Assessment of the aerosolization potential for fungal spores in moldy homes

The airborne fungal concentration measured with air samplers during specific time intervals may not adequately represent the indoor air quality because of the sporadic nature of spore release from sources. The conventional source evaluation (e.g. swab and tape sampling) characterizes the mold source but does not relate to the fraction of spores that can be aerosolized from a contaminated material. As an alternative to these methods, we have recently developed and laboratory-tested a novel Fungal Spore Source Strength Tester (FSSST). It allows assessing the potential of aerosolization of fungal spores from contaminated surfaces under the most favorable release conditions. In this study, the FSSST was used to characterize the release of spores from four building materials in mold-problem homes. The spores of different species were efficiently aerosolized by the FSSST, exhibiting a total spore release rate ranging approximately from 10(2) to 10(3) cm2/min. For all tested materials, <2% of the spores on the contaminated surface were released during the tests. The airborne spore concentration estimated from the release rate data was found in most cases to be significantly greater than the concentration actually measured in these environments with simultaneous air sampling. The results suggest that the FSSST can be used for the assessment of maximum potential exposure to airborne spores released from identified sources in homes. PRACTICAL IMPLICATIONS: A recently developed FSSST was found to be suitable to measure the aerosolization potential of indoor fungal sources at the most favorable release conditions. The FSSST generates the data that allows assessing the strength of mold sources in homes with respect to their maximum ability to contaminate indoor air with fungi. The novel approach bridges two conventional methods, the air sampling and the direct source evaluation (e.g. swab sampling), thus providing a better representation of the airborne fungal exposure than these methods individually. The device prototype can be used for evaluating the effectiveness of environmental interventions by taking samples before and after the intervention. As a broader application, the FSSST can be utilized for assessing the release of various hazardous biological and non-biological particles from contaminated surfaces.     

Determination of fungal spore release from wet building materials

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 of fungal spores was induced by well-defined jets of air impacting from rotating nozzles. The spores and other particles released from the surface were transported by the air flowing from the chamber through a top outlet to a particle counter and sizer. For two of the fungi (Penicillium chrysogenum and Trichoderma harzianum), the number of spores produced on the gypsum board and subsequently released was quantified. Also the relationship between air velocities from 0.3 to 3 m/s over the surface and spore release has been measured. The method was found to give very reproducible results for each fungal isolate, whereas the spore release is very different for different fungi under identical conditions. Also, the relationship between air velocity and spore release depends on the fungus. For some fungi a significant number of particles smaller than the spore size were released. The method applied in the study may also be useful for field studies and for generation of spores for exposure studies.     

甘肃省文县石坊群孢子化石的发现及地质意义

在测制甘肃文县地区石坊群地层剖面中发现了孢子化石,共计14个属,56个种,将其与贵州独山、都匀地 区的早、中泥盆世孢子化石组合对比,发现组合特征基本一致。因而确定了石坊群地层时代属早、中泥盆世,为 石坊群地层划分、时代确定及地质构造发展史研究提供了可靠的依据。     

Strategy to inactivate Clostridium perfringens spores in meat products

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 °C, 10 min); spore germination at high temperatures (55, 60 or 65 °C); and inactivation of germinated spores with elevated temperatures (80 and 90 °C, 10 and 20 min) and high pressure (586 MPa, at 23 and 73 °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 °C, 10 min), and germinated at temperatures lethal for vegetative cells (≥55 °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 (∼4 decimal reduction) in meat by elevated temperatures (80–90 °C for 20 min) required a long germination period (55 °C for 60 min). However, similar inactivation level was reached with shorter germination period (55 °C for 15 min) when spore contaminated-meat was treated with pressure-assisted thermal processing (568 MPa, 73 °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 °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 °C in about 20 min and further incubation at 55 °C for about 15 min for spore germination; and (iii) inactivation of germinated spores by pressure-assisted thermal processing (586 MPa at 73 °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. perfringens.     

Phylogenetic and toxinogenic characteristics of Bacillus cereus group members isolated from spices and herbs

The foodborne pathogen Bacillus (B.) cereus is a common contaminant in spices and herbs. To further characterise B. cereus and its closely related group members present in spices and herbs, we analysed presumptive B. cereus strains isolated from six different condiments with view to B. cereus group species, phylogenetic affiliation and toxinogenic potential.Of a total of 59 isolates 44 were identified as B. cereus sensu stricto (s.s.), four as B. toyonensis-like, five as B. thuringiensis, one as B. weihenstephanensis, two as B. pseudomycoides/B. mycoides and three as undefined B. cereus group species. A maximum of three different species occurred simultaneously in the same spice sample. The isolates comprised 33 multilocus (ML) sequence types (STs), which can be assigned to three different phylogenetic groups. Except two B. pseudomycoides/B. mycoides strains, all isolates were able to produce enterotoxins and one strain the emetic toxin cereulide as detected by an immunoassay and LC-MS, respectively. The prevalence of toxin genes was 96.6% for nheA, 94.9% for hblD, 50.8% for cytK-2 and 1.7% for ces. The emetic strain was characterised by ST 869, which for the first time was assigned to an emetic B. cereus (s.s.) strain and is not part of the previously known two emetic MLST clusters.Our results demonstrate that not only B. cereus (s.s.) but also toxin producing B. thuringiensis, B. weihenstephanensis and B. toyonensis-like strains could be detected in condiments. For some isolates MLST revealed disagreements between phylogenetic relationship and the classification as B. weihenstephanensis and B. mycoides based on previously described species markers.     

Antagonistic action of Bacillus subtilis strain fmbj on the postharvest pathogen Rhizopus stolonifer

Abstract: The antagonistic activities of Bacillus subtilis fmbj against the Rhizopus stolonifer pathogen causing peach soft rot disease were studied in this paper. Bacillus subtilis fmbj exhibited a high antifungal effect on the mycelium growth, sportulation, and germ tube elongation of R. stolonifer with the inhibition rate of 86.5% and 97.42%, respectively, spore germination rate of 20% under antagonist strain concentration of 10⁸ CFU/mL. By using of scanning electron microscope (SEM) and transmission electron microscope (TEM), it was observed that B. subtilis fmbj strain strongly induced morphological abnormalities of R. stolonifer and destroyed structure of hypha and spore. When hypha cell wall of R. stolonifer was damaged, the organelles and cytoplasms inside cell would exude, which led to cell death.     

Efficient production of fungal spores by the combination of reduction of nitrogen source content and embedding of hydrophobic polymer in an agar plate

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Effect of Ellman's Reagent and Dithiothreitol on the Curing of the Spore Adhesive Glycoprotein of the Green Alga Ulva

Green algae of the genus Ulva (syn. Enteromorpha) are common, green macroalgae found throughout the world in the upper intertidal zone of seashores and as a fouling organism on a variety of man-made structures including ships' hulls. Adhesion of motile spores is achieved via the secretion of an adhesive, which is present in spores in highly condensed form within membrane-bound vesicles. The adhesive is initially liquid and displays a hydrogel-like behavior on release. It then starts to undergo “curing reactions,” becoming progressively less soluble with time in anionic detergents, less sensitive to proteolysis, and less viscoelastic, which suggests that extensive cross-linking occurs. Spores also become progressively more difficult to detach from a surface. However, the nature of this adhesive curing process is totally unknown. In the present article we have tested the hypothesis that thiol cross-linking may be involved. We show that nontoxic concentrations of the thiol-capping reagent (Ellman's reagent) or thiol-reducing agent (dithiothreitol) effectively inhibit the time-dependent development of adhesive spore strength after attachment to a surface. Furthermore, we show by SDS-PAGE immunoblot analysis of extracted adhesive proteins that the major adhesive antigen retains solubility in the presence of these reagents, after release from spores, which suggests that cross-linking had been inhibited.     

The impact of microwave heating of infant formula model on neo-formed contaminant formation, nutrient degradation and spore destruction

Heat treatment of milk results in the development of a large range of Maillard products, especially in infant formulas due to their high content in Maillard reaction substrates. The Maillard reaction should be minimized because some of those products, called neo-formed contaminants (NFC), are currently suspected of promoting deleterious health effects. The objective of this work was to study the impact of microwave heating on the formation of Maillard compounds, degradation of nutrients and destruction of spores of an infant formula model. An infant formula model based on pure-whey proteins was used as experimental model. A spore suspension of Geobacillus stearothermophilus was prepared and inoculated in the formulated milk before microwave treatment. The treatment was performed on a laboratory device running at 2450 MHz and equipped with a magnetron (power range 0-1860 W) and the temperature was controlled by three optical fibers. Heating kinetics were established at several microwave specific powers (5, 7, 10, 12 W/mL). The impact of the treatment on NFC formation was evaluated by 3 indicators: the FAST index (indicator of global advancement of Maillard reaction), carboxymethyllysine (CML) and furosine. All these NFC indicators seemed to follow an exponential model as a function of time. The thermal sensitive nutrient vitamin C was chosen as indicator of the nutritional quality. Like in conventional heating, an exponential model was observed for spore inactivation and vitamin C degradation. Furthermore, laws similar to those used for conventional heating could be used to express heat resistance regarding germ viability, vitamin C concentration and Maillard products. As a result, D and z-values relative to specific microwave powers were determined for each indicator. Finally, it was found that the best way to minimize NFC formation and nutrient degradation was to use high specific power for a short time, similar to what is obtained with the UHT process for conventional heating.     

Concentrations of viable airborne fungal spores and trichloroanisole in wine cellars

In wineries, unwanted microorganisms present not only hygienic problems but also have a negative influence on wine quality. An evaluation of Austrian/Styrian wine cellars with regard to the volume and the composition of the mycoflora is very important both for the process of wine production and for occupational safety.Thirty-six wine cellars of 20 vintners were investigated with regard to microorganisms in the air and on material surfaces. Moreover, the presence of trichloroanisole in the air was determined by means of solid-phase micro-extraction. Microorganisms were sampled using the six-stage Andersen-Cascade impactor.The results showed that the concentrations of xerophilic fungi in the air of cellars with large visible mold areas (> 80%) reached values up to 1.4 × 10⁴ colony forming units per m³. In the wine cellars fourteen predominant fungal genera were found in the indoor air, the most frequent was Penicillium. Trichloroanisole was detected in the air of wine cellars with large visible moldy patches. The spore concentrations in the cellar air were two times higher in cellars with Zasmidium cellare growth than in cellars without Z. cellare.These results will serve as a database for further studies.