hydrostatic Pressure.

Science.gov (United States)

Sokołowska, Barbara; Skapska, Sylwia; Fonberg-Broczek, Monika; Niezgoda, Jolanta; Porebska, Izabela; Dekowska, Agnieszka; Rzoska, Sylwester J

2015-01-01

Given the importance of spoilage caused by Alicyclobacillus acidoterrestris for the fruit juice industry, the objective of this work was to study the germination and inactivation of A. acidoterrestris spores induced by moderate hydrostatic pressure. Hydrostatic pressure treatment can induce the germination and inactivation of A. acidoterrestris spores. At low pH, spore germination of up to 3.59-3.75 log and inactivation of 1.85-2.04 log was observed in a low pressure window (200-300 MPa) applied at 50 degrees C for 20 min. Neutral pH suppressed inactivation, the number of spores inactivated at pH 7.0 was only 0.24-1.06 log. The pressurization temperature significantly affected spore germination and inactivation. The degree of germination in apple juice after pressurization for 30 min with 200 MPa at 20 degrees C was 2.04 log, with only 0.61 log of spores being inactivated, while at 70 degrees C spore germination was 5.94 log and inactivation 4.72 log. This temperature strongly stimulated germination and inactivation under higher (500 MPa) than lower (200 MPa) pressure. When the oscillatory mode was used, the degree of germination and inactivation was slightly higher than at continuous mode. The degree of germination and inactivation was inversely proportional to the soluble solids content and was lowest in concentrated apple juice.

Biosorption of radionuclide Americium-241 by A. niger spore and hyphae

International Nuclear Information System (INIS)

Yang Yuanyou; Liu Ning; Jin Jiannan; Hua Xinfeng; Zhang Taiming; Luo Shunzhong; Sun Qiling

2002-01-01

The biosorption of radionuclide 241 Am from solution was studied by a. niger spore and hyphae, and the effects of the operational conditions on the treatment were investigated. The results showed the treatment by A. niger spore and hyphae were very efficient. An average of 96% of the total 241 Am was removed from 241 Am solutions of 5.6-111 MBq/L (C 0 ), with adsorption capacities (W) of 7.2-142.4 MBq/g biomass, 5.2-106.5 MBq/g, respectively. The biosorption equilibrium was achieved within 1 h and the optimum pH value ranged 3-0.1 mol/L HNO 3 and 3-2 for spore and hyphae of A. niger, respectively. No significant effects on 241 Am biosorption were observed at 15 degree C-45 degree C, or challenged with containing Au 3+ or Ag + , even 2000 times above 241 Am amount. the index relationship between concentrations and adsorption capacities of 241 Am indicated that the 241 Am biosorption by A. niger spore and hyphae obey to Freundlich adsorption equation. The adsorption behavior of A. niger spore and hyphae were basically coincident

Analysis of Canadian and Irish forage, oats and commercially available equine concentrate feed for pathogenic fungi and mycotoxins

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Buckley Thomas

2007-04-01

Full Text Available Abstract Respiratory infections, recurrent airway obstruction (RAO and exercise induced pulmonary haemorrhage (EIPH are major causes of poor performance in horses. Fungi and mycotoxins are now recognised as a major cause of these conditions. The most notable fungi are Aspergillus and Fusarium. Fungal spores can originate from forage, bedding and feed and, in turn, these fungal spores can produce a series of mycotoxins as secondary metabolites. This study set out to ascertain the degree of fungal and mycotoxin contamination in feed and fodder used in Irish racing yards over a one-year period. Weather conditions in forage producing areas were sampled by Met Eireann and the Canadian Meteorological Service. Fifty per cent of Irish hay, 37% of haylage and 13% of Canadian hay contained pathogenic fungi. Of the mycotoxins, T2 and zearalenone were most prominent. Twenty-one per cent of Irish hay and 16% of pelleted feed contained zearalenone. Forty per cent of oats and 54% of pelleted feed contained T2 toxins.

Analysis of Canadian and Irish forage, oats and commercially available equine concentrate feed for pathogenic fungi and mycotoxins.

Science.gov (United States)

Buckley, Thomas; Creighton, Alan; Fogarty, Ursula

2007-04-01

Respiratory infections, recurrent airway obstruction (RAO) and exercise induced pulmonary haemorrhage (EIPH) are major causes of poor performance in horses. Fungi and mycotoxins are now recognised as a major cause of these conditions. The most notable fungi are Aspergillus and Fusarium. Fungal spores can originate from forage, bedding and feed and, in turn, these fungal spores can produce a series of mycotoxins as secondary metabolites.This study set out to ascertain the degree of fungal and mycotoxin contamination in feed and fodder used in Irish racing yards over a one-year period. Weather conditions in forage producing areas were sampled by Met Eireann and the Canadian Meteorological Service.Fifty per cent of Irish hay, 37% of haylage and 13% of Canadian hay contained pathogenic fungi. Of the mycotoxins, T2 and zearalenone were most prominent. Twenty-one per cent of Irish hay and 16% of pelleted feed contained zearalenone. Forty per cent of oats and 54% of pelleted feed contained T2 toxins.

Maternal parentage influences spore production but not spore pigmentation in the anisogamous and hermaphroditic fungus Neurospora crassa

DEFF Research Database (Denmark)

Zimmerman, Kolea; Levitis, Daniel; Pringle, Anne

2014-01-01

. In this fungus, pigmented spores are viable and unpigmented spores are inviable. These results show that while both parents influence all these traits, maternal influence is strongest on both fertility and mortality traits until the spores are physiologically independent of the maternal cytoplasm.......In this study, we tested the hypothesis that maternal effects on offspring production and quality are greater than paternal effects in both offspring number (fertility) and offspring viability (mortality). We used the model filamentous fungus Neurospora crassa. This fungus is anisogamous......, and various ascospore characteristics. Mixed effects models of these data show that the female parent accounts for the majority of variation in perithecial production, number of spores produced, and spore germination. Surprisingly, both sexes equally influence the percentage of spores that are pigmented...

Morphologic and molecular data help adopting the insect-pathogenic nephridiophagids (Nephridiophagidae among the early diverging fungal lineages, close to the Chytridiomycota

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Renate Radek

2017-07-01

Full Text Available Nephridiophagids are poorly known unicellular eukaryotes, previously of uncertain systematic position, that parasitize the Malpighian tubules of insects. Their life cycle includes merogony with multinucleate plasmodia and sporogony leading to small, uninucleate spores. We examined the phylogenetic affiliations of three species of Nephridiophaga, including one new species, Nephridiophaga maderae, from the Madeira cockroach (Leucophaea maderae. In addition to the specific host, the new species differs from those already known by the size of the spores and by the number of spores within the sporogenic plasmodium. The inferred phylogenetic analyses strongly support a placement of the nephridiophagids in the fungal kingdom near its root and with a close, but unresolved, relationship to the chytids (Chytridiomycota. We found evidence for the nephridiophagidean speciation as being strongly coupled to host speciation.

Effects of steam autoclave treatment on Geobacillus stearothermophilus spores.

Science.gov (United States)

Huesca-Espitia, L C; Suvira, M; Rosenbeck, K; Korza, G; Setlow, B; Li, W; Wang, S; Li, Y-Q; Setlow, P

2016-11-01

To determine the mechanism of autoclave killing of Geobacillus stearothermophilus spores used in biological indicators (BIs) for steam autoclave sterilization, and rates of loss of spore viability and a spore enzyme used in BIs. Spore viability, dipicolinic acid (DPA) release, nucleic acid staining, α-glucosidase activity, protein structure and mutagenesis were measured during autoclaving of G. stearothermophilus spores. Loss of DPA and increases in spore core nucleic acid staining were slower than loss of spore viability. Spore core α-glucosidase was also lost more slowly than spore viability, although soluble α-glucosidase in spore preparations was lost more rapidly. However, spores exposed to an effective autoclave sterilization lost all viability and α-glucosidase activity. Apparently killed autoclaved spores were not recovered by artificial germination in supportive media, much spore protein was denatured during autoclaving, and partially killed autoclave-treated spore preparations did not acquire mutations. These results indicate that autoclave-killed spores cannot be revived, spore killing by autoclaving is likely by protein damage, and spore core α-glucosidase activity is lost more slowly than spore viability. This work provides insight into the mechanism of autoclave killing of spores of an organism used in BIs, and that a spore enzyme in a BI is more stable to autoclaving than spore viability. © 2016 The Society for Applied Microbiology.

Simultaneous Chronic Invasive Fungal Infection and Tracheal Fungus Ball Mimicking Cancer in an Immunocompetent Patient

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Erdoğan Çetinkaya

2016-01-01

Full Text Available Fungal infections of the lung are uncommon and mainly affect people with immune deficiency. There are crucial problems in the diagnosis and treatment of this condition. Invasive pulmonary aspergillosis and candidiasis are the most common opportunistic fungal infections. Aspergillus species (spp. are saprophytes molds that exist in nature as spores and rarely cause disease in immunocompetent individuals. In patients with immune deficiency or chronic lung disease, such as cavitary lung disease or bronchiectasis, Aspergillus may cause a variety of aspergillosis infections. Here we present a case of a 57-year-old patient without immunodeficiency or chronic lung disease who was diagnosed with endotracheal fungus ball and chronic fungal infection, possibly due to Aspergillus. Bronchoscopic examination showed a paralyzed right vocal cord and vegetating mass that was yellow in color, at the posterior wall of tracheal lumen. After 3 months, both the parenchymal and tracheal lesions were completely resolved.

Handling technique of spore-forming bacteria in radiation sterilization. 2. Determination of numbers and radiation resistance of spores

International Nuclear Information System (INIS)

Koshikawa, Tomihiko

1994-01-01

Stepwise ten-fold dilution of bacterial solution is required in the determination of bacterial spores. For this, the selection of diluted solution is important according to the purpose of experiment. First, the preparation of suspension of bacterial spores and selection of diluted solution are presented. Then, a method for determining the number of bacterial spores in materials is outlined in terms of dilution methods of bacterial solution (shaking and homogenization) and application method of diluted solution to the plating medium. Finally, a method for determining radiation resistance of spore-forming bacteria is explained according to the measurement conditions (suspension of bacterial spores and filters applied with bacterial spores). (N.K.)

Handling technique of spore-forming bacteria in radiation sterilization. 2. Determination of numbers and radiation resistance of spores

Energy Technology Data Exchange (ETDEWEB)

Koshikawa, Tomihiko [Japan Radioisotope Association, Shiga (Japan). Koka Laboratory

1994-12-01

Stepwise ten-fold dilution of bacterial solution is required in the determination of bacterial spores. For this, the selection of diluted solution is important according to the purpose of experiment. First, the preparation of suspension of bacterial spores and selection of diluted solution are presented. Then, a method for determining the number of bacterial spores in materials is outlined in terms of dilution methods of bacterial solution (shaking and homogenization) and application method of diluted solution to the plating medium. Finally, a method for determining radiation resistance of spore-forming bacteria is explained according to the measurement conditions (suspension of bacterial spores and filters applied with bacterial spores). (N.K.).

A Study on the Frequency of Fungal Rhinosinusitis and to Determine the Sensitivity of the Isolates to Antifungal Drugs in Shiraz, Iran 2012-2013

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Parisa Badiei

2015-06-01

Full Text Available Abstract Background: Fungal spores are present in large amounts in the nature which may cause diseases in the susceptible individuals. Fungal rhinosinusitis has been increased during recent decades. This study aims to investigate fungal rhinosinusitis in the patients with chronic needing endoscopic sinus surgery aims and to determine the sensitivity of the isolates to rhin sinusitis antifungal agents. Materials and Methods: In cross sectional study, specimens were obtained in sterile plates containing normal saline. from the suspected cases of chronic rhinosinusitis who had undergone endoscopic sinus surgery. Microscopic experiments and Cultures were done in Sabouraud Dextrose Agar medium. Minimum inhibitory concentration (MIC of the growth of fungi for seven antifungal agents against the isolates was determined using E-Test. Results: In total, specimens from 102 immunocompetent patients were examined. Nine positive specimens were presented, consisting of 3 for Aspergillus fumigatus, 3 for Penicillium, 2 for Aspergillus flavus and 1 for Alternaria. The mean age of the patients was 38 years, with 59 males and 43 females. The isolates were most sensitive to the voriconazole and amphotericin B. Conclusion: Since the prevalence of fungal rhinosinusitis is low, unfortunately, the clinicians are not attentive enough to the condition, and they initiate antifungal therapy if the patients are not responsive to antibiotics. Early diagnosis and suitable treatment can help more efficient management of patients.

Absence of transient elevated uv resistance during germination of Bacillus subtilis spores lacking small, acid-soluble spore proteins α and β

International Nuclear Information System (INIS)

Setlow, B.; Setlow, P.

1988-01-01

Dormant spores of various Bacillus species are much more resistant to UV irradiation than are the corresponding vegetative cells. This elevated spore UV resistance appears to have two causes. First, UV irradiation of spores does not produce the pyrimidine dimers formed in vegetative-cell DNA, but rather produces several other photoproducts, the most predominant of which is termed the spore photoproduct, a 5-thyminyl-5,6-dihydrothymine adduct (1, 10). Second, spores have at least two mechanisms which efficiently repair this spore photoproduct during spore germination, including one which monomerizes the adduct back to two thymines. This study shows that germinating spores of bacillus subtilis mutants which lack small, acid-soluble spore proteins α and β did not exhibit the transient elevated UV resistance seen during germination of wild-type spores

Root Fungal Endophytes Enhance Heavy-Metal Stress Tolerance of Clethra barbinervis Growing Naturally at Mining Sites via Growth Enhancement, Promotion of Nutrient Uptake and Decrease of Heavy-Metal Concentration.

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Keiko Yamaji

Full Text Available Clethra barbinervis Sieb. et Zucc. is a tree species that grows naturally at several mine sites and seems to be tolerant of high concentrations of heavy metals, such as Cu, Zn, and Pb. The purpose of this study is to clarify the mechanism(s underlying this species' ability to tolerate the sites' severe heavy-metal pollution by considering C. barbinervis interaction with root fungal endophytes. We measured the heavy metal concentrations of root-zone soil, leaves, branches, and fine roots collected from mature C. barbinervis at Hitachi mine. We isolated fungal endophytes from surface-sterilized root segments, and we examined the growth, and heavy metal and nutrient absorption of C. barbinervis seedlings growing in sterilized mine soil with or without root fungal endophytes. Field analyses showed that C. barbinervis contained considerably high amounts of Cu, Zn, and Pb in fine roots and Zn in leaves. The fungi, Phialocephala fortinii, Rhizodermea veluwensis, and Rhizoscyphus sp. were frequently isolated as dominant fungal endophyte species. Inoculation of these root fungal endophytes to C. barbinervis seedlings growing in sterilized mine soil indicated that these fungi significantly enhanced the growth of C. barbinervis seedlings, increased K uptake in shoots and reduced the concentrations of Cu, Ni, Zn, Cd, and Pb in roots. Without root fungal endophytes, C. barbinervis could hardly grow under the heavy-metal contaminated condition, showing chlorosis, a symptom of heavy-metal toxicity. Our results indicate that the tree C. barbinervis can tolerate high heavy-metal concentrations due to the support of root fungal endophytes including P. fortinii, R. veluwensis, and Rhizoscyphus sp. via growth enhancement, K uptake promotion and decrease of heavy metal concentrations.

Fungal endophytes – the hidden inducers of volatile terpene biosynthesis in tomato plants

DEFF Research Database (Denmark)

Ntana, Fani; Jensen, Birgit; Jørgensen, Hans Jørgen Lyngs

mycorrhizal spores in the Indian Thar desert, colonizes the root cortex of a wide range of plants, enhancing plant growth and modulating plant specialized metabolism. The effect of S. indica colonization on the metabolism of the host can be potentially used in improving plant defence against pathogens...... and herbivores. Tomato (Solanum lycopersicum) is an important crop, often challenged by fungal pathogens and insect pests. The wide variety of secondary metabolites produced by the plant, and especially terpenes, play a crucial role in plant defence, helping in repelling possible enemies. This project is focused....... indica-inoculated and S. indica-free tomato plants. Preliminary data suggest that fungal colonization results in increased production of specific volatile terpenes. A transcriptome analysis on fungus-associated and fungus-free plant tissues is currently ongoing to elucidate in depth the mechanisms...

Mate Limitation in Fungal Plant Parasites Can Lead to Cyclic Epidemics in Perennial Host Populations.

Science.gov (United States)

Ravigné, Virginie; Lemesle, Valérie; Walter, Alicia; Mailleret, Ludovic; Hamelin, Frédéric M

2017-03-01

Fungal plant parasites represent a growing concern for biodiversity and food security. Most ascomycete species are capable of producing different types of infectious spores both asexually and sexually. Yet the contributions of both types of spores to epidemiological dynamics have still to been fully researched. Here we studied the effect of mate limitation in parasites which perform both sexual and asexual reproduction in the same host. Since mate limitation implies positive density dependence at low population density, we modeled the dynamics of such species with both density-dependent (sexual) and density-independent (asexual) transmission rates. A first simple SIR model incorporating these two types of transmission from the infected compartment, suggested that combining sexual and asexual spore production can generate persistently cyclic epidemics in a significant part of the parameter space. It was then confirmed that cyclic persistence could occur in realistic situations by parameterizing a more detailed model fitting the biology of the Black Sigatoka disease of banana, for which literature data are available. We discuss the implications of these results for research on and management of Sigatoka diseases of banana.

Significant influence of fungi on coarse carbonaceous and potassium aerosols in a tropical rainforest

International Nuclear Information System (INIS)

Zhang, Zhisheng; Tao, Jun; Engling, Guenter; Zhang, Leiming; Kawamura, Kimitaka; Yang, Yihong; Zhang, Renjian; Chan, Chuen-yu; Li, Yide

2015-01-01

Fungal spores are ubiquitous in the Earth’s atmosphere, especially in the environment of tropical rainforests with intense biological activities. To assess the impact of fungi on chemical components of atmospheric aerosols at a Chinese tropical rainforest site, size-segregated fungal spore tracers (i.e. arabitol and mannitol) were measured along with major aerosol components, including carbonaceous species and water-soluble inorganic ions. The fungal spore tracers were found to be predominately associated with coarse particles, in which organic carbon (OC) and potassium (K + ) were also present at significant levels. Enhanced amounts of fungal spore tracers were closely linked to rainfall events. Moreover, fungal spore tracers exhibited positive correlations with relative humidity and negative correlations with wind speed, temperature or radiation. The relationships between fungal spore tracers and meteorological factors are consistent with the emission features of actively discharged fungal spores, which are generally associated with sugar alcohols and by-products such as the inorganic ion K + . The excellent correlations between fungal spore tracers and OC or K + in the coarse particles further suggested their common emission sources. Absolute principal factor analysis further identified fungi as the largest contributor to coarse OC and K + (both at ∼66%) in this rainforest. (letter)

Pollen and spores as a passive monitor of ultraviolet radiation

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Wesley Toby Fraser

2014-04-01

Full Text Available Sporopollenin is the primary component of the outer walls of pollen and spores. The chemical composition of sporopollenin is responsive to levels of ultraviolet (UV radiation exposure, via a concomitant change in the concentration of phenolic compounds. This relationship offers the possibility of using fossil pollen and spore chemistry as a novel proxy for past UV flux. Phenolic compounds in sporopollenin can be quantified using Fourier Transform infrared spectroscopy. The high potential for preservation of pollen and spores in the geologic record, and the conservative nature of sporopollenin chemistry across the land plant phylogeny, means that this new proxy has the potential to reconstruct UV flux over much longer timescales than has previously been possible. This new tool has important implications for understanding the relationship between UV flux, solar insolation and climate in the past, as well as providing a possible means of assessing paleoaltitude, and ozone thickness.

Biodegradable and bioactive CGP/PVA film for fungal growth inhibition.

Science.gov (United States)

Silva, Bárbara Dumas S; Ulhoa, Cirano J; Batista, Karla A; Di Medeiros, Maria Carolina; Da Silva Filho, Rômulo Roosevelt; Yamashita, Fabio; Fernandes, Kátia F

2012-07-01

In this study, chitinolytic enzymes produced by Trichoderma asperellum were immobilized on a biodegradable film manufactured with a blend of cashew gum polysaccharide (CGP) and polyvinyl alcohol (PVA), and tested as a fungal growth inhibitor. The film was produced by casting a blend of CGP and PVA solution on glass molds. The CGP/PVA film showed 68% water solubility, tensile strength of 23.7 MPa, 187.2% elongation and 52% of mass loss after 90 days in soil. The presence of T-CWD enzymes immobilized by adsorption or covalent attachment resulted in effective inhibition of fungal growth. Sclerotinia sclerotiorum was the most sensitive organism, followed by Aspergillus niger and Penicillium sp. SEM micrograph showed that the presence of immobilized T-CWD enzymes on CGP/PVA film produced morphological modifications on vegetative and germinative structures of the microorganisms, particularly hyphae disruption and changes of spores shape. Copyright © 2012 Elsevier Ltd. All rights reserved.

A Spore Counting Method and Cell Culture Model for Chlorine Disinfection Studies of Encephalitozoon syn. Septata intestinalis

OpenAIRE

Wolk, D. M.; Johnson, C. H.; Rice, E. W.; Marshall, M. M.; Grahn, K. F.; Plummer, C. B.; Sterling, C. R.

2000-01-01

The microsporidia have recently been recognized as a group of pathogens that have potential for waterborne transmission; however, little is known about the effects of routine disinfection on microsporidian spore viability. In this study, in vitro growth of Encephalitozoon syn. Septata intestinalis, a microsporidium found in the human gut, was used as a model to assess the effect of chlorine on the infectivity and viability of microsporidian spores. Spore inoculum concentrations were determine...

Minimizing the level of Bacillus cereus spores in farm tank milk

NARCIS (Netherlands)

Vissers, M.M.M.; Giffel, M.C.T.; Driehuis, F.; Jong, de P.; Lankveld, J.M.G.

2007-01-01

In a year-long survey on 24 Dutch farms, Bacillus cereus spore concentrations were measured in farm tank milk (FTM), feces, bedding material, mixed grass and corn silage, and soil from the pasture. The aim of this study was to determine, in practice, factors affecting the concentration of B. cereus

Image Cytometric Analysis of Algal Spores for Evaluation of Antifouling Activities of Biocidal Agents.

Science.gov (United States)

Il Koo, Bon; Lee, Yun-Soo; Seo, Mintae; Seok Choi, Hyung; Leng Seah, Geok; Nam, Taegu; Nam, Yoon Sung

2017-07-31

Chemical biocides have been widely used as marine antifouling agents, but their environmental toxicity impose regulatory restriction on their use. Although various surrogate antifouling biocides have been introduced, their comparative effectiveness has not been well investigated partly due to the difficulty of quantitative evaluation of their antifouling activity. Here we report an image cytometric method to quantitatively analyze the antifouling activities of seven commercial biocides using Ulva prolifera as a target organism, which is known to be a dominant marine species causing soft fouling. The number of spores settled on a substrate is determined through image analysis using the intrinsic fluorescence of chlorophylls in the spores. Pre-determined sets of size and shape of spores allow for the precise determination of the number of settled spores. The effects of biocide concentration and combination of different biocides on the spore settlement are examined. No significant morphological changes of Ulva spores are observed, but the amount of adhesive pad materials is appreciably decreased in the presence of biocides. It is revealed that the growth rate of Ulva is not directly correlated with the antifouling activities against the settlement of Ulva spores. This work suggests that image cytometric analysis is a very convenient, fast-processable method to directly analyze the antifouling effects of biocides and coating materials.

Evaluation of surface sampling method performance for Bacillus Spores on clean and dirty outdoor surfaces.

Energy Technology Data Exchange (ETDEWEB)

Wilson, Mollye C.; Einfeld, Wayne; Boucher, Raymond M.; Brown, Gary Stephen; Tezak, Matthew Stephen

2011-06-01

Recovery of Bacillus atrophaeous spores from grime-treated and clean surfaces was measured in a controlled chamber study to assess sampling method performance. Outdoor surfaces investigated by wipe and vacuum sampling methods included stainless steel, glass, marble and concrete. Bacillus atrophaeous spores were used as a surrogate for Bacillus anthracis spores in this study designed to assess whether grime-coated surfaces significantly affected surface sampling method performance when compared to clean surfaces. A series of chamber tests were carried out in which known amounts of spores were allowed to gravitationally settle onto both clean and dirty surfaces. Reference coupons were co-located with test coupons in all chamber experiments to provide a quantitative measure of initial surface concentrations of spores on all surfaces, thereby allowing sampling recovery calculations. Results from these tests, carried out under both low and high humidity conditions, show that spore recovery from grime-coated surfaces is the same as or better than spore recovery from clean surfaces. Statistically significant differences between method performance for grime-coated and clean surfaces were observed in only about half of the chamber tests conducted.

A Novel Protocol for Decoating and Permeabilizing Bacterial Spores for Epifluorescent Microscopy

Science.gov (United States)

LaDuc, Myron T.; Mohapatra, Bidyut

2014-01-01

Based on previously reported procedures for permeabilizing vegetative bacterial cells, and numerous trial-and-error attempts with bacterial endospores, a protocol was developed for effectively permeabilizing bacterial spores, which facilitated the applicability of fluorescent in situ hybridization (FISH) microscopy. Bacterial endospores were first purified from overgrown, sporulated suspensions of B. pumilus SAFR-032. Purified spores at a concentration of approx equals 10 million spores/mL then underwent proteinase-K treatment, in a solution of 468.5 µL of 100 mM Tris-HCl, 30 µL of 10% SDS, and 1.5 microL of 20 mg/mL proteinase-K for ten minutes at 35 ºC. Spores were then harvested by centrifugation (15,000 g for 15 minutes) and washed twice with sterile phosphate-buffered saline (PBS) solution. This washing process consisted of resuspending the spore pellets in 0.5 mL of PBS, vortexing momentarily, and harvesting again by centrifugation. Treated and washed spore pellets were then resuspended in 0.5 mL of decoating solution, which consisted of 4.8 g urea, 3 mL Milli-Q water, 1 mL 0.5M Tris, 1 mL 1M dithiothreitol (DTT), and 2 mL 10% sodium-dodecylsulfate (SDS), and were incubated at 65 ºC for 15 minutes while being shaken at 165 rpm. Decoated spores were then, once again, washed twice with sterile PBS, and subjected to lysozyme/mutanolysin treatment (7 mg/mL lysozyme and 7U mutanolysin) for 15 minutes at 35 C. Spores were again washed twice with sterile PBS, and spore pellets were resuspended in 1-mL of 2% SDS. This treatment, facilitating inner membrane permeabilization, lasted for ten minutes at room temperature. Permeabilized spores were washed two final times with PBS, and were resuspended in 200 mkcroL of sterile PBS. At this point, the spores were permeable and ready for downstream processing, such as oligonucleotideprobe infiltration, hybridization, and microscopic evaluation. FISH-microscopic imagery confirmed the effective and efficient (˜50

A spore counting method and cell culture model for chlorine disinfection studies of Encephalitozoon syn. Septata intestinalis.

Science.gov (United States)

Wolk, D M; Johnson, C H; Rice, E W; Marshall, M M; Grahn, K F; Plummer, C B; Sterling, C R

2000-04-01

The microsporidia have recently been recognized as a group of pathogens that have potential for waterborne transmission; however, little is known about the effects of routine disinfection on microsporidian spore viability. In this study, in vitro growth of Encephalitozoon syn. Septata intestinalis, a microsporidium found in the human gut, was used as a model to assess the effect of chlorine on the infectivity and viability of microsporidian spores. Spore inoculum concentrations were determined by using spectrophotometric measurements (percent transmittance at 625 nm) and by traditional hemacytometer counting. To determine quantitative dose-response data for spore infectivity, we optimized a rabbit kidney cell culture system in 24-well plates, which facilitated calculation of a 50% tissue culture infective dose (TCID(50)) and a minimal infective dose (MID) for E. intestinalis. The TCID(50) is a quantitative measure of infectivity and growth and is the number of organisms that must be present to infect 50% of the cell culture wells tested. The MID is as a measure of a system's permissiveness to infection and a measure of spore infectivity. A standardized MID and a standardized TCID(50) have not been reported previously for any microsporidian species. Both types of doses are reported in this paper, and the values were used to evaluate the effects of chlorine disinfection on the in vitro growth of microsporidia. Spores were treated with chlorine at concentrations of 0, 1, 2, 5, and 10 mg/liter. The exposure times ranged from 0 to 80 min at 25 degrees C and pH 7. MID data for E. intestinalis were compared before and after chlorine disinfection. A 3-log reduction (99.9% inhibition) in the E. intestinalis MID was observed at a chlorine concentration of 2 mg/liter after a minimum exposure time of 16 min. The log(10) reduction results based on percent transmittance-derived spore counts were equivalent to the results based on hemacytometer-derived spore counts. Our data

Imaging bacterial spores by soft-x-ray microscopy

International Nuclear Information System (INIS)

Stead, A.D.; Ford, T.W.; Judge, J.

1997-01-01

Bacterial spores are able to survive dehydration, but neither the physiological nor structural basis of this have been fully elucidated. Furthermore, once hydrated, spores often require activation before they will germinate. Several treatments can be used to activate spores, but in the case of Bacillus subtlis the most effective is heat treatment. The physiological mechanism associated with activation is also not understood, but some workers suggest that the loss of calcium from the spores may be critical. However, just prior to germination, the spores change from being phase bright to phase dark when viewed by light microscopy. Imaging spores by soft x-ray microscopy is possible without fixation. Thus, in contrast to electron microscopy, it is possible to compare the structure of dehydrated and hydrated spores in a manner not possible previously. A further advantage is that it is possible to monitor individual spores by phase contrast light microscopy immediately prior to imaging with soft x-rays; whereas, with both electron microscopy and biochemical studies, it is a population of spores being studied without knowledge of the phase characteristics of individual spores. This study has therefore tried to compare dehydrated and hydrated spores and to determine if there is a mass loss from individual spores as they pass the transition from being phase bright to phase dark

Use of yeast spores for microencapsulation of enzymes.

Science.gov (United States)

Shi, Libing; Li, Zijie; Tachikawa, Hiroyuki; Gao, Xiao-Dong; Nakanishi, Hideki

2014-08-01

Here, we report a novel method to produce microencapsulated enzymes using Saccharomyces cerevisiae spores. In sporulating cells, soluble secreted proteins are transported to the spore wall. Previous work has shown that the spore wall is capable of retaining soluble proteins because its outer layers work as a diffusion barrier. Accordingly, a red fluorescent protein (RFP) fusion of the α-galactosidase, Mel1, expressed in spores was observed in the spore wall even after spores were subjected to a high-salt wash in the presence of detergent. In vegetative cells, however, the cell wall cannot retain the RFP fusion. Although the spore wall prevents diffusion of proteins, it is likely that smaller molecules, such as sugars, pass through it. In fact, spores can contain much higher α-galactosidase activity to digest melibiose than vegetative cells. When present in the spore wall, the enzyme acquires resistance to environmental stresses including enzymatic digestion and high temperatures. The outer layers of the spore wall are required to retain enzymes but also decrease accessibility of the substrates. However, mutants with mild spore wall defects can retain and stabilize the enzyme while still permitting access to the substrate. In addition to Mel1, we also show that spores can retain the invertase. Interestingly the encapsulated invertase has significantly lower activity toward raffinose than toward sucrose.This suggests that substrate selectivity could be altered by the encapsulation.

Imaging bacterial spores by soft-x-ray microscopy

Energy Technology Data Exchange (ETDEWEB)

Stead, A.D.; Ford, T.W. [Univ. of London, Surrey (United Kingdom); Judge, J. [Unilever plc, Sharnbrook (United Kingdom)] [and others

1997-04-01

Bacterial spores are able to survive dehydration, but neither the physiological nor structural basis of this have been fully elucidated. Furthermore, once hydrated, spores often require activation before they will germinate. Several treatments can be used to activate spores, but in the case of Bacillus subtlis the most effective is heat treatment. The physiological mechanism associated with activation is also not understood, but some workers suggest that the loss of calcium from the spores may be critical. However, just prior to germination, the spores change from being phase bright to phase dark when viewed by light microscopy. Imaging spores by soft x-ray microscopy is possible without fixation. Thus, in contrast to electron microscopy, it is possible to compare the structure of dehydrated and hydrated spores in a manner not possible previously. A further advantage is that it is possible to monitor individual spores by phase contrast light microscopy immediately prior to imaging with soft x-rays; whereas, with both electron microscopy and biochemical studies, it is a population of spores being studied without knowledge of the phase characteristics of individual spores. This study has therefore tried to compare dehydrated and hydrated spores and to determine if there is a mass loss from individual spores as they pass the transition from being phase bright to phase dark.

Survival of Spores of Trichoderma longibrachiatum in Space: data from the Space Experiment SPORES on EXPOSE-R

Science.gov (United States)

Neuberger, Katja; Lux-Endrich, Astrid; Panitz, Corinna

2015-01-01

In the space experiment `Spores in artificial meteorites' (SPORES), spores of the fungus Trichoderma longibrachiatum were exposed to low-Earth orbit for nearly 2 years on board the EXPOSE-R facility outside of the International Space Station. The environmental conditions tested in space were: space vacuum at 10-7-10-4 Pa or argon atmosphere at 105 Pa as inert gas atmosphere, solar extraterrestrial ultraviolet (UV) radiation at λ > 110 nm or λ > 200 nm with fluences up to 5.8 × 108 J m-2, cosmic radiation of a total dose range from 225 to 320 mGy, and temperature fluctuations from -25 to +50°C, applied isolated or in combination. Comparable control experiments were performed on ground. After retrieval, viability of spores was analysed by two methods: (i) ethidium bromide staining and (ii) test of germination capability. About 30% of the spores in vacuum survived the space travel, if shielded against insolation. However, in most cases no significant decrease was observed for spores exposed in addition to the full spectrum of solar UV irradiation. As the spores were exposed in clusters, the outer layers of spores may have shielded the inner part. The results give some information about the likelihood of lithopanspermia, the natural transfer of micro-organisms between planets. In addition to the parameters of outer space, sojourn time in space seems to be one of the limiting parameters.

Sensitizing Clostridium difficile Spores With Germinants on Skin and Environmental Surfaces Represents a New Strategy for Reducing Spores via Ambient Mechanisms

Directory of Open Access Journals (Sweden)

Michelle Marie Nerandzic

2017-10-01

Full Text Available Background: Clostridium difficile is a leading cause of healthcare-associated infections worldwide. Prevention of C. difficile transmission is challenging because spores are not killed by alcohol-based hand sanitizers or many commonly used disinfectants. One strategy to control spores is to induce germination, thereby rendering the spores more susceptible to benign disinfection measures and ambient stressors. Methods/Results: C. difficile spores germinated on skin after a single application of cholic acid-class bile salts and co-germinants; for 4 C. difficile strains, recovery of viable spores from skin was reduced by ~0.3 log10CFU to 2 log10CFU after 2 hours and ~1 log10CFU to >2.5 log 10CFU after 24 hours. The addition of taurocholic acid to 70% and 30% ethanol significantly enhanced reduction of viable spores on skin and on surfaces. Desiccation, and to a lesser extent the presence of oxygen, were identified as the stressors responsible for reductions of germinated spores on skin and surfaces. Additionally, germinated spores became susceptible to killing by pH 1.5 hydrochloric acid, suggesting that germinated spores that remain viable on skin and surfaces might be killed by gastric acid after ingestion. Antibiotic-treated mice did not become colonized after exposure to germinated spores, whereas 100% of mice became colonized after exposure to the same quantity of dormant spores. Conclusions: Germination could provide a new approach to reduce C. difficile spores on skin and in the environment and to render surviving spores less capable of causing infection. Our findings suggest that it may be feasible to develop alcohol-based hand sanitizers containing germinants that reduce spores on hands.

Direct Surface Analysis of Fungal Species by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

Energy Technology Data Exchange (ETDEWEB)

Valentine, Nancy B.(BATTELLE (PACIFIC NW LAB)); Wahl, Jon H.(BATTELLE (PACIFIC NW LAB)); Kingsley, Mark T.(BATTELLE (PACIFIC NW LAB)); Wahl, Karen L.(BATTELLE (PACIFIC NW LAB))

2001-12-01

Intact spores and/or hyphae of Aspergillus niger, Rhizopus oryzae, Trichoderma reesei and Phanerochaete chrysosporium are analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). This study investigates various methods of sample preparation and matrices to determine optimum collection and analysis criteria for fungal analysis by MALDI-MS. Fungi are applied to the MALDI sample target as untreated, sonicated, acid/heat treated, or blotted directly from the fungal culture with double-stick tape. Ferulic acid or sinapinic acid matrix solution is layered over the dried samples and analyzed by MALDI-MS. Statistical analysis of the data show that simply using double stick tape to collect and transfer to a MALDI sample plate typically worked as well as the other preparation methods, but requires the least sample handling.

Altered immunological response in mice subjected to stress and exposed to fungal spores

Science.gov (United States)

Kurup, Viswanath P.; Choi, Hongyung; Kumar, Anoopa; Murali, Pazhayannur S.; Mishra, S. K.; Pierson, Duane L.

1992-01-01

Space flight and related factors such as stress appear to have an adverse effect on astronauts' immune systems. The presence of potentially pathogenic microbes including several genera of fungi reported from spacecraft environment may be a cause of concern in such situations. In order to study the role of such organisms in causing opportunistic or allergic diseases in crewmembers, we have tried to develop an animal model. BALB/c mice were suspended upside down for varying periods of time to induce stress, and their lymphocyte functions were evaluated. These studies indicate that the stress resulted in lowered mitogen induced lymphocyte stimulation as represented by 3H-thymidine uptake. We have also studied the ability of these animals to respond to Aspergillus fumigatus spores. The results of the study clearly demonstrate a definite down-regulation in T-cell proliferation and a higher incidence of infection with A. fumigatus.

Sorption of 241Am by Aspergillus niger spore and hyphae

International Nuclear Information System (INIS)

Yuanyou Yang; Ning Liu; Jiali Liao; Jiannan Jin; Shunzhong Luo; Taiming Zhang; Pengji Zhao

2004-01-01

Biosorption of 241 Am by a fungus A. niger, including the spore and hyphae, was investigated. The preliminary results showed that the adsorption of 241 Am by the microorganism was efficient. More than 96% of the total 241 Am could be removed from 241 Am solutions of 5.6-111 MBq/l (C 0 ) by spore and hyphae of A. niger, with adsorbed 241 Am metal (Q) of 7.2-142.4 MBq/g biomass, and 5.2-106.5 MBq/g, respectively. The biosorption equilibrium was achieved within 1 hour and the optimum pH range was pH 1-3. No obvious effects on 241 Am adsorption by the fungus were observed at 10-45 deg C, or in solutions containing Au 3+ or Ag + , even 2000 times above the 241 Am concentration. The 241 Am biosorption by the fungus obeys the Freundlich adsorption equation. There was no significant difference between the adsorption behavior of A. niger spore and hyphae. (author)

Effect of medium components and culture conditions in Bacillus subtilis EA-CB0575 spore production.

Science.gov (United States)

Posada-Uribe, Luisa F; Romero-Tabarez, Magally; Villegas-Escobar, Valeska

2015-10-01

Bacillus subtilis spores have important biotechnological applications; however, achieving both, high spore cell densities and sporulation efficiencies in fermentation, is poorly reported. In this study, medium components and culture conditions were optimized with different statistical methods to increase spore production of the plant growth promoting rhizobacteria B. subtilis EA-CB0575. Key medium components were determined with Plackett-Burman (PB) design, and the optimum concentration levels of two components (glucose, MgSO4·7H2O) were optimized with a full factorial and central composite design, achieving 1.37 × 10(9) CFU/mL of spore cell density and 93.5 % of sporulation efficiency in shake flask. The optimized medium was used to determine the effect of culture conditions on spore production at bioreactor level, finding that maintaining pH control did not affect significantly spore production, while the interaction of agitation and aeration rates had a significant effect on spore cell density. The overall optimization generated a 17.2-fold increase in spore cell density (8.78 × 10(9) CFU/mL) and 1.9-fold increase in sporulation efficiency (94.2 %) compared to that of PB design. These results indicate the potential of B. subtilis EA-CB0575 to produce both, high spore cell densities and sporulation efficiencies, with very low nutrient requirements and short incubation period which can represent savings of process production.

Effect of Coat Layers in Bacillus Subtilis Spores Resistance to Photo-Catalytic Inactivation

Directory of Open Access Journals (Sweden)

Luz del Carmen Huesca-Espitia

2017-10-01

Full Text Available Different water treatment processes (physical and chemical exist to obtain safe water for human or food industry supply. The advanced oxidation technologies are rising as a new alternative to eliminate undesirable chemicals and waterborne diseases. In this work, we analyze the power of the photo-assisted Fenton process using Fe(II/H2O2 and UV radiation (365 nm to inactivate Bacillus subtilis spores, considered among the most resistant biological structures known. Different concentrations of Fe(II, H2O2 and UV radiation (365 nm were used to inactivate wt and some coat spore mutants of B. subtilis. Wt spores of B. subtilis were inactivated after 60 min using this process. In general, all defective coat mutants were more sensitive than the wt spores and, particularly, the double mutant was 10 folds more sensitive than others being inactivated during the first 10 minutes using soft reaction conditions. Presence of Fe(II ions was found essential for spore inactivating process and, for those spores inactivated using the Fe(II/H2O2 under UV radiation process, it is suggested that coat structures are important to their resistance to the treatment process. The photo-assisted Fenton process using Fe(II, H2O2 and UV radiation (365 nm can be used to inactivate any water microorganisms with the same or less resistance that B. subtilis spores to produce safe drinking water in relatively short treatment time.

Particle size distribution of airborne Aspergillus fumigatus spores emitted from compost using membrane filtration

Science.gov (United States)

Deacon, L. J.; Pankhurst, L. J.; Drew, G. H.; Hayes, E. T.; Jackson, S.; Longhurst, P. J.; Longhurst, J. W. S.; Liu, J.; Pollard, S. J. T.; Tyrrel, S. F.

Information on the particle size distribution of bioaerosols emitted from open air composting operations is valuable in evaluating potential health impacts and is a requirement for improved dispersion simulation modelling. The membrane filter method was used to study the particle size distribution of Aspergillus fumigatus spores in air 50 m downwind of a green waste compost screening operation at a commercial facility. The highest concentrations (approximately 8 × 10 4 CFU m -3) of culturable spores were found on filters with pore diameters in the range 1-2 μm which suggests that the majority of spores are emitted as single cells. The findings were compared to published data collected using an Andersen sampler. Results were significantly correlated ( p < 0.01) indicating that the two methods are directly comparable across all particles sizes for Aspergillus spores.

Spores of Mucor ramosissimus, Mucor plumbeus and Mucor circinelloides and their ability to activate human complement system in vitro.

Science.gov (United States)

Granja, Luiz Fernando Zmetek; Pinto, Lysianne; Almeida, Cátia Amancio; Alviano, Daniela Sales; Da Silva, Maria Helena; Ejzemberg, Regina; Alviano, Celuta Sales

2010-03-01

Complement activation by spores of Mucor ramosissimus, Mucor plumbeus and Mucor circinelloides was studied using absorbed human serum in the presence or absence of chelators (EGTA or EDTA). We found that the spore caused full complement activation when incubated with EGTA-Mg2+ or without chelators, indicating that the alternative pathway is mainly responsible for this response. In order to compare activation profiles from each species, ELISAs for C3 and C4 fragments, mannan binding lectin (MBL), C-reactive protein (CRP) and IgG studies were carried out. All proteins were present on the species tested. Immunofluorescence tests demonstrated the presence of C3 fragments on the surface of all samples, which were confluent throughout fungal surfaces. The same profile of C3, C4, MBL, CRP and IgG deposition, observed in all species, suggests a similar activation behavior for these species.

Extracellular proteases from Streptomyces phaeopurpureus ExPro138 inhibit spore adhesion, germination and appressorium formation in Colletotrichum coccodes.

Science.gov (United States)

Palaniyandi, S A; Yang, S H; Suh, J-W

2013-07-01

To study the antifungal mechanism of proteases from Streptomyces phaeopurpureus strain ExPro138 towards Colletotrichum coccodes and to evaluate its utilization as biofungicide. We screened proteolytic Streptomyces strains from the yam rhizosphere with antifungal activity. Forty proteolytic Streptomyces were isolated, among which eleven isolates showed gelatinolytic activity and antagonistic activity on C. coccodes. Of the 11 isolates, protease preparation from an isolate designated ExPro138 showed antifungal activity. 16S rDNA sequence analysis of the strain showed 99% similarity with Streptomyces phaeopurepureus (EU841588.1). Zymography analysis of the ExPro138 culture filtrate revealed that the strain produced several extracellular proteases. The protease preparation inhibited spore germination, spore adhesion to polystyrene surface and appressorium formation. Microscopic study of the interaction between ExPro138 and C. coccodes revealed that ExPro138 was mycoparasitic on C. coccodes. The protease preparation also reduced anthracnose incidence on tomato fruits compared with untreated control. This study demonstrates possibility of utilizing antifungal proteases derived from antagonistic microbes as biofungicide. Microbial proteases having the ability to inhibit spore adhesion and appressorium formation could be used to suppress infection establishment by foliar fungal pathogens at the initial stages of the infection process. Journal of Applied Microbiology © 2013 The Society for Applied Microbiology.

Administration of Brevibacillus laterosporus spores as a poultry feed additive to inhibit house fly development in feces: a new eco-sustainable concept.

Science.gov (United States)

Ruiu, L; Satta, A; Floris, I

2014-03-01

The success of a microbial pesticide application against house flies developing in manure should accomplish the uniform mixing of active ingredients with this breeding medium, thus enhancing residual effects. The oral administration of the entomopathogenic bacterium Brevibacillus laterosporus to caged poultry species allows the homogeneous incorporation of its active ingredients with fly breeding media. Feces from treated broilers or hens show toxicity against exposed fly adults and larvae. Insecticidal effects are concentration-dependent with a lethal median concentration (LC50) value of 1.34 × 10(8) and 0.61 × 10(8) spores/g of feces for adults and larvae, respectively. Manure toxicity against flies was maintained as long as chickens were fed a diet containing adequate concentrations of B. laterosporus spores. Toxicity significantly decreased after spore administration to birds was interrupted. When poultry diet contained 10(10) spores/g, mortality of flies reared on feces exceeded 80%. The use of B. lateroporus spores as a feed additive in poultry production systems fostering a more integrated approach to farming is discussed.

Dynamics of Spore Coat Morphogenesis in Bacillus subtilis

Science.gov (United States)

McKenney, Peter T.; Eichenberger, Patrick

2011-01-01

SUMMARY Spores of Bacillus subtilis are encased in a protective coat made up of at least 70 proteins. The structure of the spore coat has been examined using a variety of genetic, imaging and biochemical techniques, however, the majority of these studies have focused on mature spores. In this study we use a library of 41 spore coat proteins fused to the Green Fluorescent Protein (GFP) to examine spore coat morphogenesis over the time-course of sporulation. We found considerable diversity in the localization dynamics of coat proteins and were able to establish 6 classes based on localization kinetics. Localization dynamics correlate well with the known transcriptional regulators of coat gene expression. Previously, we described the existence of multiple layers in the mature spore coat. Here, we find that the spore coat initially assembles a scaffold that is organized into multiple layers on one pole of the spore. The coat then encases the spore in multiple coordinated waves. Encasement is driven, at least partially, by transcription of coat genes and deletion of sporulation transcription factors arrests encasement. We also identify the trans-compartment SpoIIIAH-SpoIIQ channel as necessary for encasement. This is the first demonstration of a forespore contribution to spore coat morphogenesis. PMID:22171814

Instrumental neutron activation analysis of wheat bunt spores

Energy Technology Data Exchange (ETDEWEB)

Liu, Y G; Schmitt, R A [Oregon State Univ., Corvallis (USA). Dept. of Chemistry; Oregon State Univ., Corvallis (USA). Radiation Center); Trione, E J [Oregon State Univ., Corvallis (USA). Dept. of Botany; Laul, J C [Battelle Pacific Northwest Labs., Richland, WA (USA)

1982-01-01

The concentrations of seventeen elements (Na, Mg, Al, Cl, K, Ca, Sc, V, Cr, Mn, Fe, Co, Zn, Br, Rb, La, Sm) in two species of fungus which cause wheat bunt disease, Tilletia caries (DC.) Tul. and Tilletia controversa Kuehn, were determined by instrumental neutron activation analysis. A standard sequential INAA procedure was used. Differences in the K and Cl concentrations between these two species of spores are large and therefore can be used as a criterion of distinguishing between the two species of fungus.

The influence of sporulation conditions on the spore coat protein composition of Bacillus subtilis spores.

OpenAIRE

Wishwas R. Abhyankar; Wishwas R. Abhyankar; Kiki Kamphorst; Bhagyashree N. Swarge; Bhagyashree N. Swarge; Henk van Veen; Nicole N. van der Wel; Stanley Brul; Chris G. de Koster; Leo J. de Koning

2016-01-01

Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has also been shown that the more mature a spore is, the higher is its heat resistance most likely mediated, at least in part, by the progressive cross-linking of coat proteins. The current study for t...

The Influence of Sporulation Conditions on the Spore Coat Protein Composition of Bacillus subtilis Spores

OpenAIRE

Abhyankar, Wishwas R.; Kamphorst, Kiki; Swarge, Bhagyashree N.; van Veen, Henk; van der Wel, Nicole N.; Brul, Stanley; de Koster, Chris G.; de Koning, Leo J.

2016-01-01

Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has also been shown that the more mature a spore is, the higher is its heat resistance most likely mediated, at least in part, by the progressive cross-linking of coat proteins. The current study for t...

Growth and sporulation of a pyrimidine spore color mutant of Sordaria fimicola.

Science.gov (United States)

el-Ani, A S

1967-04-07

A nonautonomous spore color mutant of Sordaria fimicola is a pyrimidine auxotroph that produces hyaline nonviable ascospores. Uracil, uridine, and cytidine are more effective growth factors than cytosine and thymine and, in high concentrations, render the mutant self-fertile by inducing the ascospores to resume development and maturation. Crosses with the unlinked arginine non-autonomus spore color mutant st-59 yielded the double mutant st-59 pyr that requires both arginine and a pyrimidine for growth, which indicates a lack of suppression of the pyrimidine requirement by the arginine locus.

Dynamic phase microscopy, a new method to detect viable and killed spores and to estimate the heterogeneity of spore populations

Science.gov (United States)

Tychinsky, Vladimir P.; Mulyukin, Andrey L.; Lisovskii, Vitalii V.; Nikolaev, Yury A.; Kretushev, Aleksander V.; Vyshenskaya, Tatyana V.; Suzina, Nataliya E.; Duda, Vitalii I.; El-Registan, Galina I.

One of the challenging tasks in monitoring studies is to estimate heterogeneity of microbial populations by the physiological state and potential viability of individual cells, especially with regard of their ability to withstand various environmental assaults. Previously, we described some approaches based on electron microscopy methods to discriminate vegetative, dormant, and dead cells in both aged microbial cultures and environmental samples, including permafrost. We propose to extend the arsenal of microscopy methods for monitoring studies by a new non-invasive and informative method - dynamic phase microscopy (DPM). The substantial advantage of DPM is that it gives quantitative (digitized) data of undestroyed (living) microscopic objects, exemplified in our work by Bacillus licheniformis spores. Using DPM made it possible to record interference images of objects (spores) and to produce picture of their "phase thickness" (PT) that is the optical path difference in nm. Thus, it was demonstrated the remarkable difference in the PT of spores at different physiological states: dormant, germinating, and heat-killed spores had PT values of 80, 40-50, and 20 nm, respectively. The other found criterion to distinguish between spores was the PT fluctuations. In contrast to dormant and killed spores, the PT of germinating spores oscillated with amplitude of up to 7 nm, with typical frequencies of 1.3 and 3.4 Hz. A combination of the recorded PT values and PT fluctuations gave a key to detect viable and dead cells. Under the conditions that did not support germination (the lack of nutrients), we were able to follow the response of a single dormant spore and a spore population to heating from 25 °C to 70 °C. Thus, a very small temperature change (from 40 °C to 42 °C) under conditions non-favorable for germination, caused a drastic decrease in the spores' PT; the second drop in the PT values was observed during heating from 60 °C to 70 °C. These changes were

Evaluation of airborne particulates and fungi during hospital renovation.

Science.gov (United States)

Overberger, P A; Wadowsky, R M; Schaper, M M

1995-07-01

This study was conducted over 30 weeks on a hospital floor undergoing partial renovation. Some patients housed on the floor were immunosuppressed, including bone marrow transplant recipients. The construction zone was placed under negative pressure and was separated from patient rooms by existing hospital walls and via erection of a temporary barrier. Other control measures minimized patient exposure to airborne materials. Air sampling was done for 3 weeks prior to construction, 24 weeks during construction, and 3 weeks after renovation was completed. Airborne particulate concentrations, total spore counts, particle size, and fungal species were assessed. At the beginning of the renovation there were increases in airborne particulates (from 0.2 to 2.0 mg/m3) and fungal spores (from 3.5 to 350 colony forming units (CFU/m3), but only in the construction zone. Throughout the remainder of the renovation, particulate and fungal spore levels fluctuated inside the construction zone but remained close to baseline values in the patient area. When renovation was completed, particulates and spore counts inside the construction zone decreased to preconstruction levels. The primary fungus isolated from air samples was Penicillium. This study demonstrated that control measures were effective in reducing exposures of hospitalized patients to airborne particulates and spores and in reducing the increased risk of aspergillosis and other fungal infections associated with hospital construction projects. The data from this study may be useful in establishing exposure guidelines for other health care settings.

Monitoring of clinical strains and environmental fungal aerocontamination to prevent invasive aspergillosis infections in hospital during large deconstruction work: a protocol study.

Science.gov (United States)

Loeffert, Sophie Tiphaine; Melloul, Elise; Dananché, Cédric; Hénaff, Laetitia; Bénet, Thomas; Cassier, Pierre; Dupont, Damien; Guillot, Jacques; Botterel, Françoise; Wallon, Martine; Gustin, Marie-Paule; Vanhems, Philippe

2017-11-25

Monitoring fungal aerocontamination is an essential measure to prevent severe invasive aspergillosis (IA) infections in hospitals. One central block among 32 blocks of Edouard Herriot Hospital (EHH) was entirely demolished in 2015, while care activities continued in surrounding blocks. The main objective was to undertake broad environmental monitoring and clinical surveillance of IA cases to document fungal dispersion during major deconstruction work and to assess clinical risk. A daily environmental survey of fungal loads was conducted in eight wards located near the demolition site. Air was collected inside and outside selected wards by agar impact samplers. Daily spore concentrations were monitored continuously by volumetric samplers at a flow rate of 10 L.min -1 . Daily temperature, wind direction and speed as well as relative humidity were recorded by the French meteorological station Meteociel. Aspergillus fumigatus strains stored will be genotyped by multiple-locus, variable-number, tandem-repeat analysis. Antifungal susceptibility will be assessed by E-test strips on Roswell Park Memorial Institute medium supplemented with agar. Ascertaining the adequacy of current environmental monitoring techniques in hospital is of growing importance, considering the rising impact of fungal infections and of curative antifungal costs. The present study could improve the daily management of IA risk during major deconstruction work and generate new data to ameliorate and redefine current guidelines. This study was approved by the clinical research and ethics committees of EHH. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Lethal and pre-lethal effects of a fungal biopesticide contribute to substantial and rapid control of malaria vectors.

Directory of Open Access Journals (Sweden)

Simon Blanford

Full Text Available Rapidly emerging insecticide resistance is creating an urgent need for new active ingredients to control the adult mosquitoes that vector malaria. Biopesticides based on the spores of entomopathogenic fungi have shown considerable promise by causing very substantial mortality within 7-14 days of exposure. This mortality will generate excellent malaria control if there is a high likelihood that mosquitoes contact fungi early in their adult lives. However, where contact rates are lower, as might result from poor pesticide coverage, some mosquitoes will contact fungi one or more feeding cycles after they acquire malaria, and so risk transmitting malaria before the fungus kills them. Critics have argued that 'slow acting' fungal biopesticides are, therefore, incapable of delivering malaria control in real-world contexts. Here, utilizing standard WHO laboratory protocols, we demonstrate effective action of a biopesticide much faster than previously reported. Specifically, we show that transient exposure to clay tiles sprayed with a candidate biopesticide comprising spores of a natural isolate of Beauveria bassiana, could reduce malaria transmission potential to zero within a feeding cycle. The effect resulted from a combination of high mortality and rapid fungal-induced reduction in feeding and flight capacity. Additionally, multiple insecticide-resistant lines from three key African malaria vector species were completely susceptible to fungus. Thus, fungal biopesticides can block transmission on a par with chemical insecticides, and can achieve this where chemical insecticides have little impact. These results support broadening the current vector control paradigm beyond fast-acting chemical toxins.

The major Alternaria alternata allergen, Alt a 1: A reliable and specific marker of fungal contamination in citrus fruits.

Science.gov (United States)

Gabriel, M F; Uriel, N; Teifoori, F; Postigo, I; Suñén, E; Martínez, J

2017-09-18

The ubiquitously present spores of Alternaria alternata can spoil a wide variety of foodstuffs, including a variety of fruits belonging to the Citrus genus. The major allergenic protein of A. alternata, Alt a 1, is a species-specific molecular marker that has been strongly associated with allergenicity and phytopathogenicity of this fungal species. This study aimed to evaluate the potential of the detection of Alt a 1 as a reliable indicator of A. alternata contamination in citrus fruits. To accomplish this aim, sixty oranges were artificially infected with a spore suspension of A. alternata. Internal fruit material was collected at different incubation times (one, two and three weeks after the fungal inoculation) and used for both total RNA extraction and protein extraction. Alt a 1 detection was then performed by polymerase chain reaction (PCR) amplification using Alt a 1 specific primers and by enzyme-linked immunosorbent assay (ELISA). The experimental model presented in this work was effective to simulate the typical Alternaria black rot phenotype and its progression. Although both PCR and ELISA techniques have been successfully carried out for detecting Alt a 1 allergen in A. alternata infected oranges, the PCR method was found to be more sensitive than ELISA. Nevertheless, ELISA results were highly valuable to demonstrate that considerable amounts of Alt a 1 are produced during A. alternata fruit infection process, corroborating the recently proposed hypothesis that this protein plays a role in the pathogenicity and virulence of Alternaria species. Such evidence suggests that the detection of Alt a 1 by PCR-based assay may be used as a specific indicator of the presence of pathogenic and allergenic fungal species, A. alternata, in fruits. This knowledge can be employed to control the fungal infection and mitigate agricultural losses as well as human exposure to A. alternata allergens and toxins. Copyright © 2017 Elsevier B.V. All rights reserved.

Sphagnum moss disperses spores with vortex rings.

Science.gov (United States)

Whitaker, Dwight L; Edwards, Joan

2010-07-23

Sphagnum spores, which have low terminal velocities, are carried by turbulent wind currents to establish colonies many kilometers away. However, spores that are easily kept aloft are also rapidly decelerated in still air; thus, dispersal range depends strongly on release height. Vascular plants grow tall to lift spores into sufficient wind currents for dispersal, but nonvascular plants such as Sphagnum cannot grow sufficiently high. High-speed videos show that exploding capsules of Sphagnum generate vortex rings to efficiently carry spores high enough to be dispersed by turbulent air currents. Spores launched ballistically at similar speeds through still air would travel a few millimeters and not easily reach turbulent air. Vortex rings are used by animals; here, we report vortex rings generated by plants.

Optimisation of a direct plating method for the detection and enumeration of Alicyclobacillus acidoterrestris spores.

Science.gov (United States)

Henczka, Marek; Djas, Małgorzata; Filipek, Katarzyna

2013-01-01

A direct plating method for the detection and enumeration of Alicyclobacillus acidoterrestris spores has been optimised. The results of the application of four types of growth media (BAT agar, YSG agar, K agar and SK agar) regarding the recovery and enumeration of A. acidoterrestris spores were compared. The influence of the type of applied growth medium, heat shock conditions, incubation temperature, incubation time, plating technique and the presence of apple juice in the sample on the accuracy of the detection and enumeration of A. acidoterrestris spores was investigated. Among the investigated media, YSG agar was the most sensitive medium, and its application resulted in the highest recovery of A. acidoterrestris spores, while K agar and BAT agar were the least suitable media. The effect of the heat shock time on the recovery of spores was negligible. When there was a low concentration of spores in a sample, the membrane filtration method was superior to the spread plating method. The obtained results show that heat shock carried out at 80°C for 10 min and plating samples in combination with membrane filtration on YSG agar, followed by incubation at 46°C for 3 days provided the optimal conditions for the detection and enumeration of A. acidoterrestris spores. Application of the presented method allows highly efficient, fast and sensitive identification and enumeration of A. acidoterrestris spores in food products. This methodology will be useful for the fruit juice industry for identifying products contaminated with A. acidoterrestris spores, and its practical application may prevent economic losses for manufacturers. Copyright © 2012 Elsevier B.V. All rights reserved.

Differential modification of oxic and anoxic components of radiation damage in Bacillus megaterium spores by caffeine

International Nuclear Information System (INIS)

Kesavan, P.C.; Powers, E.L.

1985-01-01

Studies were carried out on the effect of caffeine on the X-irradiation sensitivity of B. megaterium spores with the following results: Caffeine exerts a concentration-dependent modifying action on oxygen-dependent components of X-ray-induced damage in B. megaterium spore suspensions causing an 'over-O 2 effect' at about 1 x 10 -4 mol dm -3 , and as the concentration is increased to 1 10 -3 mol dm -3 or above, a small but consistent protection is seen. In the absence of O 2 , at a wide range of concentrations (8.5 x 10 -5 to 1 x 10 -1 mol dm -3 ), caffeine enhances the inactivation constant, k, from 1.17 to about 1.50 kGy -1 . Both ethanol and t-butanol (5 x 10 -2 mol dm -3 ) remove the 'over O 2 -effect' produced by 1 x 10 -4 mol dm -3 caffeine in O 2 ; such an effect, however, is not accompanied by reduction in the H 2 O 2 concentrations in the spore suspensions. Ethanol prevents caffeine-induced anoxic sensitization, as well as H 2 O 2 buildup. t-BuOH has no influence on either the low dose part of the log fraction survival curve or on the H 2 O 2 yield in the spore suspensions. Caffeine reacts with radiation-induced e - sub(aq) and radicalOH with rate constants of 1.5 x 10 10 and 6.9 x 10 9 dm 3 mol -1 s -1 , respectively. (author)

Sensitivity of thermally treated Bacillus subtilis spores to subsequent irradiation

International Nuclear Information System (INIS)

Mostafa, S.A.; El-Zawahry, Y.A.; Awny, N.M.

1986-01-01

B. subtilis spores exposed to thermal treatment at 70 or 80 0 C for 1 hr were more sensitive to subsequent radiation exposure than non-heated spores. Deactivation of previously heated spores by increasing dose of 0-radiation followed an exponential function while, for non-heated spores a shoulder followed by exponential deactivation was noticed. Combined heat-radiation treatment exhibited a synergistic effect on spore deactivation at low irradiation doses, while at high irradiation doses, the effect was more or less additive. Added values of spore injury was higher for B. subtilis spores that received heat and radiation separately than the observed injury for spores that received combined treatment (heat followed by radiation). Results of spore deactivation and injury due to heat followed by radiation treatment are discussed in comparison to those of spores that received radiation-heat sequence

The Influence of Sporulation Conditions on the Spore Coat Protein Composition of Bacillus subtilis Spores

NARCIS (Netherlands)

Abhyankar, Wishwas R.; Kamphorst, Kiki; Swarge, Bhagyashree N.; van Veen, Henk; van der Wel, Nicole N.; Brul, Stanley; de Koster, Chris G.; de Koning, Leo J.

2016-01-01

Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has

Elastic and inelastic light scattering from single bacterial spores in an optical trap allows the monitoring of spore germination dynamics

Science.gov (United States)

Peng, Lixin; Chen, De; Setlow, Peter; Li, Yong-qing

2009-01-01

Raman scattering spectroscopy and elastic light scattering intensity (ESLI) were used to simultaneously measure levels of Ca-dipicolinic acid (CaDPA) and changes in spore morphology and refractive index during germination of individual B. subtilis spores with and without the two redundant enzymes (CLEs), CwlJ and SleB, that degrade spores’ peptidoglycan cortex. Conclusions from these measurements include: 1) CaDPA release from individual wild-type germinating spores was biphasic; in a first heterogeneous slow phase, Tlag, CaDPA levels decreased ∼15% and in the second phase ending at Trelease, remaining CaDPA was released rapidly; 2) in L-alanine germination of wild-type spores and spores lacking SleB: a) the ESLI rose ∼2-fold shortly before Tlag at T1; b) following Tlag, the ESLI again rose ∼2-fold at T2 when CaDPA levels had decreased ∼50%; and c) the ESLI reached its maximum value at ∼Trelease and then decreased; 3) in CaDPA germination of wild-type spores: a) Tlag increased and the first increase in ESLI occurred well before Tlag, consistent with different pathways for CaDPA and L-alanine germination; b) at Trelease the ESLI again reached its maximum value; 4) in L-alanine germination of spores lacking both CLEs and unable to degrade their cortex, the time ΔTrelease (Trelease–Tlag) for excretion of ≥75% of CaDPA was ∼15-fold higher than that for wild-type or sleB spores; and 5) spores lacking only CwlJ exhibited a similar, but not identical ESLI pattern during L-alanine germination to that seen with cwlJ sleB spores, and the high value for ΔTrelease. PMID:19374431

The Survival and Recovery of Irradiated Bacterial Spores as Affected by Population Density and Some External Factors

International Nuclear Information System (INIS)

Farkas, J.; Kiss, I.; Andrássy, E.

1967-01-01

The radiation resistance of Bacillus cereus spores as affected by the pH-value and cell density of the irradiated spore suspensions was investigated. The portions of the survival curves of suspensions of 10 8 , 4 x 10 3 and 5 x 10 1 per millilitre viable cell counts, respectively, were compared for a three-orders-of-magnitude decrease in viable cell count. It was established that the initial cell density did not affect radiation resistance of spores. Radiation resistance as affected by pH-value in the range of 3 to 8 was investigated. In the range of pH 5 to 8, the radiation resistance of B. cereus spores was not affected. By lowering the pH-value to below 5, the radiation resistance decreased below that observed in the neutral region. The colony-forming capacity of B. cereus, B. coagulans and B. pumilus as a function of the pH-value in the nutrient medium, and the pH-sensitivity of bacterial spores as affected by radiation, were also investigated. It was established that irradiation increased the pH-sensitivity of surviving bacterial spores in all three strains. The initial phase of spore germination (the phase accompanied by decrease of refractivity of the spores) and the division stage of vegetative cells proved to be the most sensitive to the value of the hydrogen ion concentration. (author)

Direct high-pressure NMR observation of dipicolinic acid leaking from bacterial spore: A crucial step for thermal inactivation.

Science.gov (United States)

Akasaka, Kazuyuki; Maeno, Akihiro; Yamazaki, Akira

2017-12-01

A bacterial spore protects itself with an unusually high concentration (~10% in dry weight of spore) of dipicolinic acid (DPA), the release of which is considered the crucial step for inactivating it under mild pressure and temperature conditions. However, the process of how the spore releases DPA in response to pressure remains obscure. Here we apply 1 H high-resolution high-pressure NMR spectroscopy, for the first time, to the spore suspension of Bacillus subtilis natto and monitor directly and in real-time the leaking process of DPA in response to pressure of 200MPa at 20°C. We find that about one third of the total DPA leaks immediately upon applying pressure, but that the rest leaks slowly in hrs upon decreasing the pressure. Once DPA is fully released from the spore, the proteins of the spore become easily denatured at a mild temperature, e.g., 80°C, much below the temperature commonly used to inactivate spores (121°C). The success of the present experiment opens a new avenue for studying bacterial spores and cells at the molecular level in response to pressure, temperature and other perturbations. Copyright © 2017 Elsevier B.V. All rights reserved.

Protection of Bacillus pumilus spores by catalases.

Science.gov (United States)

Checinska, Aleksandra; Burbank, Malcolm; Paszczynski, Andrzej J

2012-09-01

Bacillus pumilus SAFR-032, isolated at spacecraft assembly facilities of the National Aeronautics and Space Administration Jet Propulsion Laboratory, is difficult to kill by the sterilization method of choice, which uses liquid or vapor hydrogen peroxide. We identified two manganese catalases, YjqC and BPUM_1305, in spore protein extracts of several B. pumilus strains by using PAGE and mass spectrometric analyses. While the BPUM_1305 catalase was present in six of the B. pumilus strains tested, YjqC was not detected in ATCC 7061 and BG-B79. Furthermore, both catalases were localized in the spore coat layer along with laccase and superoxide dismutase. Although the initial catalase activity in ATCC 7061 spores was higher, it was less stable over time than the SAFR-032 enzyme. We propose that synergistic activity of YjqC and BPUM_1305, along with other coat oxidoreductases, contributes to the enhanced resistance of B. pumilus spores to hydrogen peroxide. We observed that the product of the catalase reaction, gaseous oxygen, forms expanding vesicles on the spore surface, affecting the mechanical integrity of the coat layer, resulting in aggregation of the spores. The accumulation of oxygen gas and aggregations may play a crucial role in limiting further exposure of Bacilli spore surfaces to hydrogen peroxide or other toxic chemicals when water is present.

The Use of Germinants to Potentiate the Sensitivity of Bacillus anthracis Spores to Peracetic Acid.

Science.gov (United States)

Celebi, Ozgur; Buyuk, Fatih; Pottage, Tom; Crook, Ant; Hawkey, Suzanna; Cooper, Callum; Bennett, Allan; Sahin, Mitat; Baillie, Leslie

2016-01-01

Elimination of Bacillus anthracis spores from the environment is a difficult and costly process due in part to the toxicity of current sporicidal agents. For this reason we investigated the ability of the spore germinants L-alanine (100 mM) and inosine (5 mM) to reduce the concentration of peracetic acid (PAA) required to inactivate B. anthracis spores. While L-alanine significantly enhanced (p = 0.0085) the bactericidal activity of 500 ppm PAA the same was not true for inosine suggesting some form of negative interaction. In contrast the germinant combination proved most effective at 100 ppm PAA (p = 0.0009). To determine if we could achieve similar results in soil we treated soil collected from the burial site of an anthrax infected animal which had been supplemented with spores of the Sterne strain of B. anthracis to increase the level of contamination to 10(4) spores/g. Treatment with germinants followed 1 h later by 5000 ppm PAA eliminated all of the spores. In contrast direct treatment of the animal burial site using this approach delivered using a back pack sprayer had no detectable effect on the level of B. anthracis contamination or on total culturable bacterial numbers over the course of the experiment. It did trigger a significant, but temporary, reduction (p < 0.0001) in the total spore count suggesting that germination had been triggered under real world conditions. In conclusion, we have shown that the application of germinants increase the sensitivity of bacterial spores to PAA. While the results of the single field trial were inconclusive, the study highlighted the potential of this approach and the challenges faced when attempting to perform real world studies on B. anthracis spores contaminated sites.

The use of germinants to potentiate the sensitivity of Bacillus anthracis spores to peracetic acid

Directory of Open Access Journals (Sweden)

Ozgur eCelebi

2016-01-01

Full Text Available Elimination of Bacillus anthracis spores from the environment is a difficult and costly process due in part to the toxicity of current sporicidal agents. For this reason we investigated the ability of the spore germinants L-alanine (100 mM and inosine (5 mM to reduce the concentration of peracetic acid (PAA required to inactivate B.anthracis spores. While L-alanine significantly enhanced (p=0.0085 the bactericidal activity of 500 ppm PAA the same was not true for inosine suggesting some form of negative interaction. In contrast the germinant combination proved most effective at 100 ppm PAA (p=0.0009. To determine if we could achieve similar results in soil we treated soil collected from the burial site of an anthrax infected animal which had been supplemented with spores of the Sterne strain of B.anthracis to increase the level of contamination to 104 spores/g. Treatment with germinants followed one hour later by 5000 ppm PAA eliminated all of the spores. In contrast direct treatment of the animal burial site using this approach delivered using a back pack sprayer had no detectable effect on the level of B.anthracis contamination or on total culturable bacterial numbers over the course of the experiment. It did trigger a significant, but temporary, reduction (p<0.0001 in the total spore count suggesting that germination had been triggered under real world conditions. In conclusion, we have shown that the application of germinants increase the sensitivity of bacterial spores to PAA. While the results of the single field trial were inconclusive, the study highlighted the potential of this approach and the challenges faced when attempting to perform real world studies on B.anthracis spores contaminated sites.

[Survival of Bacillus anthracis spores in various tannery baths].

Science.gov (United States)

Mendrycka, M; Mierzejewski, J

2000-01-01

The influence of tannery baths: liming, deliming, bating, pickling, tanning, retannage on the survival and on the germination dynamism of B. anthracis spores (Sterne strain) was investigated. The periods and the conditions of this influence were established according to technological process of cow hide tannage. Practically after every bath some part of the spores remained vital. The most effective killing of spores occurred after pickling, liming and deliming. Inversely, the most viable spores remained after bating and retannage process. The lack of correlation that was observed between survival and germination of spores after retannage bath can be explained by different mechanism of spores germination inhibition and their killing.

Genetic diversity, virulence and fitness evolution in an obligate fungal parasite of bees.

Science.gov (United States)

Evison, S E F; Foley, K; Jensen, A B; Hughes, W O H

2015-01-01

Within-host competition is predicted to drive the evolution of virulence in parasites, but the precise outcomes of such interactions are often unpredictable due to many factors including the biology of the host and the parasite, stochastic events and co-evolutionary interactions. Here, we use a serial passage experiment (SPE) with three strains of a heterothallic fungal parasite (Ascosphaera apis) of the Honey bee (Apis mellifera) to assess how evolving under increasing competitive pressure affects parasite virulence and fitness evolution. The results show an increase in virulence after successive generations of selection and consequently faster production of spores. This faster sporulation, however, did not translate into more spores being produced during this longer window of sporulation; rather, it appeared to induce a loss of fitness in terms of total spore production. There was no evidence to suggest that a greater diversity of competing strains was a driver of this increased virulence and subsequent fitness cost, but rather that strain-specific competitive interactions influenced the evolutionary outcomes of mixed infections. It is possible that the parasite may have evolved to avoid competition with multiple strains because of its heterothallic mode of reproduction, which highlights the importance of understanding parasite biology when predicting disease dynamics. © 2014 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2014 European Society For Evolutionary Biology.

Ptaquiloside in bracken spores from Britain

DEFF Research Database (Denmark)

Rasmussen, Lars Holm; Schmidt, Bjørn; Sheffield, Elizabeth

2013-01-01

Secondary metabolites from bracken fern (Pteridium aquilinum (L.) Kuhn) are suspected of causing cancer in humans. The main carcinogen is the highly water-soluble norsesquiterpene glucoside ptaquiloside, which may be ingested by humans through food, e.g. via contaminated water, meat or milk. It has...... been postulated that carcinogens could also be ingested through breathing air containing bracken spores. Ptaquiloside has not previously been identified in bracken spores. The aim of the study was to determine whether ptaquiloside is present in bracken spores, and if so, to estimate its content...

Asynchronous spore germination in isogenic natural isolates of Saccharomyces paradoxus.

Science.gov (United States)

Stelkens, Rike B; Miller, Eric L; Greig, Duncan

2016-05-01

Spores from wild yeast isolates often show great variation in the size of colonies they produce, for largely unknown reasons. Here we measure the colonies produced from single spores from six different wild Saccharomyces paradoxus strains. We found remarkable variation in spore colony sizes, even among spores that were genetically identical. Different strains had different amounts of variation in spore colony sizes, and variation was not affected by the number of preceding meioses, or by spore maturation time. We used time-lapse photography to show that wild strains also have high variation in spore germination timing, providing a likely mechanism for the variation in spore colony sizes. When some spores from a laboratory strain make small colonies, or no colonies, it usually indicates a genetic or meiotic fault. Here, we demonstrate that in wild strains spore colony size variation is normal. We discuss and assess potential adaptive and non-adaptive explanations for this variation. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Strategy to inactivate Clostridium perfringens spores in meat products.

Science.gov (United States)

Akhtar, Saeed; Paredes-Sabja, Daniel; Torres, J Antonio; Sarker, Mahfuzur R

2009-05-01

The current study aimed to develop an inactivation strategy for Clostridium perfringens spores in meat through a combination of spore activation at low pressure (100-200 MPa, 7 min) and elevated temperature (80 degrees C, 10 min); spore germination at high temperatures (55, 60 or 65 degrees C); and inactivation of germinated spores with elevated temperatures (80 and 90 degrees C, 10 and 20 min) and high pressure (586 MPa, at 23 and 73 degrees C, 10 min). Low pressures (100-200 MPa) were insufficient to efficiently activate C. perfringens spores for germination. However, C. perfringens spores were efficiently activated with elevated temperature (80 degrees C, 10 min), and germinated at temperatures lethal for vegetative cells (>or= 55 degrees C) when incubated for 60 min with a mixture of L-asparagine and KCl (AK) in phosphate buffer (pH 7) and in poultry meat. Inactivation of spores (approximately 4 decimal reduction) in meat by elevated temperatures (80-90 degrees C for 20 min) required a long germination period (55 degrees C for 60 min). However, similar inactivation level was reached with shorter germination period (55 degrees C for 15 min) when spore contaminated-meat was treated with pressure-assisted thermal processing (568 MPa, 73 degrees C, 10 min). Therefore, the most efficient strategy to inactivate C. perfringens spores in poultry meat containing 50 mM AK consisted: (i) a primary heat treatment (80 degrees C, 10 min) to pasteurize and denature the meat proteins and to activate C. perfringens spores for germination; (ii) cooling of the product to 55 degrees C in about 20 min and further incubation at 55 degrees C for about 15 min for spore germination; and (iii) inactivation of germinated spores by pressure-assisted thermal processing (586 MPa at 73 degrees C for 10 min). Collectively, this study demonstrates the feasibility of an alternative and novel strategy to inactivate C. perfringens spores in meat products formulated with germinants specific for C

Spore coat protein of Bacillus subtilis. Structure and precursor synthesis.

Science.gov (United States)

Munoz, L; Sadaie, Y; Doi, R H

1978-10-10

The coat protein of Bacillus subtilis spores comprises about 10% of the total dry weight of spores and 25% of the total spore protein. One protein with a molecular weight of 13,000 to 15,000 comprises a major portion of the spore coat. This mature spore coat protein has histidine at its NH2 terminus and is relatively rich in hydrophobic amino acids. Netropsin, and antibiotic which binds to A-T-rich regions of DNA and inhibits sporulation, but not growth, decreased the synthesis of this spore coat protein by 75%. A precursor spore coat protein with a molecular weight of 25,000 is made initially at t1 of sporulation and is converted to the mature spore coat protein with a molecular weight of 13,500 at t2 - t3. These data indicate that the spore coat protein gene is expressed very early in sporulation prior to the modifications of RNA polymerase which have been noted.

Identification of some factors affecting pharmaceutical active compounds (PhACs) removal in real wastewater. Case study of fungal treatment of reverse osmosis concentrate.

Science.gov (United States)

Badia-Fabregat, Marina; Lucas, Daniel; Gros, Meritxell; Rodríguez-Mozaz, Sara; Barceló, Damià; Caminal, Glòria; Vicent, Teresa

2015-01-01

Many technologies are being developed for the efficient removal of micropollutants from wastewater and, among them, fungal degradation is one of the possible alternative biological treatments. In this article, some factors that might affect pharmaceutically active compounds (PhACs) removal in a fungal treatment of real wastewater were identified in batch bioreactor treating reverse osmosis concentrate (ROC) from urban wastewater treatment plant (WWTP). We found that degradation of PhACs by Trametes versicolor was enhanced by addition of external nutrients (global removal of 44%). Moreover, our results point out that high aeration might be involved in the increase in the concentration of some PhACs. In fact, conjugation and deconjugation processes (among others) affect the removal assessment of emerging contaminants when working with real concentrations in comparison to experiments with spiked samples. Moreover, factors that could affect the quantification of micropollutants at lab-scale experiments were studied. Copyright © 2014 Elsevier B.V. All rights reserved.

Effect of arbuscular mycorrhizal fungal inoculation in combination with different organic fertilizers on maize crop in eroded soils

International Nuclear Information System (INIS)

Sharif, M.; Saud, S.; Khan, F.

2012-01-01

A pot experiment was conducted to study the effect of inoculating maize (Zea mays L. Azam) with Arbuscular mycorrhizal (AM) fungi in 2 different series of North West Pakistan during the year 2007. Data showed significant increase in shoots and roots yield of maize with the inoculation of AM fungi alone and in combination with farm yard manure (FYM), poultry manure (PM) and humic acid (HA) over control and N-P-K treatments. Accumulation of N by maize shoots increased significantly by the addition of HA, PM and FYM plus N-P-K with or without inoculation of AM fungi over the treatments of N-P-K and control. Plants P accumulation increased significantly over control and N-P-K treatments with the inoculation of AM fungi alone and in combination with FYM, PM and HA in missa soil series. In missa gullied soil series, significantly increased plants P accumulation was noted by the treatments of AM inoculation with PM followed by HA. Accumulation of Mn by maize shoots increased significantly with AM inoculation with HA and PM over all other treatments, Fe increased with PM, HA and FYM. Plants Cu accumulation in missa series increased significantly over control and N-P-K treatments by AM alone and in combination with PM, FYM and HA and by AM fungi with PM, FYM and HA in missa gullied series. Maximum Mycorrhizal root infection rate of 51 % was recorded in the treatment of AM fungal inoculation with HA followed by the treatment inoculated with AM fungi with FYM. In missa gullied soil series, Maximum (59 %) and significantly increased roots infection rates over all treatments were observed in the treatment of AM fungal inoculation with HA followed by PM. Spores concentrations of AM fungi increased significantly with AM inoculation alone and with FYM, PM and HA. Maximum spores numbers of 50 in 20 g soil were recorded by the inoculation of AM fungi alone and with HA. (author)

The infrared spectral transmittance of Aspergillus niger spore aggregated particle swarm

Science.gov (United States)

Zhao, Xinying; Hu, Yihua; Gu, Youlin; Li, Le

2015-10-01

Microorganism aggregated particle swarm, which is quite an important composition of complex media environment, can be developed as a new kind of infrared functional materials. Current researches mainly focus on the optical properties of single microorganism particle. As for the swarm, especially the microorganism aggregated particle swarm, a more accurate simulation model should be proposed to calculate its extinction effect. At the same time, certain parameters deserve to be discussed, which helps to better develop the microorganism aggregated particle swarm as a new kind of infrared functional materials. In this paper, take Aspergillus Niger spore as an example. On the one hand, a new calculation model is established. Firstly, the cluster-cluster aggregation (CCA) model is used to simulate the structure of Aspergillus Niger spore aggregated particle. Secondly, the single scattering extinction parameters for Aspergillus Niger spore aggregated particle are calculated by using the discrete dipole approximation (DDA) method. Thirdly, the transmittance of Aspergillus Niger spore aggregated particle swarm is simulated by using Monte Carlo method. On the other hand, based on the model proposed above, what influences can wavelength causes has been studied, including the spectral distribution of scattering intensity of Aspergillus Niger spore aggregated particle and the infrared spectral transmittance of the aggregated particle swarm within the range of 8-14μm incident infrared wavelengths. Numerical results indicate that the scattering intensity of Aspergillus Niger spore aggregated particle reduces with the increase of incident wavelengths at each scattering angle. Scattering energy mainly concentrates on the scattering angle between 0-40°, forward scattering has an obvious effect. In addition, the infrared transmittance of Aspergillus Niger spore aggregated particle swarm goes up with the increase of incident wavelengths. However, some turning points of the trend are

Magnetic Ganoderma lucidum spore microspheres: A novel material to immobilize CotA multicopper oxidase for dye decolorization

International Nuclear Information System (INIS)

Fan, Lili; Wang, Yan; Zhao, Min; Song, Jinzhu; Wang, Jueyu; Jin, Zijing

2016-01-01

Highlights: • Hollow microspheres were obtained from Ganoderma lucidum spores. • Novel magnetic microspheres were prepared by load hollow spore microspheres with Fe_3O_4 nanoparticles. • CotA multicopper oxidase was immobilized on the magnetic spore microspheres for indigo carmine decolorization. • The immobilized CotA displayed higher decolorization capability and reusability. - Abstract: In this study, hollow microspheres were obtained from Ganoderma lucidum spores. Then the hollow microspheres were loaded with Fe_3O_4 nanoparticles to prepare novel magnetic spore microspheres. TEM images and X-ray diffractometry demonstrated that the Fe_3O_4 nanoparticles were incorporated throughout the spore microsphere. CotA multicopper oxidase was chosen as biomacromolecule to study the loading ability of the magnetic spore microspheres. The combination of the CotA enzyme with the microsphere was observed by laser scanning confocal microscope. The loaded amount of CotA on the microspheres was 75 mg/g when the CotA concentration was 1.2 mg/mL and the activity recovery of the immobilized CotA was 81%. The magnetic microspheres loaded with CotA, which can be easily and quickly recovered by an external magnetic field, were used for dye decolorization. After 1 h decolorization, 99% of the indigo carmine has been removed by 10 mg microspheres. In addition, the immobilized CotA retained 75% of activity after 10 consecutive cycles, which indicated that the magnetic spore microspheres are good support material for immobilization of the enzyme.

Nanoceria and bulk cerium oxide effects on the germination of asplenium adiantum-nigrum spores

Directory of Open Access Journals (Sweden)

Aranzazu Gomez-Garay

2016-12-01

Full Text Available Aim of study: The effect of cerium oxide engineered nanoparticles on the spore germination of the fern. Asplenium adiantum-nigrum. Area of study: France, Britanny Region, Finistére Department, Plougonvelin, in rocks near the sea. Material and methods: Asplenium spores were cultured in vitro on agar medium with Nano-CeO2 (less than 25 nm particle size and bulk-CeO2. The addition of each nano- and bulk particles ranged from 0 to 3000 mg L-1. Observations on rhizoidal and prothallial cells during first stages of gametophyte development were made. The No-Observed-Adverse-Effect concentration (NOAEC and Lowest-Observed-Adverse-Effect-Concentration (LOEC values for spore germination rate data were analyzed.  Main results: Germination was speeded up by 100 to 2000 mg L-1 nanoceria, while bulk cerium oxide had the same effect for 500 to 200 mg L-1 concentrations. Present results showed cellular damage in the protonema while rhizoid cells seemed not to be affected, as growth and membrane integrity remained. Research highlights: Both nanosized and bulk cerium oxide are toxic for the fern Asplenium adiantum-nigrum, although diverse toxicity patterns were shown for both materials. Diverse toxic effects have been observed: chloroplast membrane damage and lysis, cell wall and membrane disruption which leads to cell lysis; and alterations in morphology and development. Keywords: Nanoparticles; rhizoid; prothallus; chloroplast; fern.

Nanoceria and bulk cerium oxide effects on the germination of asplenium adiantum-nigrum spores

Energy Technology Data Exchange (ETDEWEB)

Gomez-Garay, A.; Pintos, B.; Manzanera, J.A.; Prada, C.; Martin, L.; Gabriel y Galan, J.M.

2016-07-01

Aim of the study: The effect of cerium oxide engineered nanoparticles on the spore germination of the fern. Asplenium adiantum-nigrum. Area of study: France, Britanny Region, Finistére Department, Plougonvelin, in rocks near the sea. Material and methods: Asplenium spores were cultured in vitro on agar medium with Nano-CeO2 (less than 25 nm particle size) and bulk-CeO2. The addition of each nano- and bulk particles ranged from 0 to 3000 mg L-1. Observations on rhizoidal and prothallial cells during first stages of gametophyte development were made. The No-Observed-Adverse-Effect concentration (NOAEC) and Lowest-Observed-Adverse-Effect-Concentration (LOEC) values for spore germination rate data were analyzed. Main results: Germination was speeded up by 100 to 2000 mg L-1 nanoceria, while bulk cerium oxide had the same effect for 500 to 200 mg L-1 concentrations. Present results showed cellular damage in the protonema while rhizoid cells seemed not to be affected, as growth and membrane integrity remained. Research highlights: Both nanosized and bulk cerium oxide are toxic for the fern Asplenium adiantum-nigrum, although diverse toxicity patterns were shown for both materials. Diverse toxic effects have been observed: chloroplast membrane damage and lysis, cell wall and membrane disruption which leads to cell lysis; and alterations in morphology and development. (Author)

Inhibiting Inosine Hydrolase and Alanine Racemase to Enhance the Germination of Bacillus anthracis Sterne Spores: Potential Spore Decontamination Strategies

Science.gov (United States)

2015-06-19

decontamination strategies>> Maryline DEFEZ 1𔃼, Melissa HUNTER3J Susan WELKOS :~J Christopher COTE3 1 University Grenoble-Alpes, Grenoble, France. 1...inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores potential spore decontamination strategies 5a...8217 • Accidentally in Humans • Natural reservoir is soil • Anthrax Disease Cycle: - animals infected by soilborne spores in food and water or bites from certain

Intra-abdominal fungal pseudomycetoma in two cats.

Science.gov (United States)

Bianchi, Matheus V; Laisse, Cláudio J M; Vargas, Thainã P; Wouters, Flademir; Boabaid, Fabiana M; Pavarini, Saulo P; Ferreiro, Laerte; Driemeier, David

Pseudomycetomas are deep cutaneous to subcutaneous lesions caused by Microsporum canis mainly described in Persian cats, with few reports of intra-abdominal location. This report describes the clinical signs and lesions of intra-abdominal pseudomycetomas caused by M. canis in two Persian cats. Two Persian cats with a history of previous laparotomy (ovariohysterectomy and nephrostomy) and fecal impaction were examined. Cat #1 was euthanized and subjected to necropsy, histopathology and mycological evaluation. Cat #2 presented with chronic dermatophytosis, and an intra-abdominal mass, that was subjected to histopathology evaluation. Cat #1 presented at necropsy a white-grayish, firm mass (6cm×3.5cm×2.8cm) in the uterine cervix. Cat #2 presented a firm whitish mass (6.5cm×1.5cm×0.5cm) located close to the left kidney. Histologically, both masses contained multifocal granules with hyphae and spores surrounded by Splendore-Hoeppli reaction, with a pyogranulomatous inflammatory infiltrate and fibrous connective tissue proliferation in the periphery. Hyphae and spores exhibited marked Grocott and periodic acid-Schiff staining. M. canis was identified by fungal isolation in cat #1. Pseudomycetoma should be considered as a differential diagnosis in cats, especially in Persian cats presenting with an intra-abdominal mass. Entrance of the agent into the cavity can occur during laparotomy. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

Mate Finding, Sexual Spore Production, and the Spread of Fungal Plant Parasites.

Science.gov (United States)

Hamelin, Frédéric M; Castella, François; Doli, Valentin; Marçais, Benoît; Ravigné, Virginie; Lewis, Mark A

2016-04-01

Sexual reproduction and dispersal are often coupled in organisms mixing sexual and asexual reproduction, such as fungi. The aim of this study is to evaluate the impact of mate limitation on the spreading speed of fungal plant parasites. Starting from a simple model with two coupled partial differential equations, we take advantage of the fact that we are interested in the dynamics over large spatial and temporal scales to reduce the model to a single equation. We obtain a simple expression for speed of spread, accounting for both sexual and asexual reproduction. Taking Black Sigatoka disease of banana plants as a case study, the model prediction is in close agreement with the actual spreading speed (100 km per year), whereas a similar model without mate limitation predicts a wave speed one order of magnitude greater. We discuss the implications of these results to control parasites in which sexual reproduction and dispersal are intrinsically coupled.

Spore analysis and tetrad dissection of Schizosaccharomyces pombe

DEFF Research Database (Denmark)

Ekwall, Karl; Thon, Genevieve

2017-01-01

Here we describe the processing of Schizosaccharomyces pombe spores in batches (random spore analysis) or through tetrad dissections. Spores are usually prepared from matings between haploid strains (producing zygotic asci) or from sporulating diploids (producing azygotic asci). In random spore...

Dispersal of spores following a persistent random walk.

Science.gov (United States)

Bicout, D J; Sache, I

2003-03-01

A model of a persistent random walk is used to describe the transport and deposition of the spore dispersal process. In this model, the spore particle flies along straight line trajectories, with constant speed v, which are interrupted by scattering, originating from interaction of spores with the field and wind variations, which randomly change its direction. To characterize the spore dispersal gradients, we have derived analytical expressions of the deposition probability epsilon (r|v) of airborne spores as a function of the distance r from the spore source in an infinite free space and in a disk of radius R with an absorbing edge that mimics an agricultural field surrounded with fields of nonhost plants and bare land. It is found in the free space that epsilon (r|v) approximately e(-alphar/l), with alpha a function of l(d)/l, where l and l(d) are the scattering and deposition mean free paths, respectively. In the disk, however, epsilon (r|v) is an infinite series of Bessel functions and, exhibits three regimes: absorbing (Rl(d)).

Effect of supplementing lactating goats fed on aflatoxin ...

African Journals Online (AJOL)

ACSS

The fungal spores are found worldwide, in air and soil, and infest both living and dead plants and animals. An experiment was conducted to investigate the concentration, total excretion and .... nutritionally inert adsorbents (calcium bentonite or activated charcoal) reduced the concentration of aflatoxin M1 in milk. (Table 1).

Effect of Selenium on Control of Postharvest Gray Mold of Tomato Fruit and the Possible Mechanisms Involved

Science.gov (United States)

Wu, Zhilin; Yin, Xuebin; Bañuelos, Gary S.; Lin, Zhi-Qing; Zhu, Zhu; Liu, Ying; Yuan, Linxi; Li, Miao

2016-01-01

Selenium (Se) has important benefits for crop growth and stress tolerance at low concentrations. However, there is very little information on antimicrobial effect of Se against the economically important fungus Botrytis cinerea. In the present study, using sodium selenite as Se source, we investigated the effect of Se salts on spore germination and mycelial growth of the fungal pathogen in vitro and gray mold control in harvested tomato fruit. Se treatment at 24 mg/L significantly inhibited spore germination of the fungal pathogen and effectively controlled gray mold in harvested tomato fruit. Se treatment at 24 mg/L seems to induce the generation of intracellular reactive oxygen species in the fungal spores. The membrane integrity damage was observed with fluorescence microscopy following staining with propidium iodide after treatment of the spores with Se. These results suggest that Se has the potential for controlling gray mold rot of tomato fruits and might be useful in integrated control against gray mold disease of postharvest fruits and vegetables caused by B. cinerea. The mechanisms by which Se decreased gray mold decay of tomato fruit may be directly related to the severe damage to the conidia plasma membrane and loss of cytoplasmic materials from the hyphae. PMID:26779128

Effect of selenium on control of postharvest gray mould of tomato fruit and the possible mechanisms involved

Directory of Open Access Journals (Sweden)

Zhilin eWu

2016-01-01

Full Text Available Selenium (Se has important benefits for crop growth and stress tolerance at low concentrations. However, there is very little information on antimicrobial effect of selenium against the economically important fungus Botrytis cinerea. In the present study, using sodium selenite as Se source, we investigated the effect of Se salts on spore germination and mycelial growth of the fungal pathogen in vitro and gray mould control in harvested tomato fruit. Se treatment at 24 mg/L significantly inhibited spore germination of the fungal pathogen and effectively controlled gray mould in harvested tomato fruit. Se treatment at 24 mg/L seems to induce the generation of intracellular reactive oxygen species in the fungal spores. The membrane integrity damage was observed with fluorescence microscopy following staining with propidium iodide after treatment of the spores with Se. These results suggest that Se has the potential for controlling gray mould rot of tomato fruits and might be useful in integrated control against gray mould disease of postharvest fruits and vegetables caused by B. cinerea. The mechanisms by which Se decreased gray mould decay of tomato fruit may be directly related to the severe damage to the conidia plasma membrane and loss of cytoplasmic materials from the hyphae.

Examination of Mechanisms Responsible for Organic Dust-related Diseases: Mediator Release induced by Microorgansims. A review

DEFF Research Database (Denmark)

Norn, Svend; Clementsen, Paul; Kristensen, K.S.

1994-01-01

Farmakologi, org. dust-related diseases, bacteria, pathogenic mechanisms, mediator release, entoxins - fungal spores......Farmakologi, org. dust-related diseases, bacteria, pathogenic mechanisms, mediator release, entoxins - fungal spores...

Fungal pathogens of Miconia calvescens (Melastomataceae) from Brazil, with reference to classical biological control.

Science.gov (United States)

Seixas, Claudine D S; Barreto, Robert W; Killgore, Eloise

2007-01-01

A survey of fungal pathogens of Miconia calvescens was carried out in Brazil aimed at finding potential classical biocontrol agents for management of this invasive alien weed in Hawaii. Coccodiella miconiae, Glomerella cingulata (= Colletotrichum gloeosporioides f. sp. miconiae) and the new species Guignardia miconiae and Korunomyces prostratus were found associated with foliar diseases and are described herein. Two previously undescribed spore stages of Coccodiella miconiae also were obtained allowing a complete description of this species. Pseudocercospora tamonae associated with leaf spots of other species of Miconia also was collected and also was proven to be pathogenic to M. calvescens.

Comparison of hand hygiene procedures for removing Bacillus cereus spores.

Science.gov (United States)

Sasahara, Teppei; Hayashi, Shunji; Hosoda, Kouichi; Morisawa, Yuji; Hirai, Yoshikazu

2014-01-01

Bacillus cereus is a spore-forming bacterium. B. cereus occasionally causes nosocomial infections, in which hand contamination with the spores plays an important role. Therefore, hand hygiene is the most important practice for controlling nosocomial B. cereus infections. This study aimed to determine the appropriate hand hygiene procedure for removing B. cereus spores. Thirty volunteers' hands were experimentally contaminated with B. cereus spores, after which they performed 6 different hand hygiene procedures. We compared the efficacy of the procedures in removing the spores from hands. The alcohol-based hand-rubbing procedures scarcely removed them. The soap washing procedures reduced the number of spores by more than 2 log10. Extending the washing time increased the spore-removing efficacy of the washing procedures. There was no significant difference in efficacy between the use of plain soap and antiseptic soap. Handwashing with soap is appropriate for removing B. cereus spores from hands. Alcohol-based hand-rubbing is not effective.

Characterization and control of fungal morphology for improved production performance in biotechnology.

Science.gov (United States)

Krull, Rainer; Wucherpfennig, Thomas; Esfandabadi, Manely Eslahpazir; Walisko, Robert; Melzer, Guido; Hempel, Dietmar C; Kampen, Ingo; Kwade, Arno; Wittmann, Christoph

2013-01-20

Filamentous fungi have been widely applied in industrial biotechnology for many decades. In submerged culture processes, they typically exhibit a complex morphological life cycle that is related to production performance--a link that is of high interest for process optimization. The fungal forms can vary from dense spherical pellets to viscous mycelia. The resulting morphology has been shown to be influenced strongly by process parameters, including power input through stirring and aeration, mass transfer characteristics, pH value, osmolality and the presence of solid micro-particles. The surface properties of fungal spores and hyphae also play a role. Due to their high industrial relevance, the past years have seen a substantial development of tools and techniques to characterize the growth of fungi and obtain quantitative estimates on their morphological properties. Based on the novel insights available from such studies, more recent studies have been aimed at the precise control of morphology, i.e., morphology engineering, to produce superior bio-processes with filamentous fungi. Copyright © 2012 Elsevier B.V. All rights reserved.

DNA repair in ultraviolet-irradiated spores of Bacillus subtilis

International Nuclear Information System (INIS)

Wang, T.C.V.

1976-01-01

It has been shown previously by others that at least two independent repair mechanisms are present in Bacillus subtilis for removing ''spore photoproduct'' from DNA of ultraviolet (254 nm)-irradiated spores after germination. One of these, designated as ''spore repair,'' is shown in this study to restore ''spore photoproduct'' to two thymine residues, leaving the DNA backbone intact at the end of the process in vivo. The circumstances under which this repair can occur and some characteristics of its energy requirements have been clarified. The second repair process is identified as excision repair, which can excise both ''spore photoproduct'' from DNA of irradiated spores and cyclobutane-type pyrimidine dimers from DNA of irradiated vegetative cells. In this study it is shown that the gene hcr 1 affects an enzyme activity for the incision step initiating this repair, while the gene hcr 42 affects a step subsequent to incision in the mechanism. In addition a third, independent repair system, termed ''germinative excision repair,'' is discovered and shown to be specific for excising only cyclobutane-type pyrimidine dimers but not ''spore photoproduct.'' This repair system is responsible for the observed high ultraviolet-resistance and temporary capacity for host cell reactivation on recently germinated spores of Bacillus subtilis HCR - strains

The Role of the Electrostatic Force in Spore Adhesion

Energy Technology Data Exchange (ETDEWEB)

Chung, Eunhyea [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Lee, Ida [University of Tennessee, Knoxville (UTK); Tsouris, Costas [ORNL

2010-01-01

Electrostatic force is investigated as one of the components of the adhesion force between Bacillus thuringiensis (Bt) spores and planar surfaces. The surface potentials of a Bt spore and a mica surface are experimentally obtained using a combined atomic force microscopy (AFM)-scanning surface potential microscopy technique. On the basis of experimental information, the surface charge density of the spores is estimated at 0.03 {micro}C/cm{sup 2} at 20% relative humidity and decreases with increasing humidity. The Coulombic force is introduced for the spore-mica system (both charged, nonconductive surfaces), and an electrostatic image force is introduced to the spore-gold system because gold is electrically conductive. The Coulombic force for spore-mica is repulsive because the components are similarly charged, while the image force for the spore-gold system is attractive. The magnitude of both forces decreases with increasing humidity. The electrostatic forces are added to other force components, e.g., van der Waals and capillary forces, to obtain the adhesion force for each system. The adhesion forces measured by AFM are compared to the estimated values. It is shown that the electrostatic (Coulombic and image) forces play a significant role in the adhesion force between spores and planar surfaces.

Cave Entrance dependent Spore Dispersion of Filamentous Fungi Isolated from Various Sediments of Iron Ore Cave in Brazil: a colloquy on human threats while caving

Directory of Open Access Journals (Sweden)

Erika Linzi Silva Taylor

2014-04-01

Full Text Available Caves are stable environments with characteristics favoring the development of fungi. The fungal community present in a cave also includes pathogenic and opportunistic species out of which some are also served as energy sources in such energy stared ecosystems. Studies on microbial diversity and their role on such energy starved ecosystem are scarce. The present study was aimed to identify the cultivable filamentous fungi present in the various sediment of an iron ore cave and to recognize them as pathogenic and/or opportunistic species. Further the impact of cave entrance on the spore depositions on various distances dependent sediments were analyzed. The results suggest a diverse microbial community inhabiting the cave and an influence of cave entrance over spore deposition on various sediments. We counted a total of 4,549 filamentous fungi that included 34 species of 12 genera: Acremonium, Aspergillus, Cladosporium, Fusarium, Geotrichum, Paecilomyces, Purpureocillium, Penicillium, Torula, Trichoderma, Mucor and Rhizopus. A positive significant relation was observed between spore deposition and distance from cave entrance (p= 0.001. Areas of potential mycoses risks were recognized. This is the first study on microbiological community of an iron ore cave in the country.

Ultra high pressure homogenization (UHPH) inactivation of Bacillus amyloliquefaciens spores in phosphate buffered saline (PBS) and milk

Science.gov (United States)

Dong, Peng; Georget, Erika S.; Aganovic, Kemal; Heinz, Volker; Mathys, Alexander

2015-01-01

Ultra high pressure homogenization (UHPH) opens up new areas for dynamic high pressure assisted thermal sterilization of liquids. Bacillus amyloliquefaciens spores are resistant to high isostatic pressure and temperature and were suggested as potential surrogate for high pressure thermal sterilization validation. B. amyloliquefaciens spores suspended in PBS buffer (0.01 M, pH 7.0), low fat milk (1.5%, pH 6.7), and whole milk (3.5%, pH 6.7) at initial concentration of ~106 CFU/mL were subjected to UHPH treatments at 200, 300, and 350 MPa with an inlet temperature at ~80°C. Thermal inactivation kinetics of B. amyloliquefaciens spores in PBS and milk were assessed with thin wall glass capillaries and modeled using first-order and Weibull models. The residence time during UHPH treatments was estimated to determine the contribution of temperature to spore inactivation by UHPH. No sublethal injury was detected after UHPH treatments using sodium chloride as selective component in the nutrient agar medium. The inactivation profiles of spores in PBS buffer and milk were compared and fat provided no clear protective effect for spores against treatments. Treatment at 200 MPa with valve temperatures lower than 125°C caused no reduction of spores. A reduction of 3.5 log10CFU/mL of B. amyloliquefaciens spores was achieved by treatment at 350 MPa with a valve temperature higher than 150°C. The modeled thermal inactivation and observed inactivation during UHPH treatments suggest that temperature could be the main lethal effect driving inactivation. PMID:26236296

Magnetic Ganoderma lucidum spore microspheres: A novel material to immobilize CotA multicopper oxidase for dye decolorization

Energy Technology Data Exchange (ETDEWEB)

Fan, Lili [School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin 150001 (China); Wang, Yan, E-mail: wangy_msn@hit.edu.cn [School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin 150001 (China); Zhao, Min [College of Life Science, Northeast Forestry University, Harbin 150040 (China); Song, Jinzhu [School of Life Science and Technology, Harbin Institute of Technology, Harbin 150001 (China); Wang, Jueyu; Jin, Zijing [College of Life Science, Northeast Forestry University, Harbin 150040 (China)

2016-08-05

Highlights: • Hollow microspheres were obtained from Ganoderma lucidum spores. • Novel magnetic microspheres were prepared by load hollow spore microspheres with Fe{sub 3}O{sub 4} nanoparticles. • CotA multicopper oxidase was immobilized on the magnetic spore microspheres for indigo carmine decolorization. • The immobilized CotA displayed higher decolorization capability and reusability. - Abstract: In this study, hollow microspheres were obtained from Ganoderma lucidum spores. Then the hollow microspheres were loaded with Fe{sub 3}O{sub 4} nanoparticles to prepare novel magnetic spore microspheres. TEM images and X-ray diffractometry demonstrated that the Fe{sub 3}O{sub 4} nanoparticles were incorporated throughout the spore microsphere. CotA multicopper oxidase was chosen as biomacromolecule to study the loading ability of the magnetic spore microspheres. The combination of the CotA enzyme with the microsphere was observed by laser scanning confocal microscope. The loaded amount of CotA on the microspheres was 75 mg/g when the CotA concentration was 1.2 mg/mL and the activity recovery of the immobilized CotA was 81%. The magnetic microspheres loaded with CotA, which can be easily and quickly recovered by an external magnetic field, were used for dye decolorization. After 1 h decolorization, 99% of the indigo carmine has been removed by 10 mg microspheres. In addition, the immobilized CotA retained 75% of activity after 10 consecutive cycles, which indicated that the magnetic spore microspheres are good support material for immobilization of the enzyme.

Decreased UV light resistance of spores of Bacillus subtilis strains deficient in pyrimidine dimer repair and small, acid-soluble spore proteins

International Nuclear Information System (INIS)

Setlow, B.; Setlow, P.

1988-01-01

Loss of small, acid-soluble spore protein alpha reduced spore UV resistance 30- to 50-fold in Bacillus subtilis strains deficient in pyrimidine dimer repair, but gave only a 5- to 8-fold reduction in UV resistance in repair-proficient strains. However, both repair-proficient and -deficient spores lacking this protein had identical heat and gamma-radiation resistance

Identification of a New Fungal Pathogen Causing White Villous Disease on the Fruiting Body of the Culinary-Medicinal Mushroom Auricularia auricula-judae (Agaricomycetes) in China.

Science.gov (United States)

Zhang, Jie-Chi; Kong, Xiang-Hui; Zhang, Pi-Qi; Liu, Jia-Ning; Ma, Yin-Peng; Dai, Xiao-Dong; Han, Zeng-Hua; Ma, Qing-Fang; Wang, Xiao-Yong; Yu, Li-Ping

2017-01-01

Auricularia auricula-judae is an edible and medicinal fungus ranking fourth in production among the edible fungi cultivated worldwide. White villous disease is rampant in Northeast China; it infects the fruiting bodies of A. auricula-judae by forming a white mycelial layer on its ventral side. The disease not only causes an unacceptable morphological appearance and a poor-quality product, but it also significantly reduces the yield. In this study, based on fungal morphology, ribosomal DNA internal transcribed spacer sequences, identification of species-specific primers, and the pathogenicity of the mycelia and spores, 2 fungal pathogens were isolated and identified as Fusarium equiseti and F. sporotrichioides.

The cellulose-binding activity of the PsB multiprotein complex is required for proper assembly of the spore coat and spore viability in Dictyostelium discoideum.

Science.gov (United States)

Srinivasan, S; Griffiths, K R; McGuire, V; Champion, A; Williams, K L; Alexander, S

2000-08-01

The terminal event of spore differentiation in the cellular slime mould Dictyostelium discoideum is the assembly of the spore coat, which surrounds the dormant amoeba and allows the organism to survive during extended periods of environmental stress. The spore coat is a polarized extracellular matrix composed of glycoproteins and cellulose. The process of spore coat formation begins by the regulated secretion of spore coat proteins from the prespore vesicles (PSVs). Four of the major spore coat proteins (SP96, PsB/SP85, SP70 and SP60) exist as a preassembled multiprotein complex within the PSVs. This complete complex has an endogenous cellulose-binding activity. Mutant strains lacking either the SP96 or SP70 proteins produce partial complexes that do not have cellulose-binding activity, while mutants lacking SP60 produce a partial complex that retains this activity. Using a combination of immunofluorescence microscopy and biochemical methods we now show that the lack of cellulose-binding activity in the SP96 and SP70 mutants results in abnormally assembled spore coats and spores with greatly reduced viability. In contrast, the SP60 mutant, in which the PsB complex retains its cellulose-binding activity, produces spores with apparently unaltered structure and viability. Thus, it is the loss of the cellulose-binding activity of the PsB complex, rather than the mere loss of individual spore coat proteins, that results in compromised spore coat structure. These results support the idea that the cellulose-binding activity associated with the complete PsB complex plays an active role in the assembly of the spore coat.

Interactions of liposome carriers with infectious fungal hyphae reveals the role of β-glucans.

Science.gov (United States)

Chavan, Neelam L; Young, Joseph K; Drezek, Rebekah A; Lewis, Russell; Bikram, Malavosklish

2012-09-04

Relatively little is known about how liposomal formulations modulate drug delivery to fungal pathogens. We compared patterns of hyphal cell wall binding for empty rhodmine-labeled liposomes and the clinically available amphotericin B-containing liposomal formulation (AmBisome) in Aspergillus fumigatus and Candida albicans. Following 0.5 h of coincubation with A. fumigatus , empty liposomes concentrated primarily in fungal septae along at the surface of the cell wall, suggesting that liposome uptake is concentrated in areas of the cell wall where linear glucan is exposed on the cell surface, which was confirmed by aniline blue staining. Consistent with this hypothesis, pretreatment of liposomes with soluble linear glucan (laminarin) decreased liposome binding in both Aspergillus and Candida fungal hyphae, while growth of Aspergillus hyphae in the presence of an agent that increases fungal cell wall surface exposure of linear β-glucans without cell death (caspofungin) increased liposome uptake throughout the Aspergillus fungal cell wall. Increasing the polyethylene glycol (PEG) concentration in liposomes from 0 to 30% significantly increased fungal uptake of liposomes that was only modestly attenuated when fungal cells were incubated in serum concentrations ranging from 10 to 100%. The presence of β-glucans on the fungal hyphae cell walls of Aspergillus fumigatus is one of the factors responsible for mediating the binding of liposome carriers to the hyphae and could explain possible synergy reported between liposomal amphotericin B and echinocanins.

The role of heat resistance in thermorestoration of hydrated bacterial spores

International Nuclear Information System (INIS)

Friedman, Y.S.; Grecz, N.

1973-01-01

This study for the first time presents evidence of the distinct role played in thermorestoration by cellular determinants such as the resistance to heat and radiation, and the ionic state of spores. In the past only radiochemical determinants associated with radical annealment have been studied in hydrated systems. The basic heat resistance of spores plays a significant role in the precipitous drop in spore survival due to 0.45 Mrad radiation plus heat above 65-75 0 C for B.cereus and 75-95 0 C for B.stearothermophilus. The effect of the spores radiation resistance was not distinct except in the frozen state and at the saturation plateau of thermorestoration where the radiation resistant B.cereus showed ca. 1 log cycle higher survival than the radiation sensitive B.stearothermophilus. When spores are chemically converted into their H + and Ca ++ ionic forms, the H + spores are distinctly more responsive than Ca ++ spores to processes of radical annealment responsible for thermorestoration in hydrated spore systems. At temperatures of extensive thermorestoration of water radicals, H + spores showed higher survival than Ca ++ spores. (F.J.)

A next generation sequencing approach with a suitable bioinformatics workflow to study fungal diversity in bioaerosols released from two different types of composting plants.

Science.gov (United States)

Mbareche, Hamza; Veillette, Marc; Bonifait, Laetitia; Dubuis, Marie-Eve; Benard, Yves; Marchand, Geneviève; Bilodeau, Guillaume J; Duchaine, Caroline

2017-12-01

Composting is used all over the world to transform different types of organic matter through the actions of complex microbial communities. Moving and handling composting material may lead to the emission of high concentrations of bioaerosols. High exposure levels are associated with adverse health effects among compost industry workers. Fungal spores are suspected to play a role in many respiratory illnesses. There is a paucity of information related to the detailed fungal diversity in compost as well as in the aerosols emitted through composting activities. The aim of this study was to analyze the fungal diversity of both organic matter and aerosols present in facilities that process domestic compost and facilities that process pig carcasses. This was accomplished using a next generation sequencing approach that targets the ITS1 genomic region. Multivariate analyses revealed differences in the fungal community present in samples coming from compost treating both raw materials. Furthermore, results show that the compost type affects the fungal diversity of aerosols emitted. Although 8 classes were evenly distributed in all samples, Eurotiomycetes were more dominant in carcass compost while Sordariomycetes were dominant in domestic compost. A large diversity profile was observed in bioaerosols from both compost types showing the presence of a number of pathogenic fungi newly identified in bioaerosols emitted from composting plants. Members of the family Herpotrichiellaceae and Gymnoascaceae which have been shown to cause human diseases were detected in compost and air samples. Moreover, some fungi were identified in higher proportion in air compared to compost. This is the first study to identify a high level of fungal diversity in bioaerosols present in composting plants suggesting a potential exposure risk for workers. This study suggests the need for creating guidelines that address human exposure to bioaerosols. The implementation of technical and organizational

Live cell imaging of germination and outgrowth of individual Bacillus subtilis spores; the effect of heat stress quantitatively analyzed with SporeTracker

NARCIS (Netherlands)

Pandey, R.; ter Beek, A.; Vischer, N.O.E.; Smelt, J.P.P.M.; Brul, S.; Manders, E.M.M.

2013-01-01

Spore-forming bacteria are a special problem for the food industry as some of them are able to survive preservation processes. Bacillus spp. spores can remain in a dormant, stress resistant state for a long period of time. Vegetative cells are formed by germination of spores followed by a more

Inhibition of spore germination of Alternaria tenuis by sulfur dioxide

Energy Technology Data Exchange (ETDEWEB)

Couey, H.M.

1962-08-01

As a part of a continuing study of SO/sub 2/ fumigation of table grapes, the effect of SO/sub 2/ on spores of an isolate of A. tenuis Auct. causing decay of table grapes was determined. The amount of SO/sub 2/ required to inhibit completely spore germination depended on availability of moisture and the temperature. At 20/sup 0/C, wet spores required 20-min exposure to 100 ppm SO/sub 2/ to prevent germination, but spores equilibrated at 90% relative humidity (RH) required 10-min exposure to 1000 ppm SO/sub 2/. Dry spores at 60% RH were unaffected by a 20-min exposure to 4000 ppm SO/sub 2/. Increasing the temperature in the range 5-20/sup 0/C increased effectiveness of the SO/sub 2/ treatment. A comparison of Alternaria with Botrytis cinerea Fr. (studied earlier) showed that wet spores of these organisms were about equally sensitive to SO/sub 2/, but that dry Alternaria spores were more resistant to SO/sub 2/ than dry Botrytis spores under comparable conditions.

Macroalgal spore dysfunction: ocean acidification delays and weakens adhesion.

Science.gov (United States)

Guenther, Rebecca; Miklasz, Kevin; Carrington, Emily; Martone, Patrick T

2018-04-01

Early life stages of marine organisms are predicted to be vulnerable to ocean acidification. For macroalgae, reproduction and population persistence rely on spores to settle, adhere and continue the algal life cycle, yet the effect of ocean acidification on this critical life stage has been largely overlooked. We explicitly tested the biomechanical impact of reduced pH on early spore adhesion. We developed a shear flume to examine the effect of reduced pH on spore attachment time and strength in two intertidal rhodophyte macroalgae, one calcified (Corallina vancouveriensis) and one noncalcified (Polyostea robusta). Reduced pH delayed spore attachment of both species by 40%-52% and weakened attachment strength in C. vancouveriensis, causing spores to dislodge at lower flow-induced shear forces, but had no effect on the attachment strength of P. robusta. Results are consistent with our prediction that reduced pH disrupts proper curing and gel formation of spore adhesives (anionic polysaccharides and glycoproteins) via protonation and cation displacement, although experimental verification is needed. Our results demonstrate that ocean acidification negatively, and differentially, impacts spore adhesion in two macroalgae. If results hold in field conditions, reduced ocean pH has the potential to impact macroalgal communities via spore dysfunction, regardless of the physiological tolerance of mature thalli. © 2017 Phycological Society of America.

Acid and bile tolerance of spore-forming lactic acid bacteria.

Science.gov (United States)

Hyronimus, B; Le Marrec, C; Sassi, A H; Deschamps, A

2000-11-01

Criteria for screening probiotics such as bile tolerance and resistance to acids were studied with 13 spore-forming lactic acid producing bacteria. Different strains of Sporolactobacillus, Bacillus laevolacticus, Bacillus racemilacticus and Bacillus coagulans grown in MRS broth were subjected to low pH conditions (2, 2.5 and 3) and increasing bile concentrations. Among these microorganisms, Bacillus laevolacticus DSM 6475 and all Sporolactobacillus strains tested except Sporolactobacillus racemicus IAM 12395, were resistant to pH 3. Only Bacillus racemilacticus and Bacillus coagulans strains were tolerant to bile concentrations over 0.3% (w/v).

Arbuscular mycorrhizal fungal community composition affected by original elevation rather than translocation along an altitudinal gradient on the Qinghai-Tibet Plateau

Science.gov (United States)

Yang, Wei; Zheng, Yong; Gao, Cheng; Duan, Ji-Chuang; Wang, Shi-Ping; Guo, Liang-Dong

2016-11-01

Elucidating arbuscular mycorrhizal (AM) fungal responses to elevation changes is critical to improve understanding of microbial function in ecosystems under global asymmetrical climate change scenarios. Here we examined AM fungal community in a two-year reciprocal translocation of vegetation-intact soil blocks along an altitudinal gradient (3,200 m to 3,800 m) in an alpine meadow on the Qinghai-Tibet Plateau. AM fungal spore density was significantly higher at lower elevation than at higher elevation regardless of translocation, except that this parameter was significantly increased by upward translocation from original 3,200 m to 3,400 m and 3,600 m. Seventy-three operational taxonomic units (OTUs) of AM fungi were recovered using 454-pyrosequencing of 18S rDNA sequences at a 97% sequence similarity. Original elevation, downward translocation and upward translocation did not significantly affect AM fungal OTU richness. However, with increasing altitude the OTU richness of Acaulosporaceae and Ambisporaceae increased, but the OTU richness of Gigasporaceae and Glomeraceae decreased generally. The AM fungal community composition was significantly structured by original elevation but not by downward translocation and upward translocation. Our findings highlight that compared with the short-term reciprocal translocation, original elevation is a stronger determinant in shaping AM fungal community in the Qinghai-Tibet alpine meadow.

Back-trajectories show export of airborne fungal spores (Ganoderma sp.) from forests to agricultural and urban areas in England

Science.gov (United States)

Sadyś, M.; Skjøth, C. A.; Kennedy, R.

2014-02-01

We propose here the hypothesis that all of United Kingdom (UK) is likely to be affected by Ganoderma sp. spores, an important plant pathogen. We suggest that the main sources of this pathogen, which acts as a bioaerosol, are the widely scattered woodlands in the country, although remote sources must not be neglected. The hypothesis is based on related studies on bioaerosols and supported by new observations from a non-forest site and model calculations to support our hypothesis.

Elastic and inelastic light scattering from single bacterial spores in an optical trap allows the monitoring of spore germination dynamics

OpenAIRE

Peng, Lixin; Chen, De; Setlow, Peter; Li, Yong-qing

2009-01-01

Raman scattering spectroscopy and elastic light scattering intensity (ESLI) were used to simultaneously measure levels of Ca-dipicolinic acid (CaDPA) and changes in spore morphology and refractive index during germination of individual B. subtilis spores with and without the two redundant enzymes (CLEs), CwlJ and SleB, that degrade spores’ peptidoglycan cortex. Conclusions from these measurements include: 1) CaDPA release from individual wild-type germinating spores was biphasic; in a first h...

Influence of Heat Shock Temperatures and Fast Freezing on Viability of Probiotic Sporeformers and the Issue of Spore Plate Count Versus True Numbers

Directory of Open Access Journals (Sweden)

Mojtaba Jafari

2016-02-01

Full Text Available Background and Objectives: The purpose of the present study was to investigate effects of various heat shock conditions and fast freezing and subsequent thawing on the viability and recovery of Bacillus coagulans and Bacillus subtilis as probiotic sporeformers, and also to compare spore plate and microscopic counts. Materials and Methods: After preparing the final suspensions of B. coagulans and Bacillus subtilis subsp. Natto spores, they were spread-plated before and after fast freezing treatment (-70°C for about 1 min. Heat shock treatments of the spores were carried out at 68oC for 15, 20, and 30 min as well as at 80oC for 10 and 15 min. Concentrations of the examined probiotic sporeformers were determined simultaneously by plate enumerations and microscopically determined counts. Student’s t-test and one-way analysis of variance (ANOVA of SPSS were used for statistical analysis of the data. Analysis of DoE results was carried out using Minitab. Results: The results presented here show that the highest recovery rates for B. coagulans (14.75 log CFU/mL and B. subtilis spores (14.80 log CFU/mL were under a heat shock condition of 68°C for 20 min in nutrient agar (p<0.05. In addition, the survival rates of B. coagulans and B. subtilis spores under the fast freezing and subsequent thawing condition were about 90% and 88%, respectively. Plate counts differed significantly from counts determined microscopically, with differences of almost 0.5 and 0.8 log for B. coagulans and B. subtilis spores, respectively (p<0.05. In addition, DoE results of the study revealed that both factors of spore count method and only freezing factor in fast freezing treatment have a significant effect on concentrations of the spores examined (p<0.05. Conclusions: Heat shock conditions, freezing and subsequent thawing circumstances, and plate counts or enumerations determined microscopically have significant influences on the viability of probiotic sporeformers and

Evaluation of fungicidal and fungistatic activity of plant essential oils towards plant pathogenic and saprophytic fungi

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Zia BANIHASHEMI

2011-09-01

Full Text Available   The contact and vapor effects of essential oils from different plants were studied in vitro for fungicidal and fungistatic activity towards different Basidiomycete, Ascomycete, Zygomycete and Oomycete taxa. Of nine essential oils tested, most were fungicidal at very low concentrations to most of the fungi. Hyphae were more sensitive than spores to the formulations. The essential oils citral, β-citronellol, geraniol and oil of lavender, at 1 μL mL-1 medium or 12 μL L-1 of air, inhibited growth and germination in the fungal species examined. Different species of fungal genera reacted differently to the formulations. Some of the formulations were fungistatic to spore germination.

A study of Ganoderma lucidum spores by FTIR microspectroscopy

Science.gov (United States)

Wang, Xin; Chen, Xianliang; Qi, Zeming; Liu, Xingcun; Li, Weizu; Wang, Shengyi

2012-06-01

In order to obtain unique information of Ganoderma lucidum spores, FTIR microspectroscopy was used to study G. lucidum spores from Anhui Province (A), Liaoning Province (B) and Shangdong Province (C) of China. IR micro-spectra were acquired with high-resolution and well-reproducibility. The IR spectra of G. lucidum spores from different areas were similar and mainly made up of the absorption bands of polysaccharide, sterols, proteins, fatty acids, etc. The results of curve fitting indicated the protein secondary structures were dissimilar among the above G. lucidum spores. To identify G. lucidum spores from different areas, the H1078/H1640 value might be a potentially useful factor, furthermore FTIR microspectroscopy could realize this identification efficiently with the help of hierarchical cluster analysis. The result indicates FTIR microspectroscopy is an efficient tool for identification of G. lucidum spores from different areas. The result also suggests FTIR microspectroscopy is a potentially useful tool for the study of TCM.

Validated modified Lycopodium spore method development for ...

African Journals Online (AJOL)

Validated modified lycopodium spore method has been developed for simple and rapid quantification of herbal powdered drugs. Lycopodium spore method was performed on ingredients of Shatavaryadi churna, an ayurvedic formulation used as immunomodulator, galactagogue, aphrodisiac and rejuvenator. Estimation of ...

Fate of ingested Clostridium difficile spores in mice.

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Amber Howerton

Full Text Available Clostridium difficile infection (CDI is a leading cause of antibiotic-associated diarrhea, a major nosocomial complication. The infective form of C. difficile is the spore, a dormant and resistant structure that forms under stress. Although spore germination is the first committed step in CDI onset, the temporal and spatial distribution of ingested C. difficile spores is not clearly understood. We recently reported that CamSA, a synthetic bile salt analog, inhibits C. difficile spore germination in vitro and in vivo. In this study, we took advantage of the anti-germination activity of bile salts to determine the fate of ingested C. difficile spores. We tested four different bile salts for efficacy in preventing CDI. Since CamSA was the only anti-germinant tested able to prevent signs of CDI, we characterized CamSa's in vitro stability, distribution, and cytotoxicity. We report that CamSA is stable to simulated gastrointestinal (GI environments, but will be degraded by members of the natural microbiota found in a healthy gut. Our data suggest that CamSA will not be systemically available, but instead will be localized to the GI tract. Since in vitro pharmacological parameters were acceptable, CamSA was used to probe the mouse model of CDI. By varying the timing of CamSA dosage, we estimated that C. difficile spores germinated and established infection less than 10 hours after ingestion. We also showed that ingested C. difficile spores rapidly transited through the GI tract and accumulated in the colon and cecum of CamSA-treated mice. From there, C. difficile spores were slowly shed over a 96-hour period. To our knowledge, this is the first report of using molecular probes to obtain disease progression information for C. difficile infection.

Sterilization Resistance of Bacterial Spores Explained with Water Chemistry.

Science.gov (United States)

Friedline, Anthony W; Zachariah, Malcolm M; Middaugh, Amy N; Garimella, Ravindranath; Vaishampayan, Parag A; Rice, Charles V

2015-11-05

Bacterial spores can survive for long periods without nutrients and in harsh environmental conditions. This survival is influenced by the structure of the spore, the presence of protective compounds, and water retention. These compounds, and the physical state of water in particular, allow some species of bacterial spores to survive sterilization schemes with hydrogen peroxide and UV light. The chemical nature of the spore core and its water has been a subject of some contention and the chemical environment of the water impacts resistance paradigms. Either the spore has a glassy core, where water is immobilized along with other core components, or the core is gel-like with mobile water diffusion. These properties affect the movement of peroxide and radical species, and hence resistance. Deuterium solid-state NMR experiments are useful for examining the nature of the water inside the spore. Previous work in our lab with spores of Bacillus subtilis indicate that, for spores, the core water is in a more immobilized state than expected for the gel-like core theory, suggesting a glassy core environment. Here, we report deuterium solid-state NMR observations of the water within UV- and peroxide-resistant spores from Bacillus pumilus SAFR-032. Variable-temperature NMR experiments indicate no change in the line shape after heating to 50 °C, but an overall decrease in signal after heating to 100 °C. These results show glass-like core dynamics within B. pumilus SAFR-032 that may be the potential source of its known UV-resistance properties. The observed NMR traits can be attributed to the presence of an exosporium containing additional labile deuterons that can aid in the deactivation of sterilizing agents.

Indole and 3-indolylacetonitrile inhibit spore maturation in Paenibacillus alvei

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Cho Moo

2011-05-01

Full Text Available Abstract Background Bacteria use diverse signaling molecules to ensure the survival of the species in environmental niches. A variety of both Gram-positive and Gram-negative bacteria produce large quantities of indole that functions as an intercellular signal controlling diverse aspects of bacterial physiology. Results In this study, we sought a novel role of indole in a Gram-positive bacteria Paenibacillus alvei that can produce extracellular indole at a concentration of up to 300 μM in the stationary phase in Luria-Bertani medium. Unlike previous studies, our data show that the production of indole in P. alvei is strictly controlled by catabolite repression since the addition of glucose and glycerol completely turns off the indole production. The addition of exogenous indole markedly inhibits the heat resistance of P. alvei without affecting cell growth. Observation of cell morphology with electron microscopy shows that indole inhibits the development of spore coats and cortex in P. alvei. As a result of the immature spore formation of P. alvei, indole also decreases P. alvei survival when exposed to antibiotics, low pH, and ethanol. Additionally, indole derivatives also influence the heat resistance; for example, a plant auxin, 3-indolylacetonitrile dramatically (2900-fold decreased the heat resistance of P. alvei, while another auxin 3-indoleacetic acid had a less significant influence on the heat resistance of P. alvei. Conclusions Together, our results demonstrate that indole and plant auxin 3-indolylacetonitrile inhibit spore maturation of P. alvei and that 3-indolylacetonitrile presents an opportunity for the control of heat and antimicrobial resistant spores of Gram-positive bacteria.

ANTI-FUNGAL ACTIVITIES OF m-IODOBENZOIC ACID AND SOME ...

African Journals Online (AJOL)

The anti-fungal activities of alkali and alkaline earth metal iodobenzoates were studied. Calcium iodobenzoate exhibited the highest anti-fungal activities of 74.60% inhibition for 15 ppm while sodium iodobenzoate exhibited the least inhibition of 61.64%. An optimum concentration of all the metal complexes for inhibition ...

Small acid soluble proteins for rapid spore identification.

Energy Technology Data Exchange (ETDEWEB)

Branda, Steven S.; Lane, Todd W.; VanderNoot, Victoria A.; Jokerst, Amanda S.

2006-12-01

This one year LDRD addressed the problem of rapid characterization of bacterial spores such as those from the genus Bacillus, the group that contains pathogenic spores such as B. anthracis. In this effort we addressed the feasibility of using a proteomics based approach to spore characterization using a subset of conserved spore proteins known as the small acid soluble proteins or SASPs. We proposed developing techniques that built on our previous expertise in microseparations to rapidly characterize or identify spores. An alternative SASP extraction method was developed that was amenable to both the subsequent fluorescent labeling required for laser-induced fluorescence detection and the low ionic strength requirements for isoelectric focusing. For the microseparations, both capillary isoelectric focusing and chip gel electrophoresis were employed. A variety of methods were evaluated to improve the molecular weight resolution for the SASPs, which are in a molecular weight range that is not well resolved by the current methods. Isoelectric focusing was optimized and employed to resolve the SASPs using UV absorbance detection. Proteomic signatures of native wild type Bacillus spores and clones genetically engineered to produce altered SASP patterns were assessed by slab gel electrophoresis, capillary isoelectric focusing with absorbance detection as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection.

Effects of post-harvest treatment using chitosan from Mucor circinelloides on fungal pathogenicity and quality of table grapes during storage.

Science.gov (United States)

de Oliveira, Carlos Eduardo Vasconcelos; Magnani, Marciane; de Sales, Camila Veríssimo; Pontes, Alline Lima de Souza; Campos-Takaki, Galba Maria; Stamford, Thayza Christina Montenegro; de Souza, Evandro Leite

2014-12-01

The aim of this study was to extract chitosan (CHI) from Mucor circinelloides UCP 050 grown in a corn steep liquor (CSL)-based medium under optimized conditions and to assess the efficacy of the obtained CHI to inhibit the post-harvest pathogenic fungi Aspergillus niger URM 5162 and Rhizopus stolonifer URM 3482 in laboratory media and as a coating on table grapes (Vitis labrusca L.). The effect of CHI coating on some physical, physicochemical and sensory characteristics of the fruits during storage was assessed. The greatest amount of CHI was extracted from M. circinelloides UCP 050 grown in medium containing 7 g of CSL per 100 mL at pH 5.5 with rotation at 180 rpm. CHI from M. circinelloides UCP 050 caused morphological changes in the spores of the fungal strains tested and inhibited mycelial growth and spore germination. CHI coating delayed the growth of the assayed fungal strains in artificially infected grapes, as well as autochthonous mycoflora during storage. CHI coating preserved the quality of grapes during storage, as measured by their physical, physicochemical and sensory attributes. These results demonstrate that edible coatings derived from M. circinelloides CHI could be a useful alternative for controlling pathogenic fungi and maintaining the post-harvest quality of table grapes. Copyright © 2014 Elsevier Ltd. All rights reserved.

Carvacrol suppresses high pressure high temperature inactivation of Bacillus cereus spores.

Science.gov (United States)

Luu-Thi, Hue; Corthouts, Jorinde; Passaris, Ioannis; Grauwet, Tara; Aertsen, Abram; Hendrickx, Marc; Michiels, Chris W

2015-03-16

The inactivation of bacterial spores generally proceeds faster and at lower temperatures when heat treatments are conducted under high pressure, and high pressure high temperature (HPHT) processing is, therefore, receiving an increased interest from food processors. However, the mechanisms of spore inactivation by HPHT treatment are poorly understood, particularly at moderately elevated temperature. In the current work, we studied inactivation of the spores of Bacillus cereus F4430/73 by HPHT treatment for 5 min at 600MPa in the temperature range of 50-100°C, using temperature increments of 5°C. Additionally, we investigated the effect of the natural antimicrobial carvacrol on spore germination and inactivation under these conditions. Spore inactivation by HPHT was less than about 1 log unit at 50 to 70°C, but gradually increased at higher temperatures up to about 5 log units at 100°C. DPA release and loss of spore refractility in the spore population were higher at moderate (≤65°C) than at high (≥70°C) treatment temperatures, and we propose that moderate conditions induced the normal physiological pathway of spore germination resulting in fully hydrated spores, while at higher temperatures this pathway was suppressed and replaced by another mechanism of pressure-induced dipicolinic acid (DPA) release that results only in partial spore rehydration, probably because spore cortex hydrolysis is inhibited. Carvacrol strongly suppressed DPA release and spore rehydration during HPHT treatment at ≤65°C and also partly inhibited DPA release at ≥65°C. Concomitantly, HPHT spore inactivation was reduced by carvacrol at 65-90°C but unaffected at 95-100°C. Copyright © 2014 Elsevier B.V. All rights reserved.

Dothistroma septosporum: spore production and weather conditions

Energy Technology Data Exchange (ETDEWEB)

Dvorak, M.; Drapela, K.; Kankovsky, L.

2012-11-01

Dartmouth's septosporum, the causal agent of Dothistroma needle blight is a widespread fungus which infects more than 80 species of coniferous trees through the entire world. Spreading of the infection is strongly affected by climatic factors of each locality where it is recorded. We attempt to describe the concrete limiting climatic factors necessary for the releasing of conidia of D. septosporum and to find out the timing of its spore production within the year. For this purpose we used an automatic volumetric spore trap and an automatic meteorological station. We found that a minimum daily average temperature of 10 degree centigrade was necessary for any spore production, as well as a long period of high air humidity. The values obtained in the present study were a little bit higher than those previously published, which may arise questions about a possible changing trend of the behaviour in the development of the Dothistroma needle blight causal agent. We used autoregressive integrated moving average (ARIMA) models to predict the spore counts on the base of previous values of spore counts and dew point. For a locality from Hackerovka, the best ARIMA model was 1,0,0; and for a locality from Lanzhot, the best was 3,1,0. (Author) 19 refs.

The combined effect of high pressure and nisin or lysozyme on the inactivation of Alicyclobacillus acidoterrestris spores in apple juice

Science.gov (United States)

Sokołowska, B.; Skąpska, S.; Fonberg-Broczek, M.; Niezgoda, J.; Chotkiewicz, M.; Dekowska, A.; Rzoska, S.

2012-03-01

Alicyclobacillus acidoterrestris, a thermoacidophilic and spore-forming bacterium is one of the important target micro-organisms in the quality control of acidic canned foods. High pressure pasteurization (HPP) at 50°C was used for the inactivation of A. acidoterrestris spores in apple juice. Pressure applied both in a continuous and oscillatory mode gave the best results when 200 MPa was used. Increasing the pressure to 500 MPa, as well as lowering its value to 100 MPa, had an adverse effect on the effectiveness of the process. The best results were achieved with the use of a combined treatment, involving oscillatory pressurization at 200 MPa, followed by holding the sample for 60 min at atmospheric pressure and subsequent pressurization at 500 MPa, resulting in a reduction in the spore count of 6.15 log. Nisin significantly enhanced the effect of HPP at 300 MPa. Using pressure of 200 MPa for 45 min with a nisin concentration of 250 IU/mL enabled total spore inactivation (over 6 log). No significant effect of lysozyme at a concentration of 0.05 and 0.1 mg/L at 300 MPa was observed.

Bacteriocins: Novel Solutions to Age Old Spore-Related Problems?

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Kevin eEgan

2016-04-01

Full Text Available Bacteriocins are ribosomally synthesized antimicrobial peptides produced by bacteria, which have the ability to kill or inhibit other bacteria. Many bacteriocins are produced by food grade lactic acid bacteria (LAB. Indeed, the prototypic bacteriocin, nisin, is produced by Lactococcus lactis, and is licensed in over 50 countries. With consumers becoming more concerned about the levels of chemical preservatives present in food, bacteriocins offer an alternative, more natural, approach, while ensuring both food safety and product shelf life. Bacteriocins also show additive/synergistic effects when used in combination with other treatments, such as heating, high pressure, organic compounds, and as part of food packaging. These features are particularly attractive from the perspective of controlling sporeforming bacteria. Bacterial spores are common contaminants of food products, and their outgrowth may cause food spoilage or food-borne illness. They are of particular concern to the food industry due to their thermal and chemical resistance in their dormant state. However, when spores germinate they lose the majority of their resistance traits, making them susceptible to a variety of food processing treatments. Bacteriocins represent one potential treatment as they may inhibit spores in the post-germination/outgrowth phase of the spore cycle. Spore eradication and control in food is critical, as they are able to spoil and in certain cases compromise the safety of food by producing dangerous toxins. Thus, understanding the mechanisms by which bacteriocins exert their sporostatic/sporicidal activity against bacterial spores will ultimately facilitate their optimal use in food. This review will focus on the use of bacteriocins alone, or in combination with other innovative processing methods to control spores in food, the current knowledge and gaps therein with regard to bacteriocin-spore interactions and discuss future research approaches to enable

Spore formation and toxin production in Clostridium difficile biofilms.

Science.gov (United States)

Semenyuk, Ekaterina G; Laning, Michelle L; Foley, Jennifer; Johnston, Pehga F; Knight, Katherine L; Gerding, Dale N; Driks, Adam

2014-01-01

The ability to grow as a biofilm can facilitate survival of bacteria in the environment and promote infection. To better characterize biofilm formation in the pathogen Clostridium difficile, we established a colony biofilm culture method for this organism on a polycarbonate filter, and analyzed the matrix and the cells in biofilms from a variety of clinical isolates over several days of biofilm culture. We found that biofilms readily formed in all strains analyzed, and that spores were abundant within about 6 days. We also found that extracellular DNA (eDNA), polysaccharide and protein was readily detected in the matrix of all strains, including the major toxins A and/or B, in toxigenic strains. All the strains we analyzed formed spores. Apart from strains 630 and VPI10463, which sporulated in the biofilm at relatively low frequencies, the frequencies of biofilm sporulation varied between 46 and 65%, suggesting that variations in sporulation levels among strains is unlikely to be a major factor in variation in the severity of disease. Spores in biofilms also had reduced germination efficiency compared to spores obtained by a conventional sporulation protocol. Transmission electron microscopy revealed that in 3 day-old biofilms, the outermost structure of the spore is a lightly staining coat. However, after 6 days, material that resembles cell debris in the matrix surrounds the spore, and darkly staining granules are closely associated with the spores surface. In 14 day-old biofilms, relatively few spores are surrounded by the apparent cell debris, and the surface-associated granules are present at higher density at the coat surface. Finally, we showed that biofilm cells possess 100-fold greater resistance to the antibiotic metronidazole then do cells cultured in liquid media. Taken together, our data suggest that C. difficile cells and spores in biofilms have specialized properties that may facilitate infection.

Spore formation and toxin production in Clostridium difficile biofilms.

Directory of Open Access Journals (Sweden)

Ekaterina G Semenyuk

Full Text Available The ability to grow as a biofilm can facilitate survival of bacteria in the environment and promote infection. To better characterize biofilm formation in the pathogen Clostridium difficile, we established a colony biofilm culture method for this organism on a polycarbonate filter, and analyzed the matrix and the cells in biofilms from a variety of clinical isolates over several days of biofilm culture. We found that biofilms readily formed in all strains analyzed, and that spores were abundant within about 6 days. We also found that extracellular DNA (eDNA, polysaccharide and protein was readily detected in the matrix of all strains, including the major toxins A and/or B, in toxigenic strains. All the strains we analyzed formed spores. Apart from strains 630 and VPI10463, which sporulated in the biofilm at relatively low frequencies, the frequencies of biofilm sporulation varied between 46 and 65%, suggesting that variations in sporulation levels among strains is unlikely to be a major factor in variation in the severity of disease. Spores in biofilms also had reduced germination efficiency compared to spores obtained by a conventional sporulation protocol. Transmission electron microscopy revealed that in 3 day-old biofilms, the outermost structure of the spore is a lightly staining coat. However, after 6 days, material that resembles cell debris in the matrix surrounds the spore, and darkly staining granules are closely associated with the spores surface. In 14 day-old biofilms, relatively few spores are surrounded by the apparent cell debris, and the surface-associated granules are present at higher density at the coat surface. Finally, we showed that biofilm cells possess 100-fold greater resistance to the antibiotic metronidazole then do cells cultured in liquid media. Taken together, our data suggest that C. difficile cells and spores in biofilms have specialized properties that may facilitate infection.

Bacteriocins: Novel Solutions to Age Old Spore-Related Problems?

Science.gov (United States)

Egan, Kevin; Field, Des; Rea, Mary C; Ross, R Paul; Hill, Colin; Cotter, Paul D

2016-01-01

Bacteriocins are ribosomally synthesized antimicrobial peptides produced by bacteria, which have the ability to kill or inhibit other bacteria. Many bacteriocins are produced by food grade lactic acid bacteria (LAB). Indeed, the prototypic bacteriocin, nisin, is produced by Lactococcus lactis, and is licensed in over 50 countries. With consumers becoming more concerned about the levels of chemical preservatives present in food, bacteriocins offer an alternative, more natural approach, while ensuring both food safety and product shelf life. Bacteriocins also show additive/synergistic effects when used in combination with other treatments, such as heating, high pressure, organic compounds, and as part of food packaging. These features are particularly attractive from the perspective of controlling sporeforming bacteria. Bacterial spores are common contaminants of food products, and their outgrowth may cause food spoilage or food-borne illness. They are of particular concern to the food industry due to their thermal and chemical resistance in their dormant state. However, when spores germinate they lose the majority of their resistance traits, making them susceptible to a variety of food processing treatments. Bacteriocins represent one potential treatment as they may inhibit spores in the post-germination/outgrowth phase of the spore cycle. Spore eradication and control in food is critical, as they are able to spoil and in certain cases compromise the safety of food by producing dangerous toxins. Thus, understanding the mechanisms by which bacteriocins exert their sporostatic/sporicidal activity against bacterial spores will ultimately facilitate their optimal use in food. This review will focus on the use of bacteriocins alone, or in combination with other innovative processing methods to control spores in food, the current knowledge and gaps therein with regard to bacteriocin-spore interactions and discuss future research approaches to enable spores to be more

Detection of spore coat protein of Bacillus subtilis by immunological method

International Nuclear Information System (INIS)

Uchida, Aritsune; Kadota, Hajime

1976-01-01

The spore coat protein of Bacillus subtilis was separated, and the qualitative assay for the spore coat protein was made by use of the immunological technique. The immunological method was found to be useful for judging the maturation of spore coat in the course of sporulation. The spore coat protein antigen appeared at t 2 stage of sporulation. The addition of rifampicin at the earlier stages of sporulation inhibited the increase in content of the spore coat antigen. (auth.)

Effects of High Pressure on Bacillus licheniformis Spore Germination and Inactivation.

Science.gov (United States)

Borch-Pedersen, Kristina; Mellegård, Hilde; Reineke, Kai; Boysen, Preben; Sevenich, Robert; Lindbäck, Toril; Aspholm, Marina

2017-07-15

Bacillus and Clostridium species form spores, which pose a challenge to the food industry due to their ubiquitous nature and extreme resistance. Pressurization at 300 MPa likely triggers germination by opening dipicolinic acid (DPA) channels present in the inner membrane of the spores. In this work, we expose spores of Bacillus licheniformis , a species associated with food spoilage and occasionally with food poisoning, to high pressure (HP) for holding times of up to 2 h. By using mutant spores lacking one or several GRs, we dissect the roles of the GerA, Ynd, and GerK GRs in moderately HP (mHP; 150 MPa)-induced spore germination. We show that Ynd alone is sufficient for efficient mHP-induced spore germination. GerK also triggers germination with mHP, although at a reduced germination rate compared to that of Ynd. GerA stimulates mHP-induced germination but only in the presence of either the intact GerK or Ynd GR. These results suggests that the effectiveness of the individual GRs in mHP-induced germination differs from their effectiveness in nutrient-induced germination, where GerA plays an essential role. In contrast to Bacillus subtilis spores, treatment with very HP (vHP) of 550 MPa at 37°C did not promote effective germination of B. licheniformis spores. However, treatment with vHP in combination with elevated temperatures (60°C) gave a synergistic effect on spore germination and inactivation. Together, these results provide novel insights into how HP affects B. licheniformis spore germination and inactivation and the role of individual GRs in this process. IMPORTANCE Bacterial spores are inherently resistant to food-processing regimes, such as high-temperature short-time pasteurization, and may therefore compromise food durability and safety. The induction of spore germination facilitates subsequent inactivation by gentler processing conditions that maintain the sensory and nutritional qualities of the food. High-pressure (HP) processing is a nonthermal

The interactive effect of phosphorus and nitrogen on "in vitro" spore germination of Glomus etunicatum Becker & Gerdemann, root growth and mycorrhizal colonization

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Bressan Wellington

2001-01-01

Full Text Available The effects of P and N amendment and its interactions on spore germination, root growth and colonized root length by Glomus etunicatum Becker & Gerdemann (INVAM S329 was studied "in vitro" in RiT - DNA transformed roots of Anthylis vulneraria sub sp. Sampaiana (Kidney vetch. Three N media concentrations (5, 10 and 50 mg/l at P constant level (2 mg/l and three P media concentrations (2, 10 and 20 mg/l at N constant level (5 mg/l were utilized as a treatment. Bécard & Fortin medium was used as a basal medium for root growth and colonized root length, and water/agar (0.8% media was the control for spore germination. Spore germination of G. etunicatum at low P level was reduced by N addition in relation to the control media, and at low N level addition of P stimulated spore germination. Total root length was stimulated by N addtion at low P level, but no significant difference (p£0.05 was observed between 10 and 50 mg/l of N. P addition at low N level media also stimulated total root growth, and a significant difference (p£0.05 was observed among P concentrations. Colonized root length by G. etunicatum increased significantly (p£0.05 with P additions at low N levels. Under low P level no significant differences was found between 10 and 50 mg/l of N. These results demonstrate that the interaction between P and N affect differently spore germination, root growth and colonized root lenght.

Ultra high pressure homogenization (UHPH inactivation of Bacillus amyloliquefaciens spores in phosphate buffered saline (PBS and milk

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Peng eDong

2015-07-01

Full Text Available Ultra high pressure homogenization (UHPH opens up new areas for dynamic high pressure assisted thermal sterilization of liquids. Bacillus amyloliquefaciens spores are resistant to high isostatic pressure and temperature and were suggested as potential surrogate for high pressure thermal sterilization validation. B. amyloliquefaciens spores suspended in PBS buffer (0.01 M, pH 7.0, low fat milk (1.5%, pH 6.7 and whole milk (3.5%, pH 6.7 at initial concentration of ~106 CFU/mL were subjected to UHPH treatments at 200, 300 and 350 MPa with an inlet temperature at ~80 °C. Thermal inactivation kinetics of B. amyloliquefaciens spores in PBS and milk were assessed with thin wall glass capillaries and modeled using mechanistic linear first order and Weibull models. The residence time during UHPH treatments was estimated to determine the contribution of temperature to spore inactivation by UHPH. No sublethal injury was detected after UHPH treatments using sodium chloride as selective component in the nutrient agar medium. The inactivation profiles of spores in PBS buffer and milk were compared and fat provided no clear protective effect for spores against treatments. Treatment at 200 MPa with valve temperatures lower than 125 °C caused no reduction of spores. A reduction of 3.5 log10 CFU/mL of B. amyloliquefaciens spores was achieved by treatment at 350 MPa with a valve temperature higher than 150 °C. The modeled thermal inactivation and observed inactivation during UHPH treatments suggest that temperature could be the main lethal effect driving inactivation.

Rapid change of AM fungal community in a rain-fed wheat field with short-term plastic film mulching practice.

Science.gov (United States)

Liu, Yongjun; Mao, Lin; He, Xinhua; Cheng, Gang; Ma, Xiaojun; An, Lizhe; Feng, Huyuan

2012-01-01

Plastic film mulching (PFM) is a widely used agricultural practice in the temperate semi-arid Loess Plateau of China. However, how beneficial soil microbes, arbuscular mycorrhizal (AM) fungi in particular, respond to the PFM practice is not known. Here, a field experiment was performed to study the effects of a 3-month short-term PFM practice on AM fungi in plots planted with spring wheat (Triticum aestivum L. cv. Dingxi-2) in the Loess Plateau. AM colonization, spore density, wheat spike weight, and grain phosphorus (P) content were significantly increased in the PFM treatments, and these changes were mainly attributable to changes in soil properties such as available P and soil moisture. Alkaline phosphatase activity was significantly higher in PFM soils, but levels of AM fungal-related glomalin were similar between treatments. A total of nine AM fungal phylotypes were detected in root samples based on AM fungal SSU rDNA analyses, with six and five phylotypes in PFM and no-PFM plots, respectively. Although AM fungal phylotype richness was not statistically different between treatments, the community compositions were different, with four and three specific phylotypes in the PFM and no-PFM plots, respectively. A significant and rapid change in AM fungal, wheat, and soil variables following PFM suggested that the functioning of the AM symbiosis had been changed in the wheat field under PFM. Future studies are needed to investigate whether PFM applied over a longer term has a similar effect on the AM fungal community and their functioning in an agricultural ecosystem.

Coupled Metagenomic and Chemical Analyses of Degrading Fungal Necromass and Implications for Fungal contributions to Stable Soil Organic Carbon

Science.gov (United States)

Egerton-Warburton, L. M.; Schreiner, K. M.; Morgan, B. S. T.; Schultz, J.; Blair, N. E.

2016-12-01

Fungi comprise a significant portion of total soil biomass, the turnover of which must represent a dominant flux within the soil carbon cycle. Fungal organic carbon (OC) can turn over on time scales of days to months, but this process is poorly understood. Here, we examined temporal changes in the chemical and microbial community composition of fungal necromass during a 2-month decomposition experiment in which Fusarium avenaceum (a common saprophyte) was exposed to a natural soil microbial community. Over the course of the experiment, residual fungal necromass was harvested and analyzed using FTIR and thermochemolysis-GCMS to examine chemical changes in the tissue. In addition, genomic DNA was extracted from tissues, amplified with barcoded ITS primers, and sequenced using the high-throughput Illumina platform to examine changes in microbial community composition. Up to 80% of the fungal necromass turned over in the first week. This rapid degradation phase corresponded to colonization of the necromass by known chitinolytic soil fungi including Mortierella species. Members of the Zygomycota and Ascomycota were among the dominant fungal groups involved in degradation with very small contributions from Basidiomycota. At the end of the 2-month degradation, only 15% of the original necromass remained. The residual material was rich in amide and C-O moieties which is consistent with previous work predicting that peptidoglycans are the main residual product from microbial tissue degradation. Straight-chain fatty acids exhibited varying degradation profiles, with some fatty acids (e.g. C16, C18:1) degrading more rapidly than bulk tissue while others maintained steady concentrations relative to bulk OC (C18) or increased in concentration throughout the degradation sequence (C24). These results indicate that the turnover of fungal necromass has the potential to rapidly and significantly influence a variety of soil OC properties including C/N ratios, lipid biomarker

Expression and characterization of a novel spore wall protein from ...

African Journals Online (AJOL)

Microsporidia are obligate intracellular, eukaryotic, spore-forming parasites. The environmentally resistant spores, which harbor a rigid cell wall, are critical for their survival outside their host cells and host-to-host transmission. The spore wall comprises two major layers: the exospore and the endospore. In Nosema ...

Survival of Bacillus anthracis spores in fruit juices and wine.

Science.gov (United States)

Leishman, Oriana N; Johnson, Miranda J; Labuza, Theodore P; Diez-Gonzalez, Francisco

2010-09-01

Foods have been identified as a potential target for bioterrorism due to their essential nature and global distribution. Foods produced in bulk have the potential to have large batches of product intentionally contaminated, which could affect hundreds or thousands of individuals. Bacillus anthracis spores are one potential bioterrorism agent that may survive pasteurization and remain viable throughout the shelf life of fruit juices and cause disease if consumed. This project examined B. anthracis spore survival in orange, apple, and grape juices, as well as wine. Samples of beverages were inoculated with spores of two nonpathogenic B. anthracis strains at approximately 10(6) CFU/ml, and the spore count was determined periodically during storage for 30 days at 4°C. After this time, the counts of survival spores never declined more than 1 log CFU/ml in any of the beverage types. These results indicate that spores can survive, with little to no loss in viability, for at least a month in fruit juices and wine.

Meteorological factors, aeroallergens and asthma-related visits in Kuwait : A 12-month retrospective study

International Nuclear Information System (INIS)

Jafar A Qasem

2010-01-01

The increasing prevalence of asthma in many countries has been related to weather factors and aeroallergen concentrations , but this has not been studied in Kuwait. We evaluated the effect of meteorological factors and the occurrence of aerobiologicals on the number of asthma cases in Kuwait.The number of daily asthma visits to the allergy center and emergency department at Al-Sabha Hospital for 1 year were examined on a monthly basis for correlation with major metereological factors (temperature , relative humidity , rain , wind speed and direction). Spore and pollen counts were collected hourly. Results: A total of 4353 patients received asthma treatment during the year. The highest pollen count was in the month of September with a maximum relative humidity of 47% and no precipitation , but with a high mean temperature of 39.7 0 C. . Pollen counts were higher in the late summer (September) and occurred with a high patient visit to the allergy center. Fungal spore counts were significantly higher in early winter (December). The high fungal spore count seemed related to with high relative humidity and high precipitation with a low mean average temperature of 19.7 0 C. The increase number of patients with bronchial asthma visiting an emergency clinic during December was significantly associated with high aerial counts for fungal spores (P<.03) , and the months of September and October were more significant for pollen.This study indicates that meteorological factors , aeroallergen concentrations and asthma related visits are interrelated. The results may prove useful in the generation of hypotheses and development of designs for more comprehensive , individual-based epidemiological studies (Author).

Are bacterial volatile compounds poisonous odors to a fungal pathogen Botrytis cinerea, alarm signals to Arabidopsis seedlings for eliciting induced resistance, or both?

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Choong-Min eRyu

2016-02-01

Full Text Available Biological control (biocontrol agents act on plants via numerous mechanisms, and can be used to protect plants from pathogens. Biocontrol agents can act directly as pathogen antagonists or competitors or indirectly to promote plant induced systemic resistance (ISR. Whether a biocontrol agent acts directly or indirectly depends on the specific strain and the pathosystem type. We reported previously that bacterial volatile organic compounds (VOCs are determinants for eliciting plant ISR. Emerging data suggest that bacterial VOCs also can directly inhibit fungal and plant growth. The aim of the current study was to differentiate direct and indirect mechanisms of bacterial VOC effects against Botrytis cinerea infection of Arabidopsis. Volatile emissions from Bacillus subtilis GB03 successfully protected Arabidopsis seedlings against B. cinerea. First, we investigated the direct effects of bacterial VOCs on symptom development and different phenological stages of B. cinerea including spore germination, mycelial attachment to the leaf surface, mycelial growth, and sporulation in vitro and in planta. Volatile emissions inhibited hyphal growth in a dose-dependent manner in vitro, and interfered with fungal attachment on the hydrophobic leaf surface. Second, the optimized bacterial concentration that did not directly inhibit fungal growth successfully protected Arabidopsis from fungal infection, which indicates that bacterial VOC-elicited plant ISR has a more important role in biocontrol than direct inhibition of fungal growth on Arabidopsis. We performed qRT-PCR to investigate the priming of the defense-related genes PR1, PDF1.2, and ChiB at 0, 12, 24, and 36 hours post-infection and 14 days after the start of plant exposure to bacterial VOCs. The results indicate that bacterial VOCs potentiate expression of PR1 and PDF1.2 but not ChiB, which stimulates SA- and JA-dependent signaling pathways in plant ISR and protects plants against pathogen

Effect of high hydrostatic pressure on mycelial development, spore viability and enzyme activity of Penicillium Roqueforti.

Science.gov (United States)

Martínez-Rodríguez, Yamile; Acosta-Muñiz, Carlos; Olivas, Guadalupe I; Guerrero-Beltrán, José; Rodrigo-Aliaga, Dolores; Mujica-Paz, Hugo; Welti-Chanes, Jorge; Sepulveda, David R

2014-01-03

This study investigated the effect of high hydrostatic pressure treatments on mycelial development, spore viability, and total proteolytic and lipolytic activity of Penicillium roqueforti PV-LYO 10 D. Fungus growing in liquid medium was pressure-treated at 300, 400, and 500 MPa for 10 min at 20°C following seven days of incubation at 25°C and analyzed periodically up to day 9 after treatments to evaluate the effect on fungal growth. Mycelial mass of P. roqueforti was significantly affected at all pressure treatments evaluated, being 15.48%, 22.28%, 30.03%, and 12.53% lower than controls on day 1, 3, 6, and 9 after 300 MPa treatment, respectively. In a similar way, at 400 and 500 MPa, mycelial mass was 31.08% and 60.34% lower than controls one day after treatments and 49.74% and 80.85% lower on day 9, respectively. The viability of P. roqueforti spores decreased by 36.53% at 300 MPa, and complete inactivation took place at ≥400 MPa from an initial count of 7 log cfu/mL. Total proteolytic activity was not significantly affected at 300 MPa but was reduced by 18.22% at 400 MPa and by 43.18% at 500 MPa. Total lipolytic activity also decreased as the intensity of the pressure treatments increased. 21.69%, 39.12%, and 56.26% activity reductions were observed when treatments of 300, 400 and 500 MPa were applied, respectively. The results from this study show that pressure treatments are able to control growth, inactivate spores, and alter enzyme activity of P. roqueforti, which could be of interest in extending the shelf-life of blue-veined cheeses and other food products. © 2013.

NanoSIMS analysis of Bacillus spores for forensics

Energy Technology Data Exchange (ETDEWEB)

Weber, P K; Davisson, M L; Velsko, S P

2010-02-23

The threat associated with the potential use of radiological, nuclear, chemical and biological materials in terrorist acts has resulted in new fields of forensic science requiring the application of state-of-the-science analytical techniques. Since the anthrax letter attacks in the United States in the fall of 2001, there has been increased interest in physical and chemical characterization of bacterial spores. While molecular methods are powerful tools for identifying genetic differences, other methods may be able to differentiate genetically identical samples based on physical and chemical properties, as well as provide complimentary information, such as methods of production and approximate date of production. Microanalysis has the potential to contribute significantly to microbial forensics. Bacillus spores are highly structured, consisting of a core, cortex, coat, and in some species, an exosporium. This structure provides a template for constraining elemental abundance differences at the nanometer scale. The primary controls on the distribution of major elements in spores are likely structural and physiological. For example, P and Ca are known to be abundant in the spore core because that is where P-rich nucleic acids and Cadipicolinic acid are located, respectively. Trace elements are known to bind to the spore coat but the controls on these elements are less well understood. Elemental distributions and abundances may be directly related to spore production, purification and stabilization methodologies, which are of particular interest for forensic investigation. To this end, we are developing a high-resolution secondary ion mass spectrometry method using a Cameca NanoSIMS 50 to study the distribution and abundance of trace elements in bacterial spores. In this presentation we will review and compare methods for preparing and analyzing samples, as well as review results on the distribution and abundance of elements in bacterial spores. We use NanoSIMS to

Fungal treatment for the removal of endocrine disrupting compounds from reverse osmosis concentrate: Identification and monitoring of transformation products of benzotriazoles.

Science.gov (United States)

Llorca, Marta; Badia-Fabregat, Marina; Rodríguez-Mozaz, Sara; Caminal, Glòria; Vicent, Teresa; Barceló, Damià

2017-10-01

The removal of 27 endocrine-disrupting compounds and related compounds (suspect effect) from a reverse osmosis concentrate using an alternative decontamination method based on a fungal treatment involving Trametes versicolor was assessed. In addition to chemical analysis, the toxicity of the treated water during the treatment was monitored using a bioluminescence inhibition test and estrogenic and anti-estrogenic tests. The compounds 1H-benzotriazole (BTZ) and two tolyltriazoles (TTZs), 4-methyl-1H-benzotriazole (4-MBTZ) and 5-methyl-1H-benzotriazole (5-MBTZ), were present in the reverse osmosis concentrate at the highest concentrations (7.4 and 12.8 μg L -1 , respectively) and were partially removed by the fungal treatment under sterile conditions (58% for BTZ and 92% for TTZs) and non-sterile conditions, although to lesser extents (32% for BTZ and 50% for TTZs). Individual biotransformation studies of BTZ and the TTZs by T. versicolor in a synthetic medium and further analysis via on-line turbulent flow chromatography coupled to an HRMS-Orbitrap allowed the tentative identification of the transformation products (TPs). Six TPs were postulated for BTZ, two TPs were postulated for 4-MBTZ, and four TPs were postulated for 5-MBTZ. Most of these TPs are suggested to have been generated by conjugation with some sugars and via the methylation of the triazole group. Only TP 148 A, postulated to be derived from the biotransformation of BTZ, was observed in the effluent of the bioreactor treating the reverse osmosis concentrate. Copyright © 2017 Elsevier Ltd. All rights reserved.

Diverse microbial species survive high ammonia concentrations

Science.gov (United States)

Kelly, Laura C.; Cockell, Charles S.; Summers, Stephen

2012-04-01

Planetary protection regulations are in place to control the contamination of planets and moons with terrestrial micro-organisms in order to avoid jeopardizing future scientific investigations relating to the search for life. One environmental chemical factor of relevance in extraterrestrial environments, specifically in the moons of the outer solar system, is ammonia (NH3). Ammonia is known to be highly toxic to micro-organisms and may disrupt proton motive force, interfere with cellular redox reactions or cause an increase of cell pH. To test the survival potential of terrestrial micro-organisms exposed to such cold, ammonia-rich environments, and to judge whether current planetary protection regulations are sufficient, soil samples were exposed to concentrations of NH3 from 5 to 35% (v/v) at -80°C and room temperature for periods up to 11 months. Following exposure to 35% NH3, diverse spore-forming taxa survived, including representatives of the Firmicutes (Bacillus, Sporosarcina, Viridibacillus, Paenibacillus, Staphylococcus and Brevibacillus) and Actinobacteria (Streptomyces). Non-spore forming organisms also survived, including Proteobacteria (Pseudomonas) and Actinobacteria (Arthrobacter) that are known to have environmentally resistant resting states. Clostridium spp. were isolated from the exposed soil under anaerobic culture. High NH3 was shown to cause a reduction in viability of spores over time, but spore morphology was not visibly altered. In addition to its implications for planetary protection, these data show that a large number of bacteria, potentially including spore-forming pathogens, but also environmentally resistant non-spore-formers, can survive high ammonia concentrations.

Fungal diversity associated with verticillium wilt of cotton

International Nuclear Information System (INIS)

Khaskheli, M.I.; Sun, J.L.; Li, F.

2014-01-01

The association of fungal diversity with Verticillium wilt is rarely known, which is important to know for the control of this detrimental disease. Our study is the preliminary attempt to find the associations of fungal diversity with Verticillium wilt and provides the baseline information for biological control. About 30 different fungi from soil and 23 from cotton plants were isolated and confirmed through molecular characterization. The colony forming unit (CFU)/g dry soil of fungi before and after planting cotton showed significant variation among all the fungi. The overall frequency of all fungi for soil after sowing was significantly higher than before sowing. A. alternata, F. equiseti, F. concentricum, A. flavus, F. proliferatum, and Chaetomium sp. associated with high resistance (Arcot-1) to Verticillium wilt, whereas, V. dahliae, A.niger and Paecilomyces sp., with high susceptible (Arcot-438) germplasm. However, T. basicola, C. ramotenellum and G. intermedia were isolated from both. Soil plating was comparatively easiest than soil dilution method for the determination of frequency percentage, however, later method is useful for the screening of single spore isolation. Most of the antagonistic species were screened from soil; nevertheless, Paecilomyces and Chaetomium spp. were screened from plant and soil. In vitro test of T. longibrachiatum. T. atroviride, Paecilomyces and T. viride showed the strongest efficacy against V. dahliae. These efficient bio-agents can be used as an effective tool for other future studies regarding to Verticillium wilt of cotton. (author)

Discrimination of Spore-Forming Bacilli Using spoIVA

Science.gov (United States)

Venkateswaran, Kasthuri; LaDuc, Myron; Stuecker, Tara

2009-01-01

A method of discriminating between spore-forming and non-spore-forming bacteria is based on a combination of simultaneous sporulation-specific and non-sporulation-specific quantitative polymerase chain reactions (Q-PCRs). The method was invented partly in response to the observation that for the purposes of preventing or reducing biological contamination affecting many human endeavors, ultimately, only the spore-forming portions of bacterial populations are the ones that are problematic (or, at least, more problematic than are the non-spore-forming portions). In some environments, spore-forming bacteria constitute small fractions of the total bacterial populations. The use of sporulation-specific primers in Q-PCR affords the ability to assess the spore-forming fraction of a bacterial population present in an environment of interest. This assessment can provide a more thorough and accurate understanding of the bacterial contamination in the environment, thereby making it possible to focus contamination- testing, contamination-prevention, sterilization, and decontamination resources more economically and efficiently. The method includes the use of sporulation-specific primers in the form of designed, optimized deoxyribonucleic acid (DNA) oligonucleotides specific for the bacterial spoIVA gene (see table). [In "spoIVA," "IV" signifies Roman numeral four and the entire quoted name refers to gene A for the fourth stage of sporulation.] These primers are mixed into a PCR cocktail with a given sample of bacterial cells. A control PCR cocktail into which are mixed universal 16S rRNA primers is also prepared. ["16S rRNA" denotes a ribosomal ribonucleic acid (rRNA) sequence that is common to all organisms.] Following several cycles of heating and cooling according to the PCR protocol to amplify amounts of DNA molecules, the amplification products can be analyzed to determine the types of bacterial cells present within the samples. If the amplification product is strong

Modeling Thermal Inactivation of Bacillus Spores

Science.gov (United States)

2009-03-01

information is preserved and replicated by the Watson - Crick base pairing in which 4-3 complementary bases recognize each other. One incorrect amino acid can...hydrolysis reactions to take place with the spore’s DNA and other proteins. These chemical reactions degrade the DNA and proteins to such an extent that the... DNA cannot be repaired or replicated, thus causing spore death. We further assert that damage to a spore is based on a certain initial DNA information

Hyphal Growth from Spores of the Mycorrhizal Fungus Glomus Caledonius: Effect of Amino Acids

DEFF Research Database (Denmark)

Hepper, C.M.; Jakobsen, Iver

1983-01-01

Hyphal growth from spores of Glomus caledonius (Nicol. and Gerd.) Trappe and Gerdemann was stimulated by cystine, glycine and lysine at optimum concentrations of 4.6, 556 and 825 mg l−1 respectively. When all three amino acids were supplied together in water agar, five times more growth...

Effect of coexisting organic substances on radiation resistance of Bacillus pumilus spores suspended in water

International Nuclear Information System (INIS)

Kigawa, Akiko; Tateishi, Tsuneo; Iso, Katsuaki; Kimura, Toshio; Mamuro, Tetsuo

1987-01-01

D values of B. pumilus spores suspended in water have been shown to increase in the presence of some coexisting organic substances. For elucidation of a mechanism or mechanisms involved in such a phenomenon, D-values of B.p. spores were examined by suspending them in aqueous solutions containing various concentrations of ethanol, glycerin, inulin and PVA. All these substances showed abrupt changes in D value at a narrow concentration range of 1 - 10 weight ppm. Solutions containing these substances at their lower limit concentrations and upper limit were prepared, sealed in incubator bottles leaving no air layer and irradiated at 0.7 Mrad with γ-rays. Winkler's method was used for the determination of oxygen concentrations in these solutions. The initial concentration of dissolved oxygen was 8.2 ppm. After irradiation, 3 - 5 ppm of oxygen remained in those solutions containing the lower limit (1 ppm), whereas only less than 0.5 ppm in those containing the upper limits, 2.5 ppm of ethanol, 5 ppm of PVA and 10 ppm each of glycerin and inulin. Therefore, the observed effect of coexisting organic substances on radiation resistance of B. pumilus can be explained by the so-called ''oxygen effect''. (author)

Efficiency of peracetic acid in inactivating bacteria, viruses, and spores in water determined with ATP bioluminescence, quantitative PCR, and culture-based methods.

Science.gov (United States)

Park, Eunyoung; Lee, Cheonghoon; Bisesi, Michael; Lee, Jiyoung

2014-03-01

The disinfection efficiency of peracetic acid (PAA) was investigated on three microbial types using three different methods (filtration-based ATP (adenosine-triphosphate) bioluminescence, quantitative polymerase chain reaction (qPCR), culture-based method). Fecal indicator bacteria (Enterococcus faecium), virus indicator (male-specific (F(+)) coliphages (coliphages)), and protozoa disinfection surrogate (Bacillus subtilis spores (spores)) were tested. The mode of action for spore disinfection was visualized using scanning electron microscopy. The results indicated that PAA concentrations of 5 ppm (contact time: 5 min), 50 ppm (10 min), and 3,000 ppm (5 min) were needed to achieve 3-log reduction of E. faecium, coliphages, and spores, respectively. Scanning electron microscopy observation showed that PAA targets the external layers of spores. The lower reduction rates of tested microbes measured with qPCR suggest that qPCR may overestimate the surviving microbes. Collectively, PAA showed broad disinfection efficiency (susceptibility: E. faecium > coliphages > spores). For E. faecium and spores, ATP bioluminescence was substantially faster (∼5 min) than culture-based method (>24 h) and qPCR (2-3 h). This study suggests PAA as an effective alternative to inactivate broad types of microbial contaminants in water. Together with the use of rapid detection methods, this approach can be useful for urgent situations when timely response is needed for ensuring water quality.

Availability of websites offering to sell psilocybin spores and psilocybin.

Science.gov (United States)

Lott, Jason P; Marlowe, Douglas B; Forman, Robert F

2009-09-01

This study assesses the availability of websites offering to sell psilocybin spores and psilocybin, a powerful hallucinogen contained in Psilocybe mushrooms. Over a 25-month period beginning in March 2003, eight searches were conducted in Google using the term "psilocybin spores." In each search the first 100 nonsponsored links obtained were scored by two independent raters according to standardized criteria to determine whether they offered to sell psilocybin or psilocybin spores. No attempts were made to procure the products offered for sale in order to ascertain whether the marketed psilocybin was in fact "genuine" or "counterfeit." Of the 800 links examined, 58% led to websites offering to sell psilocybin spores. Additionally, evidence that whole Psilocybe mushrooms are offered for sale online was obtained. Psilocybin and psilocybin spores were found to be widely available for sale over the Internet. Online purchase of psilocybin may facilitate illicit use of this potent psychoactive substance. Additional studies are needed to assess whether websites offering to sell psilocybin and psilocybin spores actually deliver their products as advertised.

Inactivation of Bacillus subtilis spores by high pressure CO2 with high temperature.

Science.gov (United States)

Rao, Lei; Xu, Zhenzhen; Wang, Yongtao; Zhao, Feng; Hu, Xiaosong; Liao, Xiaojun

2015-07-16

The objective of this study was to investigate the inactivation of the Bacillus subtilis spores by high pressure CO2 combined with high temperature (HPCD+HT) and to analyze the clumping effect of the spores on their HPCD+HT resistance. The spores of B. subtilis were subjected to heat at 0.1 MPa and HPCD at 6.5-25 MPa, and 82 °C, 86 °C, and 91 °C for 0-120 min. The spores were effectively inactivated by HPCD+HT, but a protective effect on the spores was also found, which was closely correlated to the pressure, temperature and time. The spores treated by HPCD+HT at 6.5 and 10 MPa exhibited a two-stage inactivation curve of shoulder and log-linear regions whereas the spores at 15-25 MPa exhibited a three-stage inactivation curve of shoulder, log-linear and tailing regions, and these curves were well fitted to the Geeraerd model. Approximately 90% of pyridine-2,6-dicarboxylic acid (DPA) was released after HPCD+HT and the 90% DPA release time depend on the pressure and temperature. Moreover, the spore clumping in suspensions was examined by dynamic light scattering. The particle size of the spore suspensions increased with the increase of pressure, temperature and time, indicating the spore clumping. 0.1% Tween 80 as a surfactant inhibited the spore clumping and increased the inactivation ratio of the spores by HPCD+HT. These results indicated that the spore clumping enhanced the spores' resistance to HPCD+HT and induced a protective effect. Copyright © 2015 Elsevier B.V. All rights reserved.

[Distribution and spatial ordering of biopolymer molecules in resting bacterial spores].

Science.gov (United States)

Duda, V I; Korolev, Iu N; El'-Registan, G I; Duzha, M V; Telegin, N L

1978-01-01

The presence, distribution and spatial arrangement of biopolymers in situ were studied in both a total intact spore and in a certain cellular layer using a spectroscopic technique of attenuated total refraction (ATR-IR) in the IR region. In contrast to vegetative cells, intact spores were characterized by isotropic distribution of protein components. This feature can be regarded as an index of the cryptobiotic state of spores. However, the distribution of protein components among individual layers of a spore was anisotropic. Bonds characterized by amide I and amide II bands were most often ordered in a layer which comprised cellular structures from the exosporium to the inner spore membrane.

Predictive modeling of Bacillus cereus spores in farm tank milk during grazing and housing periods

NARCIS (Netherlands)

Vissers, M.M.M.; Giffel, M.C.T.; Driehuis, F.; Jong, de P.; Lankveld, J.M.G.

2007-01-01

The shelf life of pasteurized dairy products depends partly on the concentration of Bacillus cereus spores in raw milk. Based on a translation of contamination pathways into chains of unit-operations, 2 simulation models were developed to quantitatively identify factors that have the greatest effect

Micromotors to capture and destroy anthrax simulant spores.

Science.gov (United States)

Orozco, Jahir; Pan, Guoqing; Sattayasamitsathit, Sirilak; Galarnyk, Michael; Wang, Joseph

2015-03-07

Towards addressing the need for detecting and eliminating biothreats, we describe a micromotor-based approach for screening, capturing, isolating and destroying anthrax simulant spores in a simple and rapid manner with minimal sample processing. The B. globilli antibody-functionalized micromotors can recognize, capture and transport B. globigii spores in environmental matrices, while showing non-interactions with excess of non-target bacteria. Efficient destruction of the anthrax simulant spores is demonstrated via the micromotor-induced mixing of a mild oxidizing solution. The new micromotor-based approach paves a way to dynamic multifunctional systems that rapidly recognize, isolate, capture and destroy biological threats.

Thermal inactivation kinetics of Bacillus coagulans spores in tomato juice.

Science.gov (United States)

Peng, Jing; Mah, Jae-Hyung; Somavat, Romel; Mohamed, Hussein; Sastry, Sudhir; Tang, Juming

2012-07-01

The thermal characteristics of the spores and vegetative cells of three strains of Bacillus coagulans (ATCC 8038, ATCC 7050, and 185A) in tomato juice were evaluated. B. coagulans ATCC 8038 was chosen as the target microorganism for thermal processing of tomato products due to its spores having the highest thermal resistance among the three strains. The thermal inactivation kinetics of B. coagulans ATCC 8038 spores in tomato juice between 95 and 115°C were determined independently in two different laboratories using two different heating setups. The results obtained from both laboratories were in general agreement, with z-values (z-value is defined as the change in temperature required for a 10-fold reduction of the D-value, which is defined as the time required at a certain temperature for a 1-log reduction of the target microorganisms) of 8.3 and 8.7°C, respectively. The z-value of B. coagulans 185A spores in tomato juice (pH 4.3) was found to be 10.2°C. The influence of environmental factors, including cold storage time, pH, and preconditioning, upon the thermal resistance of these bacterial spores is discussed. The results obtained showed that a storage temperature of 4°C was appropriate for maintaining the viability and thermal resistance of B. coagulans ATCC 8038 spores. Acidifying the pH of tomato juice decreased the thermal resistance of these spores. A 1-h exposure at room temperature was considered optimal for preconditioning B. coagulans ATCC 8038 spores in tomato juice.

Genetic Factors and Host Traits Predict Spore Morphology for a Butterfly Pathogen

Directory of Open Access Journals (Sweden)

Jacobus C. de Roode

2013-08-01

Full Text Available Monarch butterflies (Danaus plexippus throughout the world are commonly infected by the specialist pathogen Ophryocystis elektroscirrha (OE. This protozoan is transmitted when larvae ingest infectious stages (spores scattered onto host plant leaves by infected adults. Parasites replicate internally during larval and pupal stages, and adult monarchs emerge covered with millions of dormant spores on the outsides of their bodies. Across multiple monarch populations, OE varies in prevalence and virulence. Here, we examined geographic and genetic variation in OE spore morphology using clonal parasite lineages derived from each of four host populations (eastern and western North America, South Florida and Hawaii. Spores were harvested from experimentally inoculated, captive-reared adult monarchs. Using light microscopy and digital image analysis, we measured the size, shape and color of 30 replicate spores per host. Analyses examined predictors of spore morphology, including parasite source population and clone, parasite load, and the following host traits: family line, sex, wing area, and wing color (orange and black pigmentation. Results showed significant differences in spore size and shape among parasite clones, suggesting genetic determinants of morphological variation. Spore size also increased with monarch wing size, and monarchs with larger and darker orange wings tended to have darker colored spores, consistent with the idea that parasite development depends on variation in host quality and resources. We found no evidence for effects of source population on variation in spore morphology. Collectively, these results provide support for heritable variation in spore morphology and a role for host traits in affecting parasite development.

Mite fauna and fungal flora in house dust from homes of asthmatic children.

Science.gov (United States)

Ishii, A; Takaoka, M; Ichinoe, M; Kabasawa, Y; Ouchi, T

1979-12-01

Mite fauna and fungal flora in the house dust from homes of asthmatic children with positive and negative skin test to house dust allergen and non-asthmatic controls were examined. There was no conspicuous difference in mite species distribution among the three groups. Pyroglyphid mites dominate the mite fauna in house dust more than half of which being Dermatophagoides: D. pteronyssinus and D. farinae. There was no statistically significant difference in numbers between the two species and either species could dominate depending on the conditions of the individual houses. The average number of acarina in 0.5 g of fine dust did not differ statistically among the three groups; however, mite number per square meter floor differed between patients with positive skin test and negative skin test. The results suggest that house-cleaning might influence the possible sensitization of children. The genetic distribution of mould fungi in house dust was largely similar to that of airborne fungi. The average number of fungal colonies detected in 0.5 g of dust did not differ statistically among the three groups. Wallemia with its minute spores may cause sensitization but has so far been insufficiently investigated.

Presence survival spores of Bacillus thuringiensis varieties in grain warehouse

Directory of Open Access Journals (Sweden)

Sánchez-Yáñez Juan Manuel

2016-08-01

Full Text Available Genus Bacillus thuringiensis (Bt synthesized spores and crystals toxic to pest-insects in agriculture. Bt is comospolitan then possible to isolate some subspecies or varieties from warehouse. The aims of study were: i to isolate Bt varieties from grain at werehouse ii to evaluate Bt toxicity on Spodoptera frugiperda and Shit-ophilus zeamaisese iii to analyze Bt spores persistence in Zea mays grains at werehouse compared to same Bt on grains exposed to sun radiation. Results showed that at werehouse were recovered more than one variety of Bt spores. According to each isolate Bt1 o Bt2 were toxic to S. frugiperda or S. zeamaisese. One those Bt belong to var morrisoni. At werehouse these spores on Z. mays grains surviving more time, while the same spores exposed to boicide sun radiation they died.

Combination treatment of clostridium perfringens spores to freezing and/or gamma irradiation

International Nuclear Information System (INIS)

El-Fouly, M.Z.; El-Zawahry, Y.A.; Aziz, N.H.

1985-01-01

Freezing process alone caused relatively low decrease in viable count of suspended spores in minced meat while it decreased the spore numbers suspended in saline solution by more than one log cycle especially in case of the Egyptian strain. An abrupt decrease in viable counts of clostridium spores was observed by application dose of 1KGY either before or after freezing followed by gradual decrease of viable counts up to 15 KGY. The synergestic effect of combined treatment was clearly obvious for spores suspended in minced meat, which usually contains protective agents which increase the resistance of microorganisms against the separate treatment of radiation of freezing especially with spores of NCTC 8798 strain. Freezing the saline suspending medium before or after irradiation after the sensitivity of clostridium spores by only small extent and gave negative synergestic effect in some treatment. The percentages of injured spores due to the combined treatment were ranged between 15-100% of the viable counts. The percentage of injured spores tended to increase as the radiation dose levels increased

Testing the sensitivity of Nested PCR method to detect Aspergillus fumigates in experimentally infected Sputum samples

International Nuclear Information System (INIS)

Ramadan, A.; Soukkaria, S.

2013-01-01

Fungal infections caused by Aspergillus species generally are occupying a second place among invasive fungal infections in the world, especially A. fumigatus, which is considered the main cause of invasive Aspergillosis (IA). Although IA rarely infects immunocompetent individuals, however, it can lead to death in immunocompromised patients. Therefore, it is necessary to diagnose the infection early in order to treat the disease efficiently. However, the conventional diagnostic tools, currently used to detect infections, has low sensitivity and reliability. Polymerase chain reaction (PCR) technology distribution as a molecular and high sensitive technology has allowed us to make comparative study between sensitivity of traditional currently used diagnostic method and Nested-PCR, the result of the study of sputum samples that experimentally infected with different concentrations of A.fumigatus spores ramping from 10 to10 6 spore/ml, have high sensitivity and specificity of Nested-PCR in detecting the lower concentrations, comparing with traditional diagnostic method (culture on Sabouraud media) that were negative in all concetrations. (author)

Conversion of xylan by recyclable spores of Bacillus subtilis displaying thermophilic enzymes.

Science.gov (United States)

Mattossovich, Rosanna; Iacono, Roberta; Cangiano, Giuseppina; Cobucci-Ponzano, Beatrice; Isticato, Rachele; Moracci, Marco; Ricca, Ezio

2017-11-28

The Bacillus subtilis spore has long been used to display antigens and enzymes. Spore display can be accomplished by a recombinant and a non-recombinant approach, with the latter proved more efficient than the recombinant one. We used the non-recombinant approach to independently adsorb two thermophilic enzymes, GH10-XA, an endo-1,4-β-xylanase (EC 3.2.1.8) from Alicyclobacillus acidocaldarius, and GH3-XT, a β-xylosidase (EC 3.2.1.37) from Thermotoga thermarum. These enzymes catalyze, respectively, the endohydrolysis of (1-4)-β-D-xylosidic linkages of xylans and the hydrolysis of (1-4)-β-D-xylans to remove successive D-xylose residues from the non-reducing termini. We report that both purified enzymes were independently adsorbed on purified spores of B. subtilis. The adsorption was tight and both enzymes retained part of their specific activity. When spores displaying either GH10-XA or GH3-XT were mixed together, xylan was hydrolysed more efficiently than by a mixture of the two free, not spore-adsorbed, enzymes. The high total activity of the spore-bound enzymes is most likely due to a stabilization of the enzymes that, upon adsorption on the spore, remained active at the reaction conditions for longer than the free enzymes. Spore-adsorbed enzymes, collected after the two-step reaction and incubated with fresh substrate, were still active and able to continue xylan degradation. The recycling of the mixed spore-bound enzymes allowed a strong increase of xylan degradation. Our results indicate that the two-step degradation of xylans can be accomplished by mixing spores displaying either one of two required enzymes. The two-step process occurs more efficiently than with the two un-adsorbed, free enzymes and adsorbed spores can be reused for at least one other reaction round. The efficiency of the process, the reusability of the adsorbed enzymes, and the well documented robustness of spores of B. subtilis indicate the spore as a suitable platform to display enzymes

The search and identification of the new immunodiagnostic targets of bacillus anthracis spore

International Nuclear Information System (INIS)

Biketov, S.; Dunaytsev, I.; Baranova, E.; Marinin, L.; Dyatlov, I.

2009-01-01

Spores of Bacillus anthracis have been used as bio warfare agent to bio terrorize purposes. As efficiency of anti-epidemic measures included urgent prevention and treatment is determined by terms within which the bio agent is identified. Direct and rapid spore detection by antibodies based detection system is very attractive alternative to current PCR-based assays or routine phenotyping which are the most accurate but are also complex, time-consumption and expensive. The main difficulty with respect to such kind of anthrax spores detection is a cross-reaction with spores of closely related bacteria. For development of species-specific antibodies to anthrax spores recombinant scFvs or hybridoma technique were used. In both case surface spore antigens contained species-specific epitopes are need. Among exosporium proteins only ExsF(BxpB), ExsK and SoaA are specific to B.cereus group. On the surface of B. anthracis spores, a unique tetrasaccharides containing an novel monosaccharide - anthrose, was discovered. It was shown that anthrose can be serving as species-specific target for B. anthracis spores detection. We have revealed that EA1 isolated from spore of Russians strain STI-1 contain carbohydrate which formed species-specific epitopes and determine immunogenicity of this antigen. Antibodies to this antigen specifically recognized the surface target of B. anthracis spores and do not reacted with others Bacillus spore. Based on these antibodies we developed the test-systems in different formats for rapid direct detection and identification of B. anthracis spores. The results of trial these test-systems with using more than 50 different Bacillus strains were indicated that carbohydrate of EA1 isolated from spore is effective immunodiagnostic target for anthrax spores bio detection.(author)

Effect of gamma irradiation on thermal inactivation and injury of Bacillus subtilis spores

International Nuclear Information System (INIS)

El-Zawahry, Y.A.; Mostafa, S.A.; Awny, N.M.

1986-01-01

Bacillus subtilis spores which received preliminary irradiation doses were more sensitive to subsequent heating than non-irradiated spores. The thermal inactivation increased by increasing any of exposure temperature, thermal exposure time or preliminary irradiation dose. The thermal (D T -) value was much higher for non-irradiated spores than the D TR value for the pre-thermal irradiated spores. The radiosensitizing effect was directly proportional to the preliminary irradiation dose. The pre-thermal irradiation treatment of B. subtilis spores resulted in a synergistic effect in spore deactivation. This synergistic effect increased gradually by increasing the preliminary irradiation dose and/or the thermal temperature from 60 to 80 0 C, but decreased for 90 0 C and for the longer exposure periods at any of the examined temperature. Thermal injury of B. subtilis spores was more for the non-irradiated than for the irradiated spores

Fungal dissemination by housefly (Musca domestica L.) and contamination of food commodities in rural areas of South Africa.

Science.gov (United States)

Phoku, J Z; Barnard, T G; Potgieter, N; Dutton, M F

2016-01-18

Several insects that act as vectors, including houseflies (Musca domestica L.), are often considered to be an important source of fungal contamination in human foods. Houseflies are also involved in the transmission of bacterial pathogens that may pose a serious hazard to human health. Thus, the rural population of South Africa, as typified by that in the Gauteng Province investigated in this study, is at high risk from fungal exposure disseminated by houseflies and it is therefore important to assess the role of flies in contaminating various food commodities. Eighty four samples of houseflies (captured from households and pit toilets) were studied for their potential to carry fungal spores into food commodities. The fungi occurring in samples of raw maize (15) and porridge (19) were also assessed. Fungal isolates were identified based on morphological characteristics by conventional identification methods. Fifteen genera of fungi were isolated and identified, of which Aspergillus, Fusarium, Penicillium, Cladosporium, Moniliella and Mucor were the most prevalent in all three sample types analysed. The incidence rates of fungal contamination per total fungal count isolated in houseflies, maize and porridge were recorded with mean fungal load of 2×10(8) CFU/ml, 1×10(7)CFU/g and 2×10(7)CFU/g respectively. Additionally, A. flavus, A. parasiticus, F. verticillioides, F. proliferatum, P. verrucosum, P. aurantiogriseum and M. suaveolens were the most frequent fungal isolates in houseflies with incidence rate of 34%, 11%, 27%, 21%, 22%, 17% and 32% respectively. F. verticillioides, A. flavus, A. niger and P. oslonii were the most prevalent species contaminating porridge and maize with incidence rate of 23%, 32%, 16% and 28% in maize samples, while incidence rates of 59%, 15% and 29% were recorded in porridge samples with the exception of F. verticillioides. The prevalence of these genera of fungi may pose serious health risks. Copyright © 2015 Elsevier B.V. All rights

Architecture and assembly of the Bacillus subtilis spore coat.

Science.gov (United States)

Plomp, Marco; Carroll, Alicia Monroe; Setlow, Peter; Malkin, Alexander J

2014-01-01

Bacillus spores are encased in a multilayer, proteinaceous self-assembled coat structure that assists in protecting the bacterial genome from stresses and consists of at least 70 proteins. The elucidation of Bacillus spore coat assembly, architecture, and function is critical to determining mechanisms of spore pathogenesis, environmental resistance, immune response, and physicochemical properties. Recently, genetic, biochemical and microscopy methods have provided new insight into spore coat architecture, assembly, structure and function. However, detailed spore coat architecture and assembly, comprehensive understanding of the proteomic composition of coat layers, and specific roles of coat proteins in coat assembly and their precise localization within the coat remain in question. In this study, atomic force microscopy was used to probe the coat structure of Bacillus subtilis wild type and cotA, cotB, safA, cotH, cotO, cotE, gerE, and cotE gerE spores. This approach provided high-resolution visualization of the various spore coat structures, new insight into the function of specific coat proteins, and enabled the development of a detailed model of spore coat architecture. This model is consistent with a recently reported four-layer coat assembly and further adds several coat layers not reported previously. The coat is organized starting from the outside into an outermost amorphous (crust) layer, a rodlet layer, a honeycomb layer, a fibrous layer, a layer of "nanodot" particles, a multilayer assembly, and finally the undercoat/basement layer. We propose that the assembly of the previously unreported fibrous layer, which we link to the darkly stained outer coat seen by electron microscopy, and the nanodot layer are cotH- and cotE- dependent and cotE-specific respectively. We further propose that the inner coat multilayer structure is crystalline with its apparent two-dimensional (2D) nuclei being the first example of a non-mineral 2D nucleation crystallization

Architecture and Assembly of the Bacillus subtilis Spore Coat

Science.gov (United States)

Plomp, Marco; Carroll, Alicia Monroe; Setlow, Peter; Malkin, Alexander J.

2014-01-01

Bacillus spores are encased in a multilayer, proteinaceous self-assembled coat structure that assists in protecting the bacterial genome from stresses and consists of at least 70 proteins. The elucidation of Bacillus spore coat assembly, architecture, and function is critical to determining mechanisms of spore pathogenesis, environmental resistance, immune response, and physicochemical properties. Recently, genetic, biochemical and microscopy methods have provided new insight into spore coat architecture, assembly, structure and function. However, detailed spore coat architecture and assembly, comprehensive understanding of the proteomic composition of coat layers, and specific roles of coat proteins in coat assembly and their precise localization within the coat remain in question. In this study, atomic force microscopy was used to probe the coat structure of Bacillus subtilis wild type and cotA, cotB, safA, cotH, cotO, cotE, gerE, and cotE gerE spores. This approach provided high-resolution visualization of the various spore coat structures, new insight into the function of specific coat proteins, and enabled the development of a detailed model of spore coat architecture. This model is consistent with a recently reported four-layer coat assembly and further adds several coat layers not reported previously. The coat is organized starting from the outside into an outermost amorphous (crust) layer, a rodlet layer, a honeycomb layer, a fibrous layer, a layer of “nanodot” particles, a multilayer assembly, and finally the undercoat/basement layer. We propose that the assembly of the previously unreported fibrous layer, which we link to the darkly stained outer coat seen by electron microscopy, and the nanodot layer are cotH- and cotE- dependent and cotE-specific respectively. We further propose that the inner coat multilayer structure is crystalline with its apparent two-dimensional (2D) nuclei being the first example of a non-mineral 2D nucleation crystallization

Phosphorescence In Bacillus Spores

National Research Council Canada - National Science Library

Reinisch, Lou; Swartz, Barry A; Bronk, Burt V

2003-01-01

.... Our present work attempts to build on this approach for environmental applications. We have measured a change in the fluorescence spectra of suspensions of Bacillus bacteria between the vegetative bacteria and their spores at room temperature...

Comparison of survivability of Staphylococcus aureus and spores of Aspergillus niger on commonly used floor materials.

Science.gov (United States)

Gupta, Mridula; Bisesi, Michael; Lee, Jiyoung

2017-07-01

The survivability of Staphylococcus aureus and spores of Aspergillus niger was compared on 5 common floor materials. Floor materials were inoculated with a known concentration of S aureus and spores of A niger on day 0. Their survivability was measured on days, 2, 7, 14, and 28 by bulk rinsate method and enumerated using culture-based method. The difference in change of S aureus levels was statistically significant for all tested days (P niger spores were undetected on residential broadloom carpet and rubber-backed commercial carpet after day 2 but survived on VCT, PT, and wood until day 28. Floor materials with hard and smooth surfaces, such as VCT and PT, can allow survival of S aureus and A niger for up to 4 weeks. It may imply that floor materials can play a major role in preserving microbial contaminants in the built environment. Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Interactions between the entomopathogenic nematode Heterorhabditis sonorensis (Nematoda: Heterorhabditidae) and the saprobic fungus Fusarium oxysporum (Ascomycota: Hypocreales).

Science.gov (United States)

Navarro, P D; McMullen, J G; Stock, S P

2014-01-01

In this study, we assessed the effect of the saprobic fungus, Fusarium oxysporum (Ascomycota: Hypocreales) on the fitness of the entomopathogenic nematode Heterorhabditis sonorensis (Caborca strain). Sand column assays were considered to evaluate the effect of fungal mycelia on infective juvenile (IJ) movement and host access. Additionally, we investigated the effect of fungal spores on the nematodes' ability to search for a host, its virulence, penetration efficiency and reproduction. Three application timings were considered to assess interactions between the fungus and the nematodes. In vitro assays were also considered to determine the effect of fungal extracts on the nematode's symbiotic bacteria. Our observations indicate that presence and age of fungal mycelia significantly affect IJ movement in the sand columns and their ability to establish in the host. These results were also reflected in a reduced insect mortality. In particular, treatments with the 15 days old mycelia showed a significant reduction in insect mortality and penetration efficiency. Presence of fungal spores also impacted nematode virulence and reproduction. In particular, two of the application timings tested (simultaneous [EPN and fungal spores applied at the same time] and alternate I [EPN applied first, fungus applied 24h later]) resulted in antagonistic interactions. Moreover, IJ progeny was reduced to half in the simultaneous application. In vitro assays revealed that fungal extracts at the highest concentration tested (10mg/ml) inhibited the growth of the symbiotic bacteria. Overall, these results suggest that saprobic fungi may play an important role in regulating. EPN populations in the soil, and that they may be one of the factors that impact nematode survival in the soil and their access to insect hosts. Copyright © 2013 Elsevier Inc. All rights reserved.

In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions.

Science.gov (United States)

Vargas, Isabel Beatriz de; Droste, Annette

2014-03-01

Cyathea atrovirens occurs in a wide range of habitats in Brazil, Paraguay, Uruguay and Argentina. In the Brazilian State of Rio Grande do Sul, this commonly found species is a target of intense exploitation, because of its ornamental characteristics. The in vitro culture is an important tool for propagation which may contribute toward the reduction of extractivism. However, exogenous contamination of spores is an obstacle for the success of aseptic long-term cultures. This study evaluated the influence of different sterilization methods combined with storage conditions on the contamination of the in vitro cultures and the gametophytic development of C. atrovirens, in order to establish an efficient propagation protocol. Spores were obtained from plants collected in Novo Hamburgo, State of Rio Grande do Sul, Brazil. In the first experiment, spores stored at 7 degrees C were surface sterilized with 0.5, 0.8 and 2% of sodium hypochlorite (NaClO) for 15 minutes and sown in Meyer's culture medium. The cultures were maintained in a growth room at 26 +/- 1 degrees C for a 12-h photoperiod and photon flux density of 100 micromol/m2/s provided by cool white fluorescent light. Contamination was assessed at 60 days, and gametophytic development was scored at 30, 60, 120 and 130 days of in vitro culture, analyzing 300 individuals for each treatment. There was no significant difference in culture contamination among the different sodium hypochlorite concentrations tested, and all treatments allowed for the development of cordiform gametophytes at 130 days of culture. In the second experiment, spores stored at 7 and -20 degrees C were divided into two groups. Half of the spores were surface sterilized with 2% of NaClO for 15 minutes and the other half was not sterilized. All spores were sown in Meyer's medium supplemented with one of the following antibiotics: nystatin, Micostatin and actidione. The culture conditions and the procedures used for evaluating contamination and

Radiocaesium in the fungal compartment of forest ecosystems

International Nuclear Information System (INIS)

Vinichuk, Mykhaylo

2003-01-01

Fungi in forest ecosystems are major contributors to accumulation and cycling of radionuclides, especially radiocaesium. However, relatively little is known about uptake and retention of 137 Cs by fungal mycelia. This thesis comprises quantitative estimates of manually prepared mycelia of mainly ectomycorrhizal fungi and their possible role in the retention, turnover and accumulation of radiocaesium in contaminated forest ecosystems. The studies were conducted in two forests during 1996-1998 and 2000-2003. One was in Ovruch district, Zhytomyr region of Ukraine (51 deg 30 min N, 28 deg 95 min E), and the other at two Swedish forest sites: the first situated about 35 km northwest of Uppsala (60 deg 05 min N, 17 deg 25 min E) and the second at Hille in the vicinity of Gaevle (60 deg 85 min N, 17 deg 15 min E). The 137 Cs activity concentration was measured in prepared mycelia and corresponding soil layers. Various extraction procedures were used to study the retention and binding of 137 Cs in Of/Oh and Ah/B horizons of forest soil. 137 Cs was also extracted from the fruit bodies and mycelia of fungi. The fungal mycelium biomass was estimated and the percentage of the total inventory of 137 Cs bound in mycelia in the Ukrainian and Swedish forests was calculated. The estimated fungal biomass in Ukrainian forests varied from 0.07 to 70.4 mg/g soil, in Swedish forests between 3.6 and 19. 4 mg/g soil. Between 0.5 to 50 % of the total 137 Cs activity in the 0-10 cm soil profile was retained in the fungal mycelia. The 137 Cs activity concentration in mycelia was thus higher than that found in soil, and 137 Cs activity concentrations in the fruit bodies was higher than that in the mycelium. The survey study revealed that a major part, around 50 % of the plant-available 137 Cs in forest soil, was retained in the fungal mycelium. The most probable sources of 137 Cs for fungal mycelia and fruit bodies of fungi were found to be water soluble substances, humic matter

The distribution of Paenibacillus larvae spores in adult bees and honey and larval mortality, following the addition of American foulbrood diseased brood or spore-contaminated honey in honey bee (Apis mellifera) colonies.

Science.gov (United States)

Lindström, Anders; Korpela, Seppo; Fries, Ingemar

2008-09-01

Within colony transmission of Paenibacillus larvae spores was studied by giving spore-contaminated honey comb or comb containing 100 larvae killed by American foulbrood to five experimental colonies respectively. We registered the impact of the two treatments on P. larvae spore loads in adult bees and honey and on larval mortality by culturing for spores in samples of adult bees and honey, respectively, and by measuring larval survival. The results demonstrate a direct effect of treatment on spore levels in adult bees and honey as well as on larval mortality. Colonies treated with dead larvae showed immediate high spore levels in adult bee samples, while the colonies treated with contaminated honey showed a comparable spore load but the effect was delayed until the bees started to utilize the honey at the end of the flight season. During the winter there was a build up of spores in the adult bees, which may increase the risk for infection in spring. The results confirm that contaminated honey can act as an environmental reservoir of P. larvae spores and suggest that less spores may be needed in honey, compared to in diseased brood, to produce clinically diseased colonies. The spore load in adult bee samples was significantly related to larval mortality but the spore load of honey samples was not.

Mechanism of Bacillus subtilis spore inactivation by and resistance to supercritical CO2 plus peracetic acid.

Science.gov (United States)

Setlow, B; Korza, G; Blatt, K M S; Fey, J P; Setlow, P

2016-01-01

Determine how supercritical CO2 (scCO2 ) plus peracetic acid (PAA) inactivates Bacillus subtilis spores, factors important in spore resistance to scCO2 -PAA, and if spores inactivated by scCO2 -PAA are truly dead. Spores of wild-type B. subtilis and isogenic mutants lacking spore protective proteins were treated with scCO2 -PAA in liquid or dry at 35°C. Wild-type wet spores (aqueous suspension) were more susceptible than dry spores. Treated spores were examined for viability (and were truly dead), dipicolinic acid (DPA), mutations, permeability to nucleic acid stains, germination under different conditions, energy metabolism and outgrowth. ScCO2 -PAA-inactivated spores retained DPA, and survivors had no notable DNA damage. However, DPA was released from inactivated spores at a normally innocuous temperature (85°C), and colony formation from treated spores was salt sensitive. The inactivated spores germinated but did not outgrow, and these germinated spores had altered plasma membrane permeability and defective energy metabolism. Wet or dry coat-defective spores had increased scCO2 -PAA sensitivity, and dry spores but not wet spores lacking DNA protective proteins were more scCO2 -PAA sensitive. These findings suggest that scCO2 -PAA inactivates spores by damaging spores' inner membrane. The spore coat provided scCO2 -PAA resistance for both wet and dry spores. DNA protective proteins provided scCO2 -PAA resistance only for dry spores. These results provide information on mechanisms of spore inactivation of and resistance to scCO2 -PAA, an agent with increasing use in sterilization applications. © 2015 The Society for Applied Microbiology.

Implementation of bio-fungicides and seed treatment in organic rice cv. KDML 105 farming.

Science.gov (United States)

Thobunluepop, Pitipong

2009-08-15

This study was aimed to evaluate the several chemical compounds of relatively composite structure with antifungal activity from Thai local medical plants. The antifungal activity of Stemona curtisii HK. f., Stemona tuberose L., Acorus calamus L., Eugenia caryophyllus, Memmea siamensis Kost. and an eugenol active compound were studied in vitro. Four pathogenic seed borne fungi, Alternaria solani, Colletotrichum sp., Fusarium moniliforme and Rhizoctonia solani were used as target organisms. The agar overlay technique and spore inhibition techniques were applied for the determination of their essential oil and active compound antifungal activity at various concentration; 0.10, 0.25, 0.50 and 1.00% (v/v) and untreated as control (0% v/v). Eugenol active compound showed the strongest antifungal activity on all species of tested fungal species. On the other hand, the antifungal activity of those bio-fungicides was lined up into a series from strong to low, as follows: Eugenia caryophyllus > Acorus calamus Linn. > Stemona tuberosa L. > Stemona curtisii Hk.f, while Mammea siamensis Kost. could not control any fungal species. Moreover, after eugenol application, lysis of spore and inhibition of mycelium growth were detected. Microscopic analysis exhibited complete lysis of spores after 24 h at a concentration of 1.00% v/v. Moreover, at the same concentration and 96 h incubation the mycelia growth was completely inhibited.

Gene discovery in EST sequences from the wheat leaf rust fungus Puccinia triticina sexual spores, asexual spores and haustoria, compared to other rust and corn smut fungi

Science.gov (United States)

2011-01-01

Background Rust fungi are biotrophic basidiomycete plant pathogens that cause major diseases on plants and trees world-wide, affecting agriculture and forestry. Their biotrophic nature precludes many established molecular genetic manipulations and lines of research. The generation of genomic resources for these microbes is leading to novel insights into biology such as interactions with the hosts and guiding directions for breakthrough research in plant pathology. Results To support gene discovery and gene model verification in the genome of the wheat leaf rust fungus, Puccinia triticina (Pt), we have generated Expressed Sequence Tags (ESTs) by sampling several life cycle stages. We focused on several spore stages and isolated haustorial structures from infected wheat, generating 17,684 ESTs. We produced sequences from both the sexual (pycniospores, aeciospores and teliospores) and asexual (germinated urediniospores) stages of the life cycle. From pycniospores and aeciospores, produced by infecting the alternate host, meadow rue (Thalictrum speciosissimum), 4,869 and 1,292 reads were generated, respectively. We generated 3,703 ESTs from teliospores produced on the senescent primary wheat host. Finally, we generated 6,817 reads from haustoria isolated from infected wheat as well as 1,003 sequences from germinated urediniospores. Along with 25,558 previously generated ESTs, we compiled a database of 13,328 non-redundant sequences (4,506 singlets and 8,822 contigs). Fungal genes were predicted using the EST version of the self-training GeneMarkS algorithm. To refine the EST database, we compared EST sequences by BLASTN to a set of 454 pyrosequencing-generated contigs and Sanger BAC-end sequences derived both from the Pt genome, and to ESTs and genome reads from wheat. A collection of 6,308 fungal genes was identified and compared to sequences of the cereal rusts, Puccinia graminis f. sp. tritici (Pgt) and stripe rust, P. striiformis f. sp. tritici (Pst), and poplar

Gene discovery in EST sequences from the wheat leaf rust fungus Puccinia triticina sexual spores, asexual spores and haustoria, compared to other rust and corn smut fungi

Directory of Open Access Journals (Sweden)

Wynhoven Brian

2011-03-01

Full Text Available Abstract Background Rust fungi are biotrophic basidiomycete plant pathogens that cause major diseases on plants and trees world-wide, affecting agriculture and forestry. Their biotrophic nature precludes many established molecular genetic manipulations and lines of research. The generation of genomic resources for these microbes is leading to novel insights into biology such as interactions with the hosts and guiding directions for breakthrough research in plant pathology. Results To support gene discovery and gene model verification in the genome of the wheat leaf rust fungus, Puccinia triticina (Pt, we have generated Expressed Sequence Tags (ESTs by sampling several life cycle stages. We focused on several spore stages and isolated haustorial structures from infected wheat, generating 17,684 ESTs. We produced sequences from both the sexual (pycniospores, aeciospores and teliospores and asexual (germinated urediniospores stages of the life cycle. From pycniospores and aeciospores, produced by infecting the alternate host, meadow rue (Thalictrum speciosissimum, 4,869 and 1,292 reads were generated, respectively. We generated 3,703 ESTs from teliospores produced on the senescent primary wheat host. Finally, we generated 6,817 reads from haustoria isolated from infected wheat as well as 1,003 sequences from germinated urediniospores. Along with 25,558 previously generated ESTs, we compiled a database of 13,328 non-redundant sequences (4,506 singlets and 8,822 contigs. Fungal genes were predicted using the EST version of the self-training GeneMarkS algorithm. To refine the EST database, we compared EST sequences by BLASTN to a set of 454 pyrosequencing-generated contigs and Sanger BAC-end sequences derived both from the Pt genome, and to ESTs and genome reads from wheat. A collection of 6,308 fungal genes was identified and compared to sequences of the cereal rusts, Puccinia graminis f. sp. tritici (Pgt and stripe rust, P. striiformis f. sp

Cohort Study of Airway Mycobiome in Adult Cystic Fibrosis Patients: Differences in Community Structure between Fungi and Bacteria Reveal Predominance of Transient Fungal Elements

Science.gov (United States)

Sauer-Heilborn, Annette; Welte, Tobias; Guzman, Carlos A.; Abraham, Wolf-Rainer; Höfle, Manfred G.

2015-01-01

The respiratory mycobiome is an important but understudied component of the human microbiota. Like bacteria, fungi can cause severe lung diseases, but their infection rates are much lower. This study compared the bacterial and fungal communities of sputum samples from a large cohort of 56 adult patients with cystic fibrosis (CF) during nonexacerbation periods and under continuous antibiotic treatment. Molecular fingerprinting based on single-strand conformation polymorphism (SSCP) analysis revealed fundamental differences between bacterial and fungal communities. Both groups of microorganisms were taxonomically classified by identification of gene sequences (16S rRNA and internal transcript spacer), and prevalences of single taxa were determined for the entire cohort. Major bacterial pathogens were frequently observed, whereas fungi of known pathogenicity in CF were detected only in low numbers. Fungal species richness increased without reaching a constant level (saturation), whereas bacterial richness showed saturation after 50 patients were analyzed. In contrast to bacteria, a large number of fungal species were observed together with high fluctuations over time and among patients. These findings demonstrated that the mycobiome was dominated by transient species, which strongly suggested that the main driving force was their presence in inhaled air rather than colonization. Considering the high exposure of human airways to fungal spores, we concluded that fungi have low colonization abilities in CF, and colonization by pathogenic fungal species may be considered a rare event. A comprehensive understanding of the conditions promoting fungal colonization may offer the opportunity to prevent colonization and substantially reduce or even eliminate fungus-related disease progression in CF. PMID:26135861

Scanning Surface Potential Microscopy of Spore Adhesion on Surfaces

Energy Technology Data Exchange (ETDEWEB)

Lee, Ida [University of Tennessee, Knoxville (UTK); Chung, Eunhyea [Georgia Institute of Technology; Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Tsouris, Costas [ORNL

2012-01-01

The adhesion of spores of Bacillus anthracis - the cause of anthrax and a likely biological threat - to solid surfaces is an important consideration in cleanup after an accidental or deliberate release. However, because of safety concerns, directly studying B. anthracis spores with advanced instrumentation is problematic. As a first step, we are examining the electrostatic potential of Bacillus thuringiensis (Bt), which is a closely related species that is often used as a simulant to study B. anthracis. Scanning surface potential microscopy (SSPM), also known as Kelvin probe force microscopy (KPFM), was used to investigate the influence of relative humidity (RH) on the surface electrostatic potential of Bt that had adhered to silica, mica, or gold substrates. AFM/SSPM side-by-side images were obtained separately in air, at various values of RH, after an aqueous droplet with spores was applied on each surface and allowed to dry before measurements. In the SSPM images, a negative potential on the surface of the spores was observed compared with that of the substrates. The surface potential decreased as the humidity increased. Spores were unable to adhere to a surface with an extremely negative potential, such as mica.

Detection of Bacillus spores using PCR and FTA filters.

Science.gov (United States)

Lampel, Keith A; Dyer, Deanne; Kornegay, Leroy; Orlandi, Palmer A

2004-05-01

Emphasis has been placed on developing and implementing rapid detection systems for microbial pathogens. We have explored the utility of expanding FTA filter technology for the preparation of template DNA for PCR from bacterial spores. Isolated spores from several Bacillus spp., B. subtilis, B. cereus, and B. megaterium, were applied to FTA filters, and specific DNA products were amplified by PCR. Spore preparations were examined microscopically to ensure that the presence of vegetative cells, if any, did not yield misleading results. PCR primers SRM86 and SRM87 targeted a conserved region of bacterial rRNA genes, whereas primers Bsub5F and Bsub3R amplified a product from a conserved sequence of the B. subtilis rRNA gene. With the use of the latter set of primers for nested PCR, the sensitivity of the PCR-based assay was increased. Overall, 53 spores could be detected after the first round of PCR, and the sensitivity was increased to five spores by nested PCR. FTA filters are an excellent platform to remove PCR inhibitors and have universal applications for environmental, clinical, and food samples.

[Microbial resistance to formaldehyde. I. Comparative quantitative studies in some selected species of vegetative bacteria, bacterial spores, fungi, bacteriophages and viruses].

Science.gov (United States)

Spicher, G; Peters, J

1976-12-01

The resistence of different microorganisms to formaldehyde was determined. As test objects served gram-negative and gram-positive vegetative germs (Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella paratyphi-B, Staphylococcus aureus, Streptococcus faecalis), bacterial spores (Bacillus cereus, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis), fungi (Aspergillus niger, Candida albicans), bacteriophages (Escherichia coli phages, T1, T2, T3), and viruses (adenovirus, poliomyelitis virus, vaccinia virus). For the studies, suspensions of germs were exposed at identical temperature (20 degrees C) and pH (7.0). The microbicidal effect of formaldehyde was measured by the decrease of the proportion of germs capable of multiplication in the suspension (lg (N/N0); where: N0 equals initial number of germs capable of multiplication; N equals number of germs capable of multiplication after exposure to formaldehyde). For all germs the dependence of the microbicidal effect on the concentration of formaldehyde was determined. In all experiments, the duration of exposure was two hours. Pseudomonas aeruginosa, Klebsiella pneumoniae, and Salmonella paratyphi-B were found to be more susceptible than Staphylococcus aureus (vf. Fig. 1 A). The strains of Pseudomonas aeruginosa used were widely varying as to their susceptibility. To obtain equal microbicidal effects, concentrations of formaldehyde almost three times as high had to be used for the most resistant strain than were necessary for the most susceptible strain of Pseudomonas aeruginosa. All strains of Klebsiella pneumoniae examined were found to have an identical resistence to formaldehyde. Streptococcus faecalis was even more resistant to formaldehyde than Staphylococcus aureus. In the case of Streptococcus faecalis, a concentration of formaldehyde about three times as high had to be used to obtain microbicidal effects of identical magnitude. For the killing of Candida albicans cells concentrations of

Inhibitory activity of Indian spice plant Cinnamomum zeylanicum extracts against Alternaria solani and Curvularia lunata, the pathogenic dematiaceous moulds

Directory of Open Access Journals (Sweden)

Sharma Bechan

2009-03-01

Full Text Available Abstract Background Dematiaceous moulds are pathogenic microorganisms and act as etiological agents of mycoses with different degrees of severity in humans and animals. These moulds also cause loss of food crops and storage food products. The information regarding antimicrobial efficacy of the plant preparations on these moulds is scanty. The present study reveals phytochemical characterization and the effect of bark and leaf extracts of Indian spice plant, Cinnamomum zeylanicum (Cz, against the growth of two species of dematiaceous moulds, Alternaria solani and Curvularia lunata. Methods Cz bark and leaf samples were sequentially extracted in different solvents using Soxhlet apparatus. Phytochemical analyses of extracts were done as per standard protocols. The antifungal bioassay of extracts was done by hanging drop technique. The inhibition of fungal spore germination was monitored under influence of three different concentrations of extracts. Results The lowest test concentration (50 μg/ml of extracts of Cz bark prepared into acetone and that of Cz leaf into petroleum ether and ethanol exhibited complete inhibition (100% of spore germination in both the moulds. At 100 μg/ml concentration all the extracts showed about 50 to 100% inhibition. However, the treatment of the spores of the two fungal species with highest concentration (500 μg/ml of bark and leaf extracts in all the solvents showed 100% fungicidal activity as it completely arrested the germination of spores. Relatively lower activity of aqueous extracts at 50 and 100 μg/ml concentrations suggests that the antifungal ingredients present in Cz bark and leaf are more soluble in organic solvents than water. Conclusion The results demonstrated that the Cz bark and leaves contain certain fungicidal constituents exhibiting potential antimould activity against A. solani and C. lunata.

Removal of dissolved heavy metals and radionuclides by microbial spores

International Nuclear Information System (INIS)

Revis, N.W.; Hadden, C.T.; Edenborn, H.

1997-01-01

Microbial systems have been shown to remove specific heavy metals from contaminated aqueous waste to levels acceptable to EPA for environmental release. However, systems capable of removing a variety of heavy metals from aqueous waste to environmentally acceptable levels remain to be reported. The present studies were performed to determine the specificity of spores of the bacterium Bacillus megaterium for the adsorption of dissolved metals and radionuclides from aqueous waste. The spores effectively adsorbed eight heavy metals from a prepared metal mix and from a plating rinse waste to EPA acceptable levels for waste water. These results suggest that spores have multiple binding sites for the adsorption of heavy metals. Spores were also effective in adsorbing the radionuclides 85 strontium and 197 cesium. The presence of multiple sites in spores for the adsorption of heavy metals and radionuclides makes this biosorbent a good candidate for the treatment of aqueous wastes associated with the plating and nuclear industries. 17 refs., 4 tabs

Assessment of Clostridium perfringens spore response to high hydrostatic pressure and heat with nisin.

Science.gov (United States)

Gao, Yulong; Qiu, Weifen; Wu, Ding; Fu, Qiang

2011-08-01

The elimination of spores from low-acid foods presents food-processing and food-safety challenges to high-pressure processing (HPP) developers as bacterial spores are extremely resistant to pressure. Therefore, the effects of pressure (400-800 MPa), temperature (35-95 °C), and nisin (0-496 IU/mL) on the inactivation of Clostridium perfringens AS 64701 spores at various pressure-holding times (7.5-17.5 min) were explored. A second-order polynomal equation for HPP- and nisin-induced inactivation of C. perfringens spores was constructed with response surface methodology. Experiment results showed that the experimental values were shown to be significantly in agreement with the predicted values because the adjusted determination coefficient (R (Adj)²) was 0.9708 and the level of significance was P pressure of 654 Mpa, temperature of 74 °C, pressure-holding time of 13.6 min, and nisin concentration of 328 IU/mL. The validation of the model equation for predicting the optimum response values was verified effectively by ten test points that were not used in the establishment of the model. Compared with conventional HPP techniques, the main process advantages of HPP-nisin combination sterilization in the UHT milk are, lower pressure, temperature, natural preservative (nisin), and in a shorter treatment time. The synergistic inactivation of bacteria by HPP-nisin combination is a promising and natural method to increase the efficiency and safety of high-pressure pasteurization.

Influence of food matrix on outgrowth heterogeneity of heat damaged Bacillus cereus spores.

Science.gov (United States)

Warda, Alicja K; den Besten, Heidy M W; Sha, Na; Abee, Tjakko; Nierop Groot, Masja N

2015-05-18

Spoilage of heat treated foods can be caused by the presence of surviving spore-formers. It is virtually impossible to prevent contamination at the primary production level as spores are ubiquitous present in the environment and can contaminate raw products. As a result spore inactivation treatments are widely used by food producing industries to reduce the microbial spore loads. However consumers prefer mildly processed products that have less impact on its quality and this trend steers industry towards milder preservation treatments. Such treatments may result in damaged instead of inactivated spores, and these spores may germinate, repair, and grow out, possibly leading to quality and safety issues. The ability to repair and grow out is influenced by the properties of the food matrix. In the current communication we studied the outgrowth from heat damaged Bacillus cereus ATCC 14579 spores on Anopore membrane, which allowed following outgrowth heterogeneity of individual spores on broccoli and rice-based media as well as standard and mildly acidified (pH 5.5) meat-based BHI. Rice, broccoli and BHI pH 5.5 media resulted in delayed outgrowth from untreated spores, and increased heterogeneity compared to BHI pH 7.4, with the most pronounced effect in rice media. Exposure to wet heat for 1 min at 95 °C caused 2 log inactivation and approximately 95% of the spores in the surviving fraction were damaged resulting in substantial delay in outgrowth based on the time required to reach a maximum microcolony size of 256 cells. The delay was most pronounced for heat-treated spores on broccoli medium followed by spores on rice media (both untreated and treated). Interestingly, the increase in outgrowth heterogeneity of heat treated spores on BHI pH 7.4 was more pronounced than on rice, broccoli and BHI pH 5.5 conceivably reflecting that conditions in BHI pH 7.4 better support spore damage repair. This study compares the effects of three main factors, namely heat treatment, p

Isolated Bacterial Spores at High-velocity Survive Surface Impacts in Vacuum

Science.gov (United States)

Austin, Daniel; Barney, Brandon

We present experiments in which bacterial spores were found to survive being accelerated in vacuum to velocities in the range 30-120 m/s and impacted on a dense target. In these experiments, spores of Bacillus subtilis spores were charged using electrospray at atmospheric pressure, dried, and then introduced into high vacuum. Through choice of skimmers and beam tubes, different velocity ranges were achieved. An image-charge detector observed the charged spores, providing total charge and velocity. The spores then impacted a glass target within a collection vessel. After the experiment, the collection vessel contents were extracted and cultured. Several positive and negative controls were used, including the use of antibiotic-resistant spores and antibiotic-containing (rifampicin) agar for culturing. These impact velocities are of particular interest for possible transport of bacterial spores from Mars to Phobos, and may have implications for planetary protection in a Phobos sample return mission. In addition, bacteria may reach similar velocities during a spacecraft crash (e.g., within components, or from spacecraft to surface materials during impact, etc.), raising concerns about forward contamination. The velocities of interest to transport of life between planets (panspermia) are somewhat higher, but these results complement shock-based experiments and contribute to the general discussion of impact survivability of organisms.

Nanoscale Structural and Mechanical Analysis of Bacillus anthracis Spores Inactivated with Rapid Dry Heating

Science.gov (United States)

Felker, Daniel L.; Burggraf, Larry W.

2014-01-01

Effective killing of Bacillus anthracis spores is of paramount importance to antibioterrorism, food safety, environmental protection, and the medical device industry. Thus, a deeper understanding of the mechanisms of spore resistance and inactivation is highly desired for developing new strategies or improving the known methods for spore destruction. Previous studies have shown that spore inactivation mechanisms differ considerably depending upon the killing agents, such as heat (wet heat, dry heat), UV, ionizing radiation, and chemicals. It is believed that wet heat kills spores by inactivating critical enzymes, while dry heat kills spores by damaging their DNA. Many studies have focused on the biochemical aspects of spore inactivation by dry heat; few have investigated structural damages and changes in spore mechanical properties. In this study, we have inactivated Bacillus anthracis spores with rapid dry heating and performed nanoscale topographical and mechanical analysis of inactivated spores using atomic force microscopy (AFM). Our results revealed significant changes in spore morphology and nanomechanical properties after heat inactivation. In addition, we also found that these changes were different under different heating conditions that produced similar inactivation probabilities (high temperature for short exposure time versus low temperature for long exposure time). We attributed the differences to the differential thermal and mechanical stresses in the spore. The buildup of internal thermal and mechanical stresses may become prominent only in ultrafast, high-temperature heat inactivation when the experimental timescale is too short for heat-generated vapor to efficiently escape from the spore. Our results thus provide direct, visual evidences of the importance of thermal stresses and heat and mass transfer to spore inactivation by very rapid dry heating. PMID:24375142

RNA synthesis during germination of UV-irradiated Dictyostelium discoideum spores

International Nuclear Information System (INIS)

Okaichi, Kumio

1987-01-01

UV irradiation to the spores of Dictyostelium discoideum NC4 resulted in a more prolonged delay of amoeba-emergence from swollen spores with increasing UV fluence. During the germination, an inhibition of total RNA synthesis and a shift of stage of maximum RNA synthesis to the later period were observed. The maximum poly(A) + RNA synthetic activity was found on an early stage of amoeba-emergence prior about 1 h to the beginning of rRNA synthesis in unirradiated spore germination; but, in UV-irradiated spore germination, the stage of maximum poly(A) + RNA synthesis shifted to the later stage of germination with increasing UV fluence. A decreased synthesis of poly(A) + RNA and a severe inhibition of rRNA synthesis were observed on UV-irradiated and germinated spores, but no significant inhibition of 4 - 5 S RNA synthesis was detected. Actinomycin D suppressed almost completely the rRNA synthesis of emerged amoebae but the drug apparently did not affect the emergence of amoebae at any stage of germination. It was postulated that the delay of amoeba-emergence in UV-irradiated spore must be mainly due to the shift of the stage of maximum synthesis of poly(A) + RNA to the later stage of germination. (author)

Ethanol, vinegar and Origanum vulgare oil vapour suppress the development of anthracnose rot in tomato fruit.

Science.gov (United States)

Tzortzakis, Nikos G

2010-08-15

Anthracnose rot (Colletotrichum coccodes) development in vitro or in tomato (Lycopersicon esculentum L.) fruit was evaluated after treatment with absolute ethyl alcohol (AEA), vinegar (VIN), chlorine (CHL) or origanum oil (ORI) and storage at 12 degrees C and 95% relative humidity during or following exposure to the volatiles. Fruit treated with vapours reduced fungal spore germination/production, but in the case of AEA- and VIN-treated fruits, fungal mycelium development was accelerated. Fruit lesion development was suppressed after fruit exposure to pure (100% v/v) AEA or ORI vapours which were accompanied by increased fruit cracking. Exposure to pure VIN-, CHL- and ORI vapours reduced (up to 92%) spore germination in vitro, but no differences were observed in the AEA treatment. The benefits associated with volatiles-enrichment were maintained in fruit pre-exposed to vapours, resulting in suppression in spore germination and spore production. However, studies performed on fungi grown on Potato Dextrose Agar revealed fewer direct effects of volatiles on fungal colony development and spore germination per se, implying that suppression of pathogen development was due in a large part to the impact of volatiles on fruit-pathogen interactions and/or 'memory' effects on fruit tissue. Work is currently focussing on the mechanisms underlying the impacts of volatiles on fruit quality related attributes. The results of this study indicate that volatiles may be considered as an alternative to the traditional postharvest sanitizing techniques. Each commodity needs to be individually assessed, and the volatile concentration and sanitising technique optimised, before the volatile treatment is used commercially. Copyright 2010 Elsevier B.V. All rights reserved.

LEVELS AND TYPES OF AEROBIC SPORE FORMING BACTERIA ...

African Journals Online (AJOL)

The four companies whose packaged product were studied had an average plate total spore counts as follows: Company A=6.2x 103; Company B= 3.1x 104; Company C= 6.0x 104 and Company D= 3.1x102 colony forming units per gram, respectively. Identification tests showed that among the aerobic spore formers were ...

Properties of spores of Bacillus subtilis strains which lack the major small, acid-soluble protein

International Nuclear Information System (INIS)

Hackett, R.H.; Setlow, P.

1988-01-01

Bacillus subtilis strains containing a deletion in the gene coding for the major small, acid-soluble, spore protein (SASP-gamma) grew and sporulated, and their spores initiated germination normally, but outgrowth of SASP-gamma- spores was significantly slower than that of wild-type spores. The absence of SASP-gamma had no effect on spore protoplast density or spore resistance to heat or radiation. Consequently, SASP-gamma has a different function in spores than do the other major small, acid-soluble proteins

AM fungal exudates activate MAP kinases in plant cells in dependence from cytosolic Ca(2+) increase.

Science.gov (United States)

Francia, Doriana; Chiltz, Annick; Lo Schiavo, Fiorella; Pugin, Alain; Bonfante, Paola; Cardinale, Francesca

2011-09-01

The molecular dialogue occurring prior to direct contact between the fungal and plant partners of arbuscular-mycorrhizal (AM) symbioses begins with the release of fungal elicitors, so far only partially identified chemically, which can activate specific signaling pathways in the host plant. We show here that the activation of MAPK is also induced by exudates of germinating spores of Gigaspora margarita in cultured cells of the non-leguminous species tobacco (Nicotiana tabacum), as well as in those of the model legume Lotus japonicus. MAPK activity peaked about 15 min after the exposure of the host cells to the fungal exudates (FE). FE were also responsible for a rapid and transient increase in free cytosolic Ca(2+) in Nicotiana plumbaginifolia and tobacco cells, and pre-treatment with a Ca(2+)-channel blocker (La(3+)) showed that in these cells, MAPK activation was dependent on the cytosolic Ca(2+) increase. A partial dependence of MAPK activity on the common Sym pathway could be demonstrated for a cell line of L. japonicus defective for LjSym4 and hence unable to establish an AM symbiosis. Our results show that MAPK activation is triggered by an FE-induced cytosolic Ca(2+) transient, and that a Sym genetic determinant acts to modulate the intensity and duration of this activity. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

Live-imaging of Bacillus subtilis spore germination and outgrowth

NARCIS (Netherlands)

Pandey, R.

2014-01-01

Spores of Gram-positive bacteria such as Bacillus and Clostridium cause huge economic losses to the food industry. In food products, spores survive under food preservation conditions and subsequent germination and outgrowth eventually causes food spoilage. Therefore efforts are being made to

Dendritic Cells Endocytose Bacillus Anthracis Spores: Implications for Anthrax Pathogenesis

National Research Council Canada - National Science Library

Brittingham, Katherine C; Ruthel, Gordon; Panchal, Rekha G; Fuller, Claudette L; Ribot, Wilson J

2005-01-01

Phagocytosis of inhaled Bacillus anthracis spores and subsequent trafficking to lymph nodes are decisive events in the progression of inhaled anthrax because they initiate germination and dissemination of spores...

Spore: Spawning Evolutionary Misconceptions?

Science.gov (United States)

Bean, Thomas E.; Sinatra, Gale M.; Schrader, P. G.

2010-10-01

The use of computer simulations as educational tools may afford the means to develop understanding of evolution as a natural, emergent, and decentralized process. However, special consideration of developmental constraints on learning may be necessary when using these technologies. Specifically, the essentialist (biological forms possess an immutable essence), teleological (assignment of purpose to living things and/or parts of living things that may not be purposeful), and intentionality (assumption that events are caused by an intelligent agent) biases may be reinforced through the use of computer simulations, rather than addressed with instruction. We examine the video game Spore for its depiction of evolutionary content and its potential to reinforce these cognitive biases. In particular, we discuss three pedagogical strategies to mitigate weaknesses of Spore and other computer simulations: directly targeting misconceptions through refutational approaches, targeting specific principles of scientific inquiry, and directly addressing issues related to models as cognitive tools.

Factors affecting fungus-induced larval mortality in Anopheles gambiae and Anopheles stephensi

Directory of Open Access Journals (Sweden)

Takken Willem

2010-01-01

Full Text Available Abstract Background Entomopathogenic fungi have shown great potential for the control of adult malaria vectors. However, their ability to control aquatic stages of anopheline vectors remains largely unexplored. Therefore, how larval characteristics (Anopheles species, age and larval density, fungus (species and concentration and environmental effects (exposure duration and food availability influence larval mortality caused by fungus, was studied. Methods Laboratory bioassays were performed on the larval stages of Anopheles gambiae and Anopheles stephensi with spores of two fungus species, Metarhizium anisopliae and Beauveria bassiana. For various larval and fungal characteristics and environmental effects the time to death was determined and survival curves established. These curves were compared by Kaplan Meier and Cox regression analyses. Results Beauveria bassiana and Metarhizium anisopliae caused high mortality of An. gambiae and An. stephensi larvae. However, Beauveria bassiana was less effective (Hazard ratio (HR Metarhizium anisopliae. Anopheles stephensi and An. gambiae were equally susceptible to each fungus. Older larvae were less likely to die than young larvae (HR Conclusions This study shows that both fungus species have potential to kill mosquitoes in the larval stage, and that mortality rate depends on fungus species itself, larval stage targeted, larval density and amount of nutrients available to the larvae. Increasing the concentration of fungal spores or reducing the exposure time to spores did not show a proportional increase and decrease in mortality rate, respectively, because the spores clumped together. As a result spores did not provide uniform coverage over space and time. It is, therefore, necessary to develop a formulation that allows the spores to spread over the water surface. Apart from formulation appropriate delivery methods are also necessary to avoid exposing non-target organisms to fungus.

Fighting Ebola with novel spore decontamination technologies for the military.

Science.gov (United States)

Doona, Christopher J; Feeherry, Florence E; Kustin, Kenneth; Olinger, Gene G; Setlow, Peter; Malkin, Alexander J; Leighton, Terrance

2015-01-01

Recently, global public health organizations such as Doctors without Borders (MSF), the World Health Organization (WHO), Public Health Canada, National Institutes of Health (NIH), and the U.S. government developed and deployed Field Decontamination Kits (FDKs), a novel, lightweight, compact, reusable decontamination technology to sterilize Ebola-contaminated medical devices at remote clinical sites lacking infra-structure in crisis-stricken regions of West Africa (medical waste materials are placed in bags and burned). The basis for effectuating sterilization with FDKs is chlorine dioxide (ClO2) produced from a patented invention developed by researchers at the US Army Natick Soldier RD&E Center (NSRDEC) and commercialized as a dry mixed-chemical for bacterial spore decontamination. In fact, the NSRDEC research scientists developed an ensemble of ClO2 technologies designed for different applications in decontaminating fresh produce; food contact and handling surfaces; personal protective equipment; textiles used in clothing, uniforms, tents, and shelters; graywater recycling; airplanes; surgical instruments; and hard surfaces in latrines, laundries, and deployable medical facilities. These examples demonstrate the far-reaching impact, adaptability, and versatility of these innovative technologies. We present herein the unique attributes of NSRDEC's novel decontamination technologies and a Case Study of the development of FDKs that were deployed in West Africa by international public health organizations to sterilize Ebola-contaminated medical equipment. FDKs use bacterial spores as indicators of sterility. We review the properties and structures of spores and the mechanisms of bacterial spore inactivation by ClO2. We also review mechanisms of bacterial spore inactivation by novel, emerging, and established non-thermal technologies for food preservation, such as high pressure processing, irradiation, cold plasma, and chemical sanitizers, using an array of Bacillus

Enhancing Growth of Vigna radiata in the Presence of Pseudomonas aeruginosa Biopolymer and Metarhizium anisopliae Spores

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Bhagwan N. Rekadwad

2016-01-01

Full Text Available Exopolysaccharide producing Pseudomonas aeruginosa NCIM 2945 (PANCL belonging to gamma-proteobacterium and entomopathogenic fungus Metarhizium anisopliae MCC 1129 (MAMCC belonging to Ascomycota were studied for their morphological features biochemical characteristics and plant growth promotion ability. Optimum growth of PANCL was recorded after 24 h at temperature 30°C and pH 7.0. Gram-negative PANCL appeared as white in color, one mm size, circular, opaque, and nonconsistent elevated colonies with entire margin. It has utilized dextrose, fructose, maltose, and sorbitol as carbon source and produced acid in the medium. PANCL was sensitive to Polymyxin B (300 µgm/disc followed by Neomycin (30 µgm/disc, Gentamycin (10 µgm/disc, and Chloramphenicol (30 µgm/disc. PANCL has secreted extracellular lipase, amylase, protease, and exopolysaccharides (EPS. Another fungal strain MAMCC sporulated after 168 h at temperature 30°C and pH 7.0. MAMCC has septate-white mycelium and bears dirty green colored spores. Growth of MAMCC was enhanced in the presence of Neem and Karela-Amla oil (0.1 mL each. Extracellular polysaccharide produced by PANCL and spores of MAMCC promoted growth of dicotyledon Vigna radiata (Mung individually as well as in consortium. Considerable increase in dry weight of Vigna radiata was recorded. Thus, reported PANCL and MAMCC strains have promoted growth Vigna radiata and may be a solution for sustainable agriculture.

Pathogenicity evaluation of native isolates of entomopathogenic fungí against andean weevil, Premnotrypes vorax (Hustache

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Giselle Rivera

2001-07-01

Full Text Available The main goal of this study was pathogenicity evaluation of Beauveria bassiana and Metarhizium anisopliae native isolates, obtained from natural habitats and stocked potato, against potato andean weevil, Premnotrypes vorax (Hustache (COLEOPTERA: Curculionidae, and important potato insect pest in Colombia and other Andean countries. Patogenicity was determined by laboratory bioassays, using either reared insects or field captured insects. Insect rearing data are presented. Pathogenicity evaluation was express as mortality against time, estimating LT50 and LT90 for all the fungal isolates, and mortality against spore concentration, estimating CL50 for two selected isolates. In all cases, total mortality percentils were above 45%. Differences between reared and field captured insects were evident. According to obtained data one of the fungal isolates: B. bassiana 9770, obtained from R vorax larva (TL50: 4.7 days - 9.8 days; TL90: 14.1 days - 20.8 days; CL50:7.03 x 104spores/ml appears as a promisory fungal isolate for further studies. Out of this study, differences in the andean weevil, P. vorax, adult mortality, with regard to entomopathogenic fungal isolate and insect origin were manifest.

Rapid Detection of Bacillus anthracis Spores Using Immunomagnetic Separation and Amperometry

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David F. Waller

2016-12-01

Full Text Available Portable detection and quantitation methods for Bacillus anthracis (anthrax spores in pure culture or in environmental samples are lacking. Here, an amperometric immunoassay has been developed utilizing immunomagnetic separation to capture the spores and remove potential interferents from test samples followed by amperometric measurement on a field-portable instrument. Antibody-conjugated magnetic beads and antibody-conjugated glucose oxidase were used in a sandwich format for the capture and detection of target spores. Glucose oxidase activity of spore pellets was measured indirectly via amperometry by applying a bias voltage after incubation with glucose, horseradish peroxidase, and the electron mediator 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid. Target capture was mediated by polyclonal antisera, whereas monoclonal antibodies were used for signal generation. This strategy maximized sensitivity (500 target spores, 5000 cfu/mL, while also providing a good specificity for Bacillus anthracis spores. Minimal signal deviation occurs in the presence of environmental interferents including soil and modified pH conditions, demonstrating the strengths of immunomagnetic separation. The simultaneous incubation of capture and detection antibodies and rapid substrate development (5 min result in short sample-to-signal times (less than an hour. With attributes comparable or exceeding that of ELISA and LFDs, amperometry is a low-cost, low-weight, and practical method for detecting anthrax spores in the field.

A Clostridium difficile-Specific, Gel-Forming Protein Required for Optimal Spore Germination

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M. Lauren Donnelly

2017-01-01

Full Text Available Clostridium difficile is a Gram-positive spore-forming obligate anaerobe that is a leading cause of antibiotic-associated diarrhea worldwide. In order for C. difficile to initiate infection, its aerotolerant spore form must germinate in the gut of mammalian hosts. While almost all spore-forming organisms use transmembrane germinant receptors to trigger germination, C. difficile uses the pseudoprotease CspC to sense bile salt germinants. CspC activates the related subtilisin-like protease CspB, which then proteolytically activates the cortex hydrolase SleC. Activated SleC degrades the protective spore cortex layer, a step that is essential for germination to proceed. Since CspC incorporation into spores also depends on CspA, a related pseudoprotease domain, Csp family proteins play a critical role in germination. However, how Csps are incorporated into spores remains unknown. In this study, we demonstrate that incorporation of the CspC, CspB, and CspA germination regulators into spores depends on CD0311 (renamed GerG, a previously uncharacterized hypothetical protein. The reduced levels of Csps in gerG spores correlate with reduced responsiveness to bile salt germinants and increased germination heterogeneity in single-spore germination assays. Interestingly, asparagine-rich repeat sequences in GerG’s central region facilitate spontaneous gel formation in vitro even though they are dispensable for GerG-mediated control of germination. Since GerG is found exclusively in C. difficile, our results suggest that exploiting GerG function could represent a promising avenue for developing C. difficile-specific anti-infective therapies.

The effect of Mirabilis jalapa leaves biopesticide treatment on the mycelium growth of entomopathogenic fungi Beauveria bassiana inside the larvae body Crocidolomia binotalis

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Pramita, Mia; Anggraeni, Tjandra

2015-09-01

Pest control with biological method (biopesticide and entomopathogenic fungi) is an alternative program to reduce application of chemical insecticide. Biopesticide of Mirabilis jalapa leaves has been discovered rich in secondary metabolites which has antifeedant activity that can provide physiological interference in insect larvae and the generation numbers[1]. Entomopathogenic fungi Beauveria bassiana has potential to control pest populations[2]. The growth of mycelium B. bassiana may interfere metabolism process inside the host body. Otherwise, B. bassiana produce toxins such as beauvericin that can increase mortality of pest. Combination of M. jalapa and B. bassiana reduce LT50 on C. binotalis larvae[3]. Thus, this study aims to determine influence of provision of biopesticide M. jalapa leaves on growth of mycelium entomopathogenic fungi B. bassiana inside larvae body C. binotalis and to detect the presence of beauvericin in vivo. Third instar larvae of C. binotalis were divided into a control, fungal and combination group. The combination group was given biopesticide and fungi. The concentration of biopesticide was 0.8% (w/v) and concentration of fungi spores was 107 spores/ml. Spores (vol. 5µl) done topically to larvae in interval 6 hours after treatment of biopesticide on non-pesticide cabbage leaves. Afterwards, histological observations performed at 24, 48, 72, 96 hours after treatment. The result show of emergence hyphae and mycelium growth inside lumen of larvae midgut on combination group faster than fungal group. This is thought to be caused by the influence of secondary metabolites of biopesticide M. jalapa leaves. In addition, beauviricin is detectable both of fungal and combination group. Thus, it can be concluded that treatment of biopesticide from M. jalapa leaves can accelerate on growth of mycelium entomopathogenic fungi B. bassiana inside the larvae body C. binotalis and toxic of B. bassiana such as beauvericin was detected on fungal and

In vitro propagation of Cyathea atrovirens (Cyatheaceae: spore storage and sterilization conditions

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Isabel Beatriz de Vargas

2014-03-01

Full Text Available Cyathea atrovirens occurs in a wide range of habitats in Brazil, Paraguay, Uruguay and Argentina. In the Brazilian State of Rio Grande do Sul, this commonly found species is a target of intense exploitation, because of its ornamental characteristics. The in vitro cultura is an important tool for propagation which may contribute toward the reduction of extractivism. However, exogenous contamination of spores is an obstacle for the success of aseptic long-term cultures. This study evaluated the influence of different sterilization methods combined with storage conditions on the contamination of the in vitro cultures and the gametophytic development of C. atrovirens, in order to establish an efficient propagation protocol. Spores were obtained from plants collected in Novo Hamburgo, State of Rio Grande do Sul, Brazil. In the first experiment, spores stored at 7oC were surface sterilized with 0.5, 0.8 and 2% of sodium hypochlorite (NaClO for 15 minutes and sown in Meyer’s culture medium. The cultures were maintained in a growth room at 26±1ºC for a 12-h photoperiod and photon flux density of 100μmol/m²/s provided by cool white fluorescent light. Contamination was assessed at 60 days, and gametophytic development was scored at 30, 60, 120 and 130 days of in vitro culture, analyzing 300 individuals for each treatment. There was no significant difference in culture contamination among the different sodium hypochlorite concentrations tested, and all treatments allowed for the development of cordiform gametophytes at 130 days of culture. In the second experiment, spores stored at 7 and -20°C were divided into two groups. Half of the spores were surface sterilized with 2% of NaClO for 15 minutes and the other half was not sterilized. All spores were sown in Meyer’s medium supplemented with one of the following antibiotics: nystatin, Micostatin® and actidione. The culture conditions and the procedures used for evaluating contamination and

Aerobiology of the built environment: Synergy between Legionella and fungi.

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Alum, Absar; Isaacs, Galahad Zachariah

2016-09-02

The modern built environment (BE) design creates unique ecological niches ideal for the survival and mutual interaction of microbial communities. This investigation focused on the synergistic relations between Legionella and the fungal species commonly found in BEs and the impact of these synergistic relationships on the survival and transmission of Legionella. A field study was conducted to identify the types and concentrations of fungi in BEs. The fungal isolates purified from BEs were cocultured with Legionella to study their synergistic association. Cocultured Legionella cells were aerosolized in an air-tight chamber to evaluate the efficacy of ultraviolet (UV) to inactivate these cells. Aspergillus, Alternaria, and Cladosporium were the most common fungi detected in samples that tested positive for Legionella. After coculturing, Legionella cells were detected inside fungal hyphae. The microscopic observations of Legionella internalization in fungal hyphae were confirmed by molecular analyses. UV disinfection of the aerosolized Legionella cells that were cocultured with fungi indicated that fungal spores and propagules act as a shield against UV radiation. The shield effect of fungal spores on Legionella cells was quantified at >2.5 log10. This study provides the first evidence, to our knowledge, of Legionella cell presence inside fungi detected in an indoor environment. This symbiotic relationship with fungi results in longer survival of Legionella under ambient conditions and provides protection against UV rays. Copyright © 2016. Published by Elsevier Inc.