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Sample records for cereus spore coat

  1. Architecture and High-Resolution Structure of Bacillus thuringiensis and Bacillus cereus Spore Coat Surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T; Wheeler, K; Malkin, A

    2005-02-18

    We have utilized atomic force microscopy (AFM) to visualize the native surface topology and ultrastructure of Bacillus thuringiensis and Bacillus cereus spores in water and in air. AFM was able to resolve the nanostructure of the exosporium and three distinctive classes of appendages. Removal of the exosporium exposed either a hexagonal honeycomb layer (B. thuringiensis) or a rodlet outer spore coat layer (B. cereus). Removal of the rodlet structure from B. cereus spores revealed an underlying honeycomb layer similar to that observed with B. thuringiensis spores. The periodicity of the rodlet structure on the outer spore coat of B. cereus was {approx}8 nm, and the length of the rodlets was limited to the cross-patched domain structure of this layer to {approx}200 nm. The lattice constant of the honeycomb structures was {approx}9 nm for both B. cereus and B. thuringiensis spores. Both honeycomb structures were composed of multiple, disoriented domains with distinct boundaries. Our results demonstrate that variations in storage and preparation procedures result in architectural changes in individual spore surfaces, which establish AFM as a useful tool for evaluation of preparation and processing ''fingerprints'' of bacterial spores. These results establish that high-resolution AFM has the capacity to reveal species-specific assembly and nanometer scale structure of spore surfaces. These species-specific spore surface structural variations are correlated with sequence divergences in a spore core structural protein SspE.

  2. Film coating of seeds with Bacillus cereus RS87 spores for early plant growth enhancement.

    Science.gov (United States)

    Jetiyanon, Kanchalee; Wittaya-Areekul, Sakchai; Plianbangchang, Pinyupa

    2008-10-01

    The plant growth-promoting rhizobacterium Bacillus cereus RS87 was previously reported to promote plant growth in various crops in both greenhouse and field trials. To apply as a plant growth promoting agent with practical use, it is essential to ease the burden of routine preparation of a fresh suspension of strain RS87 in laboratory. The objectives of this study were to investigate the feasibility of film-coating seeds with B. cereus RS87 spores for early plant growth enhancement and to reveal the indoleacetic acid (IAA) production released from strain RS87. The experiment consisted of the following 5 treatments: nontreated seeds, water-soaked seeds, film-coated seeds, seeds soaked with vegetative cells of strain RS87, and film-coated seeds with strain RS87 spores. Three experiments were conducted separately to assess seed emergence, root length, and plant height. Results showed that both vegetative cells and spores of strain RS87 significantly promoted (P seed emergence, root length and plant height over the control treatments. The strain RS87 also produced IAA. In conclusion, the film coating of seeds with spores of B. cereus RS87 demonstrated early plant growth enhancement as well as seeds using their vegetative cells. IAA released from strain RS87 would be one of the mechanisms for plant growth enhancement.

  3. 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.

  4. Inactivation of vegetative cells, but not spores, of Bacillus anthracis, B. cereus, and B. subtilis on stainless steel surfaces coated with an antimicrobial silver- and zinc-containing zeolite formulation.

    Science.gov (United States)

    Galeano, Belinda; Korff, Emily; Nicholson, Wayne L

    2003-07-01

    Stainless steel surfaces coated with paints containing a silver- and zinc-containing zeolite (AgION antimicrobial) were assayed in comparison to uncoated stainless steel for antimicrobial activity against vegetative cells and spores of three Bacillus species, namely, B. anthracis Sterne, B. cereus T, and B. subtilis 168. Under the test conditions (25 degrees C and 80% relative humidity), the zeolite coating produced approximately 3 log(10) inactivation of vegetative cells within a 5- to 24-h period, but viability of spores of the three species was not significantly affected.

  5. An Optical Biosensor for Bacillus Cereus Spore Detection

    Science.gov (United States)

    Li, Chengquan; Tom, Harry W. K.

    2005-03-01

    We demonstrate a new transduction scheme for optical biosensing. Bacillus cereus is a pathogen that may be found in food and dairy products and is able to produce toxins and cause food poisoning. It is related to Bacillus anthracis (anthrax). A CCD array covered with micro-structured glass coverslip is used to detect the optical resonant shift due to the binding of the antigen (bacillus cereus spore) to the antibody (polyclonal antibody). This novel optical biosensor scheme has the potential for detecting 10˜100 bioagents in a single device as well as the potential to test for antigens with multiple antibody tests to avoid ``false positives.''

  6. Bacillus cereus spores and cereulide in food-borne illness

    OpenAIRE

    Shaheen, Ranad

    2009-01-01

    B. cereus is a gram-positive bacterium that possesses two different forms of life:the large, rod-shaped cells (ca. 0.002 mm by 0.004 mm) that are able to propagate and the small (0.001 mm), oval shaped spores. The spores can survive in almost any environment for up to centuries without nourishment or water. They are insensitive towards most agents that normally kill bacteria: heating up to several hours at 90 ºC, radiation, disinfectants and extreme alkaline (≥ pH 13) and acid (≀ pH 1) e...

  7. Inhibition of Bacillus cereus spore outgrowth and multiplication by chitosan.

    Science.gov (United States)

    Mellegård, Hilde; From, Cecilie; Christensen, Bjørn E; Granum, Per E

    2011-10-03

    Bacillus cereus is an endospore-forming bacterium able to cause food-associated illness. Different treatment processes are used in the food industry to reduce the number of spores and thereby the potential of foodborne disease. Chitosan is a polysaccharide with well-documented antibacterial activity towards vegetative cells. The activity against bacterial spores, spore germination and subsequent outgrowth and growth (the latter two events hereafter denoted (out)growth), however, is poorly documented. By using six different chitosans with defined macromolecular properties, we evaluated the effect of chitosan on Bacillus cereus spore germination and (out)growth using optical density assays and a dipicolinic acid release assay. (Out)growth was inhibited by chitosan, but germination was not. The action of chitosan was found to be concentration-dependent and also closely related to weight average molecular weight (M(w)) and fraction of acetylation (F(A)) of the biopolymer. Chitosans of low acetylation (F(A)=0.01 or 0.16) inhibited (out)growth more effectively than higher acetylated chitosans (F(A)=0.48). For the F(A)=0.16 chitosans with medium (56.8kDa) and higher M(w) (98.3kDa), a better (out)growth inhibition was observed compared to low M(w) (10.6kDa) chitosan. The same trend was not evident with chitosans of 0.48 acetylation, where the difference in activity between the low (19.6kDa) and high M(w) (163.0kDa) chitosans was only minor. In a spore test concentration corresponding to 10(2)-10(3)CFU/ml (spore numbers relevant to food), less chitosan was needed to suppress (out)growth compared to higher spore numbers (equivalent to 10(8)CFU/ml), as expected. No major differences in chitosan susceptibility between three different strains of B. cereus were detected. Our results contribute to a better understanding of chitosan activity towards bacterial spore germination and (out)growth.

  8. Characterization of a spore-specific protein of the Bacillus cereus group

    NARCIS (Netherlands)

    From, C.; Voort, van der M.; Abee, T.; Granum, P.E.

    2012-01-01

    Bc1245 is a monocistronic chromosomal gene of Bacillus cereus ATCC 14579 encoding a putative protein of 143 amino acids identified in this study to have a spore-related function in B. cereus. Bc1245 is highly conserved in the genome of members of the B. cereus group, indicating an important function

  9. Soya bean tempe extracts show antibacterial activity against Bacillus cereus cells and spores

    NARCIS (Netherlands)

    Roubos-van den Hil, P.J.; Dalmas, E.; Nout, M.J.R.; Abee, T.

    2010-01-01

    Aims: Tempe, a Rhizopus ssp.-fermented soya bean food product, was investigated for bacteriostatic and/or bactericidal effects against cells and spores of the food-borne pathogen Bacillus cereus. Methods and results: Tempe extract showed a high antibacterial activity against B. cereus ATCC 14579 bas

  10. Spore coat architecture of Clostridium novyi NT spores.

    Science.gov (United States)

    Plomp, Marco; McCaffery, J Michael; Cheong, Ian; Huang, Xin; Bettegowda, Chetan; Kinzler, Kenneth W; Zhou, Shibin; Vogelstein, Bert; Malkin, Alexander J

    2007-09-01

    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. Toward this end, we have used both transmission electron microscopy and atomic force microscopy to determine the structure of both dormant and 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.

  11. Germination and outgrowth of spores of Bacillus cereus group members: diversity and role of germinant receptors.

    Science.gov (United States)

    Abee, Tjakko; Groot, Masja Nierop; Tempelaars, Marcel; Zwietering, Marcel; Moezelaar, Roy; van der Voort, Menno

    2011-04-01

    Bacillus cereus is a gram-positive, facultative anaerobic, endospore-forming toxicogenic human pathogen. Endospores are highly specialized, metabolically dormant cell types that are resistant to extreme environmental conditions, including heat, dehydration and other physical stresses. B. cereus can enter a range of environments, and can in its spore form, survive harsh conditions. If these conditions become favorable, spores can germinate and grow out and reach considerable numbers in a range of environments including processed foods. Certainly the last decade, when consumer preferences have shifted to mildly processed food, new opportunities arose for spore-forming spoilage and pathogenic organisms. Only rigorous methods have been shown to be capable of destroying all spores present in food, thus a shift toward e.g., milder heat preservation strategies, may result in low but significant amounts of viable spores in food products. Hence, the need for a mild spore destruction strategy is eminent including control of spore outgrowth. Consequently, there is a large interest in triggering spore germination in foodstuffs, since germinated spores have lost the extreme resistance of dormant spores and are relatively easy to kill. Another option could be to prevent germination so that no dangerous levels can be reached. This contribution will focus on germination and outgrowth characteristics of B. cereus and other members of the B. cereus group, providing an overview of the niches these spore-formers can occupy, the signals that trigger germination, and how B. cereus copes with these wake-up calls in different environments including foods, during food processing and upon interaction with the human host.

  12. 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.

  13. Persistence strategies of Bacillus cereus spores isolated from dairy silo tanks.

    Science.gov (United States)

    Shaheen, Ranad; Svensson, Birgitta; Andersson, Maria A; Christiansson, Anders; Salkinoja-Salonen, Mirja

    2010-05-01

    Survival of Bacillus cereus spores of dairy silo tank origin was investigated under conditions simulating those in operational dairy silos. Twenty-three strains were selected to represent all B. cereus isolates (n = 457) with genotypes (RAPD-PCR) that frequently colonised the silo tanks of at least two of the sampled eight dairies. The spores were studied for survival when immersed in liquids used for cleaning-in-place (1.0% sodium hydroxide at pH 13.1, 75 degrees C; 0.9% nitric acid at pH 0.8, 65 degrees C), for adhesion onto nonliving surfaces at 4 degrees C and for germination and biofilm formation in milk. Four groups with different strategies for survival were identified. First, high survival (log 15 min kill steel from cold water. Third, a cereulide producing group with spores characterised by slow germination in rich medium and well preserved viability when exposed to heating at 90 degrees C. Fourth, spores capable of germinating at 8 degrees C and possessing the cspA gene. There were indications that spores highly resistant to hot 1% sodium hydroxide may be effectively inactivated by hot 0.9% nitric acid. Eight out of the 14 dairy silo tank isolates possessing hot-alkali resistant spores were capable of germinating and forming biofilm in whole milk, not previously reported for B. cereus.

  14. Fate of pathogenic Bacillus cereus spores after ingestion by protist grazers

    DEFF Research Database (Denmark)

    Winding, Anne; Santos, Susana; Hendriksen, Niels Bohse

    was initially investigated in microcosms inoculated with pure cultures of the protists Acanthamoeba castellanii, Tetrahymena pyriformis and Cercomonas sp. as grazers. Individual protist cultures were fed with fluorescently labelled (CellTracker™RedCMTPX) B. cereus spores or vegetative cells as the only food...

  15. Quantification of the impact of single and multiple mild stresses on outgrowth heterogeneity of Bacillus cereus spores

    NARCIS (Netherlands)

    Melis, van C.C.J.; Besten, den H.M.W.; Nierop Groot, M.N.; Abee, T.

    2014-01-01

    Outgrowth heterogeneity of bacterial spore populations complicates both prediction and efficient control of spore outgrowth. In this study, the impact of mild preservation stresses on outgrowth of Bacillus cereus ATCC 14579 spores was quantified during the first stages of outgrowth. Heterogeneity in

  16. Growth/no growth models for heat-treated psychrotrophic Bacillus cereus spores under cold storage.

    Science.gov (United States)

    Daelman, Jeff; Vermeulen, An; Willemyns, Tine; Ongenaert, Rebecca; Jacxsens, Liesbeth; Uyttendaele, Mieke; Devlieghere, Frank

    2013-01-15

    The microbiological safety of refrigerated and processed foods of extended durability (REPFED) is linked to spore-forming pathogens, more specifically Clostridium botulinum and Bacillus cereus. In this study two sets of growth/no growth (GNG) models are presented for the spores of two B. cereus strains. The models incorporate both product (water activity (a(w)) and pH) and process parameters (pasteurization value at 90 °C (P(90)) or heating temperature). The first model evaluates the effect of four different P(90)-values (P(90)=0, 4, 7 or 10 min, all applied at 90 °C) on the germination and subsequent growth of B. cereus spores under different conditions of pH and a(w) at 10 °C. These models show that a heat treatment not only increases the time to growth (TTG), but also significantly increases the minimal a(w) and pH necessary for germination and subsequent growth: e.g. at a(w) 0.995 and without heat treatment (P(90)=0), strain FF355 B. cereus spores were predicted to germinate and grow at pH 5.3. With a P(90) of 10 min, the minimal pH increased to 5.7. The second set of models for B. cereus spores compares the effect of three heat treatments with the same P(90)-value (10 min) but applied at different temperatures (85, 87 and 90 °C), on the germination and subsequent growth at 10 °C. The second model shows that lower heating temperatures (85 and 87 °C) had less effect on the TTG and minimal a(w) and pH than a higher temperature (90 °C). Finally, the first set of models was validated in broth using spores of seven psychrotrophic B. cereus strains, to evaluate the effect of strain variability on the model predictions. The results of the validation (% growth) were compared to the predicted growth probability. The results showed that the models were prone to fail-dangerous results (i.e. predicting no growth when growth was observed: 17%-34%). Using a very low threshold for growth (0.1% predicted chance of growth was considered to be complete growth), the models

  17. Architecture and Assembly of the Bacillus subtilis Spore Coat

    Science.gov (United States)

    2014-09-26

    icandy contaminated with germinated spores and these germinat ed spores were removed by centrifugation in a one step HistodenzTM (Sigma, St. Louis...spore resistance but also because some coat proteins play significant roles in spore germination . However, much recent work on the spore coat has... germinating spores of various Bacillus [14,21 30] and Clostridium [3 1] species. H owever, this analysis has generally been conducted on wild type

  18. Efficacy of gaseous chlorine dioxide in inactivating Bacillus cereus spores attached to and in a biofilm on stainless steel.

    Science.gov (United States)

    Nam, Hyegyeong; Seo, Hyun-Sun; Bang, Jihyun; Kim, Hoikyung; Beuchat, Larry R; Ryu, Jee-Hoon

    2014-10-01

    We evaluated the lethal activity of gaseous chlorine dioxide (ClO2) against Bacillus cereus spores attached to and in biofilm formed on a stainless steel surface. Aqueous ClO2 was prepared by mixing sulfuric acid (5% w/v) with sodium chlorite (10mg/mL), and gaseous ClO2 was produced by vaporization of aqueous ClO2 in an air-tight container. The concentration of gaseous ClO2 in the air within the container increased rapidly at first but gradually decreased over time. The lethality of gaseous ClO2 against B. cereus spores attached to stainless steel coupons (SSCs) and in biofilm formed by the pathogen on SSCs was determined. The B. cereus spores attached to SSCs (5.3±0.1logCFU/coupon) were completely inactivated within 1h at 25°C when treated with gaseous ClO2 (peak concentration: 115.3±5.0 parts per million [ppm]). The total number of vegetative cells and spores in biofilm formed by B. cereus on SSCs was 5.9±0.3logCFU/coupon; the spore count was 5.3±0.1logCFU/coupon. The vegetative cells and spores in biofilm were completely inactivated within 6h (peak concentration: 115.3±5.0ppm). Results show that B. cereus spores in biofilms are more resistant to gaseous ClO2 than are attached spores not in biofilms. Gaseous ClO2 was, nevertheless, very effective in killing B. cereus spores in biofilm on the surface of stainless steel. Results show promise for application of gaseous ClO2 to enhance the microbiological safety of foods that may come in contact with stainless steel and possibly other hard surfaces on which B. cereus biofilms have formed.

  19. Study of the antibacterial effects of chitosans on Bacillus cereus (and its spores) by atomic force microscopy imaging and nanoindentation

    Energy Technology Data Exchange (ETDEWEB)

    Fernandes, Joao C. [Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Rua Dr. Antonio Bernardino de Almeida, 4200-072 Porto (Portugal); Eaton, Peter, E-mail: peter.eaton@fc.up.pt [REQUIMTE, Departamento de Quimica, Faculdade de Ciencias da Universidade do Porto, Rua do Campo Alegre, 4169-007 Porto (Portugal); Gomes, Ana M.; Pintado, Manuela E.; Xavier Malcata, F. [Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Rua Dr. Antonio Bernardino de Almeida, 4200-072 Porto (Portugal)

    2009-07-15

    Bacillus cereus is a Gram-positive, spore-forming bacterium that is widely distributed in nature. Its intrinsic thermal resistance coupled with the extraordinary resistance against common food preservation techniques makes it one of the most frequent food-poisoning microorganisms causing both intoxications and infections. In order to control B. cereus growth/sporulation, and hence minimize the aforementioned hazards, several antimicrobial compounds have been tested. The aim of this work was to assess by atomic force microscopy (AFM) the relationship between the molecular weight (MW) of chitosan and its antimicrobial activity upon both vegetative and resistance forms of B. cereus. The use of AFM imaging studies helped us to understand how chitosans with different MW act differently upon B. cereus. Higher MW chitosans (628 and 100 kDa) surrounded both forms of B. cereus cells by forming a polymer layer-which eventually led to the death of the vegetative form by preventing the uptake of nutrients yet did not affect the spores since these can survive for extended periods without nutrients. Chitooligosaccharides (COS) (<3 kDa), on the other hand, provoked more visible damages in the B. cereus vegetative form-most probably due to the penetration of the cells by the COS. The use of COS by itself on B. cereus spores was not enough for the destruction of a large number of cells, but it may well weaken the spore structure and its ability to contaminate, by inducing exosporium loss.

  20. Sporulation environment of emetic toxin-producing Bacillus cereus strains determines spore size, heat resistance and germination capacity

    NARCIS (Netherlands)

    Voort, van der M.; Abee, T.

    2013-01-01

    Aim Heat resistance, germination and outgrowth capacity of Bacillus cereus spores in processed foods are major factors in causing the emetic type of gastrointestinal disease. In this study, we aim to identify the impact of different sporulation conditions on spore properties of emetic toxin-producin

  1. Spore prevalence and toxigenicity of Bacillus cereus and Bacillus thuringiensis isolates from U.S. retail spices.

    Science.gov (United States)

    Hariram, Upasana; Labbé, Ronald

    2015-03-01

    Recent incidents of foodborne illness associated with spices as the vehicle of transmission prompted this examination of U.S. retail spices with regard to Bacillus cereus. This study focused on the levels of aerobic-mesophilic spore-forming bacteria and B cereus spores associated with 247 retail spices purchased from five states in the United States. Samples contained a wide range of aerobic-mesophilic bacterial spore counts ( 10(7) CFU/g). Using a novel chromogenic agar, B. cereus and B. thuringiensis spores were isolated from 77 (31%) and 11 (4%) samples, respectively. Levels of B. cereus were thuringiensis isolates possessed at least one type of enterotoxin gene: HBL (hemolysin BL) or nonhemolytic enterotoxin (NHE). None of the 88 isolates obtained in this study possessed the emetic toxin gene (ces). Using commercially available immunological toxin detection kits, the toxigenicity of the isolates was confirmed. The NHE enterotoxin was expressed in 98% of B. cereus and 91% of B. thuringiensis isolates that possessed the responsible gene. HBL enterotoxin was detected in 87% of B. cereus and 100% of B. thuringiensis PCR-positive isolates. Fifty-two percent of B. cereus and 54% of B. thuringiensis isolates produced both enterotoxins. Ninety-seven percent of B. cereus isolates grew at 12°C, although only two isolates grew well at 9°C. The ability of these spice isolates to form spores, produce diarrheal toxins, and grow at moderately abusive temperatures makes retail spices an important potential vehicle for foodborne illness caused by B. cereus strains, in particular those that produce diarrheal toxins.

  2. Synergistic effect of electrolyzed water and citric Acid against bacillus cereus cells and spores on cereal grains.

    Science.gov (United States)

    Park, Young Bae; Guo, Jin Yong; Rahman, S M E; Ahn, Juhee; Oh, Deog-Hwan

    2009-01-01

    The effects of acidic electrolyzed water (AcEW), alkaline electrolyzed water (AlEW), 100 ppm sodium hypochlorite (NaClO), and 1% citric acid (CA) alone, and combinations of AcEW with 1% CA (AcEW + CA) and AlEW with 1% CA (AlEW + CA) against Bacillus cereus vegetative cells and spores was evaluated as a function of temperature (25, 30, 40, 50, or 60 degrees C) and dipping time (3 or 6 h). A 3-strain cocktail of Bacillus cereus cells or spores of approximately 10(7) CFU/g was inoculated in various cereal grains (brown rice, Job's tear rice, glutinous rice, and barley rice). B. cereus vegetative cells and spores were more rapidly inactivated at 40 degrees C than at 25 degrees C. Regardless of the dipping time, all treatments reduced the numbers of B. cereus vegetative cells and spore by more than 1 log CFU/g, except the deionized water (DIW), which showed approximately 0.7 log reduction. The reductions of B. cereus cells increased with increasing dipping temperature (25 to 60 degrees C). B. cereus vegetative cells were much more sensitive to the combined treatments than spores. The effectiveness of the combined electrolyzed water (EW) and 1% CA was considerable in inhibiting B. cereus on cereal grains. The application of combined EW and CA for controlling B. cereus cells and spores on cereal grains has not been previously reported. Therefore, the synergistic effect of EW and CA may provide a valuable insight on reducing foodborne pathogens on fruits, vegetables, and cereal grains.

  3. Morphological and mechanical imaging of Bacillus cereus spore formation at the nanoscale.

    Science.gov (United States)

    Wang, Congzhou; Stanciu, Cristina; Ehrhardt, Christopher J; Yadavalli, Vamsi K

    2015-04-01

    Bacteria from the genus Bacillus are able to transform into metabolically dormant states called (endo) spores in response to nutrient deprivation and other harsh conditions. These morphologically distinct spores are fascinating constructs, amongst the most durable cells in nature, and have attracted attention owing to their relevance in food-related illnesses and bioterrorism. Observing the course of bacterial spore formation (sporulation) spatially, temporally and mechanically, from the vegetative cell to a mature spore, is critical for a better understanding of this process. Here, we present a fast and versatile strategy for monitoring both the morphological and mechanical changes of Bacillus cereus bacteria at the nanoscale using atomic force microscopy. Through a strategy of imaging and nanomechanical mapping, we show the morphogenesis of the endospore and released mature endospore. Finally, we investigate individual spores to characterize their surface mechanically. The progression in elasticity coupled with a similarity of characteristic distributions between the incipient endospores and the formed spores show these distinct stages. Taken together, our data demonstrates the power of atomic force microscopy applied in microbiology for probing this important biological process at the single cell scale.

  4. Gamma radiation effect on Bacillus cereus spores inoculated in black pepper; Efeitos da radiacao gama sobre esporos de Bacillus cereus inoculados em pimenta-do-reino

    Energy Technology Data Exchange (ETDEWEB)

    Froehlich, Angela; Axeredo, Raquel M.C.; Vanetti, Maria Cristina D. [Vicosa Univ., MG (Brazil). Dept. de Microbiologia; Villavicencio, Anna Lucia C. H. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail: villavic@net.ipen.br

    2000-07-01

    It had been analyzed 37 samples of worn out black pepper and in 85% of these samples was observed the presence of Bacillus cereus in numbers of up to 4,6 x 10{sup 4} UFC/g. The population of aerobic mesofilis bacteria varied of 2,8 x 10{sup 5} the 1,9 x 10{sup 8} UFC/g. The black pepper used during the experiment was evaluated, evidencing the aerobic presence of one aerobic mesofilis microbiota of, approximately, 2,6 x 10{sup 6} UFC/g, consisting, mainly, for species of the Bacillus sort. It was observed that the absence of B. cereus, coliforms, filamentous fungus and leavenings. The evaluation of the irradiation of the black pepper inoculated with 10{sup 6} UFC/g of B. cereus spores of with doses of gamma radiation varying between 2 and 10 kGy evidenced that doses up to 5 kGy had been enough to reduce the counting of, approximately, 10{sup 6} UFC/g of aerobic mesofilis organisms and 10{sup 4} UFC/g of B. cereus spores the not detectable numbers by the used methodology. The dose of reduction decimal (D{sub 10}) for the inoculated B. cereus spores in black pepper was of 1,78 kGy.

  5. Role of Spore Coat Proteins in the Resistance of Bacillus subtilis Spores to Caenorhabditis elegans Predation▿

    OpenAIRE

    2008-01-01

    Bacterial spores are resistant to a wide range of chemical and physical insults that are normally lethal for the vegetative form of the bacterium. While the integrity of the protein coat of the spore is crucial for spore survival in vitro, far less is known about how the coat provides protection in vivo against predation by ecologically relevant hosts. In particular, assays had characterized the in vitro resistance of spores to peptidoglycan-hydrolyzing enzymes like lysozyme that are also imp...

  6. Effects of Electrolyzed Oxidizing Water on Inactivation of Bacillus subtilis and Bacillus cereus Spores in Suspension and on Carriers.

    Science.gov (United States)

    Zhang, Chunling; Li, Baoming; Jadeja, Ravirajsinh; Hung, Yen-Con

    2016-01-01

    Spores of some Bacillus species are responsible for food spoilage and foodborne disease. These spores are highly resistant to various interventions and cooking processes. In this study, the sporicidal efficacy of acidic electrolyzed oxidizing (EO) water (AEW) and slightly acidic EO water (SAEW) with available chlorine concentration (ACC) of 40, 60, 80, 100, and 120 mg/L and treatment time for 1, 2, 3, 4, 5, and 6 min were tested on Bacillus subtilis and Bacillus cereus spores in suspension and on carrier with or without organics. The reduction of spore significantly increased with increasing ACC and treatment time (P waters containing 120 mg/L ACC, while only SAEW at 120 mg/L and 2 min treatment achieved >6 log reductions of B. subtilis spore. Both types of EO water with ACC of 60 mg/L and 6 min treatment achieved a reduction of B. subtilis and B. cereus spores to nondetectable level. EO water with ACC of 80 mg/L and treatment time of 3 min on carrier test without organics addition resulted in reductions of B. subtilis spore to nondetectable level. But, addition of 0.3% organics on carrier decreased the inactivation effect of EO water. This study indicated that EO water was highly effective in inactivation of B. subtilis and B. cereus spores in suspension or on carrier, and therefore, rendered it as a promising disinfectant to be applied in food industry.

  7. Infrared decontamination of oregano: effects on Bacillus cereus spores, water activity, color, and volatile compounds.

    Science.gov (United States)

    Eliasson, Lovisa; Libander, Patrik; Lövenklev, Maria; Isaksson, Sven; Ahrné, Lilia

    2014-12-01

    Infrared (IR) heating, a novel technology for decontaminating oregano, was evaluated by investigating the reduction of inoculated Bacillus cereus spores and the effect on water activity (a(w)), color, and headspace volatile compounds after exposure to IR treatment. Conditioned oregano (a(w) 0.88) was IR-treated in a closed heating unit at 90 and 100 °C for holding times of 2 and 10 min, respectively. The most successful reduction in B. cereus spore numbers (5.6 log units) was achieved after a holding time of 10 min at 90 °C, while treatment at 100 °C for the same time resulted in a lower reduction efficiency (4.7 log units). The lower reduction at 100 °C was probably due to a reduced aw (aw 0.76) during IR treatment or possibly to the alteration or loss of volatile compounds possessing antimicrobial properties. The green color of oregano was only slightly affected, while the composition of volatile compounds was clearly altered by IR heating. However, two of the key aroma compounds, carvacrol and thymol, were only slightly affected, compared to the effect on the other studied compounds, indicating that the typical oregano aroma can likely be preserved. In conclusion, IR heating shows potential for the successful decontamination of oregano without severe alteration of its color or the key aroma compounds, carvacrol and thymol.

  8. A procedure for estimating Bacillus cereus spores in soil and stream-sediment samples - A potential exploration technique

    Science.gov (United States)

    Watterson, J.R.

    1985-01-01

    The presence of bacterial spores of the Bacillus cereus group in soils and stream sediments appears to be a sensitive indicator of several types of concealed mineral deposits, including vein-type gold deposits. The B. cereus assay is rapid, inexpensive, and inherently reproducible. The test, currently under investigation for its potential in mineral exploration, is recommended for use on a research basis. Among the aerobic spore-forming bacilli, only B. cereus and closely related strains produce an opaque zone in egg-yolk emulsion agar. This characteristic, also known as the Nagler of lecitho-vitellin reaction, has long been used to rapidly indentify and estimate presumptive B. cereus. The test is here adapted to permit rapid estimation of B. cereus spores in soil and stream-sediment samples. Relative standard deviation was 10.3% on counts obtained from two 40-replicate pour-plate determinations. As many as 40 samples per day can be processed. Enough procedural detail is included to permit investigation of the test in conventional geochemical laboratories using standard microbiological safety precautions. ?? 1985.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Food Sensing: Aptamer-Based Trapping of Bacillus cereus Spores with Specific Detection via Real Time PCR in Milk.

    Science.gov (United States)

    Fischer, Christin; Hünniger, Tim; Jarck, Jan-Hinnerk; Frohnmeyer, Esther; Kallinich, Constanze; Haase, Ilka; Hahn, Ulrich; Fischer, Markus

    2015-09-16

    Aerobic spores pose serious problems for both food product manufacturers and consumers. Milk is particularly at risk and thus an important issue of preventive consumer protection and quality assurance. The spore-former Bacillus cereus is a food poisoning Gram-positive pathogen which mainly produces two different types of toxins, the diarrhea inducing and the emetic toxins. Reliable and rapid analytical assays for the detection of B. cereus spores are required, which could be achieved by combining in vitro generated aptamers with highly specific molecular biological techniques. For the development of routine bioanalytical approaches, already existing aptamers with high affinity to B. cereus spores have been characterized by surface plasmon resonance (SPR) spectroscopy and fluorescence microscopy in terms of their dissociation constants and selectivity. Dissociation constants in the low nanomolar range (from 5.2 to 52.4 nM) were determined. Subsequently, the characterized aptamers were utilized for the establishment and validation of an aptamer-based trapping technique in both milk simulating buffer and milk with fat contents between 0.3 and 3.5%. Thereby, enrichment factors of up to 6-fold could be achieved. It could be observed that trapping protocol and characterized aptamers were fully adaptable to the application in milk. Due to the fact that aptamer selectivity is limited, a highly specific real time PCR assay was utilized following trapping to gain a higher degree of selectivity.

  11. Bacillus atrophaeus Outer Spore Coat Assembly and Ultrastructure

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T J; Wheeler, K E; Pitesky, M E; Malkin, A J

    2005-11-21

    Our previous atomic force microscopy (AFM) studies successfully visualized native Bacillus atrophaeus spore coat ultrastructure and surface morphology. We have shown that the outer spore coat surface is formed by a crystalline array of {approx}11 nm thick rodlets, having a periodicity of {approx}8 nm. We present here further AFM ultrastructural investigations of air-dried and fully hydrated spore surface architecture. In the rodlet layer, planar and point defects, as well as domain boundaries, similar to those described for inorganic and macromolecular crystals, were identified. For several Bacillus species, rodlet structure assembly and architectural variation appear to be a consequence of species-specific nucleation and crystallization mechanisms that regulate the formation of the outer spore coat. We propose a unifying mechanism for nucleation and self-assembly of this crystalline layer on the outer spore coat surface.

  12. 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

  13. A versatile nano display platform from bacterial spore coat proteins.

    Science.gov (United States)

    Wu, I-Lin; Narayan, Kedar; Castaing, Jean-Philippe; Tian, Fang; Subramaniam, Sriram; Ramamurthi, Kumaran S

    2015-04-09

    Dormant bacterial spores are encased in a thick protein shell, the 'coat', which contains ∼70 different proteins. The coat protects the spore from environmental insults, and is among the most durable static structures in biology. Owing to extensive cross-linking among coat proteins, this structure has been recalcitrant to detailed biochemical analysis, so molecular details of how it assembles are largely unknown. Here, we reconstitute the basement layer of the coat atop spherical membranes supported by silica beads to create artificial spore-like particles. We report that these synthetic spore husk-encased lipid bilayers (SSHELs) assemble and polymerize into a static structure, mimicking in vivo basement layer assembly during sporulation in Bacillus subtilis. In addition, we demonstrate that SSHELs may be easily covalently modified with small molecules and proteins. We propose that SSHELs may be versatile display platforms for drugs and vaccines in clinical settings, or for enzymes that neutralize pollutants for environmental remediation.

  14. The Exosporium of B.cereus Contains a Binding Site for gC1qR/p33: Implication in Spore Attachment and/or Entry.

    Energy Technology Data Exchange (ETDEWEB)

    GHEBREHIWET,B.; TANTRAL, L.; TITMUS, M.A.; PANESSA-WARREN, B.J.; TORTORA, G.T.; WONG, S.S.; WARREN, J.B.

    2008-01-01

    B. cereus, is a member of a genus of aerobic, gram-positive, spore-forming rod-like bacilli, which includes the deadly, B. anthracis. Preliminary experiments have shown that gC1qR binds to B.cereus spores that have been attached to microtiter plates. The present studies were therefore undertaken, to examine if cell surface gC1qR plays a role in B.cereus spore attachment and/or entry. Monolayers of human colon carcinoma (Caco-2) and lung cells were grown to confluency on 6 mm coverslips in shell vials with gentle swirling in a shaker incubator. Then, 2 {micro}l of a suspension of strain SB460 B.cereus spores (3x10{sup 8}/ml, in sterile water), were added and incubated (1-4 h; 36{sup 0} C) in the presence or absence of anti-gC1qR mAb-carbon nanoloops. Examination of these cells by EM revealed that: (1) When B. cereus endospores contacted the apical Caco-2 cell surface, or lung cells, gClqR was simultaneously detectable, indicating upregulation of the molecule. (2) In areas showing spore contact with the cell surface, gClqR expression was often adjacent to the spores in association with microvilli (Caco-2 cells) or cytoskeletal projections (lung cells). (3) Furthermore, the exosporia of the activated and germinating spores were often decorated with mAb-nanoloops. These observations were further corroborated by experiments in which B.cereus spores were readily taken up by monocytes and neutrophils, and this uptake was partially inhibited by mAb 60.11, which recognizes the C1q binding site on gC1qR. Taken together, the data suggest a role, for gC1qR at least in the initial stages of spore attachment and/or entry.

  15. Nanosensors having dipicolinic acid imprinted nanoshell for Bacillus cereus spores detection

    Energy Technology Data Exchange (ETDEWEB)

    Gueltekin, Aytac [Trakya University, Department of Chemistry (Turkey); Ersoez, Arzu [Anadolu University, Department of Chemistry, Faculty of Science, Yunusemre Campus (Turkey); Sarioezlue, Nalan Yilmaz [Anadolu University, Department of Biology (Turkey); Denizli, Adil [Hacettepe University, Department of Chemistry (Turkey); Say, Ridvan, E-mail: rsay@anadolu.edu.t [Anadolu University, Department of Chemistry, Faculty of Science, Yunusemre Campus (Turkey)

    2010-08-15

    Molecular imprinted polymers (MIPs) as a recognition element for sensors are increasingly of interest and MIP nanoclusters have started to appear in the literature. In this study, we have proposed a novel thiol ligand-capping method with polymerizable methacryloylamido-cysteine (MAC) attached to gold-silver nanoclusters, reminiscent of a self-assembled monolayer and have reconstructed surface shell by synthetic host polymers based on molecular imprinting method for recognition. In this method, methacryloylamidoantipyrine-terbium ((MAAP){sub 2}-Tb(III)) has been used as a new metal-chelating monomer via metal coordination-chelation interactions and dipicolinic acid (DPA) which is main participant of Bacillus cereus spores used as a model. Nanoshell sensors with templates give a cavity that is selective for DPA. The DPA can simultaneously chelate to Tb(III) metal ion and fit into the shape-selective cavity. Thus, the interaction between Tb(III) ion and free coordination spheres has an effect on the binding ability of the gold-silver nanoclusters nanosensor. The binding affinity of the DPA imprinted nanoclusters has been investigated by using the Langmuir and Scatchard methods, and the respective affinity constants (K{sub affinity}) determined were found to be 1.43 x 10{sup 4} and 9.1 x 10{sup 6} mol L{sup -1}.

  16. Bacillus cereus and Bacillus thuringiensis spores in Korean rice: prevalence and toxin production as affected by production area and degree of milling.

    Science.gov (United States)

    Kim, Booyoung; Bang, Jihyun; Kim, Hoikyung; Kim, Yoonsook; Kim, Byeong-Sam; Beuchat, Larry R; Ryu, Jee-Hoon

    2014-09-01

    We determined the prevalence of and toxin production by Bacillus cereus and Bacillus thuringiensis in Korean rice as affected by production area and degree of milling. Rough rice was collected from 64 farms in 22 agricultural areas and polished to produce brown and white rice. In total, rice samples were broadly contaminated with B. cereus spores, with no effect of production area. The prevalence and counts of B. cereus spores declined as milling progressed. Frequencies of hemolysin BL (HBL) production by isolates were significantly (P ≤ 0.01) reduced as milling progressed. This pattern corresponded with the presence of genes encoding the diarrheal enterotoxins. The frequency of B. cereus isolates positive for hblC, hblD, or nheB genes decreased as milling progressed. Because most B. cereus isolates from rice samples contained six enterotoxin genes, we concluded that B. cereus in rice produced in Korea is predominantly of the diarrheagenic type. The prevalence of B. thuringiensis in rice was significantly lower than that of B. cereus and not correlated with production area. All B. thuringiensis isolates were of the diarrheagenic type. This study provides information useful for predicting safety risks associated with B. cereus and B. thuringiensis in rough and processed Korean rice.

  17. Gold nanoparticles having dipicolinic acid imprinted nanoshell for Bacillus cereus spores recognition

    Energy Technology Data Exchange (ETDEWEB)

    Gueltekin, Aytac [Department of Chemistry, Trakya University, Edirne (Turkey); Ersoez, Arzu; Huer, Deniz [Department of Chemistry, Anadolu University, Eskisehir (Turkey); Sarioezlue, Nalan Yilmaz [Department of Biology, Anadolu University, Eskisehir (Turkey); Denizli, Adil [Department of Chemistry, Hacettepe University, Ankara (Turkey); Say, Ridvan, E-mail: rsay@anadolu.edu.tr [Department of Chemistry, Anadolu University, Eskisehir (Turkey); BIBAM (Plant, Drug and Scientific Research Center) Anadolu University (Turkey)

    2009-10-15

    Taking into account the recognition element for sensors linked to molecular imprinted polymers (MIPs), a proliferation of interest has been witnessed by those who are interested in this subject. Indeed, MIP nanoparticles are theme which recently has come to light in the literature. In this study, we have proposed a novel thiol ligand-capping method with polymerizable methacryloylamidocysteine (MAC) attached to gold nanoparticles, reminiscent of a self-assembled monolayer. Furthermore, a surface shell by synthetic host polymers based on molecular imprinting method for recognition has been reconstructed. In this method, methacryloyl iminodiacetic acid-chrome (MAIDA-Cr(III)) has been used as a new metal-chelating monomer via metal coordination-chelation interactions and dipicolinic acid (DPA) which is the main participant of Bacillus cereus spores has been used as a template. Nanoshell sensors with templates produce a cavity that is selective for DPA. The DPA can simultaneously chelate to Cr(III) metal ion and fit into the shape-selective cavity. Thus, the interaction between Cr(III) ion and free coordination spheres has an effect on the binding ability of the gold nanoparticles nanosensor. The interactions between DPA and MIP particles were studied observing fluorescence measurements. DPA addition caused significant decreases in fluorescence intensity because they induced photoluminescence emission from Au nanoparticles through the specific binding to the recognition sites of the crosslinked nanoshell polymer matrix. The binding affinity of the DPA imprinted nanoparticles has been explored by using the Langmuir and Scatchard methods and the analysis of the quenching results has been performed in terms of the Stern-Volmer equation.

  18. Use of fatty acid methyl ester profiles for discrimination of Bacillus cereus T-strain spores grown on different media.

    Science.gov (United States)

    Ehrhardt, Christopher J; Chu, Vivian; Brown, TeeCie; Simmons, Terrie L; Swan, Brandon K; Bannan, Jason; Robertson, James M

    2010-03-01

    The goal of this study was to determine if cellular fatty acid methyl ester (FAME) profiling could be used to distinguish among spore samples from a single species (Bacillus cereus T strain) that were prepared on 10 different medium formulations. To analyze profile differences and identify FAME biomarkers diagnostic for the chemical constituents in each sporulation medium, a variety of statistical techniques were used, including nonmetric multidimensional scaling (nMDS), analysis of similarities (ANOSIM), and discriminant function analysis (DFA). The results showed that one FAME biomarker, oleic acid (18:1 omega9c), was exclusively associated with spores grown on Columbia agar supplemented with sheep blood and was indicative of blood supplements that were present in the sporulation medium. For spores grown in other formulations, multivariate comparisons across several FAME biomarkers were required to discern profile differences. Clustering patterns in nMDS plots and R values from ANOSIM revealed that dissimilarities among FAME profiles were most pronounced when spores grown with disparate sources of complex additives or protein supplements were compared (R > 0.8), although other factors also contributed to FAME differences. DFA indicated that differentiation could be maximized with a targeted subset of FAME variables, and the relative contributions of branched FAME biomarkers to group dissimilarities changed when different media were compared. When taken together, these analyses indicate that B. cereus spore samples grown in different media can be resolved with FAME profiling and that this may be a useful technique for providing intelligence about the production methods of Bacillus organisms in a forensic investigation.

  19. Identification of CdnL, a Putative Transcriptional Regulator Involved in Repair and Outgrowth of Heat-Damaged Bacillus cereus Spores.

    Directory of Open Access Journals (Sweden)

    Alicja K Warda

    Full Text Available Spores are widely present in the environment and are common contaminants in the food chain, creating a challenge for food industry. Nowadays, heat treatments conventionally applied in food processing may become milder to comply with consumer desire for products with higher sensory and nutritional values. Consequently subpopulations of spores may emerge that are sublethally damaged rather than inactivated. Such spores may germinate, repair damage, and eventually grow out leading to uncontrolled spoilage and safety issues. To gain insight into both the behaviour of damaged Bacillus cereus spores, and the process of damage repair, we assessed the germination and outgrowth performance using OD595 measurements and microscopy combined with genome-wide transcription analysis of untreated and heat-treated spores. The first two methods showed delayed germination and outgrowth of heat-damaged B. cereus ATCC14579 spores. A subset of genes uniquely expressed in heat-treated spores was identified with putative roles in the outgrowth of damaged spores, including cdnL (BC4714 encoding the putative transcriptional regulator CdnL. Next, a B. cereus ATCC14579 cdnL (BC4714 deletion mutant was constructed and assessment of outgrowth from heat-treated spores under food relevant conditions showed increased damage compared to wild type spores. The approach used in this study allows for identification of candidate genes involved in spore damage repair. Further identification of cellular parameters and characterisation of the molecular processes contributing to spore damage repair may provide leads for better control of spore outgrowth in foods.

  20. Quantification of the impact of single and multiple mild stresses on outgrowth heterogeneity of Bacillus cereus spores.

    Science.gov (United States)

    van Melis, C C J; den Besten, H M W; Nierop Groot, M N; Abee, T

    2014-05-02

    Outgrowth heterogeneity of bacterial spore populations complicates both prediction and efficient control of spore outgrowth. In this study, the impact of mild preservation stresses on outgrowth of Bacillus cereus ATCC 14579 spores was quantified during the first stages of outgrowth. Heterogeneity in outgrowth of heat-treated (90°C for 10 min) and non-heat-treated germinated single spores to the maximum micro-colony stage of 256 cells was assessed by direct imaging on Anopore strips, placed on BHI plates at pH7 and pH5.5, without and with added NaCl or sorbic acid (HSA). At pH7 non-heated and heat-treated germinated spores required 6h to reach the maximum microcolony stage with limited heterogeneity, and these parameters were only slightly affected with both types of spores when incubated at pH7 with added NaCl. Notably, the most pronounced effects were observed during outgrowth of spores at pH5.5 without and with added NaCl or HSA. Non-heat-treated germinated spores showed again efficient outgrowth with limited heterogeneity reaching the maximum microcolony stage after 6h at pH5.5, which increased to 12h and 16 h with added NaCl and HSA, respectively. In contrast, heat-treated spores displayed a strong delay between initial germination and swelling and further outgrowth at pH5.5, resulting in large heterogeneity and low numbers of fastest growers reaching the maximum microcolony stage after 10, 12 and 24h, without and with added NaCl or HSA, respectively. This work shows that Anopore technology provides quantitative information on the impact of combined preservation stresses on outgrowth of single spores, showing that outgrowth of germinated heat-treated spores is significantly affected at pH5.5 with a large fraction of spores arrested in the early outgrowth stage, and with outgrowing cells showing large heterogeneity with only a small fraction committed to relatively fast outgrowth.

  1. Survival of foodborne pathogenic bacteria (Bacillus cereus, Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Listeria monocytogenes) and Bacillus cereus spores in fermented alcoholic beverages (beer and refined rice wine).

    Science.gov (United States)

    Kim, S A; Kim, N H; Lee, S H; Hwang, I G; Rhee, M S

    2014-03-01

    Only limited information is available on the microbiological safety of fermented alcoholic beverages because it is still a common belief that such beverages do not provide a favorable environment for bacterial growth and survival. Thus, in this study, we examined the survival of major foodborne pathogens and spores in fermented alcoholic beverages. Foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Staphylococcus aureus) and B. cereus spores (initial population, 3 to 4 log CFU/ml) were inoculated separately into three types of beer and refined rice wine, which were then stored at 5 and 22°C. Bacterial counts were assayed periodically for up to 28 days. Vegetative B. cereus counts decreased rapidly, whereas B. cereus spore counts remained constant (P > 0.05) for a long period of time in all beverages. Vegetative B. cereus cells formed spores in beer at 5 and 22°C, and the spores survived for long periods. Among vegetative cells, E. coli O157:H7 had the highest survival (only 1.49 to 1.56 log reduction during 28 days in beer at 5°C). Beer and refined rice wine supported microbial survival from several days to several weeks. Our results appear to contradict the common belief that pathogens cannot survive in alcoholic beverages. Long-term survival of pathogens (especially B. cereus and E. coli O157:H7) in beer and refined rice wine should be taken into consideration by the manufacturers of these beverages. This study provides basic information that should help further research into microbial survival in alcoholic beverages and increase the microbiological safety regulation of fermented alcoholic beverages.

  2. Studies on the bacterial spore coat 6 effects of alkali extraction on the spore of Bacillus thiaminolyticus.

    Science.gov (United States)

    Minami, J; Ichikawa, T; Kondo, M

    1977-01-01

    Thin sections of the spore of Bacillus thiaminolyticus Matsukawa and Misawa show a characteristic surface structure with five ridges, and a series of three district layers. The outer layer of the spore coat was peeled off by SDS sonic treatment, and than the middle layer was solubilized by alkali extraction of the SDS sonic-treated spore. The spores subjected to these treatments were still refractile, heat resistant, and contained dipicolinic acid, but lost their resistance to mechanical shock.

  3. Impact of sorbic acid on germination and outgrowth heterogeneity of Bacillus cereus ATCC 14579 spores

    NARCIS (Netherlands)

    Besten, den H.M.W.; Melis, van C.C.J.; Sanders, J.W.; Nierop Groot, M.N.; Abee, T.

    2012-01-01

    Population heterogeneity complicates the predictability of the outgrowth kinetics of individual spores. Flow cytometry sorting and monitoring of the germination and outgrowth of single dormant spores allowed the quantification of acid-induced spore population heterogeneity at pH 5.5 and in the prese

  4. Influence of food matrix on outgrowth heterogeneity of heat damaged Bacillus cereus spores

    NARCIS (Netherlands)

    Warda, A.K.; Besten, den H.M.W.; Sha, N.; Abee, T.; Nierop Groot, M.N.

    2015-01-01

    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

  5. Influence of sporulation medium composition on transcription of ger operons and the germination response of spores of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Hornstra, L.M.; Vries, de Y.P.; Vos, de W.M.; Abee, T.

    2006-01-01

    Bacillus cereus ATCC 14579 endospores were produced in Y1 medium, a nutrient-rich, chemically defined sporulation medium, and in modified G medium, containing low amounts of nutrients. The average transcription level of the seven ger operons per cell was 3.5 times higher in Y1 medium, and the spores

  6. Deletion in sigB in Bacillus cereus affects spore properties

    NARCIS (Netherlands)

    Vries, de Y.P.; Hornstra, L.M.; Atmadja, R.D.; Schaik, van W.; Vos, de W.M.; Abee, T.

    2005-01-01

    In Bacillus cereus and other gram-positive bacteria the alternative sigma factor ¿B is an important regulator of the stress response. Deletion of the sigB gene generally leads to a stress-sensitive phenotype of vegetative cells. In this study, we describe the effect of the deletion of the sigB gene

  7. Drosophila melanogaster Selection for Survival after Infection with Bacillus cereus Spores: Evolutionary Genetic and Phenotypic Investigations of Respiration and Movement

    Directory of Open Access Journals (Sweden)

    Junjie Ma

    2013-01-01

    Full Text Available Laboratory populations of D. melanogaster have been subjected to selection for survival after live spores of B. cereus were introduced as a pathogenic agent. The present study was designed to investigate correlated traits: respiration as a metabolic trait and movement as a behavioral trait. An underlying hypothesis was that the evolution of increased survival after B. cereus infection exerts a metabolic cost associated with elevated immunity and this would be detected by increased respiration rates. There was support for this hypothesis in the male response to selection, but not for selected-line females. Two phenotypic effects were also observed in the study. Females especially showed a marked increase in respiration after mating compared to the other assay stages regardless of whether respiration was measured per fly or adjusted by lean mass or dry weight. Given that mating stimulates egg production, it is feasible that elevated metabolism was needed to provision oocytes with yolk. Females also moved less than males, perhaps due to behaviors related to oviposition whereas elevated male activity might be due to behaviors associated with seeking females and courtship. Relatively low movement of females indicated that their elevated respiration after mating was not due to a change in locomotion.

  8. Maximum shields: the assembly and function of the bacterial spore coat.

    Science.gov (United States)

    Driks, Adam

    2002-06-01

    Spores produced by bacilli and clostridia are surrounded by a multilayered protein shell called the coat. As the armor-like appearance of the coat suggests, this structure, along with others within the spore, confers the remarkable resistance properties that make Bacillus anthracis spores such potent biological weapons. Here, I review recent studies of coat assembly in the model organism Bacillus subtilis, and explore the implications of these findings for coat assembly in B. anthracis and for defense against biological weapons.

  9. Effects of Mn2+ Levels on the Resistance Properties of Bacillus cereus Spores

    Science.gov (United States)

    2013-01-01

    In contrast, Bacillus subtilis spores with over a 200-fold range of protoplast Mn levels exhibited no significant differences in resistance to...Bacillus megaterium by wet heat. Lett. Appl . Microbiol. 50:507-514. Daly MJ (2012) Death by protein damage in irradiated cells. DNA Repair 11:12-21...levels on resistance of Bacillus megaterium spores to heat, radiation and hydrogen peroxide. J. Appl . Microbiol. 111:663-670. Ghosh S, Setlow P (2010

  10. Fate and effect of ingested Bacillus cereus spores and vegetative cells in the intestinal tract of human-flora-associated rats

    DEFF Research Database (Denmark)

    Wilcks, Andrea; Hansen, Bjarne Munk; Hendriksen, Niels Bohse;

    2006-01-01

    The fate and effect of Bacillus cereus F4433/73R in the intestine of human-flora-associated rats was studied using bacteriological culturing techniques and PCR-denaturing gradient gel electrophoresis in combination with cell assays and immunoassays for detection of enterotoxins. In faecal samples...... gradient gel electrophoresis analysis with universal 16S rRNA gene primers revealed significant changes in the intestinal microbiota of animals dosed with spores. Vero cell assays and a commercial kit (BCET-RPLA) did not reveal any enterotoxin production from B. cereus F4433/73R in the intestinal tract....

  11. Decontamination of Streptococci biofilms and Bacillus cereus spores on plastic surfaces with DC and pulsed corona discharges

    Science.gov (United States)

    Koval'ová, Zuzana; Tarabová, Kataŕna; Hensel, Karol; Machala, Zdenko

    2013-02-01

    Cold air plasmas of DC and pulsed corona discharges: positive streamers and negative Trichel pulses were used for bio-decontamination of Streptococci biofilm and Bacillus cereus spores on polypropylene plastic surfaces. The reduction of bacterial population (evaluated as log10) in the biofilm on plastic surfaces treated by DC corona reached 2.4 logs with 10 min treatment time and 3.3 logs with 2 min treatment time with water spraying. The enhancement of plasma biocidal effects on the biofilm by electro-spraying of water through a hollow needle high-voltage electrode was investigated. No significant polarity effect was found with DC corona. Pulsed corona was demonstrated slightly more bactericidal for spores, especially in the negative polarity where the bacterial population reduction reached up to 2.2 logs at 10 min exposure time. Contribution to the Topical Issue "13th International Symposium on High Pressure Low Temperature Plasma Chemistry (Hakone XIII)", Edited by Nicolas Gherardi, Henryca Danuta Stryczewska and Yvan Ségui.

  12. 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.

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

    Directory of Open Access Journals (Sweden)

    Knols Bart GJ

    2010-01-01

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

  14. Field evaluation of a bioregulator containing live Bacillus cereus spores on health status and performance of sows and their litters.

    Science.gov (United States)

    Alexopoulos, C; Karagiannidis, A; Kritas, S K; Boscos, C; Georgoulakis, I E; Kyriakis, S C

    2001-04-01

    The efficacy of Paciflor, a bioregulator containing live Bacillus cereus CIP 5832 spores, was assessed in sows during late pregnancy and lactation, as well as in their piglets up to the growing phase. Two groups each of 30 pregnant gilts and sows received normal feed (T1 group), or feed with 85 g Paciflor per ton feed (T2 group), from 15 days prior to farrowing up to the end of the lactation period. Furthermore, 15 litters of the T1 group and 15 litters of the T2 group, were offered normal feed from the 5th to the 70th days of life (T1.1 and T2.1 groups, respectively), while the remaining 15 litters each of the T1 and T2 groups received the same feed but including Paciflor at a dose of 100 g/ton (from day 5 to day 49) and 50 g/ton (from day 50 to day 70). These pig litters were T1.2 and T2.2, respectively. No differences were seen between the T1 and T2 groups with respect to the clinical observations (loss of appetite, fever, mastitis, metritis and returns to oestrus, treatments applied, deaths, or removals to the slaughterhouse), gestation length, bodyweight of sows at farrowing or litter-size at birth. However, during lactation, the fat content of the dam's milk was increased (0.46% more fat), the body weight loss of sows was reduced and the number of weaned pigs per sow was increased (0.6 more pigs per litter) after administration of Paciflor (P pigs originating from Paciflor-treated dams (T2.2 group) (P < 0.05). Despite the fact that no difference was seen between groups with regard to the amount of feed consumed, the feed conversion ratio of Paciflor-treated piglets (T2.2 and T1.2) was significantly improved compared to that of the untreated piglets (T2.1 and T1.1) (P < 0.05). With respect to weight gain, for the Paciflor-treated piglets, those born to Paciflor-treated mothers (T2.2) were 0.56 kg heavier than those born to untreated dams (T1.2) (P < 0.05). It is concluded that administration of Paciflor in dams during the end of pregnancy and during lactation

  15. Understanding of the importance of the spore coat structure and pigmentation in the Bacillus subtilis spore resistance to low-pressure plasma sterilization

    Science.gov (United States)

    Raguse, Marina; Fiebrandt, Marcel; Denis, Benjamin; Stapelmann, Katharina; Eichenberger, Patrick; Driks, Adam; Eaton, Peter; Awakowicz, Peter; Moeller, Ralf

    2016-07-01

    Low-pressure plasmas have been evaluated for their potential in biomedical and defense purposes. The sterilizing effect of plasma can be attributed to several active agents, including (V)UV radiation, charged particles, radical species, neutral and excited atoms and molecules, and the electric field. Spores of Bacillus subtilis were used as a bioindicator and a genetic model system to study the sporicidal effects of low-pressure plasma decontamination. Wild-type spores, spores lacking the major protective coat layers (inner, outer, and crust), pigmentation-deficient spores or spore impaired in encasement (a late step in coat assembly) were systematically tested for their resistance to low-pressure argon, hydrogen, and oxygen plasmas with and without admixtures. We demonstrate that low-pressure plasma discharges of argon and oxygen discharges cause significant physical damage to spore surface structures as visualized by atomic force microscopy. Spore resistance to low-pressure plasma was primarily dependent on the presence of the inner, and outer spore coat layers as well as spore encasement, with minor or less importance of the crust and spore pigmentation, whereas spore inactivation itself was strongly influenced by the gas composition and operational settings.

  16. Spore Coat Architecture of Clostridium novyi NT Spores▿

    OpenAIRE

    Plomp, Marco; McCaffery, J. Michael; Cheong, Ian; Huang, Xin; Bettegowda, Chetan; Kinzler, Kenneth W.; Zhou, Shibin; Vogelstein, Bert; Malkin, Alexander J.

    2007-01-01

    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. Toward this end, we have used both transmission electron microscopy and atomic force microscopy to determine the structure of both dormant and germinating spores. We found th...

  17. Spores from mesophilic Bacillus cereus strains germinate better and grow faster in simulated gastro-intestinal conditions than spores from psychrotrophic strains.

    NARCIS (Netherlands)

    Wijnands, L M; Dufrenne, J B; Zwietering, M H; Leusden, F M van

    2006-01-01

    The species Bacillus cereus, known for its ability to cause food borne disease, consists of a large variety of strains. An important property for discrimination of strains is their growth temperature range. Psychrotrophic strains can grow well at refrigerator temperatures but grow at 37 degrees C wi

  18. Spores from mesophilic Bacillus cereus strains germinate better and grow faster in simulated gastro-intestinal conditions than spores from psychrotrophic strains

    NARCIS (Netherlands)

    Wijnands, L.M.; Dufrenne, J.B.; Zwietering, M.H.; Leusden, van F.M.

    2006-01-01

    The species Bacillus cereus, known for its ability to cause food borne disease, consists of a large variety of strains. An important property for discrimination of strains is their growth temperature range. Psychrotrophic strains can grow well at refrigerator temperatures but grow at 37 °C with diff

  19. Involvement of Coat Proteins in Bacillus subtilis Spore Germination in High-Salinity Environments.

    Science.gov (United States)

    Nagler, Katja; Setlow, Peter; Reineke, Kai; Driks, Adam; Moeller, Ralf

    2015-10-01

    The germination of spore-forming bacteria in high-salinity environments is of applied interest for food microbiology and soil ecology. It has previously been shown that high salt concentrations detrimentally affect Bacillus subtilis spore germination, rendering this process slower and less efficient. The mechanistic details of these salt effects, however, remained obscure. Since initiation of nutrient germination first requires germinant passage through the spores' protective integuments, the aim of this study was to elucidate the role of the proteinaceous spore coat in germination in high-salinity environments. Spores lacking major layers of the coat due to chemical decoating or mutation germinated much worse in the presence of NaCl than untreated wild-type spores at comparable salinities. However, the absence of the crust, the absence of some individual nonmorphogenetic proteins, and the absence of either CwlJ or SleB had no or little effect on germination in high-salinity environments. Although the germination of spores lacking GerP (which is assumed to facilitate germinant flow through the coat) was generally less efficient than the germination of wild-type spores, the presence of up to 2.4 M NaCl enhanced the germination of these mutant spores. Interestingly, nutrient-independent germination by high pressure was also inhibited by NaCl. Taken together, these results suggest that (i) the coat has a protective function during germination in high-salinity environments; (ii) germination inhibition by NaCl is probably not exerted at the level of cortex hydrolysis, germinant accessibility, or germinant-receptor binding; and (iii) the most likely germination processes to be inhibited by NaCl are ion, Ca(2+)-dipicolinic acid, and water fluxes.

  20. In pursuit of protein targets: proteomic characterization of bacterial spore outer layers.

    Science.gov (United States)

    Abhyankar, Wishwas; Hossain, Abeer H; Djajasaputra, André; Permpoonpattana, Patima; Ter Beek, Alexander; Dekker, Henk L; Cutting, Simon M; Brul, Stanley; de Koning, Leo J; de Koster, Chris G

    2013-10-04

    Bacillus cereus, responsible for food poisoning, and Clostridium difficile, the causative agent of Clostridium difficile-associated diarrhea (CDAD), are both spore-forming pathogens involved in food spoilage, food intoxication, and other infections in humans and animals. The proteinaceous coat and the exosporium layers from spores are important for their resistance and pathogenicity characteristics. The exosporium additionally provides an ability to adhere to surfaces eventually leading to spore survival in food. Thus, studying these layers and identifying suitable protein targets for rapid detection and removal of spores is of the utmost importance. In this study, we identified 100 proteins from B. cereus spore coat, exosporium and 54 proteins from the C. difficile coat insoluble protein fraction. In an attempt to define a universal set of spore outer layer proteins, we identified 11 superfamily domains common to the identified proteins from two Bacilli and one Clostridium species. The evaluated orthologue relationships of identified proteins across different spore formers resulted in a set of 13 coat proteins conserved across the spore formers and 12 exosporium proteins conserved in the B. cereus group, which could be tested for quick and easy detection or targeted in strategies aimed at removal of spores from surfaces.

  1. Development of a powder formulation based on Bacillus cereus sensu lato strain B25 spores for biological control of Fusarium verticillioides in maize plants.

    Science.gov (United States)

    Martínez-Álvarez, Juan C; Castro-Martínez, Claudia; Sánchez-Peña, Pedro; Gutiérrez-Dorado, Roberto; Maldonado-Mendoza, Ignacio E

    2016-05-01

    Maize is an economically important crop in northern Mexico. Different fungi cause ear and root rot in maize, including Fusarium verticillioides (Sacc.) Nirenberg. Crop management of this pathogen with chemical fungicides has been difficult. By contrast, the recent use of novel biocontrol strategies, such as seed bacterization with Bacillus cereus sensu lato strain B25, has been effective in field trials. These approaches are not without their problems, since insufficient formulation technology, between other factors, can limit success of biocontrol agents. In response to these drawbacks, we have developed a powder formulation based on Bacillus B25 spores and evaluated some of its characteristics, including shelf life and efficacy against F. verticillioides, in vitro and in maize plants. A talc-based powder formulation containing 1 × 10(9) c.f.u. g(-1) was obtained and evaluated for seed adherence ability, seed germination effect, shelf life and antagonism against F. verticillioides in in vitro and in planta assays. Seed adherence of viable bacterial spores ranged from 1.0 to 1.41 × 10(7) c.f.u. g(-1). Bacteria did not display negative effects on seed germination. Spore viability for the powder formulation slowly decreased over time, and was 53 % after 360 days of storage at room temperature. This formulation was capable of controlling F. verticillioides in greenhouse assays, as well as eight other maize phytopathogenic fungi in vitro. The results suggest that a talc-based powder formulation of Bacillus B25 spores may be sufficient to produce inoculum for biocontrol of maize ear and root rots caused by F. verticillioides.

  2. Diverse supramolecular structures formed by self-assembling proteins of the B acillus subtilis spore coat

    OpenAIRE

    2015-01-01

    Summary Bacterial spores (endospores), such as those of the pathogens C lostridium difficile and B acillus anthracis, are uniquely stable cell forms, highly resistant to harsh environmental insults. B acillus subtilis is the best studied spore‐former and we have used it to address the question of how the spore coat is assembled from multiple components to form a robust, protective superstructure. B . subtilis coat proteins (CotY, CotE, CotV and CotW) expressed in E scherichia coli can arrange...

  3. Detection of Bacillus anthracis spores in water using biosensors based on magnetostrictive microcantilever coated with phage

    Science.gov (United States)

    Fu, Liling; Li, Suiqiong; Zhang, Kewei; Cheng, Z.-Y.; Barbaree, J. M.

    2007-04-01

    Microcantilevers (MCs) as state-of-art sensor platforms have been widely investigated. We recently introduced a new type of MC, magnetostrictive microcantilever (MSMC), as high performance sensor platform. The MSMC is a remote/wireless sensor platform and exhibits a high quality merit factor in liquid. In this paper, a MSMC-based biosensor is developed for detecting B. anthracis spores in liquid, a potential biothreaten agent. The results demonstrated the advantages of MSMCs as a sensor platform. MSMCs with different sizes were fabricated and utilized in the experiments. The MSMCs were coated with the filamentous phage as a bio-recognition element to capture the B. anthracis spores. The phage-coated MSMCs as biosensors were exposed to cultures containing target spores with concentrations ranging from 5 * 10 4 spores/mL to 5 * 10 8 spores/mL. The resonance frequency of the MSMC sensors in cultures was monitored in a real-time manner. The results showed that for MSMCs of 2.8 mm * 1.0 mm * 35 μm and with 1.4 mm * 0.8 mm * 35 μm have a detection limit of 10 5 and 10 4 spores/mL, respectively.

  4. Comparing the mannitol-egg yolk-polymyxin agar plating method with the three-tube most-probable-number method for enumeration of Bacillus cereus spores in raw and high-temperature, short-time pasteurized milk.

    Science.gov (United States)

    Harper, Nigel M; Getty, Kelly J K; Schmidt, Karen A; Nutsch, Abbey L; Linton, Richard H

    2011-03-01

    The U.S. Food and Drug Administration's Bacteriological Analytical Manual recommends two enumeration methods for Bacillus cereus: (i) standard plate count method with mannitol-egg yolk-polymyxin (MYP) agar and (ii) a most-probable-number (MPN) method with tryptic soy broth (TSB) supplemented with 0.1% polymyxin sulfate. This study compared the effectiveness of MYP and MPN methods for detecting and enumerating B. cereus in raw and high-temperature, short-time pasteurized skim (0.5%), 2%, and whole (3.5%) bovine milk stored at 4°C for 96 h. Each milk sample was inoculated with B. cereus EZ-Spores and sampled at 0, 48, and 96 h after inoculation. There were no differences (P > 0.05) in B. cereus populations among sampling times for all milk types, so data were pooled to obtain overall mean values for each treatment. The overall B. cereus population mean of pooled sampling times for the MPN method (2.59 log CFU/ml) was greater (P plate count method (1.89 log CFU/ml). B. cereus populations in the inoculated milk samples ranged from 2.36 to 3.46 and 2.66 to 3.58 log CFU/ml for inoculated milk treatments for the MYP plate count and MPN methods, respectively, which is below the level necessary for toxin production. The MPN method recovered more B. cereus, which makes it useful for validation research. However, the MYP plate count method for enumeration of B. cereus also had advantages, including its ease of use and faster time to results (2 versus 5 days for MPN).

  5. Impact of sorbic acid and other mild preservation stresses on germination and outgrowth of Bacillus cereus spores

    NARCIS (Netherlands)

    Melis, van C.C.J.

    2013-01-01

      Weak organic acids such as sorbic acid, lactate, and acetic acid are widely used by the food industry as preservatives to control growth of micro-organisms. With the current trend towards milder processing of food products, opportunities arise for spore-forming spoilage and pathogenic microo

  6. Production and stability of chlorine dioxide in organic acid solutions as affected by pH, type of acid, and concentration of sodium chlorite, and its effectiveness in inactivating Bacillus cereus spores.

    Science.gov (United States)

    Kim, Hoikyung; Kang, Youngjee; Beuchat, Larry R; Ryu, Jee-Hoon

    2008-12-01

    We studied the production and stability of chlorine dioxide (ClO(2)) in organic acid solutions and its effectiveness in killing Bacillus cereus spores. Sodium chlorite (5000, 10,000, or 50,000 microg/ml) was added to 5% acetic, citric, or lactic acid solution, adjusted to pH 3.0, 4.0, 5.0, or 6.0, and held at 21 degrees C for up to 14 days. The amount of ClO(2) produced was higher as the concentration of sodium chlorite was increased and as the pH of the acid solutions was decreased. However, the stability in production of ClO(2) was enhanced by increasing the pH of the organic acid solutions. To evaluate the lethal activity of ClO(2) produced in various acid solutions as affected by acidulant and pH, suspensions of B. cereus spores were treated at 21 degrees C for 1, 3, 5, or 10 min in hydrochloric acid or organic acid solutions (pH 3.0, 4.0, 5.0, or 6.0) containing ClO(2) at concentrations of 100, 50, or 25 microg/ml. Populations of viable spores treated with ClO(2) at concentrations of 100 or 50 microg/ml in organic acid solutions decreased more rapidly than populations treated with the same concentrations of ClO(2) in HCl. Rates of inactivation tended to increase with higher pH of ClO(2) solutions. Results show that ClO(2) formed in organic acid solutions has higher stability and is more lethal to B. cereus spores than ClO(2) formed at the same concentration in HCl solution. This finding emphasizes the benefits of using organic acid solutions to prepare ClO(2) intended for use as an antimicrobial.

  7. In pursuit of protein targets: proteomic characterization of bacterial spore outer layers

    NARCIS (Netherlands)

    Abhyankar, W.; Hossain, A.H.; Djajasaputra, A.; Permpoonpattana, P.; ter Beek, A.; Dekker, H.L.; Cutting, S.M.; Brul, S.; de Koning, L.J.; de Koster, C.G.

    2013-01-01

    Bacillus cereus, responsible for food poisoning, and Clostridium difficile, the causative agent of Clostridium difficile-associated diarrhea (CDAD), are both spore-forming pathogens involved in food spoilage, food intoxication, and other infections in humans and animals. The proteinaceous coat and t

  8. Investigation of bacterial spore structure by high resolution solid-state nuclear magnetic resonance spectroscopy and transmission electron microscopy.

    Science.gov (United States)

    Leuschner, R G; Lillford, P J

    2001-01-22

    High resolution solid-state nuclear magnetic resonance spectroscopy (NMR) in combination with transmission electron microscopy (TEM) of spores of Bacillus cereus, an outer coatless mutant B. subtilis 322, an inner coatless mutant B. subtilis 325 and of germinated spores of B. subtilis CMCC 604 were carried out. Structural differences in the coats, mainly protein of spores were reflected by NMR spectra which indicated also differences in molecular mobility of carbohydrates which was partially attributed to the cortex. Dipicolinic acid (DPA) of spores of B. cereus displayed a high degree of solid state order and may be crystalline. Heat activation was studied on spores of B. subtilis 357 lux + and revealed a structural change when analysed by TEM but this was not associated with increases in molecular mobility since no effects were measured by NMR.

  9. Chemical and morphological studies of bacterial spore formation. IV. The development of spore refractility.

    Science.gov (United States)

    YOUNG, I E; JAMES, P C

    1962-01-01

    From the stage of a completed membranous forespore to that of a fully ripened free spore, synchronously sporulating cells of a variant Bacillus cereus were studied by cytological and chemical methods. Particular attention was paid to the development of the three spore layers-cortex, coat, and exosporium-in relation to the forespore membrane. First, the cortex is laid down between the recently described (5) double layers of the forespore membrane. Then when the cortex is (1/3) fully formed, the spore coat and exosporium are laid down peripheral to the outer membrane layer covering the cortex. As these latter layers appear, the spores, previously dense by dark phase contrast, gradually "whiten" or show an increase in refractive index. With this whitening, calcium uptake commences, closely followed by the synthesis of dipicolinic acid and the process is terminated, an hour later, with the formation of a fully refractile spore. In calcium-deficient media, final refractility is lessened and dipicolinic acid is formed only in amounts proportional to the available calcium. If calcium is withheld during the period of uptake beyond a critical point, sporulating cells lose the ability to assimilate calcium and to form normal amounts of dipicolinic acid. The resulting deficient spores are liberated from the sporangia but are unstable in water suspensions. Unlike ripe spores, they do not react violently to acid hydrolysis and, in thin sections, their cytoplasmic granules continue to stain with lead solutions.

  10. Inhibitory effects of nisin-coated multi-walled carbon nanotube sheet on biofilm formation from Bacillus anthracis spores

    Institute of Scientific and Technical Information of China (English)

    Xiuli Dong; Eric McCoy; Mei Zhang; Liju Yang

    2014-01-01

    Multi-walled carbon nanotube (MWCNT) sheet was fabricated from a drawable MWCNT forest and then deposited on poly(methyl methacrylate) film.The film was further coated with a natural antimicrobial peptide nisin.We studied the effects of nisin coating on the attachment of Bacillus anthracis spores,the germination of attached spores,and the subsequent biofilm formation from attached spores.It was found that the strong adsorptivity and the super hydrophobicity of MWCNTs provided an ideal platform for nisin coating.Nisin coating on MWCNT sheets decreased surface hydrophobicity,reduced spore attachment,and reduced the germination of attached spores by 3.5 fold,and further inhibited the subsequent biofilm formation by 94.6% compared to that on uncoated MWCNT sheet.Nisin also changed the morphology of vegetative cells in the formed biofilm.The results of this study demonstrated that the anti-adhesion and antimicrobial effect of nisin in combination with the physical properties of carbon nanotubes had the potential in producing effective anti-biofilm formation surfaces.

  11. Inhibitory effects of nisin-coated multi-walled carbon nanotube sheet on biofilm formation from Bacillus anthracis spores.

    Science.gov (United States)

    Dong, Xiuli; McCoy, Eric; Zhang, Mei; Yang, Liju

    2014-12-01

    Multi-walled carbon nanotube (MWCNT) sheet was fabricated from a drawable MWCNT forest and then deposited on poly(methyl methacrylate) film. The film was further coated with a natural antimicrobial peptide nisin. We studied the effects of nisin coating on the attachment of Bacillus anthracis spores, the germination of attached spores, and the subsequent biofilm formation from attached spores. It was found that the strong adsorptivity and the super hydrophobicity of MWCNTs provided an ideal platform for nisin coating. Nisin coating on MWCNT sheets decreased surface hydrophobicity, reduced spore attachment, and reduced the germination of attached spores by 3.5 fold, and further inhibited the subsequent biofilm formation by 94.6% compared to that on uncoated MWCNT sheet. Nisin also changed the morphology of vegetative cells in the formed biofilm. The results of this study demonstrated that the anti-adhesion and antimicrobial effect of nisin in combination with the physical properties of carbon nanotubes had the potential in producing effective anti-biofilm formation surfaces.

  12. Diverse supramolecular structures formed by self‐assembling proteins of the B acillus subtilis spore coat

    Science.gov (United States)

    Jiang, Shuo; Wan, Qiang; Krajcikova, Daniela; Tang, Jilin; Tzokov, Svetomir B.; Barak, Imrich

    2015-01-01

    Summary Bacterial spores (endospores), such as those of the pathogens C lostridium difficile and B acillus anthracis, are uniquely stable cell forms, highly resistant to harsh environmental insults. B acillus subtilis is the best studied spore‐former and we have used it to address the question of how the spore coat is assembled from multiple components to form a robust, protective superstructure. B . subtilis coat proteins (CotY, CotE, CotV and CotW) expressed in E scherichia coli can arrange intracellularly into highly stable macro‐structures through processes of self‐assembly. Using electron microscopy, we demonstrate the capacity of these proteins to generate ordered one‐dimensional fibres, two‐dimensional sheets and three‐dimensional stacks. In one case (CotY), the high degree of order favours strong, cooperative intracellular disulfide cross‐linking. Assemblies of this kind could form exquisitely adapted building blocks for higher‐order assembly across all spore‐formers. These physically robust arrayed units could also have novel applications in nano‐biotechnology processes. PMID:25872412

  13. Atomic force microscopy imaging and single molecule recognition force spectroscopy of coat proteins on the surface of Bacillus subtilis spore.

    Science.gov (United States)

    Tang, Jilin; Krajcikova, Daniela; Zhu, Rong; Ebner, Andreas; Cutting, Simon; Gruber, Hermann J; Barak, Imrich; Hinterdorfer, Peter

    2007-01-01

    Coat assembly in Bacillus subtilis serves as a tractable model for the study of the self-assembly process of biological structures and has a significant potential for use in nano-biotechnological applications. In the present study, the morphology of B. subtilis spores was investigated by magnetically driven dynamic force microscopy (MAC mode atomic force microscopy) under physiological conditions. B. subtilis spores appeared as prolate structures, with a length of 0.6-3 microm and a width of about 0.5-2 microm. The spore surface was mainly covered with bump-like structures with diameters ranging from 8 to 70 nm. Besides topographical explorations, single molecule recognition force spectroscopy (SMRFS) was used to characterize the spore coat protein CotA. This protein was specifically recognized by a polyclonal antibody directed against CotA (anti-CotA), the antibody being covalently tethered to the AFM tip via a polyethylene glycol linker. The unbinding force between CotA and anti-CotA was determined as 55 +/- 2 pN. From the high-binding probability of more than 20% in force-distance cycles it is concluded that CotA locates in the outer surface of B. subtilis spores.

  14. YwdL in Bacillus cereus: its role in germination and exosporium structure.

    Directory of Open Access Journals (Sweden)

    Cassandra Terry

    Full Text Available In members of the Bacillus cereus group the outermost layer of the spore is the exosporium, which interacts with hosts and the environment. Efforts have been made to identify proteins of the exosporium but only a few have so far been characterised and their role in determining spore architecture and spore function is still poorly understood. We have characterised the exosporium protein, YwdL. ΔywdL spores have a more fragile exosporium, subject to damage on repeated freeze-thawing, although there is no evidence of altered resistance properties, and coats appear intact. Immunogold labelling and Western blotting with anti-YwdL antibodies identified YwdL to be located exclusively on the inner surface of the exosporium of B. cereus and B. thuringiensis. We conclude that YwdL is important for formation of a robust exosporium but is not required to maintain the crystalline assembly within the basal layer or for attachment of the hairy nap structure. ΔywdL spores are unable to germinate in response to CaDPA, and have altered germination properties, a phenotype that confirms the expected defect in localization of the cortex lytic enzyme CwlJ in the coat.

  15. Spore Resistance Properties.

    Science.gov (United States)

    Setlow, Peter

    2014-10-01

    Spores of various Bacillus and Clostridium species are among the most resistant life forms known. Since the spores of some species are causative agents of much food spoilage, food poisoning, and human disease, and the spores of Bacillus anthracis are a major bioweapon, there is much interest in the mechanisms of spore resistance and how these spores can be killed. This article will discuss the factors involved in spore resistance to agents such as wet and dry heat, desiccation, UV and γ-radiation, enzymes that hydrolyze bacterial cell walls, and a variety of toxic chemicals, including genotoxic agents, oxidizing agents, aldehydes, acid, and alkali. These resistance factors include the outer layers of the spore, such as the thick proteinaceous coat that detoxifies reactive chemicals; the relatively impermeable inner spore membrane that restricts access of toxic chemicals to the spore core containing the spore's DNA and most enzymes; the low water content and high level of dipicolinic acid in the spore core that protect core macromolecules from the effects of heat and desiccation; the saturation of spore DNA with a novel group of proteins that protect the DNA against heat, genotoxic chemicals, and radiation; and the repair of radiation damage to DNA when spores germinate and return to life. Despite their extreme resistance, spores can be killed, including by damage to DNA, crucial spore proteins, the spore's inner membrane, and one or more components of the spore germination apparatus.

  16. Identification of CdnL, a putative transcriptional regulator involved in repair and outgrowth of heat-damaged bacillus cereus spores

    NARCIS (Netherlands)

    Warda, Alicja K.; Tempelaars, Marcel H.; Boekhorst, Jos; Abee, Tjakko; Groot, Masja Nierop N.

    2016-01-01

    Spores are widely present in the environment and are common contaminants in the food chain, creating a challenge for food industry. Nowadays, heat treatments conventionally applied in food processing may become milder to comply with consumer desire for products with higher sensory and nutritional

  17. Comparative genome analysis of Bacillus cereus group genomes withBacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Iain; Sorokin, Alexei; Kapatral, Vinayak; Reznik, Gary; Bhattacharya, Anamitra; Mikhailova, Natalia; Burd, Henry; Joukov, Victor; Kaznadzey, Denis; Walunas, Theresa; D' Souza, Mark; Larsen, Niels; Pusch,Gordon; Liolios, Konstantinos; Grechkin, Yuri; Lapidus, Alla; Goltsman,Eugene; Chu, Lien; Fonstein, Michael; Ehrlich, S. Dusko; Overbeek, Ross; Kyrpides, Nikos; Ivanova, Natalia

    2005-09-14

    Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1,381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-layer proteins suggesting differences in their phenotype were identified. The B. cereus group has signal transduction systems including a tyrosine kinase related to two-component system histidine kinases from B. subtilis. A model for regulation of the stress responsive sigma factor sigmaB in the B. cereus group different from the well studied regulation in B. subtilis has been proposed. Despite a high degree of chromosomal synteny among these genomes, significant differences in cell wall and spore coat proteins that contribute to the survival and adaptation in specific hosts has been identified.

  18. Gel-free proteomic identification of the Bacillus subtilis insoluble spore coat protein fraction

    NARCIS (Netherlands)

    W. Abhyankar; A. ter Beek; H. Dekker; R. Kort; S. Brul; C.G. de Koster

    2011-01-01

    Species from the genus Bacillus have the ability to form endospores, dormant cellular forms that are able to survive heat and acid preservation techniques commonly used in the food industry. Resistance characteristics of spores towards various environmental stresses are in part attributed to their c

  19. Bacillus globigii bugbeads: a model simulant of a bacterial spore.

    Science.gov (United States)

    Farrell, Svetlana; Halsall, H Brian; Heineman, William R

    2005-01-15

    Nonpathogenic microorganisms are often used as simulants of biological pathogens during the initial phase of detection method development. While these simulants approximate the size, shape, and cellular organization of the microorganism of interest, they do not resemble its surface protein content, a factor particularly important in methods based on immunorecognition. Here, we develop and detect an artificial bacterial spore--B. globigii (BG) Bugbead-a particle mimicking the antigenic surface of BG spores. Two methods of spore protein extraction were compared both quantitatively (by protein concentration assay) and qualitatively (by SDS-PAGE and Western blot): extraction by mechanical disruption and extraction by chemical decoating. The former method was more efficient in producing more protein and a greater number of antigens. BG Bugbeads were made by conjugating the extracted proteins to 0.8-microm carboxyl-coated polystyrene particles via carbodiimide coupling. BG Bugbeads were successfully detected by a bead-based enzyme-labeled immunoassay with fluorescence detection with a detection limit of 6.9 x 10(3) particles/mL. Formation of the Bugbead-capture bead complex was confirmed by ESEM. The concept of a harmless artificial spore can be applied to developing improved simulants for pathogenic spore-forming microorganisms such as B. anthracis, C. botulinum, and B. cereus, which can to be used for method validation, instrument calibration, and troubleshooting.

  20. Anthrax Spores under a microscope

    Science.gov (United States)

    2003-01-01

    Anthrax spores are inactive forms of Bacillus anthracis. They can survive for decades inside a spore's tough protective coating; they become active when inhaled by humans. A result of NASA- and industry-sponsored research to develop small greenhouses for space research is the unique AiroCide TiO2 system that kills anthrax spores and other pathogens.

  1. Heat stress leads to superoxide formation in Bacillus cereus detected using the fluorescent probe MitoSOX

    NARCIS (Netherlands)

    Mols, J.M.; Ceragioli, M.; Abee, T.

    2011-01-01

    Bacillus cereus is a food-borne human pathogen and food spoilage organism. Spores and vegetative cells of B. cereus can be found almost everywhere and therefore often end up in food processing equipment and food products. To remove spores and vegetative cells from food or equipment, harsh treatments

  2. Binding Affinity of Glycoconjugates to BACILLUS Spores and Toxins

    Science.gov (United States)

    Rasol, Aveen; Eassa, Souzan; Tarasenko, Olga

    2010-04-01

    Early recognition of Bacillus cereus group species is important since they can cause food-borne illnesses and deadly diseases in humans. Glycoconjugates (GCs) are carbohydrates covalently linked to non-sugar moieties including lipids, proteins or other entities. GCs are involved in recognition and signaling processes intrinsic to biochemical functions in cells. They also stimulate cell-cell adhesion and subsequent recognition and activation of receptors. We have demonstrated that GCs are involved in Bacillus cereus spore recognition. In the present study, we have investigated whether GCs possess the ability to bind and recognize B. cereus spores and Bacillus anthracis recombinant single toxins (sTX) and complex toxins (cTX). The affinity of GCs to spores + sTX and spores + cTX toxins was studied in the binding essay. Our results demonstrated that GC9 and GC10 were able to selectively bind to B. cereus spores and B. anthracis toxins. Different binding affinities for GCs were found toward Bacillus cereus spores + sTX and spores + cTX. Dilution of GCs does not impede the recognition and binding. Developed method provides a tool for simultaneous recognition and targeting of spores, bacteria toxins, and/or other entities.

  3. Bacillus cereus, a volatile human pathogen.

    Science.gov (United States)

    Bottone, Edward J

    2010-04-01

    Bacillus cereus is a Gram-positive aerobic or facultatively anaerobic, motile, spore-forming, rod-shaped bacterium that is widely distributed environmentally. While B. cereus is associated mainly with food poisoning, it is being increasingly reported to be a cause of serious and potentially fatal non-gastrointestinal-tract infections. The pathogenicity of B. cereus, whether intestinal or nonintestinal, is intimately associated with the production of tissue-destructive exoenzymes. Among these secreted toxins are four hemolysins, three distinct phospholipases, an emesis-inducing toxin, and proteases. The major hurdle in evaluating B. cereus when isolated from a clinical specimen is overcoming its stigma as an insignificant contaminant. Outside its notoriety in association with food poisoning and severe eye infections, this bacterium has been incriminated in a multitude of other clinical conditions such as anthrax-like progressive pneumonia, fulminant sepsis, and devastating central nervous system infections, particularly in immunosuppressed individuals, intravenous drug abusers, and neonates. Its role in nosocomial acquired bacteremia and wound infections in postsurgical patients has also been well defined, especially when intravascular devices such as catheters are inserted. Primary cutaneous infections mimicking clostridial gas gangrene induced subsequent to trauma have also been well documented. B. cereus produces a potent beta-lactamase conferring marked resistance to beta-lactam antibiotics. Antimicrobials noted to be effective in the empirical management of a B. cereus infection while awaiting antimicrobial susceptibility results for the isolate include ciprofloxacin and vancomycin.

  4. Effects of Bacillus cereus Endospores on Free-Living Protist Growth

    DEFF Research Database (Denmark)

    Santos, Susana S; Hendriksen, Niels Bohse; Jakobsen, Hans Henrik;

    2016-01-01

    We studied the predator-prey interactions between heterotrophic protists and endospores of Bacillus cereus group bacteria, in order to gain insight on survival and dispersal of B. cereus endospores in the environment. It has been hypothesised that the spore stage protects against digestion...

  5. Incidence, diversity and characteristics of spores of psychrotolerant spore formers in various REPFEDS produced in Belgium.

    Science.gov (United States)

    Samapundo, S; Devlieghere, F; Xhaferi, R; Heyndrickx, M

    2014-12-01

    The major objectives of this study were to determine the incidence of psychrotolerant spore formers from REPFEDS marketed in Belgium, and their diversity and characteristics. Spore formers in general were found as spores on 38.3% of the food samples and in 85% food products types evaluated. 76% of the food samples containing spore formers had spores before enrichment. A total of 86 spore formers were isolated from the samples. 28 of 86 bacterial spore formers (32.6%) were capable of vegetative growth at 7 °C. 96% (27/28) of these psychrotolerant spore formers were determined to belong to Bacillus or related genera. According to a (GTG)5-PCR analysis, 24 of these 28 isolates were genetically distinct from each other. 10.7% (3/28) of the bacilli were determined to belong to the Bacillus cereus group, namely B. cereus (chicken curry and Edam cheese) and Bacillus mycoides (Emmental cheese). Almost half of the bacilli (12/27) were putatively identified as Bacillus pumilus, which occurs ubiquitously in nature and has been associated with outbreaks of foodborne disease. Only one psychrotolerant clostridium, Clostridium tyrobutyricum, was isolated in the study. The results of this study show the highly diverse ecology and spoilage potential of psychrotolerant spore formers in REPFEDs marketed in Belgium.

  6. 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.

    Science.gov (United States)

    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.

  7. Sampling and quantitative analysis of clean B. subtilis spores at sub-monolayer coverage by reflectance fourier transform infrared microscopy using gold-coated filter substrates.

    Science.gov (United States)

    Brooke, Heather; Perkins, David L; Setlow, Barbara; Setlow, Peter; Bronk, Burt V; Myrick, Michael L

    2008-08-01

    A study was conducted to determine the concentration dependency of the mid-infrared (MIR) absorbance of bacterial spores. A range of concentrations of Bacillus subtilis endospores filtered across gold-coated filter membranes were analyzed by Fourier transform infrared (FT-IR) reflectance microscopy. Calibration curves were derived from the peak absorbances associated with Amide A, Amide I, and Amide II vibrational frequencies by automatic baseline fitting to remove most of the scattering contribution. Linear relationships (R2 >or= 0.99) were observed between the concentrations of spores and the baseline-corrected peak absorbance for each frequency studied. Detection limits for our sampled area of 100 x100 microm2 were determined to be 79, 39, and 184 spores (or 7.92 x 10(5), 3.92 x 10(5), and 1.84 x 10(6) spores/cm2) for the Amide A, Amide I, and Amide II peaks, respectively. Absorbance increased linearly above the scattering baseline with particle surface concentration up to 0.9 monolayer (ML) coverage, with the monolayer density calculated to be approximately 1.17 x 10(8) spores/cm2. Scattering as a function of surface concentration, as estimated from extinction values at wavelengths exhibiting low absorbance, becomes nonlinear at a much lower surface concentration. The apparent scattering cross-section per spore decreased monotonically as concentrations increased toward 1.2 ML, while the absolute scattering decreased between 0.9 ML and 1.2 ML coverage. Calculations suggest that transverse spatial coherence effects are the origin of this nonlinearity, while the onset of nonlinearity in the baseline-corrected absorption is probably due to multiple scattering effects, which appear at a high surface concentration. Absorption cross-sections at peaks of the three bands were measured to be (2.15 +/- 0.05) x 10(-9), (1.48 +/- 0.03) x 10(-9), and (0.805 +/- 0.023) x 10(-9) cm2, respectively. These values are smaller by a factor of 2-4 than expected from the literature

  8. Bacillus cereus immune escape: a journey within macrophages.

    Science.gov (United States)

    Tran, Seav-Ly; Ramarao, Nalini

    2013-10-01

    During bacterial infection, professional phagocytes are attracted to the site of infection, where they constitute a first line of host cell defense. Their function is to engulf and destroy the pathogens. Thus, bacteria must withstand the bactericidal activity of professional phagocytes, including macrophages to counteract the host immune system. Bacillus cereus infections are characterized by bacteremia despite the accumulation of inflammatory cells at the site of infection. This implies that the bacteria have developed means of resisting the host immune system. Bacillus cereus spores survive, germinate, and multiply in contact with macrophages, eventually producing toxins that kill these cells. However, the exact mechanism by which B. cereus evades immune attack remains unclear. This review addresses the interaction between B. cereus and macrophages, highlighting, in particular, the ways in which the bacteria escape the microbicidal activities of professional phagocytes.

  9. Bacillus cereus Biofilms—Same, Only Different

    Science.gov (United States)

    Majed, Racha; Faille, Christine; Kallassy, Mireille; Gohar, Michel

    2016-01-01

    Bacillus cereus displays a high diversity of lifestyles and ecological niches and include beneficial as well as pathogenic strains. These strains are widespread in the environment, are found on inert as well as on living surfaces and contaminate persistently the production lines of the food industry. Biofilms are suspected to play a key role in this ubiquitous distribution and in this persistency. Indeed, B. cereus produces a variety of biofilms which differ in their architecture and mechanism of formation, possibly reflecting an adaptation to various environments. Depending on the strain, B. cereus has the ability to grow as immersed or floating biofilms, and to secrete within the biofilm a vast array of metabolites, surfactants, bacteriocins, enzymes, and toxins, all compounds susceptible to act on the biofilm itself and/or on its environment. Within the biofilm, B. cereus exists in different physiological states and is able to generate highly resistant and adhesive spores, which themselves will increase the resistance of the bacterium to antimicrobials or to cleaning procedures. Current researches show that, despite similarities with the regulation processes and effector molecules involved in the initiation and maturation of the extensively studied Bacillus subtilis biofilm, important differences exists between the two species. The present review summarizes the up to date knowledge on biofilms produced by B. cereus and by two closely related pathogens, Bacillus thuringiensis and Bacillus anthracis. Economic issues caused by B. cereus biofilms and management strategies implemented to control these biofilms are included in this review, which also discuss the ecological and functional roles of biofilms in the lifecycle of these bacterial species and explore future developments in this important research area. PMID:27458448

  10. The Water Cycle, a Potential Source of the Bacterial Pathogen Bacillus cereus

    Directory of Open Access Journals (Sweden)

    Julien Brillard

    2015-01-01

    Full Text Available The behaviour of the sporulating soil-dwelling Bacillus cereus sensu lato (B. cereus sl which includes foodborne pathogenic strains has been extensively studied in relation to its various animal hosts. The aim of this environmental study was to investigate the water compartments (rain and soil water, as well as groundwater closely linked to the primary B. cereus sl reservoir, for which available data are limited. B. cereus sl was present, primarily as spores, in all of the tested compartments of an agricultural site, including water from rain to groundwater through soil. During rain events, leachates collected after transfer through the soil eventually reached the groundwater and were loaded with B. cereus sl. In groundwater samples, newly introduced spores of a B. cereus model strain were able to germinate, and vegetative cells arising from this event were detected for up to 50 days. This first B. cereus sl investigation in the various types of interrelated environments suggests that the consideration of the aquatic compartment linked to soil and to climatic events should provide a better understanding of B. cereus sl ecology and thus be relevant for a more accurate risk assessment of food poisoning caused by B. cereus sl pathogenic strains.

  11. Biodiversity in Bacillus cereus

    NARCIS (Netherlands)

    Pielaat A; Fricker M; Nauta MJ; van Leusden FM; MGB

    2006-01-01

    In het kader van een EU project zijn door de verschillende partners experimenten uitgevoerd om inzicht te krijgen in de variatie in eigenschappen van B. cereus-stammen welke bijdragen aan de mate van virulentie. Hiertoe zijn 100 B. cereus-stammen geselecteerd en eigenschappen zoals toxine-vorming,

  12. Biodiversity in Bacillus cereus

    NARCIS (Netherlands)

    Pielaat A; Fricker M; Nauta MJ; Leusden FM van; MGB

    2006-01-01

    Experiments have been performed by different partners to identify variability in properties of Bacillus cereus strains that contribute to the extent of their virulence as part of an EU project. To this end, 100 B. cereus strains were selected and screened for biological properties, such as toxin pro

  13. Presence and significance of Bacillus cereus in dehydrated potato products.

    Science.gov (United States)

    King, Nicola J; Whyte, Rosemary; Hudson, J Andrew

    2007-02-01

    Dehydrated potato contains Bacillus cereus at a prevalences of 10 to 40% and at numbers usually less than 10(3) CFU g(-1). B. cereus in dehydrated potato is likely to be present as spores that are able to survive drying of the raw vegetable and may represent a significant inoculum in the reconstituted (rehydrated) product where conditions favor germination of, and outgrowth from, spores. Holding rehydrated mashed potato alone, or as part of another product (e.g., potato-topped pie), at temperatures above 10 degrees C and below 60 degrees C may allow growth of vegetative B. cereus. Levels exceeding 10(4) CFU g(-1) are considered hazardous to human health and may be reached within a few hours if stored inappropriately between these temperatures. Foods incorporating mashed potato prepared from dehydrated potato flakes have been implicated in B. cereus foodborne illness. This review is a summary of the information available concerning the prevalence and numbers of B. cereus in dehydrated potato flakes and the rate at which growth might occur in the rehydrated product.

  14. Toxin producing Bacillus cereus persist in ready-to-reheat spaghetti Bolognese mainly in vegetative state.

    Science.gov (United States)

    Rajkovic, Andreja; Kljajic, Milica; Smigic, Nada; Devlieghere, Frank; Uyttendaele, Mieke

    2013-10-15

    The potential of Bacillus cereus to cause a diarrheal toxico-infection is related to its ability to perform de novo enterotoxin production in the small intestine. A prerequisite for this is presence of sufficient numbers of B. cereus that have survived gastro-intestinal passage. It is known that the percentage of survival is much smaller for vegetative cells in comparison to spores and it is therefore important to know the state in which B. cereus is ingested. The results of the current study performed on twelve B. cereus strains, comprising both diarrheal and emetic type, indicate that exposure via contaminated foods mainly concerns vegetative cells. Inoculated vegetative cells grew to high counts, with the growth dynamic depending on the storage temperature. At 28 °C growth to high counts resulted in spore formation, in general, after 1 day of storage. One strain was an exception, producing spores only after 16 days. At 12 °C obtained high counts did not result in spore formation for 11 of 12 tested strains after two weeks of storage. The highest counts and time to sporulation were different between strains, but no difference was observed on the group level of diarrheal and emetic strains. The spore counts were always lower than vegetative cell counts and occurred only when food was obviously sensory spoiled (visual and odor evaluation). Similar observations were made with food inoculated with B. cereus spores instead of vegetative cells. Although the prospect of consuming spores was found very weak, the numbers of vegetative B. cereus cells were high enough, without obvious sensory deviation, to survive in sufficient level to cause diarrheal toxico-infection.

  15. The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host.

    Science.gov (United States)

    Stewart, George C

    2015-12-01

    Much of what we know regarding bacterial spore structure and function has been learned from studies of the genetically well-characterized bacterium Bacillus subtilis. Molecular aspects of spore structure, assembly, and function are well defined. However, certain bacteria produce spores with an outer spore layer, the exosporium, which is not present on B. subtilis spores. Our understanding of the composition and biological functions of the exosporium layer is much more limited than that of other aspects of the spore. Because the bacterial spore surface is important for the spore's interactions with the environment, as well as being the site of interaction of the spore with the host's innate immune system in the case of spore-forming bacterial pathogens, the exosporium is worthy of continued investigation. Recent exosporium studies have focused largely on members of the Bacillus cereus family, principally Bacillus anthracis and Bacillus cereus. Our understanding of the composition of the exosporium, the pathway of its assembly, and its role in spore biology is now coming into sharper focus. This review expands on a 2007 review of spore surface layers which provided an excellent conceptual framework of exosporium structure and function (A. O. Henriques and C. P. Moran, Jr., Annu Rev Microbiol 61:555-588, 2007, http://dx.doi.org/10.1146/annurev.micro.61.080706.093224). That review began a process of considering outer spore layers as an integrated, multilayered structure rather than simply regarding the outer spore components as independent parts.

  16. Analysis of germination capacity and germinant receptor (sub)clusters of genomesequenced Bacillus cereus environmental isolates and model strains

    NARCIS (Netherlands)

    Warda, Alicja K.; Xiao, Yinghua; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Nierop Groot, Masja N.; Abee, Tjakko

    2017-01-01

    Spore germination of 17 Bacillus cereus food isolates and reference strains was evaluated using flow cytometry analysis in combination with fluorescent staining at a single-spore level. This approach allowed for rapid collection of germination data under more than 20 conditions, including heat ac

  17. Evaluation of the Sporicidal Activity of Ethanol Extract of Arctium lappa Root against Bacillus cereus

    OpenAIRE

    Vajihe Karbasizade; Arezoo Dabiri

    2014-01-01

    Background: Bacillus cereus is one of the most common causes of food spoilage, keratitis, endophthalmitis, and panophthalmitis. These bacteria produce spores which are resistant to chemical and physical agents. Nowadays, the sporicidal properties of plants have been considered as alternatives to chemical sporicidal agents. Materials and Methods: In this empirical-experimental study the effect of ethanol extract of edible burdock (Arctium lappa) root has been studied on Bacillus cereus spo...

  18. A cluster of Bacillus cereus bacteremia cases among injection drug users

    OpenAIRE

    Benusic, Michael A; Press, Natasha M; Linda MN Hoang; Romney, Marc G.

    2015-01-01

    Bacillus cereus is a ubiquitous spore-forming organism that is infrequently implicated in extraintestinal infections. The authors report three cases of B cereus bacteremia among injection drug users presenting within one month to an urban tertiary care hospital. Treatment with intravenous vancomycin was successful in all three cases. While temporal association suggested an outbreak, molecular studies of patient isolates using pulsed-field gel electrophoresis did not suggest a common source. A...

  19. The effect of selected factors on the survival of Bacillus cereus in the human gastrointestinal tract.

    Science.gov (United States)

    Berthold-Pluta, Anna; Pluta, Antoni; Garbowska, Monika

    2015-05-01

    Bacillus cereus is a Gram-positive bacterium widely distributed in soil and vegetation. This bacterial species can also contaminate raw or processed foods. Pathogenic B. cereus strains can cause a range of infections in humans, as well as food poisoning of an emetic (intoxication) or diarrheal type (toxico-infection). Toxico-infections are due to the action of the Hbl toxin, Nhe toxin, and cytotoxin K produced by the microorganism in the gastrointestinal tract. This occurs once the spores or vegetative B. cereus cells survive the pH barrier of the stomach and reach the small intestine where they produce toxins in sufficient amounts. This article discusses the effect of various factors on the survival of B. cereus in the gastrointestinal tract, including low pH and the presence of digestive enzymes in the stomach, bile salts in the small intestine, and indigenous microflora in the lower parts of the gastrointestinal tract. Additional aspects also reported to affect B. cereus survival and virulence in the gastrointestinal tract include the interaction of the spores and vegetative cells with enterocytes. In vitro studies revealed that both vegetative B. cereus and spores can survive in the gastrointestinal tract suggesting that the biological form of the microorganism may have less influence on the occurrence of the symptoms of infection than was once believed. It is most likely the interaction between the pathogen and enterocytes that is necessary for the diarrheal form of B. cereus food poisoning to develop. The adhesion of B. cereus to the intestinal epithelium allows the bacterium to grow and produce enterotoxins in the proximity of the epithelium. Recent studies suggest that the human intestinal microbiota inhibits the growth of vegetative B. cereus cells considerably.

  20. Initiation of bacterial spore germination.

    Science.gov (United States)

    Vary, J C; Halvorson, H O

    1968-04-01

    To investigate the problem of initiation in bacterial spore germination, we isolated, from extracts of dormant spores of Bacillus cereus strain T and B. licheniformis, a protein that initiated spore germination when added to a suspension of heat-activated spores. The optimal conditions for initiatory activity of this protein (the initiator) were 30 C in 0.01 to 0.04 m NaCl and 0.01 m tris(hydroxymethyl)aminomethane (pH 8.5). The initiator was inhibited by phosphate but required two co-factors, l-alanine (1/7 of K(m) for l-alanine-inhibited germination) and nicotinamide adenine dinucleotide (1.25 x 10(-4)m). In the crude extract, the initiator activity was increased 3.5-fold by heating the extract at 65 C for 10 min, but the partially purified initiator preparation was completely heat-sensitive (65 C for 5 min). Heat stability could be conferred on the purified initiator by adding 10(-3)m dipicolinic acid. A fractionation of this protein that excluded l-alanine dehydrogenase and adenosine deaminase from the initiator activity was developed. The molecular weight of the initiator was estimated as 7 x 10(4). The kinetics of germination in the presence of initiator were examined at various concentrations of l-alanine and nicotinamide adenine dinucleotide.

  1. BACILLUS CEREUS: ISOLATION IN JENNET MILK

    Directory of Open Access Journals (Sweden)

    M.L. Scatassa

    2011-01-01

    Full Text Available Jennet milk as human food is hypoallergenic for patients affected by Cow Milk Protein Allergy and multiple food allergies. For these pathologies, jennet milk represents the best alternative to other types of milk. Therefore, jennet milk consumers are very sensible to the effects of pathogens' contaminations, and several hygienic practices during the milk production need to be adopted. During regular monitoring in one Sicilian jennet farm, Bacillus cereus in the milk was detected. In 3 bulk milk samples (maximum concentration: 1.2 x 103 ufc/ml, in 3 individual milk samples (10, 20 e 60 ufc/ml, in the milk filter (5 ufc/cm2, in the soil (maximum concentration: 1.5 x 103 ufc/g, on the hands and the gloves of two milkers, on the animal hide (from 1 to 3 ufc/cm2. No spores were detected. A total of 8 Bacillus cereus s.s. strains were analyzed for diarrhoic toxin, and 6 strains producing enterotoxins resulted. The improvement of environmental and milking hygienic conditions reduced Bacillus cereus concentration.

  2. A Cluster of Bacillus cereus Bacteremia Cases among Injection Drug Users

    Directory of Open Access Journals (Sweden)

    Michael A Benusic

    2015-01-01

    Full Text Available Bacillus cereus is a ubiquitous spore-forming organism that is infrequently implicated in extraintestinal infections. The authors report three cases of B cereus bacteremia among injection drug users presenting within one month to an urban tertiary care hospital. Treatment with intravenous vancomycin was successful in all three cases. While temporal association suggested an outbreak, molecular studies of patient isolates using pulsed-field gel electrophoresis did not suggest a common source. A review of the association of B cereus infections with heroin use and treatment of this pathogen is provided.

  3. A cluster of Bacillus cereus bacteremia cases among injection drug users.

    Science.gov (United States)

    Benusic, Michael A; Press, Natasha M; Hoang, Linda Mn; Romney, Marc G

    2015-01-01

    Bacillus cereus is a ubiquitous spore-forming organism that is infrequently implicated in extraintestinal infections. The authors report three cases of B cereus bacteremia among injection drug users presenting within one month to an urban tertiary care hospital. Treatment with intravenous vancomycin was successful in all three cases. While temporal association suggested an outbreak, molecular studies of patient isolates using pulsed-field gel electrophoresis did not suggest a common source. A review of the association of B cereus infections with heroin use and treatment of this pathogen is provided.

  4. gerR, a novel ger operon involved in L-alanine- and inosine-initiated germination of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Hornstra, L.M.; Vries, de Y.P.; Vos, de W.M.; Abee, T.; Wells-Bennik, M.H.J.

    2005-01-01

    Bacillus cereus endospores germinate in response to particular nutrients. Spores are able to sense these nutrients in the environment by receptors encoded by the gerA family of operons. Analysis of the Bacillus cereus ATCC 14579 genome revealed seven gerA family homologues. Using a transposon Tn917-

  5. Genomics, evolution, and crystal structure of a new family of bacterial spore kinases

    OpenAIRE

    2009-01-01

    Bacterial spore formation is a complex process of fundamental relevance to biology and human disease. The spore coat structure is complex and poorly understood, and the roles of many of the protein components remain unclear. We describe a new family of spore coat proteins, the bacterial spore kinases (BSKs), and the first crystal structure of a BSK, YtaA (CotI) from Bacillus subtilis. BSKs are widely distributed in spore-forming Bacillus and Clostridium species, and have a dynamic evolutionar...

  6. Characterization of Bacillus cereus

    NARCIS (Netherlands)

    Wijnands LM; Dufrenne JB; Leusden FM; MGB

    2002-01-01

    Bacillus cereus is a ubiquitary microorganism that may cause food borne disease. Pathogenicity, however, depends on various characteristics such as the ability to form (entero)-toxin(s) that can not be detected by microbiological methods. Further characterization of pathogenic properties is not only

  7. Morphology and physico-chemical properties of Bacillus spores surrounded or not with an exosporium: consequences on their ability to adhere to stainless steel.

    Science.gov (United States)

    Faille, Christine; Lequette, Yannick; Ronse, Annette; Slomianny, Christian; Garénaux, Estelle; Guerardel, Yann

    2010-10-15

    This study was designed to elucidate the influence of spore properties such as the presence of an exosporium, on their ability to adhere to materials. This analysis was performed on 17 strains belonging to the B. cereus group and to less related Bacillus species. We first demonstrated that spores of the B. cereus group, surrounded by an exosporium, differed in their morphological features such as exosporium size, number of appendages or hair-like nap length. We also found that the saccharidic composition of exosporium differed among strains, e.g. concerning a newly identified rhamnose derivative: the 2,4-O-dimethyl-rhamnose. Conversely, spores of distant Bacillus species shared morphological and physico-chemical properties with B. cereus spores. Some external features were also observed on these spores, such as a thin loose-fitting layer, whose nature is still to be determined, or a thick saccharidic layer (mainly composed of rhamnose and quinovose). The ability of spores to adhere to stainless steel varied among strains, those belonging to the B. cereus group generally being the most adherent. However, the presence of an exosporium is not sufficient to explain the ability of spores to adhere to inanimate surfaces. Indeed, when the 17 strains were compared, hydrophobicity and the number of appendages were the only significant adhesion parameters. Furthermore, the differences in spore adhesion observed within the B. cereus group were related to differences in the number of appendages, the exosporium length and to a lesser extent, the zeta potential.

  8. Characterization of bacterial spore germination using integrated phase contrast microscopy, Raman spectroscopy, and optical tweezers.

    Science.gov (United States)

    Kong, Lingbo; Zhang, Pengfei; Setlow, Peter; Li, Yong-Qing

    2010-05-01

    We present a methodology that combines external phase contrast microscopy, Raman spectroscopy, and optical tweezers to monitor a variety of changes during the germination of single Bacillus cereus spores in both nutrient (l-alanine) and non-nutrient (Ca-dipicolinic acid (DPA)) germinants with a temporal resolution of approximately 2 s. Phase contrast microscopy assesses changes in refractility of individual spores during germination, while Raman spectroscopy gives information on changes in spore-specific molecules. The results obtained include (1) the brightness of the phase contrast image of an individual dormant spore is proportional to the level of CaDPA in that spore; (2) the end of the first Stage of germination, revealed as the end of the rapid drop in spore refractility by phase contrast microscopy, precisely corresponds to the completion of the release of CaDPA as revealed by Raman spectroscopy; and (3) the correspondence between the rapid drop in spore refractility and complete CaDPA release was observed not only for spores germinating in the well-controlled environment of an optical trap but also for spores germinating when adhered on a microscope coverslip. Using this latter method, we also simultaneously characterized the distribution of the time-to-complete-CaDPA release (T(release)) of hundreds of individual B. cereus spores germinating with both saturating and subsaturating concentrations of l-alanine and with CaDPA.

  9. One-day pulsed-field gel electrophoresis protocol for rapid determination of emetic Bacillus cereus isolates.

    Science.gov (United States)

    Kaminska, Paulina S; Fiedoruk, Krzysztof; Jankowska, Dominika; Mahillon, Jacques; Nowosad, Karol; Drewicka, Ewa; Zambrzycka, Monika; Swiecicka, Izabela

    2015-04-01

    Bacillus cereus, the Gram-positive and spore-forming ubiquitous bacterium, may cause emesis as the result of food intoxication with cereulide, a heat-stable emetic toxin. Rapid determination of cereulide-positive B. cereus isolates is of highest importance due to consequences of this intoxication for human health and life. Here we present a 1-day pulsed-field gel electrophoresis for emetic B. cereus isolates, which allows rapid and efficient determination of their genomic relatedness and helps determining the source of intoxication in case of outbreaks caused by these bacilli.

  10. Evaluation of the Sporicidal Activity of Ethanol Extract of Arctium lappa Root against Bacillus cereus

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    Vajihe Karbasizade

    2014-10-01

    Full Text Available Background: Bacillus cereus is one of the most common causes of food spoilage, keratitis, endophthalmitis, and panophthalmitis. These bacteria produce spores which are resistant to chemical and physical agents. Nowadays, the sporicidal properties of plants have been considered as alternatives to chemical sporicidal agents. Materials and Methods: In this empirical-experimental study the effect of ethanol extract of edible burdock (Arctium lappa root has been studied on Bacillus cereus spores. In this investigation, the suspensions of tested microorganisms were cultured in sporulating agar. Sporulation process was assessed by optical microscopy following the staining of spores. Then the produced spores were exposed to various concentrations (100, 150, 200, 250, 300 mg/mL of ethanol extract of edible burdock (Arctium lappa root and finally the remaining spores were counted. With increasing concentrations of ethanol extract, the number of spores declined. Results: Pearson correlation showed inverse relation between the spores count and concentration of ethanol extract of edible burdock (Arctium lappa root (r=-0.765, p<0.001. The most effective extract concentration was 300 mg /mL. Conclusion: Ethanol extract of edible burdock (Arctium lappa root, has sporicidal activity. Only, the sporicidal nature of ethanol extract has been evaluated by this study; therefore, the assessment of other extracts and essences is necessary.

  11. Current Physical and SDS Extraction Methods Do Not Efficiently Remove Exosporium Proteins from Bacillus anthracis spores

    Science.gov (United States)

    Thompson, Brian M.; Binkley, Jana M.; Stewart, George C.

    2011-01-01

    Biochemical studies of the outermost spore layers of the Bacillus cereus family are hindered by difficulties in efficient dispersal of the external spore layers and difficulties in dissociating protein complexes that comprise the exosporium layer. Detergent and physical methods have been utilized to disrupt the exosporium layer. Herein we compare commonly used SDS extraction buffers used to extract spore proteins and demonstrate the incomplete extractability of the exosporium layer by these methods. Sonication and bead beating methods for exosporium layer removal were also examined. A combination of genetic and physical methods is the most effective for isolating proteins found in the spore exosporium. PMID:21338631

  12. Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific.

    Science.gov (United States)

    Maes, Emmanuel; Krzewinski, Frederic; Garenaux, Estelle; Lequette, Yannick; Coddeville, Bernadette; Trivelli, Xavier; Ronse, Annette; Faille, Christine; Guerardel, Yann

    2016-04-29

    The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets.

  13. Bacillus cereus food poisoning: international and Indian perspective.

    Science.gov (United States)

    Tewari, Anita; Abdullah, Swaid

    2015-05-01

    Food borne illnesses result from eating food or drinking beverages that are contaminated with chemical matter, heavy metals, parasites, fungi, viruses and Bacteria. Bacillus cereus is one of the food-borne disease causing Bacteria. Species of Bacillus and related genera have long been troublesome to food producers on account of their resistant endospores. Their spores may be present on various types of raw and cooked foods, and their ability to survive high cooking temperatures requires that cooked foods be served hot or cooled rapidly to prevent the growth of this bacteria. Bacillus cereus is well known as a cause of food poisoning, and much more is now known about the toxins produced by various strains of this species, so that its significance in such episodes are clearer. However, it is still unclear why such cases are so rarely reported worldwide.

  14. Toxigenic potential and heat survival of spore-forming bacteria isolated from bread and ingredients.

    Science.gov (United States)

    De Bellis, Palmira; Minervini, Fiorenza; Di Biase, Mariaelena; Valerio, Francesca; Lavermicocca, Paola; Sisto, Angelo

    2015-03-16

    Fifty-four spore-forming bacterial strains isolated from bread ingredients and bread, mainly belonging to the genus Bacillus (including Bacillus cereus), together with 11 reference strains were investigated to evaluate their cytotoxic potential and heat survival in order to ascertain if they could represent a risk for consumer health. Therefore, we performed a screening test of cytotoxic activity on HT-29 cells using bacterial culture filtrates after growing bacterial cells in Brain Heart Infusion medium and in the bread-based medium Bread Extract Broth (BEB). Moreover, immunoassays and PCR analyses, specifically targeting already known toxins and related genes of B. cereus, as well as a heat spore inactivation assay were carried out. Despite of strain variability, the results clearly demonstrated a high cytotoxic activity of B. cereus strains, even if for most of them it was significantly lower in BEB medium. Cytotoxic activity was also detected in 30% of strains belonging to species different from B. cereus, although, with a few exceptions (e.g. Bacillus simplex N58.2), it was low or very low. PCR analyses detected the presence of genes involved in the production of NHE, HBL or CytK toxins in B. cereus strains, while genes responsible for cereulide production were not detected. Production of NHE and HBL toxins was also confirmed by specific immunoassays only for B. cereus strains even if PCR analyses revealed the presence of related toxin genes also in some strains of other species. Viable spore count was ascertained after a heat treatment simulating the bread cooking process. Results indicated that B. amyloliquefaciens strains almost completely survived the heat treatment showing less than 2 log-cycle reductions similarly to two strains of B. cereus group III and single strains belonging to Bacillus subtilis, Bacillus mojavensis and Paenibacillus spp. Importantly, spores from strains of the B. cereus group IV exhibited a thermal resistance markedly lower than B

  15. Flow-cytometric Analysis of Bacillus anthracis Spores

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    D. V. Kamboj

    2006-11-01

    Full Text Available Flow-cytometric technique has been established as a powerful tool for detection andidentification of microbiological agents. Unambiguous and rapid detection of Bacillus anthracisspores has been reported using immunoglobulin G-fluorescein isothiocyanate conjugate againstlive spores. In addition to the high sensitivity, the present technique could differentiate betweenspores of closely related species, eg, Bacillus cereus and Bacillus subtilis using fluorescenceintensity. The technique can be used for detection of live as well as inactivated spores makingit more congenial for screening of suspected samples of bioterrorism.

  16. Inhibition of Bacillus cereus Growth and Toxin Production by Bacillus amyloliquefaciens RD7-7 in Fermented Soybean Products.

    Science.gov (United States)

    Eom, Jeong Seon; Choi, Hye Sun

    2016-01-01

    Bacillus cereus is a gram-positive, rod-shaped, spore-forming bacterium that has been isolated from contaminated fermented soybean food products and from the environment. B. cereus produces diarrheal and emetic toxins and has caused many outbreaks of foodborne diseases. In this study, we investigated whether B. amyloliquefaciens RD7-7, isolated from rice doenjang (Korean fermented soybean paste), a traditional Korean fermented soybean food, shows antimicrobial activity against B. cereus and regulates its toxin gene expression. B. amyloliquefaciens RD7-7 exhibited strong antibacterial activity against B. cereus and inhibited the expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM). We also found that addition of water extracts of soybean and buckwheat soksungjang (Korean fermented soybean paste made in a short time) fermented with B. amyloliquefaciens RD7-7 significantly reduced the growth and toxin expression of B. cereus. These results indicate that B. amyloliquefaciens RD7-7 could be used to control B. cereus growth and toxin production in the fermented soybean food industry. Our findings also provide a basis for the development of candidate biological control agents against B. cereus to improve the safety of fermented soybean food products.

  17. [The significance of some potentially pathogenic microorganisms in occurence of food toxicoinfections. Report 2. Assessment of the role of toxigenic strains of Bacillus cereus].

    Science.gov (United States)

    Efimochkina, N R; Batishcheva, S Iu; Bykova, I B; Sheveleva, S A

    2012-01-01

    The data on nomenclature, classification and taxonomy of aerobic spore-forming Bacillus cereus are summarized. The main features of the two types of diseases, caused B. cereus, and statistical information on outbreaks of food-borne disease caused by B. cereus are presented. The detailed description of emetic toxin cereulide properties are given. The analysis of existing methods for detection of the presence of B. cereus and their toxins in foods are conducted. The data on the use of different cell models for studying the cytotoxic effects and the enterotoxigenic properties of B. cereus are described. Results of own researches allowed to conclude that certain types of products, primarily made from milk and vegetable raw materials, can be a source of transmission to humans of toxins produced by B. cereus. It is shown that in the absence of competing vegetative microflora increases the risk of accumulation of toxins produced by the most stable populations, including toxigenic spore B. cereus. Tested and proposed for the practical implementation of the dry culture media on the basis of the balanced growth and selective components, dyes and buffer mixtures. The developed environment were used for the isolation and identification B. cereus during microbial control these groups of foods.

  18. Contamination of healthcare workers' hands with bacterial spores.

    Science.gov (United States)

    Sasahara, Teppei; Ae, Ryusuke; Watanabe, Michiyo; Kimura, Yumiko; Yonekawa, Chikara; Hayashi, Shunji; Morisawa, Yuji

    2016-08-01

    Clostridium species and Bacillus spp. are spore-forming bacteria that cause hospital infections. The spores from these bacteria are transmitted from patient to patient via healthcare workers' hands. Although alcohol-based hand rubbing is an important hand hygiene practice, it is ineffective against bacterial spores. Therefore, healthcare workers should wash their hands with soap when they are contaminated with spores. However, the extent of health care worker hand contamination remains unclear. The aim of this study is to determine the level of bacterial spore contamination on healthcare workers' hands. The hands of 71 healthcare workers were evaluated for bacterial spore contamination. Spores attached to subject's hands were quantitatively examined after 9 working hours. The relationship between bacterial spore contamination and hand hygiene behaviors was also analyzed. Bacterial spores were detected on the hands of 54 subjects (76.1%). The mean number of spores detected was 468.3 CFU/hand (maximum: 3300 CFU/hand). Thirty-seven (52.1%) and 36 (50.7%) subjects were contaminated with Bacillus subtilis and Bacillus cereus, respectively. Nineteen subjects (26.8%) were contaminated with both Bacillus species. Clostridium difficile was detected on only one subject's hands. There was a significant negative correlation between the hand contamination level and the frequency of handwashing (r = -0.44, P < 0.01) and a significant positive correlation between the hand contamination level and the elapsed time since last handwashing (r = 0.34, P < 0.01). Healthcare workers' hands may be frequently contaminated with bacterial spores due to insufficient handwashing during daily patient care.

  19. Diversity of commensal Bacillus cereus sensu lato isolated from the common sow bug (Porcellio scaber, Isopoda).

    Science.gov (United States)

    Swiecicka, Izabela; Mahillon, Jacques

    2006-04-01

    Although Bacillus cereus sensu lato are important both from an ecological and an economical point of view, little is known about their population structure, ecology, and relationships with other organisms. In the present work, the genotypic similarity of arthropod-borne B. cereus s.l. isolates, and their symbiotic relationship with the host are assessed. Bacilli of this group were recovered from the digestive tracts of sow bugs (Porcellio scaber) collected in three closely located sites. Their genotypic diversity was investigated using pulse-field gel electrophoresis (PFGE) following the whole-genome DNA digestions with NotI and AscI, and PCR amplification of virulence genes. The majority of the sow-bug Bacillus cereus sensu stricto isolates originating from the same but also from different sites displayed identical PFGE patterns, virulence gene content and enterotoxicity, indicating strong genetic and genomic relationships. The sow-bug Bacillus mycoides/Bacillus pseudomycoides strains displayed a higher diversity. The isopod-B. cereus s.l. relationship was also evaluated using antibiotic-resistant derivatives of B. cereus s.s., B. mycoides/B. pseudomycoides and Bacillus thuringiensis reintroduced into sow bugs. Both spores and vegetative cells of B. cereus s.l. were recovered from sow bugs over a 30-day period, strongly suggesting that these bacteria are natural residents of terrestrial isopods.

  20. Characterization of Bacillus cereus isolates from local dairy farms in China.

    Science.gov (United States)

    Cui, Yifang; Liu, Xiaoye; Dietrich, Richard; Märtlbauer, Erwin; Cao, Jie; Ding, Shuangyang; Zhu, Kui

    2016-06-01

    Bacillus cereus is an important opportunistic foodborne pathogen. In the present work, a total of 306 milk and environmental samples were collected from 10 local dairy farms in Beijing, China. Of the 92 B. cereus-like isolates, 88 and 4 belonged to B. cereus and B. thuringiensis, respectively. The prevalence of B. cereus isolates in bedding, feces, feed, liquid manure and raw milk was 93.3%, 78.9%, 41.2%, 100.0% and 9.8%, respectively. Three main toxin genes nhe, hbl and ces were detected with rates of 100.0%, 78.3% and 1.1%, but no strain harbored cytK1 The production of Nhe, Hbl and cereulide could be confirmed by specific monoclonal antibodies-based enzyme immunoassays in 94.6%, 70.7% and 1.1% of all isolates, respectively. Cytotoxicity tests were used to further corroborate the results of genetic and protein-based assays; 91.3% of the isolates showed cytotoxicity to Vero cells. All isolates were tested for antimicrobial resistance against 17 antibiotics. All isolates were resistant to lincomycin, retapamulin, tiamulin and valnemulin, while two strains were susceptible to ampicillin and ceftiofur. A total of 16 isolated strains were resistant to tetracycline. Since spores of B. cereus are not inactivated during manufacturing of most milk products, contamination of milk with B. cereus on the farm level may represent a potential hazard, particularly with respect to emetic toxin-producing strains.

  1. Bacillus cereus in Brazilian Ultra High Temperature milk Bacillus cereus em leite UHT brasileiro

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    Cristiana de Paula Pacheco-Sanchez

    2007-01-01

    Full Text Available Brazilian Ultra High Temperature (UHT milk consumption has increased during the last decade from 187 to 4,200 million liters. In the continuous UHT process, milk is submitted for 2-4 s to 130-150ºC, in a continuous flow system with immediate refrigeration and aseptical packing in hermetic packages. This research had the purpose to verify the incidence of B. cereus species from the B. cereus group, in UHT milk. In 1998 high indexes of these organisms were reported, reaching 34.14% of the analyzed samples. Beyond this fact, there was the need to establish methods and processes adjusted for correct identification of B. cereus. Thus, commercial sterility tests of 6,500 UHT milk packages were investigated in two assays, after ten days incubation at 37ºC and 7ºC to germinate all possible spores and/or to recuperate injured vegetative cells followed by pH measurement. Samples (1,300 packages each from five Brazilian UHT plants of whole UHT milk processed by direct steam injection, packaged in carton were investigated for the presence of Bacillus cereus through phenotypic and genetic (PCR tests. Values of pH were different for the samples, ranging between 6.57 and 6.73. After storage of the samples, only four packages with pH measurement below the lower limit of 6.5 were found and analyzed for the presence of B. cereus. This organism was not detected in any of the samples indicating that the five Brazilian UHT milk processors control pathogenic microorganisms and it can be said that the consumption of UHT milk does not present safety problems to consumers. Fourier Transform Infrared Spectroscopy (FTIR and PCR tests were efficient and must be adopted to confirm the biochemical series for B. cereus.O consumo de leite ultra-alta temperatura (UHT brasileiro aumentou, durante a última década, de 187 milhões de litros para 4,200 milhões de litros. No processo contínuo de leite UHT o leite é submetido por 2-4 seg a 130-150ºC, em sistemas de

  2. A bacterial spore model of pulsed electric fields on spore morphology change revealed by simulation and SEM.

    Science.gov (United States)

    Qiu, Xing; Lee, Yin Tung; Yung, Pun To

    2014-01-01

    A two-layered spore model was proposed to analyze morphological change of bacterial spores subjected under pulsed electric fields. The outer layer, i.e. spore coat, was defined by Mooney-Rivlin hyper-elastic material model. The inner layer, i.e. peptidoglycan and spore core, was modeled by applying additional adhesion forces. The effect of pulsed electric fields on surface displacement was simulated in COMSOL Multiphysics and verified by SEM. The electro-mechanical theory, considering spore coat as a capacitor, was used to explain concavity; and the thin viscoelastic film theory, considering membrane bilayer as fluctuating surfaces, was used to explain leakage forming. Mutual interaction of external electric fields, charged spores, adhesion forces and ions movement were all predicted to contribute to concavity and leakage.

  3. Ultraviolet irradiation of DNA complexed with. alpha. /. beta. -type small, acid-soluble proteins from spores of Bacillus or Clostridium species makes spore photoproduct but not thymine dimers

    Energy Technology Data Exchange (ETDEWEB)

    Nicholson, W.L.; Setlow, B.; Setlow, P. (Univ. of Connecticut Health Center, Farmington (United States))

    1991-10-01

    UV irradiation of complexes of DNA and an {alpha}/{beta}-type small, acid-soluble protein (SASP) from Bacillus subtilis spores gave decreasing amounts of pyrimidine dimers and increasing amounts of spore photoproduct as the SASP/DNA ratio was increased. The yields of pyrimidine dimers and spore photoproduct were < 0.2% and 8% of total thymine, respectively, when DNA saturated with SASP was irradiated at 254 nm with 30 kJ/m{sup 2}; in the absence of SASP the yields were reversed - 4.5% and 0.3%, respectively. Complexes of DNA with {alpha}/{beta}-type SASP from Bacillus cereus, Bacillus megaterium, or Clostridium bifermentans spores also gave spore photoproduct upon UV irradiation. However, incubation of these SASPs with DNA under conditions preventing complex formation or use of mutant SASPs that do not form complexes did not affect the photoproducts formed in vitro. These results suggest that the UV photochemistry of bacterial spore DNA in vivo is due to the binding of {alpha}/{beta}-type SASP, a binding that is known to cause a change in DNA conformation in vitro from the B form to the A form. The yields of spore photoproduct in vitro were significantly lower than in vivo, perhaps because of the presence of substances other than SASP in spores. It is suggested that as these factors diffuse out in the first minutes of spore germination, spore photoproduct yields become similar to those observed for irradiation of SASP/DNA complexes in vitro.

  4. The Pore-Forming Haemolysins of Bacillus Cereus: A Review

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    Vincent Sanchis

    2013-06-01

    Full Text Available The Bacillus cereus sensu lato group contains diverse Gram-positive spore-forming bacteria that can cause gastrointestinal diseases and severe eye infections in humans. They have also been incriminated in a multitude of other severe, and frequently fatal, clinical infections, such as osteomyelitis, septicaemia, pneumonia, liver abscess and meningitis, particularly in immuno-compromised patients and preterm neonates. The pathogenic properties of this organism are mediated by the synergistic effects of a number of virulence products that promote intestinal cell destruction and/or resistance to the host immune system. This review focuses on the pore-forming haemolysins produced by B. cereus: haemolysin I (cereolysin O, haemolysin II, haemolysin III and haemolysin IV (CytK. Haemolysin I belongs to the cholesterol-dependent cytolysin (CDC family whose best known members are listeriolysin O and perfringolysin O, produced by L. monocytogenes and C. perfringens respectively. HlyII and CytK are oligomeric ß-barrel pore-forming toxins related to the α-toxin of S. aureus or the ß-toxin of C. perfringens. The structure of haemolysin III, the least characterized haemolytic toxin from the B. cereus, group has not yet been determined.

  5. The Arthromitus stage of Bacillus cereus: intestinal symbionts of animals

    Science.gov (United States)

    Margulis, L.; Jorgensen, J. Z.; Dolan, S.; Kolchinsky, R.; Rainey, F. A.; Lo, S. C.

    1998-01-01

    In the guts of more than 25 species of arthropods we observed filaments containing refractile inclusions previously discovered and named "Arthromitus" in 1849 by Joseph Leidy [Leidy, J. (1849) Proc. Acad. Nat. Sci. Philadelphia 4, 225-233]. We cultivated these microbes from boiled intestines of 10 different species of surface-cleaned soil insects and isopod crustaceans. Literature review and these observations lead us to conclude that Arthromitus are spore-forming, variably motile, cultivable bacilli. As long rod-shaped bacteria, they lose their flagella, attach by fibers or fuzz to the intestinal epithelium, grow filamentously, and sporulate from their distal ends. When these organisms are incubated in culture, their life history stages are accelerated by light and inhibited by anoxia. Characterization of new Arthromitus isolates from digestive tracts of common sow bugs (Porcellio scaber), roaches (Gromphodorhina portentosa, Blaberus giganteus) and termites (Cryptotermes brevis, Kalotermes flavicollis) identifies these flagellated, spore-forming symbionts as a Bacillus sp. Complete sequencing of the 16S rRNA gene from four isolates (two sow bug, one hissing roach, one death's head roach) confirms these as the low-G+C Gram-positive eubacterium Bacillus cereus. We suggest that B. cereus and its close relatives, easily isolated from soil and grown on nutrient agar, enjoy filamentous growth in moist nutrient-rich intestines of healthy arthropods and similar habitats.

  6. Detection of hblA and bal Genes in Bacillus cereus Isolates From Cheese Samples Using the Polymerase Chain Reaction

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    Molayi Kohneshahri

    2016-03-01

    Full Text Available Background Bacillus cereus is a Gram-positive spore-forming bacterium, which causes food poisoning. Spores enable the persistence of B. cereus in the environment, and B. cereus strains can tolerate adverse environmental conditions, such as temperature and insufficient nutrients. B. cereus causes food poisoning via the production of two enterotoxins. Most isolates produce toxins leading to diarrhea (enterotoxins and vomiting (emetic forms. Diarrhea is caused by the production of three different heat-labile enterotoxins: HBL, NHE, and cytotoxin K. A heat-stable toxin, cereulide, is responsible for emesis. Objectives This study aimed to detect enterotoxigenic B. cereus isolates in cheese samples using the polymerase chain reaction (PCR. Materials and Methods Two-hundred pasteurized (n = 100 and nonpasteurized (n = 100 cheese samples were collected. The initial isolation was performed on PEMBA specific medium. Antibiotic susceptibility testing was performed using several antibiotic disks, according to the guidelines of the Clinical Laboratory and Standards Institute. Specific primers amplifying the hblA enterotoxin-encoding gene and bal hemolysin-encoding gene were used for the molecular detection of the toxins. Results Ten samples were positive for the presence of B. cereus, with both Gram staining and biochemical reactions. All the isolates were resistant to penicillin and ampicillin but susceptible to vancomycin, erythromycin, and ciprofloxacin. Six and three isolates were resistant to tetracycline and trimethoprim-sulfamethoxazole, respectively. The hblA and bal genes were amplified in all the B. cereus isolates. Conclusions The prevalence of B. cereus among the cheese samples was low. All the isolates were positive for genes encoding the hblA enterotoxin and bal toxin.

  7. A simple identification method for spore-forming bacteria showing high resistance against [gamma]-rays

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    Koshikawa, Tomihiko; Sone, Koji; Kobayashi, Toshikazu (Japan Radioisotope Association, Koka, Shiga (Japan). Koka Lab.)

    1993-11-01

    A simple identification method was developed for spore-forming bacteria which are highly resistant against [gamma]-rays. Among 23 species of Bacillus studied, the spores of Bacillus megaterium, B. cereus, B. thuringiensis, B. pumilus and B. aneurinolyticus showed high resistance against [gamma]-rays as compared with other spores of Bacillus species. Combination of the seven kinds of biochemical tests, namely, the citrate utilization test, nitrate reduction test, starch hydrolysis test, Voges-Proskauer reaction test, gelatine hydrolysis test, mannitol utilization test and xylose utilization test showed a characteristic pattern for each species of Bacillus. The combination pattern of each the above tests with a few supplementary test, if necessary, was useful to identify Bacillus species showing high radiation resistance against [gamma]-rays. The method is specific for B. megaterium, B. thuringiensis and B. pumilus, and highly selective for B. aneurinolyticus and B. cereus. (author).

  8. Identification and validation of specific markers of Bacillus anthracis spores by proteomics and genomics approaches.

    Science.gov (United States)

    Chenau, Jérôme; Fenaille, François; Caro, Valérie; Haustant, Michel; Diancourt, Laure; Klee, Silke R; Junot, Christophe; Ezan, Eric; Goossens, Pierre L; Becher, François

    2014-03-01

    Bacillus anthracis is the causative bacteria of anthrax, an acute and often fatal disease in humans. The infectious agent, the spore, represents a real bioterrorism threat and its specific identification is crucial. However, because of the high genomic relatedness within the Bacillus cereus group, it is still a real challenge to identify B. anthracis spores confidently. Mass spectrometry-based tools represent a powerful approach to the efficient discovery and identification of such protein markers. Here we undertook comparative proteomics analyses of Bacillus anthracis, cereus and thuringiensis spores to identify proteoforms unique to B. anthracis. The marker discovery pipeline developed combined peptide- and protein-centric approaches using liquid chromatography coupled to tandem mass spectrometry experiments using a high resolution/high mass accuracy LTQ-Orbitrap instrument. By combining these data with those from complementary bioinformatics approaches, we were able to highlight a dozen novel proteins consistently observed across all the investigated B. anthracis spores while being absent in B. cereus/thuringiensis spores. To further demonstrate the relevance of these markers and their strict specificity to B. anthracis, the number of strains studied was extended to 55, by including closely related strains such as B. thuringiensis 9727, and above all the B. cereus biovar anthracis CI, CA strains that possess pXO1- and pXO2-like plasmids. Under these conditions, the combination of proteomics and genomics approaches confirms the pertinence of 11 markers. Genes encoding these 11 markers are located on the chromosome, which provides additional targets complementary to the commonly used plasmid-encoded markers. Last but not least, we also report the development of a targeted liquid chromatography coupled to tandem mass spectrometry method involving the selection reaction monitoring mode for the monitoring of the 4 most suitable protein markers. Within a proof

  9. In vitro high-resolution structural dynamics of single germinating bacterial spores

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T; Wheeler, K; Malkin, A

    2006-11-14

    Although significant progress has been achieved in understanding the genetic and biochemical bases of the spore germination process, the structural basis for breaking the dormant spore state remains poorly understood. We have used atomic force microscopy (AFM) to probe the high-resolution structural dynamics of single Bacillus atrophaeus spores germinating under native conditions. Here we show that AFM can reveal previously unrecognized germination-induced alterations in spore coat architecture and topology as well as the disassembly of outer spore coat rodlet structures. These results and previous studies in other microorganisms suggest that the spore coat rodlets are structurally similar to amyloid fibrils. AFM analysis of the nascent surface of the emerging germ cell revealed a porous network of peptidoglycan fibers. The results are consistent with a honeycomb model structure for synthetic peptidoglycan oligomers determined by nuclear magnetic resonance. AFM is a promising experimental tool for investigating the morphogenesis of spore germination and cell wall peptidoglycan structure.

  10. In vitro high-resolution structural dynamics of single germinating bacterial spores

    Energy Technology Data Exchange (ETDEWEB)

    Lawrence Livermore National Laboratory

    2006-12-11

    Although significant progress has been achieved in understanding the genetic and biochemical bases of the spore germination process, the structural basis for breaking the dormant spore state remains poorly understood. We have used atomic force microscopy (AFM) to probe the high-resolution structural dynamics of single Bacillus atrophaeus spores germinating under native conditions. Here we show that AFM can reveal previously unrecognized germination-induced alterations in spore coat architecture and topology as well as the disassembly of outer spore coat rodlet structures. These results and previous studies in other microorganisms suggest that the spore coat rodlets are structurally similar to amyloid fibrils. AFM analysis of the nascent surface of the emerging germ cell revealed a porous network of peptidoglycan fibers. The results are consistent with a honeycomb model structure for synthetic peptidoglycan oligomers determined by nuclear magnetic resonance. AFM is a promising experimental tool for investigating the morphogenesis of spore germination and cell wall peptidoglycan structure.

  11. Hydrazine inactivates bacillus spores

    Science.gov (United States)

    Schubert, Wayne; Plett, G. A.; Yavrouian, A. H.; Barengoltz, J.

    2005-01-01

    Planetary Protection places requirements on the maximum number of viable bacterial spores that may be delivered by a spacecraft to another solar system body. Therefore, for such space missions, the spores that may be found in hydrazine are of concern. A proposed change in processing procedures that eliminated a 0.2 um filtration step propmpted this study to ensure microbial contamination issue existed, especially since no information was found in the literature to substantiate bacterial spore inactivation by hydrazine.

  12. Detection of Bacillus spores within 15 minutes by surface-enhanced Raman spectroscopy

    Science.gov (United States)

    Shende, Chetan; Inscore, Frank; Huang, Hermes; Farquharson, Stuart; Sengupta, Atanu

    2012-06-01

    Since the distribution of Bacillus anthracis causing spores through the US Postal System, there has been a persistent fear that biological warfare agents (BWAs) will be used by terrorists against our military abroad and our civilians at home. Despite the substantial effort to develop BWA analyzers, they remain either too slow, produce high falsealarm rates, lack sensitivity, or cannot be fielded. Consequently there remains a need for a portable analyzer that can overcome these limitations as expressed at the 2011 Biological Weapons Convention. To meet this need we have been developing a sample system that selectively binds BWAs and produce surface-enhanced Raman (SER) spectra using portable Raman spectrometers. Here we describe the use of a short peptide ligand functionalized on silver nanoparticles to selectively capture Bacillus cereus spores (a surrogate of B. anthracis) and their subsequent detection by SER spectroscopy. This technique was used to specifically detect B. cereus spores over closely related species like B. subtilis belonging to the same genus within 15 minutes. Sensitivity of the method was demonstrated by detecting 104 B. cereus spores/mL of water. The technology, once developed should prove invaluable for rapid monitoring of BWAs, which will immensely help first responders and emergency personnel in implementing appropriate counter measures.

  13. Effect of storage temperatures and ingredients on growth of Bacillus cereus in coffee creamers.

    Science.gov (United States)

    Feijoo, S C; Cotton, L N; Watson, C E; Martin, J H

    1997-08-01

    Growth of Bacillus cereus ATCC 33018 was evaluated in half and half (10.5% fat), whipping cream (30% fat), and nondairy creamer (7.5% fat). Samples were inoculated with approximately 10 vegetative cells/ml or 100 spores/ml and were subsequently stored at 4, 7, 23 and 32 degrees C. Within 9 h at 32 degrees C and 11 h at 23 degrees C, in both half and half and whipping cream, vegetative cells and spores reached population levels that can cause foodborne illness. No growth occurred in any product stored at 4 or 7 degrees C. Sodium stearoyl lactylate, a fatty acid derivative that is used as an emulsifier, inhibited growth of spores and vegetative cells in the nondairy creamers stored at either 32 or 23 degrees C.

  14. Cryopreservation of fern spores

    Science.gov (United States)

    Spore banks for ferns are analogous to seed banks for angiosperms and provide a promising ex situ conservation tool because large quantities of germplasm with high genetic variation can be conserved in a small space with low economic and technical costs. Ferns produce two types of spores with very ...

  15. Properties of the Bacillus Cereus strain used in probiotic CenBiot Propriedades da cepa de Bacillus cereus utilizada no probiótico CenBiot

    Directory of Open Access Journals (Sweden)

    Carlos Gil-Turnes

    1999-01-01

    Full Text Available Bacillus cereus CenBiot fulfilled the requirements to be used as probiotic. The spores showed D80 of 14 hs, inhibited Escherichia coli and Yersinia pseudotuberculosis after 24 hs in associative culture, were innocuous for suckling and adult mice and were not inhibited by antibiotics at low concentrations.Bacillus cereus CenBiot possui as características necessárias para ser utilizada como probiótico. Os esporos apresentaram D80 de 14 hs, inibiram Escherichia coli e Yersinia pseudotuberculosis após cultivadas associativamente por 24 hs, foram inócuos para camundongos lactentes e adultos e não foram inibidos por antibióticos a baixas concentrações.

  16. A genomic region involved in the formation of adhesin fibers in Bacillus cereus biofilms

    Directory of Open Access Journals (Sweden)

    Joaquín eCaro-Astorga

    2015-01-01

    Full Text Available Bacillus cereus is a bacterial pathogen that is responsible for many recurrent disease outbreaks due to food contamination. Spores and biofilms are considered the most important reservoirs of B. cereus in contaminated fresh vegetables and fruits. Biofilms are bacterial communities that are difficult to eradicate from biotic and abiotic surfaces because of their stable and extremely strong extracellular matrix. These extracellular matrixes contain exopolysaccharides, proteins, extracellular DNA, and other minor components. Although B. cereus can form biofilms, the bacterial features governing assembly of the protective extracellular matrix are not known. Using the well-studied bacterium B. subtilis as a model, we identified two genomic loci in B. cereus, which encodes two orthologs of the amyloid-like protein TasA of B. subtilis and a SipW signal peptidase. Deletion of this genomic region in B. cereus inhibited biofilm assembly; notably, mutation of the putative signal peptidase SipW caused the same phenotype. However, mutations in tasA or calY did not completely prevent biofilm formation; strains that were mutated for either of these genes formed phenotypically different surface attached biofilms. Electron microscopy studies revealed that TasA polymerizes to form long and abundant fibers on cell surfaces, whereas CalY does not aggregate similarly. Heterologous expression of this amyloid-like cassette in a B. subtilis strain lacking the factors required for the assembly of TasA amyloid-like fibers revealed i the involvement of this B. cereus genomic region in formation of the air-liquid interphase pellicles and ii the intrinsic ability of TasA to form fibers similar to the amyloid-like fibers produced by its B. subtilis ortholog.

  17. Emetic toxin-producing strains of Bacillus cereus show distinct characteristics within the Bacillus cereus group.

    NARCIS (Netherlands)

    Carlin, Frédéric; Fricker, Martina; Pielaat, Annemarie; Heisterkamp, Simon; Shaheen, Ranad; Salonen, Mirja Salkinoja; Svensson, Birgitta; Nguyen-the, Christophe; Ehling-Schulz, Monika

    2006-01-01

    One hundred representative strains of Bacillus cereus were selected from a total collection of 372 B. cereus strains using two typing methods (RAPD and FT-IR) to investigate if emetic toxin-producing hazardous B. cereus strains possess characteristic growth and heat resistance profiles. The strains

  18. Selective detection of 1000 B. anthracis spores within 15 minutes using a peptide functionalized SERS assay.

    Science.gov (United States)

    Farquharson, Stuart; Shende, Chetan; Smith, Wayne; Huang, Hermes; Inscore, Frank; Sengupta, Atanu; Sperry, Jay; Sickler, Todd; Prugh, Amber; Guicheteau, Jason

    2014-12-21

    A surface-enhanced Raman spectroscopy (SERS) assay has been designed to detect Bacillus anthracis spores. The assay consists of silver nanoparticles embedded in a porous glass structure that have been functionalized with ATYPLPIR, a peptide developed to discriminately bind B. anthracis versus other species of Bacillus. Once bound, acetic acid was used to release the biomarker dipicolinic acid from the spores, which was detected by SERS through the addition of silver colloids. This SERS assay was used to selectively bind B. anthracis with a 100-fold selectivity versus B. cereus, and to detect B. anthracis Ames at concentrations of 1000 spores per mL within 15 minutes. The SERS assay measurements provide a basis for the development of systems that can detect spores collected from the air or from water supplies.

  19. Inactivation and ultrastructure analysis of Bacillus spp. and Clostridium perfringens spores.

    Science.gov (United States)

    Brantner, Christine A; Hannah, Ryan M; Burans, James P; Pope, Robert K

    2014-02-01

    Bacterial endospores are resistant to many environmental factors from temperature extremes to ultraviolet irradiation and are generally more difficult to inactivate or kill than vegetative bacterial cells. It is often considered necessary to treat spores or samples containing spores with chemical fixative solutions for prolonged periods of time (e.g., 1-21 days) to achieve fixation/inactivation to enable electron microscopy (EM) examination outside of containment laboratories. Prolonged exposure to chemical fixatives, however, can alter the ultrastructure of spores for EM analyses. This study was undertaken to determine the minimum amount of time required to inactivate/sterilize and fix spore preparations from several bacterial species using a universal fixative solution for EM that maintains the ultrastructural integrity of the spores. We show that a solution of 4% paraformaldehyde with 1% glutaraldehyde inactivated spore preparations of Bacillus anthracis, Bacillus cereus, Bacillus megaterium, Bacillus thuringiensis, and Clostridium perfringens in 30 min, and Bacillus subtilis in 240 min. These results suggest that this fixative solution can be used to inactivate and fix spores from several major groups of bacterial spore formers after 240 min, enabling the fixed preparations to be removed from biocontainment and safely analyzed by EM outside of biocontainment.

  20. A selective chromogenic agar that distinguishes Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis.

    Science.gov (United States)

    Juergensmeyer, Margaret A; Gingras, Bruce A; Restaino, Lawrence; Frampton, Elon W

    2006-08-01

    A selective and differential plating medium, R & F anthracis chromogenic agar (ACA), has been developed for isolating and identifying presumptive colonies of Bacillus anthracis. ACA contains the chromogenic substrate 5-bromo-4-chloro-3-indoxyl-choline phosphate that upon hydrolysis yields teal (blue green) colonies indicating the presence of phosphatidylcholine-specific phospholipase C (PC-PLC) activity. Among seven Bacillus species tested on ACA, only members of the Bacillus cereus group (B. anthracis, B. cereus, and B. thuringiensis) produced teal colonies (PC-PLC positive) having cream rings. Examination of colony morphology in 18 pure culture strains of B. anthracis (15 ATCC strains plus AMES-1-RIID, ANR-1, and AMED-RIID), with one exception, required 48 h at 35 to 37 degrees C for significant color production, whereas only 24 h was required for B. cereus and B. thuringiensis. This differential rate of PC-PLC synthesis in B. anthracis (due to the truncated plcR gene and PlcR regulator in B. anthracis) allowed for the rapid differentiation on ACA of presumptive colonies of B. anthracis from B. cereus and B. thuringiensis in both pure and mixed cultures. Effective recovery of B. anthracis from a variety of matrices having both high (soil and sewage) and low microbial backgrounds (cloth, paper, and blood) spiked with B. anthracis ANR-1 spores suggests the probable utility of ACA plating for B. anthracis recovery in a diversity of applications.

  1. Antibacterial polymer coatings.

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, Mollye C.; Allen, Ashley N.; Barnhart, Meghan; Tucker, Mark David; Hibbs, Michael R.

    2009-09-01

    A series of poly(sulfone)s with quaternary ammonium groups and another series with aldehyde groups are synthesized and tested for biocidal activity against vegetative bacteria and spores, respectively. The polymers are sprayed onto substrates as coatings which are then exposed to aqueous suspensions of organisms. The coatings are inherently biocidal and do not release any agents into the environment. The coatings adhere well to both glass and CARC-coated coupons and they exhibit significant biotoxicity. The most effective quaternary ammonium polymers kills 99.9% of both gram negative and gram positive bacteria and the best aldehyde coating kills 81% of the spores on its surface.

  2. Effects of Chlorine Dioxide on Spore Structural and Fuctional Properties

    Science.gov (United States)

    2006-05-31

    A., Price, B., Leighton, T. and K. Wheeler. 2003. Kinetics of size changes of individual Bacillus thuringiensis spores in response to changes in...vegetative growth . The germination process involves a defined temporal order of events, characterized initially by hydrolysis of the spore coat and...capable of early germination but not resumption of vegetative growth and cell division. We have explored the use of rapid spectrophotometric assays to

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

    Science.gov (United States)

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

    2013-09-02

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

  4. Lifesaving liver transplantation for multi-organ failure caused by Bacillus cereus food poisoning.

    Science.gov (United States)

    Tschiedel, Eva; Rath, Peter-Michael; Steinmann, Jörg; Becker, Heinz; Dietrich, Rudolf; Paul, Andreas; Felderhoff-Müser, Ursula; Dohna-Schwake, Christian

    2015-02-01

    Bacillus cereus is a spore-forming, gram-positive bacterium that causes food poisoning presenting with either emesis or diarrhea. Diarrhea is caused by proteinaceous enterotoxin complexes, mainly hemolysin BL, non-hemolytic enterotoxin (NHE), and cytotoxin K. In contrast, emesis is caused by the ingestion of the depsipeptide toxin cereulide, which is produced in B. cereus contaminated food, particularly in pasta or rice. In general, the illness is mild and self-limiting. However, due to cereulide intoxication, nine severe cases with rhabdomyolysis and/or liver failure, five of them lethal, are reported in literature. Here we report the first case of life-threatening liver failure and severe rhabdomyolysis in this context that could not be survived without emergency hepatectomy and consecutive liver transplantation.

  5. Requirements for the Development of Bacillus Anthracis Spore Reference Materials Used to Test Detection Systems

    Science.gov (United States)

    2006-01-01

    in some strains of Bacillus cereus and Bacillus thuringiensis [55, 56]. The Ba813 marker has been used for a real time PCR assay using Taqman-type...pXO1, the large Bacillus anthracis plasmid harboring the anthrax toxin genes, J. Bacteriol. 181, 6509-6515 (1999). [36] L.B. Price, M. Hugh-Jones, P. J...useful results. The spores of Bacillus anthracis (BA) are particular- ly dangerous because they persist in the environment, and relatively small numbers

  6. High resolution FESEM and TEM reveal bacterial spore attachment.

    Science.gov (United States)

    Panessa-Warren, Barbara J; Tortora, George T; Warren, John B

    2007-08-01

    Transmission electron microscopy (TEM) studies in the 1960s and early 1970s using conventional thin section and freeze fracture methodologies revealed ultrastructural bacterial spore appendages. However, the limited technology at that time necessitated the time-consuming process of imaging serial sections and reconstructing each structure. Consequently, the distribution and function of these appendages and their possible role in colonization or pathogenesis remained unknown. By combining high resolution field emission electron microscopy with TEM images of identical bacterial spore preparations, we have been able to obtain images of intact and sectioned Bacillus and Clostridial spores to clearly visualize the appearance, distribution, resistance (to trypsin, chloramphenicol, and heat), and participation of these structures to facilitate attachment of the spores to glass, agar, and human cell substrates. Current user-friendly commercial field emission scanning electron microscopes (FESEMs), permit high resolution imaging, with high brightness guns at lower accelerating voltages for beam sensitive intact biological samples, providing surface images at TEM magnifications for making direct comparisons. For the first time, attachment structures used by pathogenic, environmental, and thermophile bacterial spores could be readily visualized on intact spores to reveal how specific appendages and outer spore coats participated in spore attachment, colonization, and invasion.

  7. Pacemaker-associated Bacillus cereus endocarditis.

    Science.gov (United States)

    Barraud, Olivier; Hidri, Nadia; Ly, Kim; Pichon, Nicolas; Manea, Petrus; Ploy, Marie-Cécile; Garnier, Fabien

    2012-11-01

    We report the case of a pacemaker-associated Bacillus cereus endocarditis in a nonimmunocompromised patient. Antibiotic treatment was ineffective, and the pacemaker had to be removed. B. cereus was cultured from several blood samples and from the pacemaker electrodes. This case underlines the contribution of the rpoB gene for Bacillus species determination.

  8. A case report with catheter caused Bacillus cereus bacteriemia and investigating the clonal relatedness between the isolates by PFGE

    Directory of Open Access Journals (Sweden)

    Fırat Zafer Mengeloğlu

    2011-09-01

    Full Text Available Bacillus cereus is a motile and spore-forming Gram positive rod and is a microorganism species which cause infections such as food poisoning, endocarditis, bacteremia sepsis.In our case; 57 years old, acute myeloid leukemia diagnosed female patient who had bone narrow transplantation and getting chemotherapy was interned with prediagnosis of neutropenic fever and pneumonia in internal medicine clinics which she admitted with high fever. Since B.cereus growth in peripheral vein blood cultures, catheter blood cultures was performed simultaneously. B.cereus growth repeated in the cultures so catheter caused bacteremia was considered as the diagnosis and antibiotherapy was expanded. After treatment, no growth was observed except catheter blood cultures, general condition of the patient improved and her fever disappeared.Pulsed-field gel electrophoresis was performed in order to investigate the clonal relatedness between B.cereus isolates which were grown in three cultures of separate times and isolates were observed as the same clone.Blood cultures yielded B.cereus are usually considered the possibility of contamination so some clinical laboratories do not perform identification at species level. But the species which is isolated may be the cause of infection, so this approach may be time-consuming for the patient and cause a delay in necessary treatment. In such cases; repeat blood culture should be requested and a decision should be done whether to perform species identification according to the patient’s immune system condition and clinical presentation.

  9. Presence and growth of Bacillus cereus in dehydrated potato flakes and hot-held, ready-to-eat potato products purchased in New Zealand.

    Science.gov (United States)

    Turner, Nicola J; Whyte, Rosemary; Hudson, J Andrew; Kaltovei, Susan L

    2006-05-01

    Potato products prepared from dehydrated potato flakes have been implicated in foodborne illness incidents involving Bacillus cereus intoxications. B. cereus can survive as spores in potato flakes and can germinate and multiply in the rehydrated product. This study assessed the frequency and concentration of B. cereus in dehydrated potato flakes and hot-held, ready-to-eat mashed potato products. Of 50 packets of potato flakes tested, eight contained greater than 100 CFU/g B. cereus (maximum 370 CFU/g). The temperature of the potato portion of 44 hot-held food products was measured immediately after purchase, and 86% were below the safe hot-holding temperature of 60 degrees C. The potato portions were subsequently tested for B. cereus. Only two of the potato portions contained B. cereus at greater than 100 CFU/g, a potato-topped pastry (1000 CFU/g) and a container of potato and gravy (120 CFU/g). To assess multiplication of B. cereus in this food, we held rehydrated potato flakes with naturally occurring B. cereus at 37, 42, and 50 degrees C and tested them over 6 h. By 6 h, the number of B. cereus in potato stored at 37 degrees C had exceeded 10(3) CFU/g, was greater than 10(4) CFU/g at 50 degrees C, and was close to 10(6) CFU/g at 42 degrees C. Growth data were compared to predictions from the U.S. Department of Agriculture Pathogen Modeling Program (PMP 7.0). The PMP predictions were found to simulate the measured growth better at 42 degrees C than at 37 degrees C. Hot-held potato products should be safe for consumption if held at 60 degrees C or above or discarded within 2 h.

  10. 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.

  11. 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.

  12. Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers

    Energy Technology Data Exchange (ETDEWEB)

    Wig, Andrew G [ORNL; Arakawa, Edward T [ORNL; Passian, Ali [ORNL; Ferrell, Thomas L [ORNL; Thundat, Thomas George [ORNL

    2006-03-01

    Microcalorimetric optical and infrared spectroscopy is a method of determining the spectral absorption of small quantities of materials over a wide range of incident wavelengths. In this paper, the first spectroscopic results for microcantilevers coated with Bacillus anthracis (BA) are presented. These results, for B. anthracis from 2.5 to 14.5 {micro}m, are compared with results from microcantilevers coated with Bacillus cereus (BC) and standard spectroscopic absorption data. The results demonstrate strong correlation between the deflection measurements and the reference spectroscopic absorption peaks. An advantage of this microcantilever-based method over traditional spectroscopy is that much smaller amounts of material (nanogram quantities) can be detected in comparison with the milligram amounts needed for standard methods. Another advantage is that the complete system can be relatively small without sacrificing spectral resolution.

  13. 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.

  14. Protein- and DNA-based anthrax toxin vaccines confer protection in guinea pigs against inhalational challenge with Bacillus cereus G9241.

    Science.gov (United States)

    Palmer, John; Bell, Matt; Darko, Christian; Barnewall, Roy; Keane-Myers, Andrea

    2014-11-01

    In the past decade, several Bacillus cereus strains have been isolated from otherwise healthy individuals who succumbed to bacterial pneumonia presenting symptoms resembling inhalational anthrax. One strain was indistinguishable from B. cereus G9241, previously cultured from an individual who survived a similar pneumonia-like illness and which was shown to possess a complete set of plasmid-borne anthrax toxin-encoding homologs. The finding that B. cereus G9241 pathogenesis in mice is dependent on pagA1-derived protective antigen (PA) synthesis suggests that an anthrax toxin-based vaccine may be effective against this toxin-encoding B. cereus strain. Dunkin Hartley guinea pigs were immunized with protein- and DNA-based anthrax toxin-based vaccines, immune responses were evaluated and survival rates were calculated after lethal aerosol exposure with B. cereus G9241 spores. Each vaccine induced seroconversion with the protein immunization regimen eliciting significantly higher serum levels of antigen-specific antibodies at the prechallenge time-point compared with the DNA-protein prime-boost immunization schedule. Complete protection against lethal challenge was observed in all groups with a detectable prechallenge serum titer of toxin neutralizing antibodies. For the first time, we demonstrated that the efficacy of fully defined anthrax toxin-based vaccines was protective against lethal B. cereus G9241 aerosol challenge in the guinea pig animal model.

  15. Bacillus cereus endocarditis in native aortic valve.

    Science.gov (United States)

    Ngow, H A; Wan Khairina, W M N

    2013-02-01

    Bacillus cereus endocarditis is rare. It has been implicated in immunocompromised individuals, especially in intravenous drug users as well as in those with a cardiac prosthesis. The patient was a 31-year-old ex-intravenous drug addict with a past history of staphylococcal pulmonary valve endocarditis, who presented with symptoms of decompensated cardiac failure. Echocardiography showed severe aortic regurgitation with an oscillating vegetation seen on the right coronary cusp of the aortic valve. The blood cultures grew Bacillus cereus. We report this as a rare case of Bacillus cereus endocarditis affecting a native aortic valve.

  16. Separation of bacterial spores from flowing water in macro-scale cavities by ultrasonic standing waves

    CERN Document Server

    Lipkens, B; Costolo, M; Stevens, A; Rietman, Edward

    2010-01-01

    The separation of micron-sized bacterial spores (Bacillus cereus) from a steady flow of water through the use of ultrasonic standing waves is demonstrated. An ultrasonic resonator with cross-section of 0.0254 m x 0.0254 m has been designed with a flow inlet and outlet for a water stream that ensures laminar flow conditions into and out of the resonator section of the flow tube. A 0.01905-m diameter PZT-4, nominal 2-MHz transducer is used to generate ultrasonic standing waves in the resonator. The acoustic resonator is 0.0356 m from transducer face to the opposite reflector wall with the acoustic field in a direction orthogonal to the water flow direction. At fixed frequency excitation, spores are concentrated at the stable locations of the acoustic radiation force and trapped in the resonator region. The effect of the transducer voltage and frequency on the efficiency of spore capture in the resonator has been investigated. Successful separation of B. cereus spores from water with typical volume flow rates of...

  17. Intestinal carriage of Bacillus cereus: faecal isolation studies in three population groups.

    Science.gov (United States)

    Turnbull, P C; Kramer, J M

    1985-12-01

    The results of examinations of stools for Bacillus cereus among three unrelated groups of individuals are presented. The groups consisted of (1) healthy school-children aged 6-11 years in a rural region of South Africa examined during each of the four seasons of the year; (2) 15 healthy volunteers comprising staff of a London microbiology laboratory and their families examined on each of 3 consecutive weeks; (3) 75 unrelated young children, 2 months to 5 years of age, in a second rural region of South Africa examined during a pilot study of 1 week's duration on the aetiology of rural gastroenteritis. The stools of the last group were submitted as being related to present or recent diarrhoea in the respective children. In group 1, B. cereus isolation rates ranged from 24.3% at the autumn visit to 43% at the summer visit with a significantly higher rate of isolation in the summer than at other seasons of the year (P less than 0.05). B. cereus was isolated from 40% of group 2 volunteers on week 1, none on week 2 and 20% on week 3. The organism was detected in the 12 positive specimens at levels of approximately 10(2)/g and constituted 2.5-30% of the total aerobic spore-forming bacillus population in the stools. In group 3, B. cereus was recovered from 18.7% of the stool samples and was isolated consecutively with other pathogens (enteropathogenic Escherichia coli and rotavirus) on only five occasions. In groups 1 and 3, less than 5% of the stools had '3+' levels of B. cereus (greater than 10 colonies per direct plate culture). B. cereus was readily isolated from all of 10 food samples, representative of the typical diet of the group 1 individuals, and was present in substantial numbers (10(4) to 5.5 X 10(6)/g) in half of them. The isolation results, supported by serotyping, indicated that carriage of B. cereus in stools is transient and its presence at any one time reflects solely its intake with foods.

  18. Antagonism between Bacillus cereus and Pseudomonas fluorescens in planktonic systems and in biofilms.

    Science.gov (United States)

    Simões, Manuel; Simoes, Lúcia C; Pereira, Maria O; Vieira, Maria J

    2008-01-01

    In the environment, many microorganisms coexist in communities competing for resources, and they are often associated as biofilms. The investigation of bacterial ecology and interactions may help to improve understanding of the ability of biofilms to persist. In this study, the behaviour of Bacillus cereus and Pseudomonas fluorescens in the planktonic and sessile states was compared. Planktonic tests were performed with single and dual species cultures in growth medium with and without supplemental FeCl3. B. cereus and P. fluorescens single cultures had equivalent growth behaviours. Also, when in co-culture under Fe-supplemented conditions, the bacteria coexisted and showed similar growth profiles. Under Fe limitation, 8 h after co-culture and over time, the number of viable B. cereus cells decreased compared with P. fluorescens. Spores were detected during the course of the experiment, but were not correlated with the decrease in the number of viable cells. This growth inhibitory effect was correlated with the release of metabolite molecules by P. fluorescens through Fe-dependent mechanisms. Biofilm studies were carried out with single and dual species using a continuous flow bioreactor rotating system with stainless steel (SS) substrata. Steady-state biofilms were exposed to a series of increasing shear stress forces. Analysis of the removal of dual species biofilms revealed that the outer layer was colonised mainly by B. cereus. This bacterium was able to grow in the outermost layers of the biofilm due to the inhibitory effect of P. fluorescens being decreased by the exposure of the cells to fresh culture medium. B. cereus also constituted the surface primary coloniser due to its favourable adhesion to SS. P. fluorescens was the main coloniser of the middle layers of the biofilm. Single and dual species biofilm removal data also revealed that B. cereus biofilms had the highest physical stability, followed by P. fluorescens biofilms. This study highlights the

  19. VUV absorption spectroscopy of bacterial spores and DNA components

    Science.gov (United States)

    Fiebrandt, Marcel; Lackmann, Jan-Wilm; Raguse, Marina; Moeller, Ralf; Awakowicz, Peter; Stapelmann, Katharina

    2017-01-01

    Low-pressure plasmas can be used to inactivate bacterial spores and sterilize goods for medical and pharmaceutical applications. A crucial factor are damages induced by UV and VUV radiation emitted by the plasma. To analyze inactivation processes and protection strategies of spores, absorption spectra of two B. subtilis strains are measured. The results indicate, that the inner and outer coat of the spore significantly contribute to the absorption of UV-C and also of the VUV, protecting the spore against radiation based damages. As the sample preparation can significantly influence the absorption spectra due to salt residues, the cleaning procedure and sample deposition is tested for its reproducibility by measuring DNA oligomers and pUC18 plasmid DNA. The measurements are compared and discussed with results from the literature, showing a strong decrease of the salt content enabling the detection of absorption structures in the samples.

  20. Microbial Transformation of Quercetin by Bacillus cereus

    OpenAIRE

    Rao, Koppaka V.; Weisner, Nghe T.

    1981-01-01

    Biotransformation of quercetin was examined with a number of bacterial cultures. In the presence of a bacterial culture (Bacillus cereus), quercetin was transformed into two crystalline products, identified as protocatechuic acid and quercetin-3-glucoside (isoquercitrin).

  1. Spore Dispersion of Tricholoma matsutake at a Pinus densiflora Stand in Korea.

    Science.gov (United States)

    Park, Hyun; Ka, Kang-Hyeon

    2010-09-01

    The spore of Tricholoma matsutake is considered to be the starting point of the mushroom growth cycle, but the mechanism of mycelial development from the spore stage is not yet clarified. In this study, we tried to measure how far the spores of T. matsutake disperse from a fruiting body located at a Pinus densiflora stand in Korea. We established 16 slide glasses coated with glycerin near a fruiting body in four directions separated by four different distance intervals within a mushroom productive stand after removing all other fruiting bodies from three plots. The number of dispersed spores increased with time from the first day (475 spores/cm(2)) to the fourth day (836 spores/cm(2)) after the pileus opened. The number of spores dispersed downward was about 1.5 times greater than that dispersed toward the ridge. The number of dispersed spores decreased exponentially as the distance from each fruiting body increased. More than 95% of the spores dropped within a meter from the fruiting body, with 75% dropping within 0.5 m. Even so, the number of spores dispersed over 5 m from the fruiting body was more than 50 million when considering the total number of spores produced by a fruiting body is about 5 billion.

  2. Bacillus cereus Bacteremia in a Preterm Neonate

    OpenAIRE

    Hilliard, Nicholaus J.; Schelonka, Robert L.; Waites, Ken B.

    2003-01-01

    Bacillus cereus is an uncommon but potentially serious bacterial pathogen causing infections of the bloodstream, lungs, and central nervous system of preterm neonates. A case of bacteremia caused by B. cereus in a 19-day-old preterm neonate who was successfully treated with vancomycin, tobramycin, meropenem, and clindamycin is described. Implications for the diagnostic laboratory and clinicians when Bacillus species are detected in normally sterile sites are discussed, and the small numbers o...

  3. Pseudomembranous tracheobronchitis due to Bacillus cereus.

    Science.gov (United States)

    Strauss, R; Mueller, A; Wehler, M; Neureiter, D; Fischer, E; Gramatzki, M; Hahn, E G

    2001-09-01

    We present a case of a rapidly progressive pseudomembranous tracheobronchitis and pneumonia in a 52-year-old woman with severe aplastic anemia. Bacillus cereus was isolated from bronchoalveolar lavage fluids, blood cultures, and pseudomembrane biopsy specimens; despite intensive antibiotic treatment, the patient's condition deteriorated rapidly. To our knowledge, this is the first report of a B. cereus infection that has caused pseudomembranous tracheobronchitis, possibly because of the production of bacterial toxins.

  4. 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.

  5. New Rapid Spore Assay

    Science.gov (United States)

    Kminek, Gerhard; Conley, Catharine

    2012-07-01

    The presentation will detail approved Planetary Protection specifications for the Rapid Spore Assay for spacecraft components and subsystems. Outlined will be the research and studies on which the specifications were based. The research, funded by ESA and NASA/JPL, was conducted over a period of two years and was followed by limited cleanroom studies to assess the feasibility of this assay during spacecraft assembly.

  6. Biofilm Formation by Bacillus cereus Is Influenced by PlcR, a Pleiotropic Regulator

    Science.gov (United States)

    Hsueh, Yi-Huang; Somers, Eileen B.; Lereclus, Didier; Wong, Amy C. Lee

    2006-01-01

    The ΔplcR mutant of Bacillus cereus strain ATCC 14579 developed significantly more biofilm than the wild type and produced increased amounts of biosurfactant. Biosurfactant production is required for biofilm formation and may be directly or indirectly repressed by PlcR, a pleiotropic regulator. Coating polystyrene plates with surfactin, a biosurfactant from Bacillus subtilis, rescued the deficiency in biofilm formation by the wild type. PMID:16820512

  7. Specific peptides as alternative to antibody ligands for biomagnetic separation of Clostridium tyrobutyricum spores.

    Science.gov (United States)

    Lavilla, Maria; Moros, Maria; Puertas, Sara; Grazú, Valeria; Pérez, María Dolores; Calvo, Miguel; de la Fuente, Jesus M; Sánchez, Lourdes

    2012-04-01

    Nowadays, the reference method for the detection of Clostridium tyrobutyricum in milk is the most-probable-number method, a very time-consuming and non-specific method. In this work, the suitability of the use of superparamagnetic beads coated with specific antibodies and peptides for bioseparation and concentration of spores of C. tyrobutyricum has been assessed. Peptide or antibody functionalized nanoparticles were able to specifically bind C. tyrobutyricum spores and concentrate them up to detectable levels. Moreover, several factors, such as particle size (200 nm and 1 μm), particle derivatization (aminated and carboxylated beads), coating method, and type of ligand have been studied in order to establish the most appropriate conditions for spore separation. Results show that concentration of spore is favored by a smaller bead size due to the wider surface of interaction in relation to particle volume. Antibody orientation, related to the binding method, is also critical in spore recovery. However, specific peptides seem to be a better ligand than antibodies, not only due to the higher recovery ratio of spores obtained but also due to the prolonged stability over time, allowing an optimal recovery of spores up to 3 weeks after bead coating. These results demonstrate that specific peptides bound to magnetic nanoparticles can be used instead of traditional antibodies to specifically bind C. tyrobutyricum spores being a potential basis for a rapid method to detect this bacterial target.

  8. Clostridium difficile spore-macrophage interactions: spore survival.

    Directory of Open Access Journals (Sweden)

    Daniel Paredes-Sabja

    Full Text Available BACKGROUND: Clostridium difficile is the main cause of nosocomial infections including antibiotic associated diarrhea, pseudomembranous colitis and toxic megacolon. During the course of Clostridium difficile infections (CDI, C. difficile undergoes sporulation and releases spores to the colonic environment. The elevated relapse rates of CDI suggest that C. difficile spores has a mechanism(s to efficiently persist in the host colonic environment. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we provide evidence that C. difficile spores are well suited to survive the host's innate immune system. Electron microscopy results show that C. difficile spores are recognized by discrete patchy regions on the surface of macrophage Raw 264.7 cells, and phagocytosis was actin polymerization dependent. Fluorescence microscopy results show that >80% of Raw 264.7 cells had at least one C. difficile spore adhered, and that ∼60% of C. difficile spores were phagocytosed by Raw 264.7 cells. Strikingly, presence of complement decreased Raw 264.7 cells' ability to phagocytose C. difficile spores. Due to the ability of C. difficile spores to remain dormant inside Raw 264.7 cells, they were able to survive up to 72 h of macrophage infection. Interestingly, transmission electron micrographs showed interactions between the surface proteins of C. difficile spores and the phagosome membrane of Raw 264.7 cells. In addition, infection of Raw 264.7 cells with C. difficile spores for 48 h produced significant Raw 264.7 cell death as demonstrated by trypan blue assay, and nuclei staining by ethidium homodimer-1. CONCLUSIONS/SIGNIFICANCE: These results demonstrate that despite efficient recognition and phagocytosis of C. difficile spores by Raw 264.7 cells, spores remain dormant and are able to survive and produce cytotoxic effects on Raw 264.7 cells.

  9. Bacterial spores as particulate carriers for gene gun delivery of plasmid DNA.

    Science.gov (United States)

    Aps, Luana R M M; Tavares, Milene B; Rozenfeld, Julio H K; Lamy, M Teresa; Ferreira, Luís C S; Diniz, Mariana O

    2016-06-20

    Bacillus subtilis spores represent a suitable platform for the adsorption of proteins, enzymes and viral particles at physiological conditions. In the present work, we demonstrate that purified spores can also adsorb DNA on their surface after treatment with cationic molecules. In addition, we demonstrate that DNA-coated B. subtilis spores can be used as particulate carriers and act as an alternative to gold microparticles for the biolistic (gene gun) administration of plasmid DNA in mice. Gene gun delivery of spores pre-treated with DODAB (dioctadecyldimethylammonium bromide) allowed efficient plasmid DNA absorption and induced protein expression levels similar to those obtained with gold microparticles. More importantly, we demonstrated that a DNA vaccine adsorbed on spores can be loaded into biolistic cartridges and efficiently delivered into mice, which induced specific cellular and antibody responses. Altogether, these data indicate that B. subtilis spores represent a simple and low cost alternative for the in vivo delivery of DNA vaccines by the gene gun technology.

  10. Cd-Resistant Strains of B. cereus S5 with Endurance Capacity and Their Capacities for Cadmium Removal from Cadmium-Polluted Water.

    Science.gov (United States)

    Wu, Huiqing; Wu, Qingping; Wu, Guojie; Gu, Qihui; Wei, Linting

    2016-01-01

    The goal of this study was to identify Cd-resistant bacterial strains with endurance capacity and to evaluate their ability to remove cadmium ions from cadmium-polluted water. The Bacillus cereusS5 strain identified in this study had the closest genetic relationship with B. cereus sp. Cp1 and performed well in the removal of Cd2+ions from solution. The results showed that both the live and dead biomasses of the Cd2+-tolerant B. cereus S5 strain could absorb Cd2+ ions in solution but that the live biomass of the B. cereus S5 strain outperformed the dead biomass at lower Cd2+concentrations. An analysis of the cadmium tolerance genes of B. cereus S5 identified ATPase genes that were associated with cadmium tolerance and involved in the ATP pumping mechanism. The FTIR spectra revealed the presence of amino, carboxyl and hydroxyl groups on the pristine biomass and indicated that the cadmium ion removal ability was related to the structure of the strain. The maximum absorption capacity of the B. cereus S5 strain in viable spore biomass was 70.16 mg/g (dry weight) based on a pseudo-second-order kinetic model fit to the experimental data. The Langmuir and Langmuir-Freundlich isotherm adsorption models fit the cadmium ion adsorption data well, and the kinetic curves indicated that the adsorption rate was second-order. For Cd2+ concentrations (mg/L) of 1-109 mg/L, good removal efficiency (>80%) was achieved using approximately 3.48-10.3 g/L of active spore biomass of the B. cereus S5 strain. A cadmium-tolerant bacteria-activated carbon-immobilized column could be used for a longer duration and exhibited greater treatment efficacy than the control column in the treatment of cadmium-polluted water. In addition, a toxicity assessment using mice demonstrated that the biomass of the B. cereus S5 strain and its fermentation products were non-toxic. Thus, the isolated B. cereus S5 strain can be considered an alternative biological adsorbent for use in emergency responses to severe

  11. Fungal Spores Viability on the International Space Station

    Science.gov (United States)

    Gomoiu, I.; Chatzitheodoridis, E.; Vadrucci, S.; Walther, I.; Cojoc, R.

    2016-11-01

    long term experiment lost the outer layer of their coat without affecting the viability since they were still protected by the middle and the inner layer of the coating. This research highlights a new protocol to perform spaceflight experiments inside the ISS with fungal spores in microgravity conditions, under the additional effect of possible cosmic radiation. According to this protocol the results are expressed in terms of viability, microscopic and morphological changes.

  12. Fungal Spores Viability on the International Space Station.

    Science.gov (United States)

    Gomoiu, I; Chatzitheodoridis, E; Vadrucci, S; Walther, I; Cojoc, R

    2016-11-01

    long term experiment lost the outer layer of their coat without affecting the viability since they were still protected by the middle and the inner layer of the coating. This research highlights a new protocol to perform spaceflight experiments inside the ISS with fungal spores in microgravity conditions, under the additional effect of possible cosmic radiation. According to this protocol the results are expressed in terms of viability, microscopic and morphological changes.

  13. Fungal Spores Viability on the International Space Station

    Science.gov (United States)

    Gomoiu, I.; Chatzitheodoridis, E.; Vadrucci, S.; Walther, I.; Cojoc, R.

    2016-04-01

    long term experiment lost the outer layer of their coat without affecting the viability since they were still protected by the middle and the inner layer of the coating. This research highlights a new protocol to perform spaceflight experiments inside the ISS with fungal spores in microgravity conditions, under the additional effect of possible cosmic radiation. According to this protocol the results are expressed in terms of viability, microscopic and morphological changes.

  14. Linking Bacillus cereus genotypes and carbohydrate utilization capacity

    NARCIS (Netherlands)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Jong, de Anne; Kuipers, Oscar P.; Nierop Groot, Masja N.; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together wi

  15. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity

    NARCIS (Netherlands)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Jong, de Anne; Kuipers, Oscar P.; Nierop Groot, Masja N.; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with

  16. Progress in food-related research focussing on Bacillus cereus

    NARCIS (Netherlands)

    Vries, de Y.P.; Voort, van der M.; Schaik, van W.; Hornstra, L.M.; Vos, de W.M.; Abee, T.

    2004-01-01

    Bacillus cereus is a gram-positive, rod-shaped, endospore-forming bacterium that occurs ubiquitously and is frequently isolated from soil and food products. When B. cereus is present in foods, it can cause spoilage and poisoning. The work of our group is focussed on several properties of B. cereus t

  17. Arthromitus (Bacillus cereus) symbionts in the cockroach Blaberus giganteus: dietary influences on bacterial development and population density

    Science.gov (United States)

    Feinberg, L.; Jorgensen, J.; Haselton, A.; Pitt, A.; Rudner, R.; Margulis, L.

    1999-01-01

    The filamentous spore-forming bacterium Arthromitus, discovered in termites, millipedes, sow bugs and other soil-dwelling arthropods by Leidy (1850), is the intestinal stage of Bacillus cereus. We extend the range of Arthromitus habitats to include the hindgut of Blaberus giganteus, the large tropical American cockroach. The occurrence and morphology of the intestinal form of the bacillus were compared in individual cockroaches (n=24) placed on four different diet regimes: diurnally maintained insects fed (1) dog food, (2) soy protein only, (3)purified cellulose only, and (4) a dog food-fed group maintained in continuous darkness. Food quality exerted strong influence on population densities and developmental stages of the filamentous bacterium and on fecal pellet composition. The most dramatic rise in Arthromitus populations, defined as the spore-forming filament intestinal stage, occurred in adult cockroaches kept in the dark on a dog food diet. Limited intake of cellulose or protein alone reduced both the frequency of Arthromitus filaments and the rate of weight gain of the insects. Spores isolated from termites, sow bugs, cockroaches and moths, grown on various hard surfaces display a branching mobility and resistance to antibiotics characteristic to group I Bacilli whose members include B. cereus, B. circulans, B. alvei and B. macerans. DNA isolated from pure cultures of these bacilli taken from the guts of Blaberus giganteus (cockroach), Junonia coenia (moth), Porcellio scaber (sow bug) and Cryptotermes brevis (termite) and subjected to Southern hybridization with a 23S-5S B. subtilis ribosomal sequence probe verified that they are indistinguishable from laboratory strains of Bacillus cereus.

  18. Bacillus cereus bacteremia in a preterm neonate.

    Science.gov (United States)

    Hilliard, Nicholaus J; Schelonka, Robert L; Waites, Ken B

    2003-07-01

    Bacillus cereus is an uncommon but potentially serious bacterial pathogen causing infections of the bloodstream, lungs, and central nervous system of preterm neonates. A case of bacteremia caused by B. cereus in a 19-day-old preterm neonate who was successfully treated with vancomycin, tobramycin, meropenem, and clindamycin is described. Implications for the diagnostic laboratory and clinicians when Bacillus species are detected in normally sterile sites are discussed, and the small numbers of infant infections proven to be due to this organism that have been described previously are reviewed.

  19. Features of Bacillus cereus swarm cells.

    Science.gov (United States)

    Senesi, Sonia; Salvetti, Sara; Celandroni, Francesco; Ghelardi, Emilia

    2010-11-01

    When propagated on solid surfaces, Bacillus cereus can produce differentiated swarm cells under a wide range of growth conditions. This behavioural versatility is ecologically relevant, since it allows this bacterium to adapt swarming to environmental changes. Swarming by B. cereus is medically important: swarm cells are more virulent and particularly prone to invade host tissues. Characterisation of swarming-deficient mutants highlights that flagellar genes as well as genes governing different metabolic pathways are involved in swarm-cell differentiation. In this review, the environmental and genetic requirements for swarming and the role played by swarm cells in the virulence this pathogen exerts will be outlined.

  20. Bacillus cereus as a nongastrointestinal pathogen

    Directory of Open Access Journals (Sweden)

    Pavani G.

    2014-02-01

    Full Text Available The potential of Bacillus cereus to cause systemic infections is of serious concern. Apart from Gastrointestinal infections, it causes respiratory tract infections, nosocomial infections, eye infections, CNS infections, cutaneous infections, endocarditis, osteomyelitis and urinary tract infections. The potential of this bacterium to cause life threatening infections has increased. Trauma is an important predisposing factor for Bacillus cereus infections. The maintenance of skin and mucous membrane integrity limits infection by this micro-organism. [Int J Res Med Sci 2014; 2(1.000: 28-30

  1. 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.

  2. Bacillus cereus cellulitis from contaminated heroin.

    Science.gov (United States)

    Dancer, S J; McNair, D; Finn, P; Kolsto, A B

    2002-03-01

    Concern exists over recent unexplained deaths among intravenous drug users. This report describes a patient with crepitant cellulitis who was admitted complaining of severe pain in the right forearm. Ultrasonography demonstrated gas in the tissues and he was referred for early surgical debridement of the arm. He was treated with intravenous benzyl penicillin, gentamicin and metronidazole and made a full recovery. Aspirate samples grew Bacillus cereus, morphologically similar to the isolate obtained from a sample of the patient's own heroin. Antibiogram and API 50CHB profiles were also similar. Further typing included 'H' flagellar serotyping, which found both blood and heroin strains to be non-typable, and amplified fragment polymorphism analysis, which showed that the strains were indistinguishable. Genotyping of two selected genes from B. cereus confirmed almost certain identity between the two strains. This case illustrates the potential virulence of B. cereus when inoculated into tissues, and to our knowledge, is the first report to demonstrate a conclusive microbiological link between contaminated heroin and serious sepsis in a drug user due to B. cereus.

  3. Bacillus cereus infection outbreak in captive psittacines.

    Science.gov (United States)

    Godoy, S N; Matushima, E R; Chaves, J Q; Cavados, C F G; Rabinovitch, L; Teixeira, R H F; Nunes, A L V; Melville, P; Gattamorta, M A; Vivoni, A M

    2012-12-28

    This study reports an uncommon epizootic outbreak of Bacillus cereus that caused the sudden death of 12 psittacines belonging to the species Anodorhynchus hyacinthinus (1 individual), Diopsittaca nobilis (1 individual), Ara severa (1 individual) and Ara ararauna (9 individuals) in a Brazilian zoo. Post-mortem examination of the animals reveled extensive areas of lung hemorrhage, hepatic congestion, hemorrhagic enteritis and cardiac congestion. Histopathological examination of the organs showed the presence of multiple foci of vegetative cells of Gram-positive bacilli associated with discrete and moderate mononuclear inflammatory cell infiltrate. Seventeen B. cereus strains isolated from blood and sterile organs of nine A. ararauna were analyzed in order to investigate the genetic diversity (assessed by Rep-PCR) and toxigenic profiles (presence of hblA, hblC and hblD; nheA, nheB and nheC as well as cytK, ces and entFM genes) of such strains. Amplification of genomic DNA by Rep-PCR of B. cereus strains generated two closely related profiles (Rep-PCR types A and B) with three bands of difference. All strains were classified as belonging to the toxigenic profile I which contained HBL and NHE gene complexes, entFM and cytK genes. Altogether, microbiological and histopathological findings and the evidence provided by the success of the antibiotic prophylaxis, corroborate that B. cereus was the causative agent of the infection that killed the birds.

  4. Hydrazine vapor inactivates Bacillus spores

    Science.gov (United States)

    Schubert, Wayne W.; Engler, Diane L.; Beaudet, Robert A.

    2016-05-01

    NASA policy restricts the total number of bacterial spores that can remain on a spacecraft traveling to any planetary body which might harbor life or have evidence of past life. Hydrazine, N2H4, is commonly used as a propellant on spacecraft. Hydrazine as a liquid is known to inactivate bacterial spores. We have now verified that hydrazine vapor also inactivates bacterial spores. After Bacillus atrophaeus ATCC 9372 spores deposited on stainless steel coupons were exposed to saturated hydrazine vapor in closed containers, the spores were recovered from the coupons, serially diluted, pour plated and the surviving bacterial colonies were counted. The exposure times required to reduce the spore population by a factor of ten, known as the D-value, were 4.70 ± 0.50 h at 25 °C and 2.85 ± 0.13 h at 35 °C. These inactivation rates are short enough to ensure that the bioburden of the surfaces and volumes would be negligible after prolonged exposure to hydrazine vapor. Thus, all the propellant tubing and internal tank surfaces exposed to hydrazine vapor do not contribute to the total spore count.

  5. The Synergistic Effect of High Pressure CO2 and Nisin on Inactivation of Bacillus subtilis Spores in Aqueous Solutions

    Science.gov (United States)

    Rao, Lei; Wang, Yongtao; Chen, Fang; Liao, Xiaojun

    2016-01-01

    The inactivation effects of high pressure CO2 + nisin (simultaneous treatment of HPCD and nisin, HPCD + nisin), HPCD→nisin (HPCD was followed by nisin), and nisin→HPCD (nisin was followed by HPCD) treatments on Bacillus subtilis spores in aqueous solutions were compared. The spores were treated by HPCD at 6.5 or 20 MPa, 84–86°C and 0–30 min, and the concentration of nisin was 0.02%. Treated spores were examined for the viability, the permeability of inner membrane (IM) using flow cytometry method and pyridine-2, 6-dicarboxylic acid (DPA) release, and structural damage by transmission electron microscopy. A synergistic effect of HPCD + nisin treatment on inactivation of the spores was found, and the inactivation efficiency of the spores was HPCD + nisin > HPCD→nisin or nisin→HPCD. Moreover, HPCD + nisin caused higher IM permeability and DPA release of the spores than HPCD. A possible action mode of nisin-enhanced inactivation of the spores was suggested as that HPCD firstly damaged the coat and cortex of spores, and nisin penetrated into and acted on the IM of spores, which increased the damage to the IM of spores, and resulted in higher inactivation of the spores. PMID:27708639

  6. Comparative transcriptional profiling of Bacillus cereus sensu lato strains during growth in CO2-bicarbonate and aerobic atmospheres.

    Directory of Open Access Journals (Sweden)

    Karla D Passalacqua

    Full Text Available BACKGROUND: Bacillus species are spore-forming bacteria that are ubiquitous in the environment and display a range of virulent and avirulent phenotypes. This range is particularly evident in the Bacillus cereus sensu lato group; where closely related strains cause anthrax, food-borne illnesses, and pneumonia, but can also be non-pathogenic. Although much of this phenotypic range can be attributed to the presence or absence of a few key virulence factors, there are other virulence-associated loci that are conserved throughout the B. cereus group, and we hypothesized that these genes may be regulated differently in pathogenic and non-pathogenic strains. METHODOLOGY/PRINCIPAL FINDINGS: Here we report transcriptional profiles of three closely related but phenotypically unique members of the Bacillus cereus group--a pneumonia-causing B. cereus strain (G9241, an attenuated strain of B. anthracis (Sterne 34F(2, and an avirulent B. cereus strain (10987--during exponential growth in two distinct atmospheric environments: 14% CO(2/bicarbonate and ambient air. We show that the disease-causing Bacillus strains undergo more distinctive transcriptional changes between the two environments, and that the expression of plasmid-encoded virulence genes was increased exclusively in the CO(2 environment. We observed a core of conserved metabolic genes that were differentially expressed in all three strains in both conditions. Additionally, the expression profiles of putative virulence genes in G9241 suggest that this strain, unlike Bacillus anthracis, may regulate gene expression with both PlcR and AtxA transcriptional regulators, each acting in a different environment. CONCLUSIONS/SIGNIFICANCE: We have shown that homologous and even identical genes within the genomes of three closely related members of the B. cereus sensu lato group are in some instances regulated very differently, and that these differences can have important implications for virulence. This study

  7. Bacillus cereus bacteremia outbreak due to contaminated hospital linens.

    Science.gov (United States)

    Sasahara, T; Hayashi, S; Morisawa, Y; Sakihama, T; Yoshimura, A; Hirai, Y

    2011-02-01

    We describe an outbreak of Bacillus cereus bacteremia that occurred at Jichi Medical University Hospital in 2006. This study aimed to identify the source of this outbreak and to implement appropriate control measures. We reviewed the charts of patients with blood cultures positive for B. cereus, and investigated B. cereus contamination within the hospital environment. Genetic relationships among B. cereus isolates were analyzed. Eleven patients developed B. cereus bacteremia between January and August 2006. The hospital linens and the washing machine were highly contaminated with B. cereus, which was also isolated from the intravenous fluid. All of the contaminated linens were autoclaved, the washing machine was cleaned with a detergent, and hand hygiene was promoted among the hospital staff. The number of patients per month that developed new B. cereus bacteremia rapidly decreased after implementing these measures. The source of this outbreak was B. cereus contamination of hospital linens, and B. cereus was transmitted from the linens to patients via catheter infection. Our findings demonstrated that bacterial contamination of hospital linens can cause nosocomial bacteremia. Thus, blood cultures that are positive for B. cereus should not be regarded as false positives in the clinical setting.

  8. Bacterial spore components which enhance the bacteriostatic effectiveness of S-nitrosothiol.

    Science.gov (United States)

    Morris, S L; Hansen, J N

    1981-12-01

    Spore components exuded into the medium during outgrowth of Bacillus cereus T enhanced the bacteriostatic effectiveness of S-nitrosomercaptoethanol, an inhibitor which prevents outgrowth at low concentrations and germination at higher concentrations. The enhancement effect was slight with respect to outgrowth, but dramatic with respect to germination, in that the inhibitory effectiveness of nitrosothiols toward germination inhibition was enhanced by as much as 33-fold when nitrosothiols was in the presence of the exuded spore component. Exudate activity was freely dialyzable and was not measurably affected by a broad-spectrum protease (proteinase K), by autoclaving at 121 degrees C, or by freezing and thawing. Sephadex G-25 chromatography of the exudate indicated that two active species were present, a major component with a molecular weight of less than 1,000 and a minor component with a molecular weight of more than 5,000.

  9. Adsorption of β-galactosidase of Alicyclobacillus acidocaldarius on wild type and mutants spores of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Sirec Teja

    2012-08-01

    Full Text Available Abstract Background The Bacillus subtilis spore has long been used as a surface display system with potential applications in a variety of fields ranging from mucosal vaccine delivery, bioremediation and biocatalyst development. More recently, a non-recombinant approach of spore display has been proposed and heterologous proteins adsorbed on the spore surface. We used the well-characterized β-galactosidase from the thermoacidophilic bacterium Alicyclobacillus acidocaldarius as a model to study enzyme adsorption, to analyze whether and how spore-adsorption affects the properties of the enzyme and to improve the efficiency of the process. Results We report that purified β-galactosidase molecules were adsorbed to purified spores of a wild type strain of B. subtilis retaining ca. 50% of their enzymatic activity. Optimal pH and temperature of the enzyme were not altered by the presence of the spore, that protected the adsorbed β-galactosidase from exposure to acidic pH conditions. A collection of mutant strains of B. subtilis lacking a single or several spore coat proteins was compared to the isogenic parental strain for the adsorption efficiency. Mutants with an altered outermost spore layer (crust were able to adsorb 60-80% of the enzyme, while mutants with a severely altered or totally lacking outer coat adsorbed 100% of the β-galactosidase molecules present in the adsorption reaction. Conclusion Our results indicate that the spore surface structures, the crust and the outer coat layer, have an negative effect on the adhesion of the β-galactosidase. Electrostatic forces, previously suggested as main determinants of spore adsorption, do not seem to play an essential role in the spore-β-galactosidase interaction. The analysis of mutants with altered spore surface has shown that the process of spore adsorption can be improved and has suggested that such improvement has to be based on a better understanding of the spore surface structure

  10. Pathogenicity and characterization of a novel Bacillus cereus sensu lato isolate toxic to the Mediterranean fruit fly Ceratitis capitata Wied.

    Science.gov (United States)

    Ruiu, Luca; Falchi, Giovanni; Floris, Ignazio; Marche, Maria Giovanna; Mura, Maria Elena; Satta, Alberto

    2015-03-01

    The lethal and sub-lethal effects of sporulated cultures of a novel Bacillus cereus sensu lato strain lacking detectable cry genes and identified through morphological and genetic analyses, have been studied on the Mediterranean fruit fly Ceratitis capitata. The lethal effects on young larvae were concentration dependent, with a median lethal concentration (LC50) of 4.48 × 10(8)spores/g of diet. Sporulated cultures of this strain significantly extended development time and reduced immature survival, and the size of emerging fly adults. Besides spores, the toxicity has been associated to the insoluble extra-spore fraction characterized through a proteomic approach. The profile of the extra-spore protein fraction (ES) showed major protein bands within the 35-65 kDa range. The results of mass spectrometry analysis highlighted the presence of putative virulence factors, including members of protein families previously associated to the insecticidal action of other microbial entomopathogens. These proteins include metalloproteases, peptidases and other enzymes.

  11. Prevalence, isolation and characterization of Bacillus cereus strains from rice of local cultivators of Sabah, Sarawak, and Peninsular Malaysia

    Science.gov (United States)

    Sawei, Jelin; Sani, Norrakiah Abdullah

    2016-11-01

    Bacillus cereus is a spore-forming, facultative anaerobic, motile microorganism that has been identified as a causative agent of two types of gastrointestinal diseases such as emetic and diarrhea. This foodborne pathogen is found in both vegetative cells and endospores form in foods such as rice either raw or cooked. The aim of this study is to investigate and determine the prevalence, characterize and identify the isolation of vegetative cells and endospores of B. cereus in thirty varieties (n=3) of raw rice from Sabah, Sarawak and Peninsular Malaysia. A total of 90 (n=90) raw rice were examined and 84 (93.33%) samples were positive to vegetative cells of B. cereus. However, only 32 (35.56%) samples were positive for endospore cells that able to germinate after samples were heated at 75°C for 15 mins. The mean log cfu/g for vegetative cells were higher range (0.00 - 4.1533) than visible endospores (0.00 - 3.7533 mean log cfu/g). Sample of raw red rice (UKMRC9) had significantly higher contamination by both vegetative cells and endospores at p<0.05, than the other raw rice samples.

  12. Glucose 6P binds and activates HlyIIR to repress Bacillus cereus haemolysin hlyII gene expression.

    Directory of Open Access Journals (Sweden)

    Elisabeth Guillemet

    Full Text Available Bacillus cereus is a Gram-positive spore-forming bacterium causing food poisoning and serious opportunistic infections. These infections are characterized by bacterial accumulation despite the recruitment of phagocytic cells. We have previously shown that B. cereus Haemolysin II (HlyII induces macrophage cell death by apoptosis. In this work, we investigated the regulation of the hlyII gene. We show that HlyIIR, the negative regulator of hlyII expression in B. cereus, is especially active during the early bacterial growth phase. We demonstrate that glucose 6P directly binds to HlyIIR and enhances its activity at a post-transcriptional level. Glucose 6P activates HlyIIR, increasing its capacity to bind to its DNA-box located upstream of the hlyII gene, inhibiting its expression. Thus, hlyII expression is modulated by the availability of glucose. As HlyII induces haemocyte and macrophage death, two cell types that play a role in the sequestration of nutrients upon infection, HlyII may induce host cell death to allow the bacteria to gain access to carbon sources that are essential components for bacterial growth.

  13. Partial purification and characterization of protease enzyme from Bacillus subtilis and Bacillus cereus.

    Science.gov (United States)

    Orhan, Elif; Omay, Didem; Güvenilir, Yüksel

    2005-01-01

    The aim of this experimental study was to isolate and partially purify protease enzyme from Bacillus cereus and Bacillus subtilis. Protease enzyme is obtained by inducing spore genesis of bacteria from Bacillus species in suitable nutrient plates. The partial purification was realized by applying, respectively, ammonium sulfate precipitation, dialysis, and DEAE-cellulose ion-exchange chromatography to the supernatant that was produced later. Optimum pH, optimum temperature, pH stability, and temperature stability were determined, as well as the effects of pH, temperature, substrate concentration, reaction time, and inhibitors and activators on enzyme activity. In addition, the molecular mass of the obtained enzyme was investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The specific activity of partially purified enzyme from B. subtilis was determined to be 84 U/mg. The final enzyme preparation was eight-fold more pure than the crude homogenate. The molecular mass of the partially purified enzyme was found to be 45 kDa by using SDS-PAGE. The protease enzyme that was partially purified from B. cereus was purified 1.2-fold after ammonium sulfate precipitation. The molecular mass of the partially purified enzyme was determined to be 37 kDa by using SDS-PAGE.

  14. Necrotizing gastritis due to Bacillus cereus in an immunocompromised patient.

    Science.gov (United States)

    Le Scanff, J; Mohammedi, I; Thiebaut, A; Martin, O; Argaud, L; Robert, D

    2006-04-01

    Bacillus cereus is increasingly being acknowledged as a serious bacterial pathogen in immunocompromised patients. We present a case of acute necrotizing gastritis caused by B. cereus in a 37-year-old woman with acute myeloblastic leukemia, who recovered following total parenteral nutrition and treatment with imipenem and vancomycin. B. cereus was isolated from gastric mucosa and blood cultures. Up to now, no case of acute necrotizing gastritis due to this organism has been reported.

  15. Effects of meteorological conditions on spore plumes

    Science.gov (United States)

    Burch, M.; Levetin, E.

    2002-05-01

    Fungal spores are an ever-present component of the atmosphere, and have long been known to trigger asthma and hay fever symptoms in sensitive individuals. The atmosphere around Tulsa has been monitored for airborne spores and pollen with Burkard spore traps at several sampling stations. This study involved the examination of the hourly spore concentrations on days that had average daily concentrations near 50,000 spores/m3 or greater. Hourly concentrations of Cladosporium, Alternaria, Epicoccum, Curvularia, Pithomyces, Drechslera, smut spores, ascospores, basidiospores, other, and total spores were determined on 4 days at three sites and then correlated with hourly meteorological data including temperature, rainfall, wind speed, dew point, air pressure, and wind direction. On each of these days there was a spore plume, a phenomenon in which spore concentrations increased dramatically over a very short period of time. Spore plumes generally occurred near midday, and concentrations were seen to increase from lows around 20,000 total spores/m3 to highs over 170,000 total spores/m3 in 2 h. Multiple regression analysis of the data indicated that increases in temperature, dew point, and air pressure correlated with the increase in spore concentrations, but no single weather variable predicted the appearance of a spore plume. The proper combination of changes in these meteorological parameters that result in a spore plume may be due to the changing weather conditions associated with thunderstorms, as on 3 of the 4 days when spore plumes occurred there were thunderstorms later that evening. The occurrence of spore plumes may have clinical significance, because other studies have shown that sensitization to certain spore types can occur during exposure to high spore concentrations.

  16. Beetroot-pigment-derived colorimetric sensor for detection of calcium dipicolinate in bacterial spores.

    Directory of Open Access Journals (Sweden)

    Letícia Christina Pires Gonçalves

    Full Text Available In this proof-of-concept study, we describe the use of the main red beet pigment betanin for the quantification of calcium dipicolinate in bacterial spores, including Bacillus anthracis. In the presence of europium(III ions, betanin is converted to a water-soluble, non-luminescent orange 1∶1 complex with a stability constant of 1.4 × 10(5 L mol(-1. The addition of calcium dipicolinate, largely found in bacterial spores, changes the color of the aqueous solution of [Eu(Bn(+] from orange to magenta. The limit of detection (LOD of calcium dipicolinate is around 2.0 × 10(-6 mol L(-1 and the LOD determined for both spores, B. cereus and B. anthracis, is (1.1 ± 0.3× 10(6 spores mL(-1. This simple, green, fast and low cost colorimetric assay was selective for calcium dipicolinate when compared to several analogous compounds. The importance of this work relies on the potential use of betalains, raw natural pigments, as colorimetric sensors for biological applications.

  17. The role of a purine-specific nucleoside hydrolase in spore germination of Bacillus thuringiensis.

    Science.gov (United States)

    Liang, Liang; He, Xihong; Liu, Gang; Tan, Huarong

    2008-05-01

    A homologous gene (iunH) of a putative nucleoside hydrolase (NH), which had been identified from the exosporia of Bacillus cereus and Bacillus anthracis spores, was cloned from Bacillus thuringiensis subsp. kurstaki. Disruption of iunH did not affect the vegetative growth and sporulation of Bacillus thuringiensis, but promoted both inosine- and adenosine-induced spore germination. The inosine- or adenosine-induced germination rate decreased when the wild-type iunH gene was overexpressed in Bacillus thuringiensis. The iunH gene product was characterized as a purine-specific NH. The kinetic parameters of IunH with inosine as substrate were K(m)=399+/-115 microM, k(cat)=48.9+/-8.5 s(-1) and k(cat)/K(m)=1.23 x 10(5) M(-1) s(-1). The optimal pH and temperature for IunH were found to be pH 6 and 80 degrees C. Meanwhile, the specific activity of inosine hydrolase in intact spores of the wild-type strain with inosine as substrate was 2.89+/-0.23x10(-2) micromol min(-1) (mg dry wt)(-1). These results indicate that IunH is important in moderating inosine- or adenosine-induced germination of Bacillus thuringiensis spores.

  18. Bacillus cereus panophthalmitis: source of the organism.

    Science.gov (United States)

    Shamsuddin, D; Tuazon, C U; Levy, C; Curtin, J

    1982-01-01

    Serious infections with the "nonpathogenic" Bacillus species are increasingly being recognized, especially in drug abusers. Cases of panophthalmitis secondary to infection with Bacillus cereus, with and without associated bacteremia, have been reported. Three drug abusers with panophthalmitis seen in our hospitals during a three-year period are described, and the similar cases reported in the literature are reviewed. The syndrome is characterized by an acute onset with a rapid fulminating course that eventually leads to enucleation or evisceration of the eye. The pathogenic mechanism is unknown, but is probably related to the production of toxin (lecithinase) by B. cereus. Clindamycin appears to be the antibiotic of choice in the treatment of this infection. In order to identify a possible source of the organism, 59 samples of heroin and injection paraphernalia were cultured. Twenty cultures yielded organisms; Bacillus species were the predominant isolates. Thirty-eight percent of the isolates were identified as B. cereus. Thus, infections caused by Bacillus species in drug abusers can probably be associated with intravenous heroin abuse because heroin mixtures and injection paraphernalia are frequently contaminated with this organism.

  19. Inactivation of Bacillus spores inoculated in milk by Ultra High Pressure Homogenization.

    Science.gov (United States)

    Amador Espejo, Genaro Gustavo; Hernández-Herrero, M M; Juan, B; Trujillo, A J

    2014-12-01

    Ultra High-Pressure Homogenization treatments at 300 MPa with inlet temperatures (Ti) of 55, 65, 75 and 85 °C were applied to commercial Ultra High Temperature treated whole milk inoculated with Bacillus cereus, Bacillus licheniformis, Bacillus sporothermodurans, Bacillus coagulans, Geobacillus stearothermophilus and Bacillus subtilis spores in order to evaluate the inactivation level achieved. Ultra High-Pressure Homogenization conditions at 300 MPa with Ti = 75 and 85 °C were capable of a spore inactivation of ∼5 log CFU/mL. Furthermore, under these processing conditions, commercial sterility (evaluated as the complete inactivation of the inoculated spores) was obtained in milk, with the exception of G. stearothermophilus and B. subtilis treated at 300 MPa with Ti = 75 °C. The results showed that G. stearothermophilus and B. subtilis have higher resistance to the Ultra High-Pressure Homogenization treatments applied than the other microorganisms inoculated and that a treatment performed at 300 MPa with Ti = 85 °C was necessary to completely inactivate these microorganisms at the spore level inoculated (∼1 × 10(6) CFU/mL). Besides, a change in the resistance of B. licheniformis, B. sporothermodurans, G. stearothermophilus and B. subtilis spores was observed as the inactivation obtained increased remarkably in treatments performed with Ti between 65 and 75 °C. This study provides important evidence of the suitability of UHPH technology for the inactivation of spores in high numbers, leading to the possibility of obtaining commercially sterile milk.

  20. Measuring Total and Germinable Spore Populations

    Science.gov (United States)

    Noell, A.C.; Yung, P.T.; Yang, W.; Lee, C.; Ponce, A.

    2011-01-01

    It has been shown that bacterial endospores can be enumerated using a microscopy based assay that images the luminescent halos from terbium ions bound to dipicolinic acid, a spore specific chemical marker released upon spore germination. Further development of the instrument has simplified it towards automation while at the same time improving image quality. Enumeration of total spore populations has also been developed allowing measurement of the percentage of viable spores in any population by comparing the germinable/culturable spores to the total. Percentage viability will allow a more quantitative comparison of the ability of spores to survive across a wide range of extreme environments.

  1. A novel multiplex PCR discriminates Bacillus anthracis and its genetically related strains from other Bacillus cereus group species.

    Directory of Open Access Journals (Sweden)

    Hirohito Ogawa

    Full Text Available Anthrax is an important zoonotic disease worldwide that is caused by Bacillus anthracis, a spore-forming pathogenic bacterium. A rapid and sensitive method to detect B. anthracis is important for anthrax risk management and control in animal cases to address public health issues. However, it has recently become difficult to identify B. anthracis by using previously reported molecular-based methods because of the emergence of B. cereus, which causes severe extra-intestinal infection, as well as the human pathogenic B. thuringiensis, both of which are genetically related to B. anthracis. The close genetic relation of chromosomal backgrounds has led to complexity of molecular-based diagnosis. In this study, we established a B. anthracis multiplex PCR that can screen for the presence of B. anthracis virulent plasmids and differentiate B. anthracis and its genetically related strains from other B. cereus group species. Six sets of primers targeting a chromosome of B. anthracis and B. anthracis-like strains, two virulent plasmids, pXO1 and pXO2, a bacterial gene, 16S rRNA gene, and a mammalian gene, actin-beta gene, were designed. The multiplex PCR detected approximately 3.0 CFU of B. anthracis DNA per PCR reaction and was sensitive to B. anthracis. The internal control primers also detected all bacterial and mammalian DNAs examined, indicating the practical applicability of this assay as it enables monitoring of appropriate amplification. The assay was also applied for detection of clinical strains genetically related to B. anthracis, which were B. cereus strains isolated from outbreaks of hospital infections in Japan, and field strains isolated in Zambia, and the assay differentiated B. anthracis and its genetically related strains from other B. cereus group strains. Taken together, the results indicate that the newly developed multiplex PCR is a sensitive and practical method for detecting B. anthracis.

  2. Comparative analysis of biofilm formation by Bacillus cereus reference strains and undomesticated food isolates and the effect of free iron.

    Science.gov (United States)

    Hayrapetyan, Hasmik; Muller, Lisette; Tempelaars, Marcel; Abee, Tjakko; Nierop Groot, Masja

    2015-05-04

    Biofilm formation of Bacillus cereus reference strains ATCC 14579 and ATCC 10987 and 21 undomesticated food isolates was studied on polystyrene and stainless steel as contact surfaces. For all strains, the biofilm forming capacity was significantly enhanced when in contact with stainless steel (SS) as a surface as compared to polystyrene (PS). For a selection of strains, the total CFU and spore counts in biofilms were determined and showed a good correlation between CFU counts and total biomass of these biofilms. Sporulation was favoured in the biofilm over the planktonic state. To substantiate whether iron availability could affect B. cereus biofilm formation, the free iron availability was varied in BHI by either the addition of FeCl3 or by depletion of iron with the scavenger 2,2-Bipyridine. Addition of iron resulted in increased air-liquid interface biofilm on polystyrene but not on SS for strain ATCC 10987, while the presence of Bipyridine reduced biofilm formation for both materials. Biofilm formation was restored when excess FeCl3 was added in combination with the scavenger. Further validation of the iron effect for all 23 strains in microtiter plate showed that fourteen strains (including ATCC10987) formed a biofilm on PS. For eight of these strains biofilm formation was enhanced in the presence of added iron and for eleven strains it was reduced when free iron was scavenged. Our results show that stainless steel as a contact material provides more favourable conditions for B. cereus biofilm formation and maturation compared to polystyrene. This effect could possibly be linked to iron availability as we show that free iron availability affects B. cereus biofilm formation.

  3. A novel multiplex PCR discriminates Bacillus anthracis and its genetically related strains from other Bacillus cereus group species.

    Science.gov (United States)

    Ogawa, Hirohito; Fujikura, Daisuke; Ohnuma, Miyuki; Ohnishi, Naomi; Hang'ombe, Bernard M; Mimuro, Hitomi; Ezaki, Takayuki; Mweene, Aaron S; Higashi, Hideaki

    2015-01-01

    Anthrax is an important zoonotic disease worldwide that is caused by Bacillus anthracis, a spore-forming pathogenic bacterium. A rapid and sensitive method to detect B. anthracis is important for anthrax risk management and control in animal cases to address public health issues. However, it has recently become difficult to identify B. anthracis by using previously reported molecular-based methods because of the emergence of B. cereus, which causes severe extra-intestinal infection, as well as the human pathogenic B. thuringiensis, both of which are genetically related to B. anthracis. The close genetic relation of chromosomal backgrounds has led to complexity of molecular-based diagnosis. In this study, we established a B. anthracis multiplex PCR that can screen for the presence of B. anthracis virulent plasmids and differentiate B. anthracis and its genetically related strains from other B. cereus group species. Six sets of primers targeting a chromosome of B. anthracis and B. anthracis-like strains, two virulent plasmids, pXO1 and pXO2, a bacterial gene, 16S rRNA gene, and a mammalian gene, actin-beta gene, were designed. The multiplex PCR detected approximately 3.0 CFU of B. anthracis DNA per PCR reaction and was sensitive to B. anthracis. The internal control primers also detected all bacterial and mammalian DNAs examined, indicating the practical applicability of this assay as it enables monitoring of appropriate amplification. The assay was also applied for detection of clinical strains genetically related to B. anthracis, which were B. cereus strains isolated from outbreaks of hospital infections in Japan, and field strains isolated in Zambia, and the assay differentiated B. anthracis and its genetically related strains from other B. cereus group strains. Taken together, the results indicate that the newly developed multiplex PCR is a sensitive and practical method for detecting B. anthracis.

  4. Biocontrol of Aspergillus species on peanut kernels by antifungal diketopiperazine producing Bacillus cereus associated with entomopathogenic nematode.

    Science.gov (United States)

    Kumar, Sasidharan Nishanth; Sreekala, Sreerag Ravikumar; Chandrasekaran, Dileep; Nambisan, Bala; Anto, Ruby John

    2014-01-01

    The rhabditid entomopathogenic nematode associated Bacillus cereus and the antifungal compounds produced by this bacterium were evaluated for their activity in reducing postharvest decay of peanut kernels caused by Aspergillus species in in vitro and in vivo tests. The results showed that B. cereus had a significant effect on biocontrol effectiveness in in vitro and in vivo conditions. The antifungal compounds produced by the B. cereus were purified using silica gel column chromatography and their structure was elucidated using extensive spectral analyses. The compounds were identified as diketopiperazines (DKPs) [cyclo-(L-Pro-Gly), cyclo(L-Tyr-L-Tyr), cyclo-(L-Phe-Gly) and cyclo(4-hydroxy-L-Pro-L-Trp)]. The antifungal activities of diketopiperazines were studied against five Aspergillus species and best MIC of 2 µg/ml was recorded against A. flavus by cyclo(4-hydroxy-L-Pro-L-Trp). To investigate the potential application of cyclo(4-hydroxy-L-Pro-L-Trp) to eliminate fungal spoilage in food and feed, peanut kernels was used as a food model system. White mycelia and dark/pale green spores of Aspergillus species were observed in the control peanut kernels after 2 days incubation. However the fungal growth was not observed in peanut kernels treated with cyclo(4-hydroxy-L-Pro-L-Trp). The cyclo(4-hydroxy-L-Pro-L-Trp) was nontoxic to two normal cell lines [fore skin (FS) normal fibroblast and African green monkey kidney (VERO)] up to 200 µg/ml in MTT assay. Thus the cyclo(4-hydroxy-L-Pro-L-Trp) identified in this study may be a promising alternative to chemical preservatives as a potential biopreservative agent which prevent fungal growth in food and feed. To the best of our knowledge, this is the first report demonstrating that the entomopathogenic nematode associated B. cereus and cyclo(4-hydroxy-L-Pro-L-Trp) could be used as a biocontrol agents against postharvest fungal disease caused by Aspergillus species.

  5. Transcriptome analysis of Bacillus thuringiensis spore life, germination and cell outgrowth in a vegetable-based food model.

    Science.gov (United States)

    Bassi, Daniela; Colla, Francesca; Gazzola, Simona; Puglisi, Edoardo; Delledonne, Massimo; Cocconcelli, Pier Sandro

    2016-05-01

    Toxigenic species belonging to Bacillus cereus sensu lato, including Bacillus thuringiensis, cause foodborne outbreaks thanks to their capacity to survive as spores and to grow in food matrixes. The goal of this work was to assess by means of a genome-wide transcriptional assay, in the food isolate B. thuringiensis UC10070, the gene expression behind the process of spore germination and consequent outgrowth in a vegetable-based food model. Scanning electron microscopy and Energy Dispersive X-ray microanalysis were applied to select the key steps of B. thuringiensis UC10070 cell cycle to be analyzed with DNA-microarrays. At only 40 min from heat activation, germination started rapidly and in less than two hours spores transformed in active growing cells. A total of 1646 genes were found to be differentially expressed and modulated during the entire B. cereus life cycle in the food model, with most of the significant genes belonging to transport, transcriptional regulation and protein synthesis, cell wall and motility and DNA repair groups. Gene expression studies revealed that toxin-coding genes nheC, cytK and hblC were found to be expressed in vegetative cells growing in the food model.

  6. Refined multivalent display of bacterial spore-binding peptides.

    Science.gov (United States)

    Lusvarghi, Sabrina; Kim, Jenny Morana; Creeger, Yehuda; Armitage, Bruce Alan

    2009-05-07

    A multiple antigen peptide display scaffold was used to create multivalent versions of a heptapeptide selected previously by phage display to bind to Bacillus subtilis spores. A simple flow cytometric assay was developed in which a biotinylated form of the peptide was first bound to fluorescent streptavidin, then the fluorescent streptavidin-peptide complex was bound to spores before introduction into the cytometer. This assay clearly demonstrated that the tetravalent scaffold enhanced the affinity for B. subtilis spores by greater than 1 and 2 orders of magnitude when compared to divalent and monovalent analogues, respectively. However, variations in the number and flexibility of spacer residues within the scaffold did not significantly affect the binding affinity of the tetravalent peptides. Similar to prior reports, these multivalent scaffolds are effective most likely because they mimic the multivalent display of the original peptide library on the phage coat. Moreover, the tetravalent peptides can be readily integrated into a variety of heterogeneous and homogeneous spore-detection assay formats.

  7. Adhesion of Spores of Bacillus thuringiensis on a Planar Surface

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Eunhyea [Georgia Institute of Technology; Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Lee, Ida [University of Tennessee, Knoxville (UTK); Joy, David Charles [ORNL; Palumbo, Anthony Vito [ORNL; Tsouris, Costas [ORNL

    2010-01-01

    Adhesion of spores of Bacillus thuringiensis (Bt) and spherical silica particles on surfaces was experimentally and theoretically investigated in this study. Topography analysis via atomic force microscopy (AFM) and electron microscopy indicates that Bt spores are rod shaped, {approx}1.3 {mu}m in length and {approx}0.8 {mu}m in diameter. The adhesion force of Bt spores and silica particles on gold-coated glass was measured at various relative humidity (RH) levels by AFM. It was expected that the adhesion force would vary with RH because the individual force components contributing to the adhesion force depend on RH. The adhesion force between a particle and a planar surface in atmospheric environments was modeled as the contribution of three major force components: capillary, van der Waals, and electrostatic interaction forces. Adhesion force measurements for Bt spore (silica particle) and the gold surface system were comparable with calculations. Modeling results show that there is a critical RH value, which depends on the hydrophobicity of the materials involved, below which the water meniscus does not form and the contribution of the capillary force is zero. As RH increases, the van der Waals force decreases while the capillary force increases to a maximum value.

  8. Bacillus cereus: emetic toxin production and gamma hypothesis for growth

    NARCIS (Netherlands)

    Biesta-Peters, E.G.

    2011-01-01

    Bacillus cereus is a food spoilage microorganism and a pathogen. Growth of B. cereus can be prevented or delayed by adding growth limiting compounds to the food product or by altered storage conditions. Combinations of growth limiting factors

  9. Molecular Kinetics of Reviving Bacterial Spores

    OpenAIRE

    2013-01-01

    Bacterial spores can remain dormant for years, yet they possess a remarkable potential to rapidly resume a vegetative life form. Here, we identified a distinct phase at the onset of spore outgrowth, designated the ripening period. This transition phase is exploited by the germinating spore for molecular reorganization toward elongation and subsequent cell division. We have previously shown that spores of different ages, kept under various temperatures, harbor dissimilar molecular reservoirs (...

  10. Relevance of diffusion through bacterial spore coats/membranes and the associated concentration boundary layers in the initial lag phase of inactivation: a case study for Bacillus subtilis with ozone and monochloramine.

    Science.gov (United States)

    Fernando, W J N; Othman, R

    2006-02-01

    Disinfectants are generally used to inactivate microorganisms in solutions. The process of inactivation involves the disinfectant in the liquid diffusing towards the bacteria sites and thereafter reacting with bacteria at rates determined by the respective reaction rates. Such processes have demonstrated an initial lag phase followed by an active depletion phase of bacteria. This paper attempts to study the importance of the combined effects of diffusion of the disinfectant through the outer membrane of the bacteria and transport through the associated concentration boundary layers (CBLs) during the initial lag phase. Mathematical equations are developed correlating the initial concentration of the disinfectant with time required for reaching a critical concentration (C*) at the inner side of the membrane of the cell based on diffusion of disinfectant through the outer membranes of the bacteria and the formation of concentration boundary layers on both sides of the membranes. Experimental data of the lag phases of inactivation already available in the literature for inactivation of Bacillus subtilis spores with ozone and monochloramine are tested with the equations. The results seem to be in good agreement with the theoretical equations indicating the importance of diffusion process across the outer cell membranes and the resulting CBL's during the lag phase of disinfection.

  11. Preparation and evaluation of spore-specific affinity- augmented bio-imprinted beads

    Energy Technology Data Exchange (ETDEWEB)

    Harvey, Scott D.; Mong, Gary M.; Ozanich, Rich M.; Mclean, Jeffrey S.; Goodwin, Shannon M.; Valentine, Nancy B.; Fredrickson, Jim K.

    2006-09-01

    The procedures previously described for imprinting bead surfaces with bacteria were applied to create novel affinity-augmented bacterial spore-imprinted beads. The imprinted beads are intended as a front-end spore capture/concentration stage of an integrated biological detection system. Our approach involves embedding bead surfaces with Bacillus thuringiensis kurstaki (Bt) spores (as a surrogate for Bacillus anthracis) during synthesis. Subsequent steps involved lithographic deactivation using a perfluoroether, spore removal to create imprint sites, and coating imprints with the lectin, concanavalin A, to provide general affinity. The synthesis of the intended material with the desired imprints was verified by scanning electron and confocal laser-scanning microscopy. The material was evaluated using spore-binding assays with either Bt or Bacillus subtilis (Bs) spores. The binding assays indicated strong spore-binding capability and a robust imprinting effect that accounted for 25 percent additional binding over nonimprinted controls. The binding assay results also indicated that further refinement of the surface deactivation procedure would enhance the performance of the imprinted substrate.

  12. Investigating the thermodynamic stability of Bacillus subtilis spore-uranium(VI) adsorption though surface complexation modeling

    Science.gov (United States)

    Harrold, Z.; Hertel, M.; Gorman-Lewis, D.

    2012-12-01

    Dissolved uranium speciation, mobility, and remediation are increasingly important topics given continued and potential uranium (U) release from mining operations and nuclear waste. Vegetative bacterial cell surfaces are known to adsorb uranium and may influence uranium speciation in the environment. Previous investigations regarding U(VI) adsorption to bacterial spores, a differentiated and dormant cell type with a tough proteinaceous coat, include U adsorption affinity and XAFS data. We investigated the thermodynamic stability of aerobic, pH dependent uranium adsorption to bacterial spore surfaces using purified Bacillus subtilis spores in solution with 5ppm uranium. Adsorption reversibility and kinetic experiments indicate that uranium does not precipitate over the duration of the experiments and equilibrium is reached within 20 minutes. Uranium-spore adsorption edges exhibited adsorption at all pH measured between 2 and 10. Maximum adsorption was achieved around pH 7 and decreased as pH increased above 7. We used surface complexation modeling (SCM) to quantify uranium adsorption based on balanced chemical equations and derive thermodynamic stability constants for discrete uranium-spore adsorption reactions. Site specific thermodynamic stability constants provide insight on interactions occurring between aqueous uranium species and spore surface ligands. The uranium adsorption data and SCM parameters described herein, also provide a basis for predicting the influence of bacterial spores on uranium speciation in natural systems and investigating their potential as biosorption agents in engineered systems.

  13. Improved proteomic analysis following trichloroacetic acid extraction of Bacillus anthracis spore proteins.

    Science.gov (United States)

    Deatherage Kaiser, Brooke L; Wunschel, David S; Sydor, Michael A; Warner, Marvin G; Wahl, Karen L; Hutchison, Janine R

    2015-11-01

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Analysis of cellular proteins is dependent upon efficient extraction from bacterial samples, which can be challenging with increasing complexity and refractory characteristics. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrichment for certain classes of proteins. The method presented here is technically simple, does not require specialized equipment such as a mechanical disrupter, and is effective for protein extraction of the particularly challenging sample type of Bacillus anthracis Sterne spores. The ability of Trichloroacetic acid (TCA) extraction to isolate proteins from spores and enrich for spore-specific proteins was compared to the traditional mechanical disruption method of bead beating. TCA extraction improved the total average number of proteins identified within a sample as compared to bead beating (547 vs 495, respectively). Further, TCA extraction enriched for 270 spore proteins, including those typically identified by first isolating the spore coat and exosporium layers. Bead beating enriched for 156 spore proteins more typically identified from whole spore proteome analyses. The total average number of proteins identified was equal using TCA or bead beating for easily lysed samples, such as B. anthracis vegetative cells. As with all assays, supplemental methods such as implementation of an alternative preparation method may simplify sample preparation and provide additional insight to the protein biology of the organism being studied.

  14. Identification of pathogenic microbial cells and spores by electrochemical detection on a biochip

    Directory of Open Access Journals (Sweden)

    Andresen Heiko

    2004-04-01

    Full Text Available Abstract Background Bacillus cereus constitutes a significant cause of acute food poisoning in humans. Despite the recent development of different detection methods, new effective control measures and better diagnostic tools are required for quick and reliable detection of pathogenic micro-organisms. Thus, the objective of this study was to determine a simple method for rapid identification of enterotoxic Bacillus strains. Here, a special attention is given to an electrochemical biosensor since it meets the requirements of minimal size, lower costs and decreased power consumption. Results A bead-based sandwich hybridization system was employed in conjugation with electric chips for detection of vegetative cells and spores of Bacillus strains based on their toxin-encoding genes. The system consists of a silicon chip based potentiometric cell, and utilizes paramagnetic beads as solid carriers of the DNA probes. The specific signals from 20 amol of bacterial cell or spore DNA were achieved in less than 4 h. The method was also successful when applied directly to unpurified spore and cell extract samples. The assay for the haemolytic enterotoxin genes resulted in reproducible signals from B. cereus and B. thuringiensis while haemolysin-negative B. subtilis strain did not yield any signal. Conclusions The sensitivity, convenience and specificity of the system have shown its potential. In this respect an electrochemical detection on a chip enabling a fast characterization and monitoring of pathogens in food is of interest. This system can offer a contribution in the rapid identification of bacteria based on the presence of specific genes without preceding nucleic acid amplification.

  15. 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

  16. Selection of high affine peptide ligands for detection of Clostridium Tyrobutyricum spores.

    Science.gov (United States)

    Lavilla, María; De Luis, Ruth; Pérez, María Dolores; Calvo, Miguel; Sánchez, Lourdes

    2009-11-01

    Clostridium tyrobutyricum is the main agent responsible for "late blowing" in cheese, which causes severe economic losses. Nowadays, the reference method for its detection is the Most-Probable-Number (MPN); however, it is time consuming and non-specific. Thus, in order to check milk contamination with spores of C. tyrobutyricum, a more specific and rapid method would be required. The objective of this work was to obtain a ligand to establish the basis to develop a biomagnetic separation method for detection of C. tyrobutyricum spores. This study describes the selection of thirteen highly affine peptides to C. tyrobutyricum spores from a phage-display peptide library. In order to test the ability of the peptides attached to a solid support to bind the spores, the most frequent peptide was synthesised and used to coat paramagnetic beads.

  17. Absorption edge imaging of sporocide-treated and non-treated bacterial spores

    Energy Technology Data Exchange (ETDEWEB)

    Panessa-Warren, B.J.; Tortora, G.T.; Warren, J.B.

    1987-01-01

    When deprived of nutrients, spore forming bacilli produce endospores which are remarkably resistant to chemical sterilization. Little is known about the morphology and response fo these spores following exposure to sporocidal agents. Light microscopy does not provide sufficient resolution for studying the rupture of the spore coat and fate of intracellular material. Transmission and scanning electron microscopy offer superior resolution but require specimen preparation methods that induce physiologic as well as morphologic changes in the spores, thereby making accurate interpretation of micrographs difficult. To eliminate the possible artifacts induced by chemical fixation, dehydration, embeddment, staining and sectioning, treated and non-sporocide-treated endospores of B. thuringiensis and B. subtilis were imaged by x-ray contact microscopy using monochromatic x-rays. 6 refs., 2 figs.

  18. Bacteriophages and bacteriophage-derived endolysins as potential therapeutics to combat Gram-positive spore forming bacteria.

    Science.gov (United States)

    Nakonieczna, A; Cooper, C J; Gryko, R

    2015-09-01

    Since their discovery in 1915, bacteriophages have been routinely used within Eastern Europe to treat a variety of bacterial infections. Although initially ignored by the West due to the success of antibiotics, increasing levels and diversity of antibiotic resistance is driving a renaissance for bacteriophage-derived therapy, which is in part due to the highly specific nature of bacteriophages as well as their relative abundance. This review focuses on the bacteriophages and derived lysins of relevant Gram-positive spore formers within the Bacillus cereus group and Clostridium genus that could have applications within the medical, food and environmental sectors.

  19. Bacterial spore inhibition and inactivation in foods by pressure, chemical preservatives, and mild heat.

    Science.gov (United States)

    Shearer, A E; Dunne, C P; Sikes, A; Hoover, D G

    2000-11-01

    Sucrose laurates, sucrose palmitate, sucrose stearates, and monolaurin (Lauricidin) were evaluated for inhibitory effects against spores of Bacillus sp., Clostridium sporogenes PA3679, and Alicyclobacillus sp. in a model agar system. The combined treatment of sucrose laurate, high hydrostatic pressure, and mild heat was evaluated on spores of Bacillus and Alicyclobacillus in foods. The minimum inhibitory concentrations of the sucrose esters were higher than that of Lauricidin for all spores tested in the model agar system, but Lauricidin was not the most readily suspended in the test media. The sucrose laurates and sucrose palmitate were more effective and more readily suspended than the sucrose stearates. A combined treatment of sucrose laurate (<1.0%), 392 megaPascals (MPa) at 45 degrees C for 10 to 15 min provided 3- to 5.5-log10 CFU/ml reductions from initial populations of 10(6) CFU/ml for Bacillus subtilis 168 in milk, Bacillus cereus 14579 in beef, Bacillus coagulans 7050 in tomato juice (pH 4.5), Alicyclobacillus sp. N1089 in tomato juice (pH 4.5), and Alicyclobacillus sp. N1098 in apple juice. The most notable change in the appearance of the products was temporary foaming during mixing of the sucrose laurate in the foods. The effect of sucrose laurate appeared to be inhibitory rather than lethal to the spores. The inhibitory effects observed on Bacillus and Alicyclobacillus spores by the combined treatment of pressure, mild heat, and sucrose laurate appear promising for food applications where alternatives to high heat processing are desired.

  20. Antimicrobial Effects of Honey on Bacillus Cereus

    Directory of Open Access Journals (Sweden)

    This paper should be cited as: Javadzadeh M, Najafi M, Rezaei M, Dastoor M, Behzadi AS, Amiri A . [ Antimicrobial Effects of Honey on Bacillus Cereus ]. MLJ. 201 4 ; 8 ( 2 : 55 - 61 [Article in Persian] Javadzadeh, M. (MSc

    2014-05-01

    Full Text Available Background and Objective: Honey is a healthy and nutritious food that has been used for a long time as a treatment for different diseases. One of the applied properties of honey is its antimicrobial effect, which differs between different types of honey due to variation of phenolic and antioxidant compositions. This study aimed to assess antimicrobial effect of honey on Bacillus cereus, considering its chemical properties. Material and Methods: Three samples of honey (A1 and A2 of Khorasan Razavi Province and A3 of South Khorasan province (were prepared and studied in terms of chemical parameters .The antibacterial effect of honey was surveyed throughTurbidimeter using spectrometer with incubator time of 2, 4, 6, and 8hrs. the level of turbidity caused by bacterium growth was measured at different times with a wavelength of 600nm. Results: According to the study, the samples containing higher concentration of polyphenol has more antimicrobial activity. The samples of A2, A3, and A1 had the highest concentration of polyphenol, respectively. Conclusion: The results indicate the prebiotic effect of honey that can be justified by the presence of fructo-oligosacharids and vitamin B. Keywords: Honey, Bacillus Cereus, Antibacterial, Turbidimetry.

  1. Enterotoxigenic profiles and polymerase chain reaction detection of Bacillus cereus group cells and B. cereus strains from foods and food-borne outbreaks.

    Science.gov (United States)

    Hsieh, Y M; Sheu, S J; Chen, Y L; Tsen, H Y

    1999-10-01

    Bacillus cereus is one of the important food pathogens. Since B. cereus group cells, such as B. cereus, B. thuringiensis, B. anthracis and B. mycoides, share many phenotypical properties and a high level of chromosomal sequence similarity, it is interesting to investigate the virulence profiles for B. cereus group cells, including B. cereus strains isolated from foods and samples associated with food-poisoning outbreaks. For this investigation, the presence of enterotoxin genes, such as those of haemolysin BL, B. cereus enterotoxin T and enterotoxin FM, were assayed by polymerase chain reaction (PCR) methods. Meanwhile, their enterotoxin activities were assayed using the BCET-RPLA kit, haemolytic patterns on sheep blood agar and their cytotoxicity to Chinese hamster ovary (CHO) cells. Results showed that there were 12 enterotoxigenic profiles for the 98 B. cereus group strains collected. In addition, if any of the three types of enterotoxins was present in the B. cereus group cells, these cells were shown to be cytotoxic to the CHO cells. Similar enterotoxigenic profiles could be found among strains of B. cereus, B. mycoides and B. thuringiensis. Thus, all B. cereus group strains may be potentially toxigenic and the detection of these cells in foods is important. We thus designed PCR primers, termed Ph1/Ph2, from the sphingomyelinase gene of B. cereus cells. These primers were specific for all B. cereus group strains and could be used for the detection of B. cereus cells contaminated in food samples.

  2. Isolating and Purifying Clostridium difficile Spores

    Science.gov (United States)

    Edwards, Adrianne N.; McBride, Shonna M.

    2016-01-01

    Summary The ability for the obligate anaerobe, Clostridium difficile, to form a metabolically dormant spore is critical for the survival of this organism outside of the host. This spore form is resistant to a myriad of environmental stresses, including heat, desiccation and exposure to disinfectants and antimicrobials. These intrinsic properties of spores allow C. difficile to survive long-term in an oxygenated environment, to be easily transmitted from host-to-host and to persist within the host following antibiotic treatment. Because of the importance of the spore form to the C. difficile lifecycle and treatment and prevention of C. difficile infection (CDI), the isolation and purification of spores are necessary to study the mechanisms of sporulation and germination, investigate spore properties and resistances, and for use in animal models of CDI. This chapter provides basic protocols, in vitro growth conditions and additional considerations for purifying C. difficile spores for a variety of downstream applications. PMID:27507337

  3. Production, Secretion and Biological Activity of Bacillus cereus Enterotoxins

    Directory of Open Access Journals (Sweden)

    Sonia Senesi

    2010-06-01

    Full Text Available Bacillus cereus behaves as an opportunistic pathogen frequently causing gastrointestinal diseases, and it is increasingly recognized to be responsible for severe local or systemic infections. Pathogenicity of B. cereus mainly relies on the secretion of a wide array of toxins and enzymes and also on the ability to undergo swarming differentiation in response to surface-sensing. In this report, the pathogenicity exerted by B. cereus toxins is described with particular attention to the regulatory mechanisms of production and secretion of HBL, Nhe and CytK enterotoxins.

  4. Phospholipase Cδ regulates germination of Dictyostelium spores

    Directory of Open Access Journals (Sweden)

    Van Haastert Peter JM

    2001-12-01

    Full Text Available Abstract Background Many eukaryotes, including plants and fungi make spores that resist severe environmental stress. The micro-organism Dictyostelium contains a single phospholipase C gene (PLC; deletion of the gene has no effect on growth, cell movement and differentiation. In this report we show that PLC is essential to sense the environment of food-activated spores. Results Plc-null spores germinate at alkaline pH, reduced temperature or increased osmolarity, conditions at which the emerging amoebae can not grow. In contrast, food-activated wild-type spores return to dormancy till conditions in the environment allow growth. The analysis of inositol 1,4,5-trisphosphate (IP3 levels and the effect of added IP3 uncover an unexpected mechanism how PLC regulates spore germination: i deletion of PLC induces the enhanced activity of an IP5 phosphatase leading to high IP3 levels in plc-null cells; ii in wild-type spores unfavourable conditions inhibit PLC leading to a reduction of IP3 levels; addition of exogenous IP3 to wild-type spores induces germination at unfavourable conditions; iii in plc-null spores IP3 levels remain high, also at unfavourable environmental conditions. Conclusions The results imply that environmental conditions regulate PLC activity and that IP3 induces spore germination; the uncontrolled germination of plc-null spores is not due to a lack of PLC activity but to the constitutive activation of an alternative IP3-forming pathway.

  5. Inhibition of cereulide toxin synthesis by emetic Bacillus cereus via long-chain polyphosphates.

    Science.gov (United States)

    Frenzel, Elrike; Letzel, Thomas; Scherer, Siegfried; Ehling-Schulz, Monika

    2011-02-01

    Severe intoxications caused by the Bacillus cereus emetic toxin cereulide can hardly be prevented due to the ubiquitous distribution and heat resistance of spores and the extreme thermal and chemical stability of cereulide. It would therefore be desirable to inhibit cereulide synthesis during food manufacturing processes or in prepared foods, which are stored under time-temperature abuse conditions. Toward this end, the impacts of three long-chain polyphosphate (polyP) formulations on growth and cereulide production were examined. The inhibition was dependent on the concentration and the type of the polyP blend, indicating that polyPs and not the orthophosphates were effective. Quantitative PCR (qPCR) monitoring at sublethal concentrations revealed that polyPs reduced the transcription of ces nonribosomal peptide synthetase (NRPS) genes by 3- to 4-fold along with a significantly reduced toxin production level. At lower concentrations, toxin synthesis was decreased, although the growth rate was not affected. These data indicate a differential effect on toxin synthesis independent of growth inhibition. The inhibition of toxin synthesis in food was also observed. Despite the growth of B. cereus, toxin synthesis was reduced by 70 to 100% in two model food systems (reconstituted infant food and oat milk), which were analyzed with HEp-2 cell culture assays and high-performance liquid chromatography (HPLC)/electrospray ionization-time of flight mass spectrometry (ESI-TOF-MS). Accordingly, ces promoter activity was strongly downregulated, as visualized by using a lux-based reporter strain. These data illustrate the potential of polyphosphate formulations to reduce the risk of cereulide synthesis in food and may contribute to targeted hurdle concepts.

  6. Bacillus cereus panophthalmitis after intravenous heroin.

    Science.gov (United States)

    Hatem, G; Merritt, J C; Cowan, C L

    1979-03-01

    Two healthy young black men developed panophthalmitis after intravenous heroin injections. Bacillus cereus, considered to be a relatively noncommon pathogen for man, was found to be the causative agent as it was recovered from the anterior chamber and viterous cavity of both cases. The ocular findings were unilateral in each case, and neither patient had any sistemic involvement from the bacteremia. The onset of visual symptoms varied from 24 to 36 hours after the last intravenous injection with the eye becoming rapidly blind. Photographs of the early fundus lesions included preretinal hypopyon-like lesions and peculiar changes in the blood vasculature. Intracameral gentamicin and steroids did not alter the cause, and treatment was enucleation.

  7. Fatal Bacillus cereus bacteremia in a patient with diabetes.

    Science.gov (United States)

    Orrett, F A

    2000-04-01

    This report describes a fatal case of Bacillus cereus septicemia in a patient with uncontrolled diabetes and re-emphasizes the potential seriousness of Bacillus infections in patients with compromised immune function.

  8. Adaptation in Bacillus cereus: from stress to disease

    Directory of Open Access Journals (Sweden)

    Catherine Duport

    2016-10-01

    Full Text Available Bacillus cereus is a foodborne pathogen that causes diarrheal disease in humans. After ingestion B. cereus experiences in the human gastro-intestinal tract abiotic physical variables encountered in food, such as acidic pH in the stomach and changing oxygen conditions in the human intestine. B. cereus responds to environmental changing conditions (stress by reversibly adjusting its physiology to maximize resource utilization while maintaining structural and genetic integrity by repairing and minimizing damage to cellular infrastructure. As reviewed in this article, B. cereus adapts to acidic pH and changing oxygen conditions through diverse regulatory mechanisms and then exploits its metabolic flexibility to grow and produce enterotoxins. We then focus on the intricate link between metabolism, redox homeostasis and enterotoxins, which are recognized as important contributors of food-borne disease.

  9. Genomics, evolution, and crystal structure of a new family of bacterial spore kinases.

    Science.gov (United States)

    Scheeff, Eric D; Axelrod, Herbert L; Miller, Mitchell D; Chiu, Hsiu-Ju; Deacon, Ashley M; Wilson, Ian A; Manning, Gerard

    2010-05-01

    Bacterial spore formation is a complex process of fundamental relevance to biology and human disease. The spore coat structure is complex and poorly understood, and the roles of many of the protein components remain unclear. We describe a new family of spore coat proteins, the bacterial spore kinases (BSKs), and the first crystal structure of a BSK, YtaA (CotI) from Bacillus subtilis. BSKs are widely distributed in spore-forming Bacillus and Clostridium species, and have a dynamic evolutionary history. Sequence and structure analyses indicate that the BSKs are CAKs, a prevalent group of small molecule kinases in bacteria that is distantly related to the eukaryotic protein kinases. YtaA has substantial structural similarity to CAKs, but also displays distinctive features that broaden our understanding of the CAK group. Evolutionary constraint analysis of the protein surfaces indicates that members of the BSK family have distinct clade-conserved patterns in the substrate binding region, and probably bind and phosphorylate distinct targets. Several classes of BSKs have apparently independently lost catalytic activity to become pseudokinases, indicating that the family also has a major noncatalytic function.

  10. The high-resolution architecture and structural dynamics of Bacillus spores

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T J; Wheeler, K E; Malkin, A J

    2004-05-06

    The capability to image single microbial cell surfaces at nanometer scale under native conditions would profoundly impact mechanistic and structural studies of pathogenesis, immunobiology, environmental resistance and biotransformation. We report here that advances in atomic force microscopy (AFM) have allowed us to directly visualize high-resolution native structures of bacterial endospores, including the exosporium and spore coats of four Bacillus species in air and water environments. The dimensions of individual Bacillus atrophaeus spores were found to decrease reversibly by 12% in response to a change in the environment from aqueous to aerial phase. Intraspecies spore size distribution analyses revealed that spore length could vary by a factor of 2 while the absolute deviation is 7 - 13% in length and 4 - 6 % in width. AFM analysis also demonstrated that the mechanisms of spore coat self-assembly are similar to those described for inorganic and macromolecular crystallization. These results establish AFM as a powerful new tool for the analysis of molecular architecture and variability as a function of spatial, temporal and developmental organizational scales.

  11. Fulminating bacteremia and pneumonia due to Bacillus cereus.

    Science.gov (United States)

    Miller, J M; Hair, J G; Hebert, M; Hebert, L; Roberts, F J; Weyant, R S

    1997-02-01

    We present two cases of rapidly progressing, fatal pneumonia caused by Bacillus cereus. These cases are interesting in that B. cereus, even from blood or sputum specimens, may often be considered a contaminant and receive inadequate attention. Also of interest was the fact that the two patients resided in the same area of the state, were welders by trade, and became ill within a few days of each other, yet there was no epidemiologic link between them.

  12. Fulminating bacteremia and pneumonia due to Bacillus cereus.

    OpenAIRE

    Miller, J M; Hair, J G; Hebert, M.; Hebert, L; Roberts, F. J.; Weyant, R S

    1997-01-01

    We present two cases of rapidly progressing, fatal pneumonia caused by Bacillus cereus. These cases are interesting in that B. cereus, even from blood or sputum specimens, may often be considered a contaminant and receive inadequate attention. Also of interest was the fact that the two patients resided in the same area of the state, were welders by trade, and became ill within a few days of each other, yet there was no epidemiologic link between them.

  13. Spatially resolved characterization of water and ion incorporation in Bacillus spores.

    Science.gov (United States)

    Ghosal, Sutapa; Leighton, Terrance J; Wheeler, Katherine E; Hutcheon, Ian D; Weber, Peter K

    2010-05-01

    We present the first direct visualization and quantification of water and ion uptake into the core of individual dormant Bacillus thuringiensis subsp. israelensis (B. thuringiensis subsp. israelensis) endospores. Isotopic and elemental gradients in the B. thuringiensis subsp. israelensis spores show the permeation and incorporation of deuterium in deuterated water (D(2)O) and solvated ions throughout individual spores, including the spore core. Under hydrated conditions, incorporation into a spore occurs on a time scale of minutes, with subsequent uptake of the permeating species continuing over a period of days. The distribution of available adsorption sites is shown to vary with the permeating species. Adsorption sites for Li(+), Cs(+), and Cl(-) are more abundant within the spore outer structures (exosporium, coat, and cortex) relative to the core, while F(-) adsorption sites are more abundant in the core. The results presented here demonstrate that elemental abundance and distribution in dormant spores are influenced by the ambient environment. As such, this study highlights the importance of understanding how microbial elemental and isotopic signatures can be altered postproduction, including during sample preparation for analysis, and therefore, this study is immediately relevant to the use of elemental and isotopic markers in environmental microbiology and microbial forensics.

  14. Bacterial spore germination and protein mobility.

    Science.gov (United States)

    Moir, Anne

    2003-10-01

    Fluorescence recovery after photobleaching (FRAP) of green fluorescent protein (GFP) has been used to report on protein mobility in single spores. Proteins found in dormant Bacillus spores are not mobile; however, mobility is restored when germination occurs and the core rehydrates. Spores of a cwlD mutant, in which the cortex is resistant to hydrolysis, are able to complete the earliest stages of germination in response to a specific germinant stimulus; in these circumstances, the protein in the spore remains immobile. Therefore, the earliest stages of spore germination, including loss of resistance to extreme heat and the complete release of the spore component dipicolinic acid, are achieved without the restoration of protein mobility.

  15. Reagent-free and portable detection of Bacillus anthracis spores using a microfluidic incubator and smartphone microscope.

    Science.gov (United States)

    Hutchison, Janine R; Erikson, Rebecca L; Sheen, Allison M; Ozanich, Richard M; Kelly, Ryan T

    2015-09-21

    Bacillus anthracis is the causative agent of anthrax and can be contracted by humans and herbivorous mammals by inhalation, ingestion, or cutaneous exposure to bacterial spores. Due to its stability and disease potential, B. anthracis is a recognized biothreat agent and robust detection and viability methods are needed to identify spores from unknown samples. Here we report the use of smartphone-based microscopy (SPM) in combination with a simple microfluidic incubation device (MID) to detect 50 to 5000 B. anthracis Sterne spores in 3 to 5 hours. This technique relies on optical monitoring of the conversion of the ∼1 μm spores to the filamentous vegetative cells that range from tens to hundreds of micrometers in length. This distinguishing filament formation is unique to B. anthracis as compared to other members of the Bacillus cereus group. A unique feature of this approach is that the sample integrity is maintained, and the vegetative biomass can be removed from the chip for secondary molecular analysis such as PCR. Compared with existing chip-based and rapid viability PCR methods, this new approach reduces assay time by almost half, and is highly sensitive, specific, and cost effective.

  16. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity.

    Directory of Open Access Journals (Sweden)

    Alicja K Warda

    Full Text Available We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with 11 additional B. cereus reference genomes to provide an overview of the different types of carbohydrate transporters and utilization systems found in B. cereus strains. The combined application of API tests, defined growth media experiments and comparative genomics enabled us to link the carbohydrate utilisation capacity of 22 B. cereus strains with their genome content and in some cases to the panC phylogenetic grouping. A core set of carbohydrates including glucose, fructose, maltose, trehalose, N-acetyl-glucosamine, and ribose could be used by all strains, whereas utilisation of other carbohydrates like xylose, galactose, and lactose, and typical host-derived carbohydrates such as fucose, mannose, N-acetyl-galactosamine and inositol is limited to a subset of strains. Finally, the roles of selected carbohydrate transporters and utilisation systems in specific niches such as soil, foods and the human host are discussed.

  17. Spore Cortex Hydrolysis Precedes Dipicolinic Acid Release during Clostridium difficile Spore Germination

    OpenAIRE

    2015-01-01

    Bacterial spore germination is a process whereby a dormant spore returns to active, vegetative growth, and this process has largely been studied in the model organism Bacillus subtilis. In B. subtilis, the initiation of germinant receptor-mediated spore germination is divided into two genetically separable stages. Stage I is characterized by the release of dipicolinic acid (DPA) from the spore core. Stage II is characterized by cortex degradation, and stage II is activated by the DPA released...

  18. Indole and 3-indolylacetonitrile inhibit spore maturation in Paenibacillus alvei

    Directory of Open Access Journals (Sweden)

    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.

  19. Anthrax Toxins in Context of Bacillus anthracis Spores and Spore Germination.

    Science.gov (United States)

    Cote, Christopher K; Welkos, Susan L

    2015-08-17

    The interaction of anthrax toxin or toxin components with B. anthracis spores has been demonstrated. Germinating spores can produce significant amounts of toxin components very soon after the initiation of germination. In this review, we will summarize the work performed that has led to our understanding of toxin and spore interactions and discuss the complexities associated with these interactions.

  20. 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.

  1. Detection of Bacillus anthracis Spores Using Peptide Functionalized SERS-Active Substrates

    Directory of Open Access Journals (Sweden)

    Atanu Sengupta

    2012-01-01

    Full Text Available The need for portable technologies that can rapidly identify biological warfare agents (BWAs in the field remains an international priority as expressed at the 2011 Biological Weapons Convention. In recent years, the ability of surface-enhanced Raman spectroscopy (SERS to rapidly detect various BWAs at very low concentrations has been demonstrated. However, in the specific case of Bacillus anthracis, differentiation at the species level is required since other bacilli are common in the environment, representing potential false-positive responses. To overcome this limitation, we describe the use of a peptide attached to the SERS-active metal that selectively binds Bacillus anthracis-Sterne as the target analyte. Using this approach, 109  B. anthracis-Sterne spores/mL produced an intense dipicolinic acid spectrum upon the addition of acetic acid, while the same concentration and treatment of B. cereus and B. subtilis did not.

  2. Reticulate spore ornamentation in Strobilomyces

    Institute of Scientific and Technical Information of China (English)

    Ronald H.Petersen; John Dunlap; Karen W.Hughes

    2011-01-01

    Reticulate spore ornamentation in Strobilomyces (Boletaceae,Basidiomycotina) is visible under light microscopy (bright field and phase contrast) up to 1,500×.While some distinctions can be made at this magnification,ontogeny and fine structure of the ornamentation cannot be discerned.Scanning electron microscope images,conversely,reveal significant additional structure from which the ontogenetic process can be traced.Citing numerous New and Old World collections,this paper presents evidence distinguishing reticulate ornamentation ontogeny in these disjunct populations.

  3. 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

    2015): << Inhibiting inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores: potential spore...inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores potential spore decontamination strategies 5a...EASIER, SAFER, and CHEAPER Inducing spore germination should make resulting bacteria much more susceptible to decontamination methods and will be

  4. Bacillus anthracis-like bacteria and other B. cereus group members in a microbial community within the International Space Station: a challenge for rapid and easy molecular detection of virulent B. anthracis.

    Directory of Open Access Journals (Sweden)

    Sandra P van Tongeren

    Full Text Available For some microbial species, such as Bacillus anthracis, the etiologic agent of the disease anthrax, correct detection and identification by molecular methods can be problematic. The detection of virulent B. anthracis is challenging due to multiple virulence markers that need to be present in order for B. anthracis to be virulent and its close relationship to Bacillus cereus and other members of the B. cereus group. This is especially the case in environments where build-up of Bacillus spores can occur and several representatives of the B. cereus group may be present, which increases the chance for false-positives. In this study we show the presence of B. anthracis-like bacteria and other members of the B. cereus group in a microbial community within the human environment of the International Space Station and their preliminary identification by using conventional culturing as well as molecular techniques including 16S rDNA sequencing, PCR and real-time PCR. Our study shows that when monitoring the microbial hygiene in a given human environment, health risk assessment is troublesome in the case of virulent B. anthracis, especially if this should be done with rapid, easy to apply and on-site molecular methods.

  5. Bacillus anthracis-like bacteria and other B. cereus group members in a microbial community within the International Space Station: a challenge for rapid and easy molecular detection of virulent B. anthracis.

    Science.gov (United States)

    van Tongeren, Sandra P; Roest, Hendrik I J; Degener, John E; Harmsen, Hermie J M

    2014-01-01

    For some microbial species, such as Bacillus anthracis, the etiologic agent of the disease anthrax, correct detection and identification by molecular methods can be problematic. The detection of virulent B. anthracis is challenging due to multiple virulence markers that need to be present in order for B. anthracis to be virulent and its close relationship to Bacillus cereus and other members of the B. cereus group. This is especially the case in environments where build-up of Bacillus spores can occur and several representatives of the B. cereus group may be present, which increases the chance for false-positives. In this study we show the presence of B. anthracis-like bacteria and other members of the B. cereus group in a microbial community within the human environment of the International Space Station and their preliminary identification by using conventional culturing as well as molecular techniques including 16S rDNA sequencing, PCR and real-time PCR. Our study shows that when monitoring the microbial hygiene in a given human environment, health risk assessment is troublesome in the case of virulent B. anthracis, especially if this should be done with rapid, easy to apply and on-site molecular methods.

  6. Uptake of and Resistance to the Antibiotic Berberine by Individual Dormant, Germinating and Outgrowing Bacillus Spores as Monitored by Laser Tweezers Raman Spectroscopy.

    Science.gov (United States)

    Wang, Shiwei; Yu, Jing; Suvira, Milomir; Setlow, Peter; Li, Yong-qing

    2015-01-01

    Berberine, an alkaloid originally extracted from the plant Coptis chinensis and other herb plants, has been used as a pharmacological substance for many years. The therapeutic effect of berberine has been attributed to its interaction with nucleic acids and blocking cell division. However, levels of berberine entering individual microbial cells minimal for growth inhibition and its effects on bacterial spores have not been determined. In this work the kinetics and levels of berberine accumulation by individual dormant and germinated spores were measured by laser tweezers Raman spectroscopy and differential interference and fluorescence microscopy, and effects of berberine on spore germination and outgrowth and spore and growing cell viability were determined. The major conclusions from this work are that: (1) colony formation from B. subtilis spores was blocked ~ 99% by 25 μg/mL berberine plus 20 μg/mL INF55 (a multidrug resistance pump inhibitor); (2) 200 μg/mL berberine had no effect on B. subtilis spore germination with L-valine, but spore outgrowth was completely blocked; (3) berberine levels accumulated in single spores germinating with ≥ 25 μg/mL berberine were > 10 mg/mL; (4) fluorescence microscopy showed that germinated spores accumulated high-levels of berberine primarily in the spore core, while dormant spores accumulated very low berberine levels primarily in spore coats; and (5) during germination, uptake of berberine began at the time of commitment (T1) and reached a maximum after the completion of CaDPA release (Trelease) and spore cortex lysis (Tlysis).

  7. Virulence of Bacillus cereus: a multivariate analysis.

    Science.gov (United States)

    Minnaard, J; Delfederico, L; Vasseur, V; Hollmann, A; Rolny, I; Semorile, L; Pérez, P F

    2007-05-10

    Biological activity and presence of DNA sequences related to virulence genes were studied in 21 strains of the Bacillus cereus group. The activity of spent culture supernatants and the effect of infection by vegetative bacterial cells were assessed on cultured human enterocytes (Caco-2 cells). The effect of extracellular factors on the detachment, necrosis and mitochondrial dehydrogenase activity of cultured human enterocytes was studied. Hemolytic activity on rabbit red blood cells was also evaluated and the effect of direct procaryotic-eucaryotic interactions was assessed in infection assays with vegetative bacterial cells. Concerning virulence genes, presence of the DNA sequences corresponding to the genes entS, entFM, nhe (A, B and C), sph, hbl (A, B, C and D), piplC and bceT was assessed by PCR. Ribopatterns were determined by an automated riboprinting analysis after digestion of the DNA with EcoRI. Principal component analysis and biplots were used to address the relationship between variables. Results showed a wide range of biological activities: decrease in mitochondrial dehydrogenase activity, necrosis, cell detachment and hemolytic activity. These effects were strain-dependent. Concerning the occurrence of the DNA sequences tested, different patterns were found. In addition, ribotyping showed that strains under study grouped into two main clusters. One of these clusters includes all the strains that were positive for all the DNA sequences tested. Positive and negative correlations between variables under study were evidenced. Interestingly, high detaching strains were positively correlated with the presence of the sequences entS, nheC and sph. Within gene complexes, high correlation was found between sequences of the hbl complex. In contrast, sequences of the nhe complex were not correlated. Some strains clustered together in the biplots. These strains were positive for all the DNA sequences tested and they were able to detach enterocytes upon infection

  8. The ice nucleation ability of one of the most abundant types of fungal spores found in the atmosphere

    Directory of Open Access Journals (Sweden)

    R. Iannone

    2011-02-01

    Full Text Available Recent atmospheric measurements show that biological particles are a potentially important class of ice nuclei. Types of biological particles that may be good ice nuclei include bacteria, pollen and fungal spores. We studied the ice nucleation properties of water droplets containing fungal spores from the genus Cladosporium, one of the most abundant types of spores found in the atmosphere. For water droplets containing a Cladosporium spore surface area of ~217 μm2 (equivalent to ~5 spores with average diameters of 3.2 μm , 1% of the droplets froze by −28.5 °C and 10% froze by –30.1 °C. However, there was a strong dependence on freezing temperature with the spore surface area of Cladosporium within a given droplet. Mean freezing temperatures for droplets containing 1–5 spores are expected to be approximately −35.1 ± 2.3 °C (1σ S. D.. Atmospheric ice nucleation on spores of Cladosporium sp., or other spores with similar surface properties, thus do not appear to explain recent atmospheric measurements showing that biological particles participate as atmospheric ice nuclei. The poor ice nucleation ability of Cladosporium sp. may be attributed to the surface which is coated with hydrophobins (a class of hydrophobic proteins that appear to be widespread in filamentous fungi. Given the ubiquity of hydrophobins on spore surfaces, the current study may be applicable to many fungal species of atmospheric importance.

  9. Use of Yeast Spores for Microencapsulation of Enzymes

    OpenAIRE

    2014-01-01

    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...

  10. Antimicrobial coatings — obtaining and characterization

    Indian Academy of Sciences (India)

    Cornelia Guran; Alexandra Pica; Denisa Ficai; Anton Ficai; Cezar Comanescu

    2013-04-01

    In this paper, we present inorganic–organic hybrid coatings with polymer matrix (water soluble) that contain silver nanoparticles (AgNPs). The structure and morphology of coating materials were determined by infrared spectroscopy (FT–IR) and scanning electron microscopy (SEM). Therefore, the antimicrobial activities and mechanisms of coatings for several pathogenic bacteria (Bacilius cereus and Staphylococcus aureus) were investigated. It was demonstrated that the obtained material with silver nanoparticles keep their antimicrobial effect even if they are subjected to several cycles of washing with water and detergent.

  11. What can spores do for us?

    NARCIS (Netherlands)

    Wolken, W.A.M.; Tramper, J.; Werf, M.J. van der

    2003-01-01

    Many organisms have the ability to form spores, a remarkable phase in their life cycles. Compared with vegetative cells, spores have several advantages (e.g. resistance to toxic compounds, temperature, desiccation and radiation) making them well suited to various applications. The applications of sp

  12. Invasive Bacillus cereus Infection in a Renal Transplant Patient: A Case Report and Review

    Directory of Open Access Journals (Sweden)

    Susan John

    2012-01-01

    Full Text Available Bacillus cereus is a common cause of gastrointestinal diseases. The majority of individuals with B cereus-related food poisoning recover without any specific treatment. It can, however, rarely cause invasive disease in immunocompromised patients.

  13. Detecting Cortex Fragments During Bacterial Spore Germination.

    Science.gov (United States)

    Francis, Michael B; Sorg, Joseph A

    2016-06-25

    The process of endospore germination in Clostridium difficile, and other Clostridia, increasingly is being found to differ from the model spore-forming bacterium, Bacillus subtilis. Germination is triggered by small molecule germinants and occurs without the need for macromolecular synthesis. Though differences exist between the mechanisms of spore germination in species of Bacillus and Clostridium, a common requirement is the hydrolysis of the peptidoglycan-like cortex which allows the spore core to swell and rehydrate. After rehydration, metabolism can begin and this, eventually, leads to outgrowth of a vegetative cell. The detection of hydrolyzed cortex fragments during spore germination can be difficult and the modifications to the previously described assays can be confusing or difficult to reproduce. Thus, based on our recent report using this assay, we detail a step-by-step protocol for the colorimetric detection of cortex fragments during bacterial spore germination.

  14. A combination of a SEM technique and X-ray microanalysis for studying the spore germination process of Clostridium tyrobutyricum.

    Science.gov (United States)

    Bassi, Daniela; Cappa, Fabrizio; Cocconcelli, Pier Sandro

    2009-06-01

    Clostridium tyrobutyricum is an anaerobic bacterium responsible for late blowing defects during cheese ripening and it is of scientific interest for biological hydrogen production. A scanning electron microscopy (SEM) coating technique and X-ray microanalysis were developed to analyze the architecture and chemical composition of spores upon germination in response to environmental changes. In addition, we investigated the effects of different compounds on this process. Agents and environmental conditions inducing germination were characterized monitoring changes in optical density (OD). Among all tested conditions, the greatest drop in OD(625) (57.4%) was obtained when spores were incubated in l-alanine/l-lactate buffer, pH 4.6. In addition, a carbon-coating SEM technique and X-ray microanalysis were used to observe the architecture of spores and to examine calcium dipicolinate release. Conditions inducing C. tyrobutyricum spore germination were identified and SEM X-ray microanalysis clearly distinguished germinating from dormant spores. We confirmed that calcium dipicolinate release is one of the first events occurring. These microscopy methods could be considered sensitive tools for evaluating morphological and chemical changes in spores of C. tyrobutyricum during the initial phase of germination. Information gathered from this work may provide new data for further research on germination.

  15. Bacillus cereus bacteremia and meningitis in immunocompromised children.

    Science.gov (United States)

    Gaur, A H; Patrick, C C; McCullers, J A; Flynn, P M; Pearson, T A; Razzouk, B I; Thompson, S J; Shenep, J L

    2001-05-15

    Two cases of Bacillus cereus meningitis in immunocompromised children at our hospital within a 2-month period prompted us to review B. cereus--related invasive disease. We identified 12 patients with B. cereus isolated in blood cultures from September 1988 through August 2000 at our institution. Three of these patients also had B. cereus isolated from CSF specimens; 1 additional patient had possible CNS involvement (33%, group A), whereas 8 patients had no evidence of CNS involvement (67%, group B). Patients in group A were more likely to have neutropenia at the onset of sepsis and were more likely to have an unfavorable outcome. They were also more likely to have received intrathecal chemotherapy in the week before the onset of their illness. Two patients from group A died. One survived with severe sequelae. The fourth patient had mild sequelae at follow-up. No sequelae or deaths occurred among patients in group B. In patients with unfavorable outcomes, the interval from the time of recognition of illness to irreversible damage or death was short, which demonstrates a need for increased awareness, early diagnosis, and more-effective therapy, particularly that which addresses B. cereus toxins.

  16. Bacillus cereus bacteraemia: comparison between haematologic and nonhaematologic patients

    Directory of Open Access Journals (Sweden)

    S. Tusgul

    2017-01-01

    Full Text Available Bacillus cereus bacteraemia can be severe, especially among patients with haematologic malignancy. We retrospectively reviewed first episodes of true B. cereus bacteraemia (more than one positive bottle plus signs of infection at our institution between 1997 and 2013 with the aim to compare haematologic versus nonhaematologic patients and analyse episodes with complicated outcome. Among 56 episodes of positive-blood cultures for B. cereus, 21 were considered significant. Median age was 54 years (range 23–82 years. Ten patients (48% had a haematologic malignancy; all were neutropenic at the time of B. cereus bacteraemia. Nonhaematologic patients were either intravenous drug users (n=3, 14%, polytraumatized (n=3, 14% or had multiple chronic comorbidities (n=5, 24%. Most episodes were hospital acquired (15, 71%. Sources of bacteraemia were intravascular catheter (n=11, 52%, digestive tract (n=6, 29%, drug injection (n=3, 14% and wound (n=1, 5%. Adequate antibiotic therapy was provided to 18 patients (86% during a median of 17 days (range 2–253 days. The intravascular catheter was removed in eight cases (42%. Three haematologic patients had a complicated course with neurologic complications (meningoencephalitis and cerebral abscesses. Complications appeared to be associated with catheter infection (100% of complicated cases vs. 29% of noncomplicated cases. In conclusion, B. cereus bacteraemia can have a complicated course in a subset of patients, mainly those with haematologic malignancy. Catheter infection may be associated with a worse outcome with frequent neurologic complications.

  17. Biocontrol of Aspergillus species on peanut kernels by antifungal diketopiperazine producing Bacillus cereus associated with entomopathogenic nematode.

    Directory of Open Access Journals (Sweden)

    Sasidharan Nishanth Kumar

    Full Text Available The rhabditid entomopathogenic nematode associated Bacillus cereus and the antifungal compounds produced by this bacterium were evaluated for their activity in reducing postharvest decay of peanut kernels caused by Aspergillus species in in vitro and in vivo tests. The results showed that B. cereus had a significant effect on biocontrol effectiveness in in vitro and in vivo conditions. The antifungal compounds produced by the B. cereus were purified using silica gel column chromatography and their structure was elucidated using extensive spectral analyses. The compounds were identified as diketopiperazines (DKPs [cyclo-(L-Pro-Gly, cyclo(L-Tyr-L-Tyr, cyclo-(L-Phe-Gly and cyclo(4-hydroxy-L-Pro-L-Trp]. The antifungal activities of diketopiperazines were studied against five Aspergillus species and best MIC of 2 µg/ml was recorded against A. flavus by cyclo(4-hydroxy-L-Pro-L-Trp. To investigate the potential application of cyclo(4-hydroxy-L-Pro-L-Trp to eliminate fungal spoilage in food and feed, peanut kernels was used as a food model system. White mycelia and dark/pale green spores of Aspergillus species were observed in the control peanut kernels after 2 days incubation. However the fungal growth was not observed in peanut kernels treated with cyclo(4-hydroxy-L-Pro-L-Trp. The cyclo(4-hydroxy-L-Pro-L-Trp was nontoxic to two normal cell lines [fore skin (FS normal fibroblast and African green monkey kidney (VERO] up to 200 µg/ml in MTT assay. Thus the cyclo(4-hydroxy-L-Pro-L-Trp identified in this study may be a promising alternative to chemical preservatives as a potential biopreservative agent which prevent fungal growth in food and feed. To the best of our knowledge, this is the first report demonstrating that the entomopathogenic nematode associated B. cereus and cyclo(4-hydroxy-L-Pro-L-Trp could be used as a biocontrol agents against postharvest fungal disease caused by Aspergillus species.

  18. Mapping of Proteomic Composition on the Surfaces of Bacillus spores by Atomic Force Microscopy-based Immunolabeling

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Malkin, A J

    2008-06-02

    Atomic force microscopy provides a unique capability to image high-resolution architecture and structural dynamics of pathogens (e.g. viruses, bacteria and bacterial spores) at near molecular resolution in native conditions. Further development of atomic force microscopy in order to enable the correlation of pathogen protein surface structures with specific gene products is essential to understand the mechanisms of the pathogen life cycle. We have applied an AFM-based immunolabeling technique for the proteomic mapping of macromolecular structures through the visualization of the binding of antibodies, conjugated with nanogold particles, to specific epitopes on Bacillus spore surfaces. This information is generated while simultaneously acquiring the surface morphology of the pathogen. The immunospecificity of this labeling method was established through the utilization of specific polyclonal and monoclonal antibodies that target spore coat and exosporium epitopes of Bacillus atrophaeus and Bacillus anthracis spores.

  19. [Incidence of Bacillus cereus in powdered dehydrated food].

    Science.gov (United States)

    Iacona, V A; Simonetta, A; Basílico, J C

    1987-01-01

    Bacillus cereus incidence on dehydrated powdered foods on sale in supermarkets of Santa Fe city was studied. Two hundred and fifty samples of five different foods: desserts, soups, mousses, pre-cooked "polenta" and mashed potatoes, were examined. Toxinogenic activity of strains confirmed as B. cereus by means of the test of lethality in rats, was analyzed. The ratio between contaminated samples and total analyzed samples was always greater than 6% (Table 1). Besides, none of the analyzed foods exceeded acceptability maximum limit (10(5) UFC/g), established by I.C.M.S.F. (Table 2). It was checked in all cases that no simple lineal correlation existed between B. cereus and total aerobic bacteria enumerations. On the other hand, the percentage of strains with lethal effects was in all cases greater than 42.8% (Table 3).

  20. The Bacillus cereus spoIIS programmed cell death system

    Directory of Open Access Journals (Sweden)

    Jana eMelnicakova

    2015-08-01

    Full Text Available Programmed cell death in bacteria is generally associated with two¬ component toxin antitoxin systems. The SpoIIS toxin-antitoxin system, consisting of a membrane bound SpoIISA toxin and a small, cytosolic antitoxin SpoIISB, was originally identified in Bacillus subtilis. In this work we describe the Bacillus cereus SpoIIS system which is a three-component system, harbouring an additional gene spoIISC. Its protein product serves as an antitoxin, and similarly as SpoIISB, is able to bind SpoIISA and abolish its toxic effect. Our results indicate that SpoIISC seems to be present not only in B. cereus but also in other Bacilli containing a SpoIIS toxin antitoxin system. In addition, we show that B. cereus SpoIISA can form higher oligomers and we discuss the possible role of this multimerization for the protein’s toxic function.

  1. Genotyping of Bacillus cereus strains by microarray-based resequencing.

    Directory of Open Access Journals (Sweden)

    Michael E Zwick

    Full Text Available The ability to distinguish microbial pathogens from closely related but nonpathogenic strains is key to understanding the population biology of these organisms. In this regard, Bacillus anthracis, the bacterium that causes inhalational anthrax, is of interest because it is closely related and often difficult to distinguish from other members of the B. cereus group that can cause diverse diseases. We employed custom-designed resequencing arrays (RAs based on the genome sequence of Bacillus anthracis to generate 422 kb of genomic sequence from a panel of 41 Bacillus cereus sensu lato strains. Here we show that RAs represent a "one reaction" genotyping technology with the ability to discriminate between highly similar B. anthracis isolates and more divergent strains of the B. cereus s.l. Clade 1. Our data show that RAs can be an efficient genotyping technology for pre-screening the genetic diversity of large strain collections to selected the best candidates for whole genome sequencing.

  2. Genotyping of Bacillus cereus strains by microarray-based resequencing.

    Science.gov (United States)

    Zwick, Michael E; Kiley, Maureen P; Stewart, Andrew C; Mateczun, Alfred; Read, Timothy D

    2008-07-02

    The ability to distinguish microbial pathogens from closely related but nonpathogenic strains is key to understanding the population biology of these organisms. In this regard, Bacillus anthracis, the bacterium that causes inhalational anthrax, is of interest because it is closely related and often difficult to distinguish from other members of the B. cereus group that can cause diverse diseases. We employed custom-designed resequencing arrays (RAs) based on the genome sequence of Bacillus anthracis to generate 422 kb of genomic sequence from a panel of 41 Bacillus cereus sensu lato strains. Here we show that RAs represent a "one reaction" genotyping technology with the ability to discriminate between highly similar B. anthracis isolates and more divergent strains of the B. cereus s.l. Clade 1. Our data show that RAs can be an efficient genotyping technology for pre-screening the genetic diversity of large strain collections to selected the best candidates for whole genome sequencing.

  3. Food–bacteria interplay: pathometabolism of emetic Bacillus cereus

    Science.gov (United States)

    Ehling-Schulz, Monika; Frenzel, Elrike; Gohar, Michel

    2015-01-01

    Bacillus cereus is a Gram-positive endospore forming bacterium known for its wide spectrum of phenotypic traits, enabling it to occupy diverse ecological niches. Although the population structure of B. cereus is highly dynamic and rather panmictic, production of the emetic B. cereus toxin cereulide is restricted to strains with specific genotypic traits, associated with distinct environmental habitats. Cereulide is an ionophoric dodecadepsipeptide that is produced non-ribosomally by an enzyme complex with an unusual modular structure, named cereulide synthetase (Ces non-ribosomal peptide synthetase). The ces gene locus is encoded on a mega virulence plasmid related to the B. anthracis toxin plasmid pXO1. Cereulide, a highly thermo- and pH- resistant molecule, is preformed in food, evokes vomiting a few hours after ingestion, and was shown to be the direct cause of gastroenteritis symptoms; occasionally it is implicated in severe clinical manifestations including acute liver failures. Control of toxin gene expression in emetic B. cereus involves central transcriptional regulators, such as CodY and AbrB, thereby inextricably linking toxin gene expression to life cycle phases and specific conditions, such as the nutrient supply encountered in food matrices. While in recent years considerable progress has been made in the molecular and biochemical characterization of cereulide toxin synthesis, far less is known about the embedment of toxin synthesis in the life cycle of B. cereus. Information about signals acting on toxin production in the food environment is lacking. We summarize the data available on the complex regulatory network controlling cereulide toxin synthesis, discuss the role of intrinsic and extrinsic factors acting on toxin biosynthesis in emetic B. cereus and stress how unraveling these processes can lead to the development of novel effective strategies to prevent toxin synthesis in the food production and processing chain. PMID:26236290

  4. Food-bacteria interplay: pathometabolism of emetic Bacillus cereus.

    Science.gov (United States)

    Ehling-Schulz, Monika; Frenzel, Elrike; Gohar, Michel

    2015-01-01

    Bacillus cereus is a Gram-positive endospore forming bacterium known for its wide spectrum of phenotypic traits, enabling it to occupy diverse ecological niches. Although the population structure of B. cereus is highly dynamic and rather panmictic, production of the emetic B. cereus toxin cereulide is restricted to strains with specific genotypic traits, associated with distinct environmental habitats. Cereulide is an ionophoric dodecadepsipeptide that is produced non-ribosomally by an enzyme complex with an unusual modular structure, named cereulide synthetase (Ces non-ribosomal peptide synthetase). The ces gene locus is encoded on a mega virulence plasmid related to the B. anthracis toxin plasmid pXO1. Cereulide, a highly thermo- and pH- resistant molecule, is preformed in food, evokes vomiting a few hours after ingestion, and was shown to be the direct cause of gastroenteritis symptoms; occasionally it is implicated in severe clinical manifestations including acute liver failures. Control of toxin gene expression in emetic B. cereus involves central transcriptional regulators, such as CodY and AbrB, thereby inextricably linking toxin gene expression to life cycle phases and specific conditions, such as the nutrient supply encountered in food matrices. While in recent years considerable progress has been made in the molecular and biochemical characterization of cereulide toxin synthesis, far less is known about the embedment of toxin synthesis in the life cycle of B. cereus. Information about signals acting on toxin production in the food environment is lacking. We summarize the data available on the complex regulatory network controlling cereulide toxin synthesis, discuss the role of intrinsic and extrinsic factors acting on toxin biosynthesis in emetic B. cereus and stress how unraveling these processes can lead to the development of novel effective strategies to prevent toxin synthesis in the food production and processing chain.

  5. Food – bacteria interplay: Pathometabolism of emetic Bacillus cereus

    Directory of Open Access Journals (Sweden)

    Monika eEhling-Schulz

    2015-07-01

    Full Text Available Bacillus cereus is a gram-positive endospore forming bacterium known for its wide spectrum of phenotypic traits, enabling it to occupy diverse ecological niches. Although the population structure of B. cereus is highly dynamic and rather panmictic, production of the emetic B. cereus toxin cereulide is restricted to strains with specific genotypic traits, associated with distinct environmental habitats. Cereulide is an ionophoric dodecadepsipeptide that is produced non-ribosomally by an enzyme complex with an unusual modular structure, named cereulide synthetase (Ces NRPS. The ces gene locus is encoded on a mega virulence plasmid related to the Bacillus anthracis toxin plasmid pXO1. Cereulide, a highly thermo- and pH- resistant molecule, is preformed in food, evokes vomiting a few hours after ingestion and was shown to be the direct cause of gastroenteritis symptoms; occasionally it is implicated in severe clinical manifestations including acute liver failures. Control of toxin gene expression in emetic Bacillus cereus involves central transcriptional regulators, such as CodY and AbrB, thereby inextricably linking toxin gene expression to life cycle phases and specific conditions, such as the nutrient supply encountered in food matrices. While in recent years considerable progress has been made in the molecular and biochemical characterization of cereulide toxin synthesis, far less is known about the embedment of toxin synthesis in the life cycle of B. cereus. Information about signals acting on toxin production in the food environment is literally lacking. We summarize the data available on the complex regulatory network controlling cereulide toxin synthesis, discuss the role of intrinsic and extrinsic factors acting on toxin biosynthesis in emetic B. cereus and stress how unraveling these processes can lead to the development of novel effective strategies to prevent toxin synthesis in the food production and processing chain.

  6. Bacillus cereus bacteremia in an adult with acute leukemia.

    Science.gov (United States)

    Funada, H; Uotani, C; Machi, T; Matsuda, T; Nonomura, A

    1988-03-01

    Bacillus cereus, which used to be considered non-pathogenic, was isolated from the blood of a patient with acute leukemia who was receiving intensive chemotherapy. Fatal bacteremia developed with a clinical syndrome of acute gastroenteritis, followed by both meningoencephalitis with subarachnoid hemorrhage and multiple liver abscesses probably caused by infective vasculitis. Surveillance stool cultures revealed colonization with the organism prior to the onset of diarrhea, and repetitive blood cultures were found to be positive. Thus, this case suggested some new important clinicopathologic features of true B. cereus bacteremia complicating acute leukemia.

  7. [Can industrial laundry remove Bacillus cereus from hospital linen?].

    Science.gov (United States)

    Yoh, Myonsun; Matsuyama, Junko; Shime, Akiko; Okayama, Kana; Sakamoto, Rei; Honda, Takeshi

    2010-09-01

    Contaminated hospital linen has caused some cases of Bacillus cereus bacteremia in Japan. We analyzed the disinfection efficacy of industrial washing of hospital towels and sheets by counting the number of B. cereus on linen before and after washing. That before washing averaged 7.6 cells/cm2 on unwashed sheets, decreasing to 1.2 cells/cm2 after washing. That on unwashed towels, however, averaged 10(6) cells/cm2 before washing and 1096 cells/cm2 after washing, which was very high and suggested the possibility of causing nosocomial infection.

  8. Nanomechanical analysis of Clostridium tyrobutyricum spores.

    Science.gov (United States)

    Andreeva, N; Bassi, D; Cappa, F; Cocconcelli, P S; Parmigiani, F; Ferrini, G

    2010-12-01

    In this work we report on the measurement of the Young modulus of the external surface of Clostridium tyrobutyricum spores in air with an atomic force microscope. The Young modulus can be reliably measured despite the strong tip-spore adhesion forces and the need to immobilize the spores due to their slipping on most substrates. Moreover, we investigate the disturbing factors and consider some practical aspects that influence the measurements of elastic properties of biological objects with the atomic force microscopy indentation techniques.

  9. Quantitative immunofluorescence studies of the serology of Bacillus anthracis spores.

    OpenAIRE

    1983-01-01

    A fluorescein-conjugated antibody against formalin-inactivated spores of Bacillus anthracis Vollum reacted only weakly with a variety of Bacillus species in microfluorometric immunofluorescence assays. A conjugated antibody against spores of B. anthracis Sterne showed little affinity for spores of several B. anthracis isolates including B. anthracis Vollum, indicating that more than one anthrax spore serotype exists.

  10. Seasonal trend and clinical presentation of Bacillus cereus bloodstream infection: association with summer and indwelling catheter.

    Science.gov (United States)

    Kato, K; Matsumura, Y; Yamamoto, M; Nagao, M; Ito, Y; Takakura, S; Ichiyama, S

    2014-08-01

    Bacillus cereus, an opportunistic pathogen, can cause fatal infection. However, B. cereus bloodstream infections (BSIs) have not been well characterised. From 2008 to 2013, B. cereus isolates from all of the specimens and patients with B. cereus BSIs were identified. Environmental samples were collected to detect B. cereus contamination. We also characterised the clinical presentation of B. cereus BSI through analyses of risk factors for BSI and mortality. A total of 217 clinical B. cereus isolates was detected. Fifty-one patients with nosocomial infections were diagnosed as B. cereus BSI, and 37 had contaminated blood cultures. The number of B. cereus isolates and BSI patients was significantly greater from June to September than from January to April (4.9 vs. 1.5 per month and 1.2 vs. 0.2, respectively). All BSIs were nosocomial and related to central or peripheral vascular catheter. Urinary catheter [odds ratio (OR) 6.93, 95% confidence interval (CI) 2.40-20.0] was the independent risk factor associated with BSI patients when compared to patients regarded as contaminated. In-hospital mortality among BSI patients was 20% and was associated with urinary catheter (OR 34.7, 95 % CI 1.89-63.6) and higher Charlson index (OR 1.99, 95 % CI 1.26-3.12). The number of B. cereus isolates and BSI increased during summer. Inpatients with indwelling vascular or urinary catheters should be carefully monitored for potential B. cereus BSIs.

  11. Rapid onsite assessment of spore viability.

    Energy Technology Data Exchange (ETDEWEB)

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

    2005-12-01

    This one year LDRD addresses problems of threat assessment and restoration of facilities following a bioterror incident like the incident that closed down mail facilities in late 2001. Facilities that are contaminated with pathogenic spores such as B. anthracis spores must be shut down while they are treated with a sporicidal agent and the effectiveness of the treatment is ascertained. This process involves measuring the viability of spore test strips, laid out in a grid throughout the facility; the CDC accepted methodologies require transporting the samples to a laboratory and carrying out a 48 hr outgrowth experiment. We proposed developing a technique that will ultimately lead to a fieldable microfluidic device that can rapidly assess (ideally less than 30 min) spore viability and effectiveness of sporicidal treatment, returning facilities to use in hours not days. The proposed method will determine viability of spores by detecting early protein synthesis after chemical germination. During this year, we established the feasibility of this approach and gathered preliminary results that should fuel a future more comprehensive effort. Such a proposal is currently under review with the NIH. Proteomic signatures of Bacillus spores and vegetative cells were assessed by both slab gel electrophoresis as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection. The conditions for germination using a number of chemical germinants were evaluated and optimized and the time course of protein synthesis was ascertained. Microseparations were carried out using both viable spores and spores inactivated by two different methods. A select number of the early synthesis proteins were digested into peptides for analysis by mass spectrometry.

  12. Structural Analysis of Bacillus subtilis Spore Peptidoglycan During Sporulation

    OpenAIRE

    2000-01-01

    Structural analysis of Bacillus subtilis spore peptidoglycan during sporulation:Jennifer L. Meador-Parton:David L. Popham, Chairman:Department of Biology:(ABSTRACT):Bacterial spore peptidoglycan (PG) is very loosely cross-linked relative to vegetative PG. Theories suggest that loosely cross-linked spore PG may have a flexibility which contributes to the attainment of spore core dehydration. The structure of the PG found in fully dormant spores has previously been examined in wild type and m...

  13. Nanomechanical Characterization of Bacillus anthracis Spores by Atomic Force Microscopy

    OpenAIRE

    2016-01-01

    The study of structures and properties of bacterial spores is important to understanding spore formation and biological responses to environmental stresses. While significant progress has been made over the years in elucidating the multilayer architecture of spores, the mechanical properties of the spore interior are not known. Here, we present a thermal atomic force microscopy (AFM) study of the nanomechanical properties of internal structures of Bacillus anthracis spores. We developed a nan...

  14. Intractable Bacillus cereus bacteremia in a preterm neonate.

    Science.gov (United States)

    John, Anna B; Razak, Eissa A S A; Razak, Emad E M H; Al-Naqeeb, Niran; Dhar, Rita

    2007-04-01

    Although often regarded as a contaminant, Bacillus spp. have been implicated in serious systemic infections. The incidence of such infections is low with only a few cases reported in the literature. We describe the clinical course of early-onset Bacillus cereus bacteremia in a preterm neonate who was successfully treated with vancomycin.

  15. Characterization of germination receptors of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Hornstra, L.M.; Vries, de Y.P.; Wells-Bennik, M.H.J.; Vos, de W.M.; Abee, T.

    2006-01-01

    Specific amino acids, purine ribonucleosides, or a combination of the two is required for efficient germination of endospores of Bacillus cereus ATCC 14579. A survey including 20 different amino acids showed that L-alanine, L-cysteine, L-threonine, and L-glutamine are capable of initiating the germi

  16. Rapid detoxification of cereulide in Bacillus cereus food poisoning.

    Science.gov (United States)

    Shiota, Mitsutaka; Saitou, Keiko; Mizumoto, Hiroshi; Matsusaka, Masanori; Agata, Norio; Nakayama, Masahiro; Kage, Masayoshi; Tatsumi, Shinji; Okamoto, Akira; Yamaguchi, Seiji; Ohta, Michio; Hata, Daisuke

    2010-04-01

    Bacillus cereus is recognized as a major pathogenic bacterium that causes food poisoning and produces gastrointestinal diseases of 2 types: emetic and diarrheal. The emetic type, which is often linked to pasta and rice, arises from a preformed toxin, cereulide, in food. Rapid and accurate diagnostic methods for this emetic toxin are important but are limited. Here we describe 3 patients with B cereus food poisoning in which cereulide was detected and measured sequentially. Three family members began to vomit frequently 30 minutes after consuming reheated fried rice. After 6 hours, a 1-year-old brother died of acute encephalopathy. A 2-year-old sister who presented with unconsciousness recovered rapidly after plasma exchange and subsequent hemodialysis. Their mother recovered soon by fluid therapy. From leftover fried rice and the children's stomach contents, B cereus was isolated. Serum cereulide was detected in both children; it decreased to an undetected level in the sister. These cases highlight the importance of measuring the value of cereulide, which would reflect the severity of B cereus emetic food poisoning. The cases also suggest the possible role of blood-purification therapy in severe cases.

  17. Successful treatment of Bacillus cereus infection with ciprofloxacin.

    OpenAIRE

    Gascoigne, A.D.; Richards, J.; Gould, K.; Gibson, G. J.

    1991-01-01

    Bacillus cereus is rarely a pulmonary pathogen but may cause pneumonia in immunocompromised patients. A patient with bronchiectasis and no recognisable immunodeficiency had this organism isolated during two infective exacerbations, once from respiratory secretions and once by blood culture. Ciprofloxacin treatment was effective on both occasions.

  18. The fate of Bacillus cereus in the gastrointestinal tract

    NARCIS (Netherlands)

    Pielaat A; Wijnands LM; Takumi K; Nauta MJ; Leusden FM van; MGB

    2006-01-01

    This report presents a mathematical dynamical model for the behaviour of Bacillus cereus in the gastro-intestinal tract. Biological processes and system dynamics are simultaneously incorporated in this mechanistic model. Variability in growth characteristics and physical traits of different B. cereu

  19. Induction of natural competence in Bacillus cereus ATCC14579

    NARCIS (Netherlands)

    Mironczuk, Aleksandra M.; Kovács, Ákos T.; Kuipers, O.P.

    2008-01-01

    Natural competence is the ability of certain microbes to take up exogenous DNA from the environment and integrate it in their genome. Competence development has been described for a variety of bacteria, but has so far not been shown to occur in Bacillus cereus. However, orthologues of most proteins

  20. Bacillus cereus bacteraemia: comparison between haematologic and nonhaematologic patients.

    Science.gov (United States)

    Tusgul, S; Prod'hom, G; Senn, L; Meuli, R; Bochud, P-Y; Giulieri, S G

    2017-01-01

    Bacillus cereus bacteraemia can be severe, especially among patients with haematologic malignancy. We retrospectively reviewed first episodes of true B. cereus bacteraemia (more than one positive bottle plus signs of infection) at our institution between 1997 and 2013 with the aim to compare haematologic versus nonhaematologic patients and analyse episodes with complicated outcome. Among 56 episodes of positive-blood cultures for B. cereus, 21 were considered significant. Median age was 54 years (range 23-82 years). Ten patients (48%) had a haematologic malignancy; all were neutropenic at the time of B. cereus bacteraemia. Nonhaematologic patients were either intravenous drug users (n = 3, 14%), polytraumatized (n = 3, 14%) or had multiple chronic comorbidities (n = 5, 24%). Most episodes were hospital acquired (15, 71%). Sources of bacteraemia were intravascular catheter (n = 11, 52%), digestive tract (n = 6, 29%), drug injection (n = 3, 14%) and wound (n = 1, 5%). Adequate antibiotic therapy was provided to 18 patients (86%) during a median of 17 days (range 2-253 days). The intravascular catheter was removed in eight cases (42%). Three haematologic patients had a complicated course with neurologic complications (meningoencephalitis and cerebral abscesses). Complications appeared to be associated with catheter infection (100% of complicated cases vs. 29% of noncomplicated cases). In conclusion, B. cereus bacteraemia can have a complicated course in a subset of patients, mainly those with haematologic malignancy. Catheter infection may be associated with a worse outcome with frequent neurologic complications.

  1. Evaluating Composite Sampling Methods of Bacillus spores at Low Concentrations

    Energy Technology Data Exchange (ETDEWEB)

    Hess, Becky M.; Amidan, Brett G.; Anderson, Kevin K.; Hutchison, Janine R.

    2016-10-13

    Restoring facility operations after the 2001 Amerithrax attacks took over three months to complete, highlighting the need to reduce remediation time. The most time intensive tasks were environmental sampling and sample analyses. Composite sampling allows disparate samples to be combined, with only a single analysis needed, making it a promising method to reduce response times. We developed a statistical experimental design to test three different composite sampling methods: 1) single medium single pass composite: a single cellulose sponge samples multiple coupons; 2) single medium multi-pass composite: a single cellulose sponge is used to sample multiple coupons; and 3) multi-medium post-sample composite: a single cellulose sponge samples a single surface, and then multiple sponges are combined during sample extraction. Five spore concentrations of Bacillus atrophaeus Nakamura spores were tested; concentrations ranged from 5 to 100 CFU/coupon (0.00775 to 0.155CFU/cm2, respectively). Study variables included four clean surface materials (stainless steel, vinyl tile, ceramic tile, and painted wallboard) and three grime coated/dirty materials (stainless steel, vinyl tile, and ceramic tile). Analysis of variance for the clean study showed two significant factors: composite method (p-value < 0.0001) and coupon material (p-value = 0.0008). Recovery efficiency (RE) was higher overall using the post-sample composite (PSC) method compared to single medium composite from both clean and grime coated materials. RE with the PSC method for concentrations tested (10 to 100 CFU/coupon) was similar for ceramic tile, painted wall board, and stainless steel for clean materials. RE was lowest for vinyl tile with both composite methods. Statistical tests for the dirty study showed RE was significantly higher for vinyl and stainless steel materials, but significantly lower for ceramic tile. These results suggest post-sample compositing can be used to reduce sample analysis time when

  2. Analysis of bacterial spore permeability to water and ions using Nano-Secondary Ion Mass Spectrometry (NanoSIMS)

    Energy Technology Data Exchange (ETDEWEB)

    Ghosal, S; Fallon, S; Leighton, T; Wheeler, K; Hutcheon, I; Weber, P K

    2005-11-17

    Regulation of bacterial spore solvent and solute permeability is a fundamental feature of dormancy but is poorly understood. Here we present a new technique, nano-scale secondary ion mass spectrometry (NanoSIMS) that allows the direct visualization and quantification of chemical gradients within spores. Using NanoSIMS, we demonstrate the penetration of water and a simple ionic salt, LiF, into the core of Bacillus thuringiensis israelensis (Bti) spores. The results demonstrate chemical gradients spanning the outer coat to the inner spore core that are driven by concentration-dependent ionic fluxes. Using deuterated water (D{sub 2}O), we have shown that external water is either retained or exchanged with water contained within the spore. Hydration and exchange are rapid, on a timescale of < 1 minute. Our results suggest a permeation mechanism by which short-time scale diffusion into and out of the spore can occur along hydration pathways. Additional studies are in progress to define the flux rates and mechanisms controlling these processes.

  3. Early quantitative method for measuring germination in non-green spores of Dryopteris paleacea using an epifluorescence-microscope technique

    Science.gov (United States)

    Scheuerlein, R.; Wayne, R.; Roux, S. J.

    1988-01-01

    A method is described to determine germination by blue-light excited red fluorescence in the positively photoblastic spores of Dryopteris paleacea Sw. This fluorescence is due to chlorophyll as evidenced from 1) a fluorescence-emission spectrum in vivo, where a bright fluorescence around 675 nm is obtained only in red light (R)-irradiated spores and 2) in vitro measurements with acetone extracts prepared from homogenized spores. Significant amounts of chlorophyll can be found only in R-treated spores; this chlorophyll exhibits an emission band around 668 nm, when irradiated with 430 nm light at 21 degrees C. Compared to other criteria for germination, such as swelling of the cell, coat splitting, greening, and rhizoid formation, which require longer periods after induction for their expression, chlorophyll fluorescence can be used to quantify germination after two days. This result is confirmed by fluence-response curves for R-induced spore germination; the same relationship between applied R and germination is obtained by the evaluation with the epifluorescence method 2 days after the light treatment as compared with the evaluation with bright-field microscopy 5 days after the inducing R. Using this technique we show for the first time that Ca2+ contributes to the signal-transduction chain in phytochrome-mediated chlorophyll synthesis in spores of Dryopteris paleacea.

  4. Discrimination between bacterial spore types using time-of-flight mass spectrometry and matrix-free infrared laser desorption and ionization.

    Science.gov (United States)

    Ullom, J N; Frank, M; Gard, E E; Horn, J M; Labov, S E; Langry, K; Magnotta, F; Stanion, K A; Hack, C A; Benner, W H

    2001-05-15

    We demonstrate that molecular ions with mass-to-charge ratios (m/z) ranging from a few hundred to 19 050 can be desorbed from whole bacterial spores using infrared laser desorption and no chemical matrix. We have measured the mass of these ions using time-of-flight mass spectrometry and we observe that different ions are desorbed from spores of Bacillus cereus, Bacillus thuringiensis, Bacillus subtilis, and Bacillus niger. Our results raise the possibility of identifying microorganisms using mass spectrometry without conventional sample preparation techniques such as the addition of a matrix. We have measured the dependence of the ion yield from B. subtilis on desorption wavelength over the range 3.05-3.8 microm, and we observe the best results at 3.05 microm. We have also generated mass spectra from whole spores using 337-nm ultraviolet laser desorption, and we find that these spectra are inferior to spectra generated with infrared desorption. Since aerosol analysis is a natural application for matrix-free desorption, we have measured mass spectra from materials such as ragweed pollen and road dust that are likely to form a background to microbial aerosols. We find that these materials are readily differentiated from bacterial spores.

  5. Bacterial spores in food : how phenotypic variability complicates prediction of spore properties and bacterial behavior

    NARCIS (Netherlands)

    Eijlander, Robyn T.; Abee, Tjakko; Kuipers, Oscar P.

    2011-01-01

    Bacillus spores are a known cause of food spoilage and their increased resistance poses a major challenge in efficient elimination. Recent studies on bacterial cultures at the single cell level have revealed how minor differences in essential spore properties, such as core water content or germinant

  6. Bacterial spores in food: how phenotypic variability complicates prediction of spore properties and bacterial behavior

    NARCIS (Netherlands)

    Eijlander, R.T.; Abee, T.; Kuipers, O.P.

    2011-01-01

    Bacillus spores are a known cause of food spoilage and their increased resistance poses a major challenge in efficient elimination. Recent studies on bacterial cultures at the single cell level have revealed how minor differences in essential spore properties, such as core water content or germinant

  7. Sensitive, Rapid Detection of Bacterial Spores

    Science.gov (United States)

    Kern, Roger G.; Venkateswaran, Kasthuri; Chen, Fei; Pickett, Molly; Matsuyama, Asahi

    2009-01-01

    A method of sensitive detection of bacterial spores within delays of no more than a few hours has been developed to provide an alternative to a prior three-day NASA standard culture-based assay. A capability for relatively rapid detection of bacterial spores would be beneficial for many endeavors, a few examples being agriculture, medicine, public health, defense against biowarfare, water supply, sanitation, hygiene, and the food-packaging and medical-equipment industries. The method involves the use of a commercial rapid microbial detection system (RMDS) that utilizes a combination of membrane filtration, adenosine triphosphate (ATP) bioluminescence chemistry, and analysis of luminescence images detected by a charge-coupled-device camera. This RMDS has been demonstrated to be highly sensitive in enumerating microbes (it can detect as little as one colony-forming unit per sample) and has been found to yield data in excellent correlation with those of culture-based methods. What makes the present method necessary is that the specific RMDS and the original protocols for its use are not designed for discriminating between bacterial spores and other microbes. In this method, a heat-shock procedure is added prior to an incubation procedure that is specified in the original RMDS protocols. In this heat-shock procedure (which was also described in a prior NASA Tech Briefs article on enumerating sporeforming bacteria), a sample is exposed to a temperature of 80 C for 15 minutes. Spores can survive the heat shock, but nonspore- forming bacteria and spore-forming bacteria that are not in spore form cannot survive. Therefore, any colonies that grow during incubation after the heat shock are deemed to have originated as spores.

  8. Ultrastructure of spore development in Scutellospora heterogama.

    Science.gov (United States)

    Jeffries, Peter; Robinson-Boyer, Louisa; Rice, Paul; Newsam, Ray J; Dodd, John C

    2007-07-01

    The ultrastructural detail of spore development in Scutellospora heterogama is described. Although the main ontogenetic events are similar to those described from light microscopy, the complexity of wall layering is greater when examined at an ultrastructural level. The basic concept of a rigid spore wall enclosing two inner, flexible walls still holds true, but there are additional zones within these three walls distinguishable using electron microscopy, including an inner layer that is involved in the formation of the germination shield. The spore wall has three layers rather than the two reported previously. An outer, thin ornamented layer and an inner, thicker layer are both derived from the hyphal wall and present at all stages of development. These layers differentiate into the outer spore layer visible at the light microscope level. A third inner layer unique to the spore develops during spore swelling and rapidly expands before contracting back to form the second wall layer visible by light microscopy. The two inner flexible walls also are more complex than light microscopy suggests. The close association with the inner flexible walls with germination shield formation consolidates the preferred use of the term 'germinal walls' for these structures. A thin electron-dense layer separates the two germinal walls and is the region in which the germination shield forms. The inner germinal wall develops at least two sub-layers, one of which has an appearance similar to that of the expanding layer of the outer spore wall. An electron-dense layer is formed on the inner surface of the inner germinal wall as the germination shield develops, and this forms the wall surrounding the germination shield as well as the germination tube. At maturity, the outer germinal wall develops a thin, striate layer within its substructure.

  9. A novel and highly specific phage endolysin cell wall binding domain for detection of Bacillus cereus.

    Science.gov (United States)

    Kong, Minsuk; Sim, Jieun; Kang, Taejoon; Nguyen, Hoang Hiep; Park, Hyun Kyu; Chung, Bong Hyun; Ryu, Sangryeol

    2015-09-01

    Rapid, specific and sensitive detection of pathogenic bacteria is crucial for public health and safety. Bacillus cereus is harmful as it causes foodborne illness and a number of systemic and local infections. We report a novel phage endolysin cell wall-binding domain (CBD) for B. cereus and the development of a highly specific and sensitive surface plasmon resonance (SPR)-based B. cereus detection method using the CBD. The newly discovered CBD from endolysin of PBC1, a B. cereus-specific bacteriophage, provides high specificity and binding capacity to B. cereus. By using the CBD-modified SPR chips, B. cereus can be detected at the range of 10(5)-10(8) CFU/ml. More importantly, the detection limit can be improved to 10(2) CFU/ml by using a subtractive inhibition assay based on the pre-incubation of B. cereus and CBDs, removal of CBD-bound B. cereus, and SPR detection of the unbound CBDs. The present study suggests that the small and genetically engineered CBDs can be promising biological probes for B. cereus. We anticipate that the CBD-based SPR-sensing methods will be useful for the sensitive, selective, and rapid detection of B. cereus.

  10. 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.

  11. Farm level survey of spore-forming bacteria on four dairy farms in the Waikato region of New Zealand.

    Science.gov (United States)

    Gupta, Tanushree B; Brightwell, Gale

    2017-03-03

    The aim of our study was to determine the occurrence and diversity of economically important spore-forming bacteria in New Zealand dairy farm systems. Farm dairy effluent (FDE) collected from Waikato dairy farms were tested for the presence of spore-forming bacteria, using a new culture-based methodology followed by genomic analysis. An enrichment step in which samples were inoculated in cooked meat glucose starch broth under anaerobic conditions, aided in the differential isolation of Bacillus and Clostridium species. Furthermore, the use of molecular methods such as ERIC genotyping, 16S rRNA gene sequence analysis identified different spore-forming bacteria present in FDE. C. sporogenes signature PCR gave further information on the phylogenetic relationship of the different Clostridium spp. isolated in this study. In total 19 Bacillus spp., 5 Paenibacillus spp. and 17 Clostridium spp. were isolated from farm dairy effluent. Sequence types similar to economically important food spoilage bacteria viz: C. butyricum, C. sporogenes and members of the Paenibacillus Genus were isolated from all four farms, whereas, sequence types similar to potential toxigenic, B. cereus, C. perfringens, C. butyricum, and C. botulinum were found on at least three of the farms. Sampling of farm dairy effluent provides a good indicator of farm level prevalence of bacterial load as it is used to irrigate dairy pasture in New Zealand. This study highlights the presence of various spore-forming bacteria in dairy waste water and indicates the implementation of good hygienic farm practices and dairy waste effluent management.

  12. The Bacillus cereus Hbl and Nhe tripartite enterotoxin components assemble sequentially on the surface of target cells and are not interchangeable.

    Directory of Open Access Journals (Sweden)

    Inka Sastalla

    Full Text Available Bacillus cereus is a spore-forming, Gram-positive bacterium commonly associated with outbreaks of food poisoning. It is also known as an opportunistic pathogen causing clinical infections such as bacteremia, meningitis, pneumonia, and gas gangrene-like cutaneous infections, mostly in immunocompromised patients. B. cereus secretes a plethora of toxins of which four are associated with the symptoms of food poisoning. Two of these, the non-hemolytic enterotoxin Nhe and the hemolysin BL (Hbl toxin, are predicted to be structurally similar and are unique in that they require the combined action of three toxin proteins to induce cell lysis. Despite their dominant role in disease, the molecular mechanism of their toxic function is still poorly understood. We report here that B. cereus strain ATCC 10876 harbors not only genes encoding Nhe, but also two copies of the hbl genes. We identified Hbl as the major secreted toxin responsible for inducing rapid cell lysis both in cultured cells and in an intraperitoneal mouse toxicity model. Antibody neutralization and deletion of Hbl-encoding genes resulted in significant reductions of cytotoxic activity. Microscopy studies with Chinese Hamster Ovary cells furthermore showed that pore formation by both Hbl and Nhe occurs through a stepwise, sequential binding of toxin components to the cell surface and to each other. This begins with binding of Hbl-B or NheC to the eukaryotic membrane, and is followed by the recruitment of Hbl-L1 or NheB, respectively, followed by the corresponding third protein. Lastly, toxin component complementation studies indicate that although Hbl and Nhe can be expressed simultaneously and are predicted to be structurally similar, they are incompatible and cannot complement each other.

  13. The Bacillus cereus Hbl and Nhe tripartite enterotoxin components assemble sequentially on the surface of target cells and are not interchangeable.

    Science.gov (United States)

    Sastalla, Inka; Fattah, Rasem; Coppage, Nicole; Nandy, Poulomi; Crown, Devorah; Pomerantsev, Andrei P; Leppla, Stephen H

    2013-01-01

    Bacillus cereus is a spore-forming, Gram-positive bacterium commonly associated with outbreaks of food poisoning. It is also known as an opportunistic pathogen causing clinical infections such as bacteremia, meningitis, pneumonia, and gas gangrene-like cutaneous infections, mostly in immunocompromised patients. B. cereus secretes a plethora of toxins of which four are associated with the symptoms of food poisoning. Two of these, the non-hemolytic enterotoxin Nhe and the hemolysin BL (Hbl) toxin, are predicted to be structurally similar and are unique in that they require the combined action of three toxin proteins to induce cell lysis. Despite their dominant role in disease, the molecular mechanism of their toxic function is still poorly understood. We report here that B. cereus strain ATCC 10876 harbors not only genes encoding Nhe, but also two copies of the hbl genes. We identified Hbl as the major secreted toxin responsible for inducing rapid cell lysis both in cultured cells and in an intraperitoneal mouse toxicity model. Antibody neutralization and deletion of Hbl-encoding genes resulted in significant reductions of cytotoxic activity. Microscopy studies with Chinese Hamster Ovary cells furthermore showed that pore formation by both Hbl and Nhe occurs through a stepwise, sequential binding of toxin components to the cell surface and to each other. This begins with binding of Hbl-B or NheC to the eukaryotic membrane, and is followed by the recruitment of Hbl-L1 or NheB, respectively, followed by the corresponding third protein. Lastly, toxin component complementation studies indicate that although Hbl and Nhe can be expressed simultaneously and are predicted to be structurally similar, they are incompatible and cannot complement each other.

  14. Spore surface proteins of Brevibacillus laterosporus are involved in insect pathogenesis

    Science.gov (United States)

    Marche, Maria Giovanna; Mura, Maria Elena; Falchi, Giovanni; Ruiu, Luca

    2017-01-01

    Outer spore envelope proteins of pathogenic bacteria often present specific virulence factors and tools to evade the defence system of their hosts. Brevibacillus laterosporus, a pathogen of invertebrates and an antimicrobial-producing species, is characterised by a unique spore coat and canoe-shaped parasporal body (SC-CSPB) complex surrounding the core spore. In the present study, we identified and characterised major proteins of the SC-CSPB complex of B. laterosporus, and we investigated their entomopathogenic role. Employing a proteomic approach and a B. laterosporus-house fly study model, we found four highly conserved proteins (ExsC, CHRD, CpbA and CpbB) that function as insect virulence factors. CpbA was associated with a significantly higher mortality of flies and greater relative gene expression levels during sporulation, compared to the other SC-CSPB proteins. Taken together, we suggest that spore surface proteins are a part of a complex set of toxins and virulence factors that B. laterosporus employs in its pathogenicity against flies. PMID:28256631

  15. A Type III protein-RNA toxin-antitoxin system from Bacillus thuringiensis promotes plasmid retention during spore development.

    Science.gov (United States)

    Short, Francesca L; Monson, Rita E; Salmond, George P C

    2015-01-01

    Members of the Bacillus cereus sensu lato group of bacteria often contain multiple large plasmids, including those encoding virulence factors in B. anthracis. Bacillus species can develop into spores in response to stress. During sporulation the genomic content of the cell is heavily compressed, which could result in counterselection of extrachromosomal genomic elements, unless they have robust stabilization and segregation systems. Toxin-antitoxin (TA) systems are near-ubiquitous in prokaryotes and have multiple biological roles, including plasmid stabilization during vegetative growth. Here, we have shown that a Type III TA system, based on an RNA antitoxin and endoribonuclease toxin, from plasmid pAW63 in Bacillus thuringiensis serovar kurstaki HD-73 can dramatically promote plasmid retention in populations undergoing sporulation and germination, and we provide evidence that this occurs through the post-segregational killing of plasmid-free forespores. Our findings show how an extremely common genetic module can be used to ensure plasmid maintenance during stress-induced developmental transitions, with implications for plasmid dynamics in B. cereus s.l. bacteria.

  16. Gene detection and toxin production evaluation of hemolysin BL of Bacillus cereus isolated from milk and dairy products marketed in Brazil.

    Science.gov (United States)

    Reis, Andre L S; Montanhini, Maike T M; Bittencourt, Juliana V M; Destro, Maria T; Bersot, Luciano S

    2013-12-01

    Bacillus cereusis an ubiquitous, spore-forming bacteria that can survive pasteurization and the majority of the heating processes used in the dairy industry. Besides, it is a pathogen responsible for different types of food poisoning. One type of foodborne disease caused by B.cereusis the diarrheal syndrome, which is caused by the ingestion of vegetative cells producing toxins in the small intestine. One virulence factor for the diarrheal syndrome is the toxin hemolysin BL (HBL), a three-component protein formed by the L1, L2 and B components. In order to evaluate the presence of diarrheal strains isolated from milk and dairy products, 63 B. cereus isolates were obtained from 260 samples of UHT milk, pasteurized milk and powdered milk, sold in commercial establishments and from different brands. The isolates were subjected to the Polymerase Chain Reaction (PCR) for the detection of the encoding genes for the L1, L2 and B components and the toxin production capacity were evaluated with an immunoassay. A total of 23 [36.5%] isolates were identified carrying simultaneously the three tested genes, from which, 20 [86.9%] showed toxigenic capacity. 26 [41.3%] isolates did not carry any of genes tested and the other 14 [22.2%] were positive for one or two of them. The results showed a high toxigenic capacity among the B. cereus isolates able to produce the HBL, indicating a potential risk for consumers.

  17. Gene detection and toxin production evaluation of hemolysin BL of Bacillus cereus isolated from milk and dairy products marketed in Brazil

    Directory of Open Access Journals (Sweden)

    Andre L.S. Reis

    2013-12-01

    Full Text Available Bacillus cereusis an ubiquitous, spore-forming bacteria that can survive pasteurization and the majority of the heating processes used in the dairy industry. Besides, it is a pathogen responsible for different types of food poisoning. One type of foodborne disease caused by B.cereusis the diarrheal syndrome, which is caused by the ingestion of vegetative cells producing toxins in the small intestine. One virulence factor for the diarrheal syndrome is the toxin hemolysin BL (HBL, a three-component protein formed by the L1, L2 and B components. In order to evaluate the presence of diarrheal strains isolated from milk and dairy products, 63 B. cereus isolates were obtained from 260 samples of UHT milk, pasteurized milk and powdered milk, sold in commercial establishments and from different brands. The isolates were subjected to the Polymerase Chain Reaction (PCR for the detection of the encoding genes for the L1, L2 and B components and the toxin production capacity were evaluated with an immunoassay. A total of 23 [36.5%] isolates were identified carrying simultaneously the three tested genes, from which, 20 [86.9%] showed toxigenic capacity. 26 [41.3%] isolates did not carry any of genes tested and the other 14 [22.2%] were positive for one or two of them. The results showed a high toxigenic capacity among the B. cereus isolates able to produce the HBL, indicating a potential risk for consumers.

  18. UV resistance of Bacillus anthracis spores revisited: validation of Bacillus subtilis spores as UV surrogates for spores of B. anthracis Sterne.

    Science.gov (United States)

    Nicholson, Wayne L; Galeano, Belinda

    2003-02-01

    Recent bioterrorism concerns have prompted renewed efforts towards understanding the biology of bacterial spore resistance to radiation with a special emphasis on the spores of Bacillus anthracis. A review of the literature revealed that B. anthracis Sterne spores may be three to four times more resistant to 254-nm-wavelength UV than are spores of commonly used indicator strains of Bacillus subtilis. To test this notion, B. anthracis Sterne spores were purified and their UV inactivation kinetics were determined in parallel with those of the spores of two indicator strains of B. subtilis, strains WN624 and ATCC 6633. When prepared and assayed under identical conditions, the spores of all three strains exhibited essentially identical UV inactivation kinetics. The data indicate that standard UV treatments that are effective against B. subtilis spores are likely also sufficient to inactivate B. anthracis spores and that the spores of standard B. subtilis strains could reliably be used as a biodosimetry model for the UV inactivation of B. anthracis spores.

  19. Clinical characteristics and antimicrobial susceptibility of Bacillus cereus blood stream infections

    OpenAIRE

    Ikeda, Mahoko; Yagihara,Yuka; Tatsuno, Keita; Okazaki, Mitsuhiro; Okugawa, Shu; Moriya, Kyoji

    2015-01-01

    Background Bacillus cereus is one of the pathogens causing nosocomial bloodstream infections (BSIs). However, few reports have documented the antimicrobial susceptibility and clinical characteristics of Bacillus cereus BSI and the importance of empirical therapy. The aim of this study was to investigate the clinical characteristics and antimicrobial susceptibility of B. cereus isolates from patients with BSI and to analyze the impact of appropriate empirical therapy on the outcome of patients...

  20. Comparative genome analysis of Bacillus cereus group genomes with Bacillus subtilis

    OpenAIRE

    Anderson, Iain; Sorokin, Alexei; Kapatral, Vinayak; Reznik, Gary; Bhattacharya, Anamitra; Mikhailova, Natalia; Burd, Henry; Joukov, Victor; Kaznadzey, Denis; Walunas, Theresa; D'Souza, Mark; Larsen, Niels; Pusch, Gordon; Liolios, Konstantinos; Grechkin, Yuri

    2005-01-01

    Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1,381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-...

  1. Naphthalene degradation and biosurfactant activity by Bacillus cereus 28BN

    Energy Technology Data Exchange (ETDEWEB)

    Tuleva, B.; Christova, N. [Inst. of Microbiology, Bulgarian Academy of Sciences, Sofia (Bulgaria); Jordanov, B.; Nikolova-Damyanova, B. [Inst. of Organic Chemistry, Sofia (Bulgaria); Petrov, P. [National Center of Infectious and Parasitic Diseases, Sofia (Bulgaria)

    2005-08-01

    Biosurfactant activity and naphthalene degradation by a new strain identified as Bacillus cereus 28BN were studied. The strain grew well and produced effective biosurfactants in the presence of n-alkanes, naphthalene, crude oil and vegetable oils. The biosurfactants were detected by the surface tension lowering of the medium, thin layer chromatography and infrared spectra analysis. With (2%) naphthalene as the sole carbon source, high levels of rhamnolipids at a concentration of 2.3 g l{sup -1} were determined in the stationary growth. After 20 d of incubation 72 {+-} 4% of the initial naphthalene was degraded. This is the first report for a Bacillus cereus rhamnolipid producing strain that utilized naphthalene under aerobic conditions. The strain looks promising for application in environmental technologies. (orig.)

  2. 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...... in a collection of spores from Britain. Ptaquiloside was present in all samples, with a maximum of 29μgg−1, which is very low compared to other parts of the fern. Considering the low abundance of spores in breathing air under normal conditions, this exposure route is likely to be secondary to milk or drinking...

  3. Genome Sequences of Three Novel Bacillus cereus Bacteriophages

    OpenAIRE

    Julianne H Grose; Jensen, Jordan D.; Merrill, Bryan D.; Fisher, Joshua N. B.; Burnett, Sandra H.; Breakwell, Donald P

    2014-01-01

    The Bacillus cereus group is an assemblage of highly related firmicute bacteria that cause a variety of diseases in animals, including insects and humans. We announce three high-quality, complete genome sequences of bacteriophages we isolated from soil samples taken at the bases of fruit trees in Utah County, Utah. While two of the phages (Shanette and JL) are highly related myoviruses, the bacteriophage Basilisk is a siphovirus.

  4. Bacillus cereus bacteremia and multiple brain abscesses during acute lymphoblastic leukemia induction therapy.

    Science.gov (United States)

    Hansford, Jordan R; Phillips, Marianne; Cole, Catherine; Francis, Joshua; Blyth, Christopher C; Gottardo, Nicholas G

    2014-04-01

    Bacillus cereus can cause serious infections in immunosuppressed patients. This population may be susceptible to B. cereus pneumonia, bacteremia, cellulitis, and rarely cerebral abscess. Here we report an 8-year-old boy undergoing induction therapy for acute lymphoblastic leukemia who developed multifocal B. cereus cerebral abscesses, highlighting the propensity for B. cereus to develop cerebral abscesses. A review of the literature over the past 25 years identified another 11 cases (3 children and 8 adults) of B. cereus cerebral abscess in patients undergoing cancer therapy. B. cereus cerebral abscesses were associated with a high mortality rate (42%) and significant morbidity. Notably, B. cereus bacteremia with concomitant cerebral abscess was associated with induction chemotherapy for acute leukemia in both children and adults (10 of 12 case reports). Our case report and review of the literature highlights the propensity for B. cereus to develop cerebral abscess(es). Therefore, early consideration for neuroimaging should be given for any neutropenic cancer patient identified with B. cereus bacteremia, in particular those with acute leukemia during induction therapy.

  5. Occurrence and characterization of toxigenic Bacillus cereus in food and infant feces

    Institute of Scientific and Technical Information of China (English)

    Sameer; Rushdi; Organji; Hussein; Hasan; Abulreesh; Khaled; Elbanna; Gamal; Ebrahim; Haridy; Osman; Manal; Khider

    2015-01-01

    Objective: To investigate the true incidence of Bacillus cereus(B. cereus) in food and children diarrhea cases. Methods: A total of 110 samples of various dairy products such as raw milk, long life pasteurized milk, yoghurt and infant powdered milk formulas, raw rice, and feces were examined for the presence of B. cereus by selective plating on mannitol-egg-yolk-polymyxin agar. Confirmation of B. cereus was carried out by biochemical tests and PCR. Identification of non-B. cereus isolates was carried out by 16 S r DNA sequencing. Antimicrobial susceptibility was done by disk diffusion method.Results: Overall 35 samples(31.8%, n = 110) yielded Bacillus-like growth. Of which 19 samples(54.28%) were positive for B. cereus. All isolates were positive for enterotoxin production. No psychrotolerant B. cereus strains were detected in all samples. All B. cereus isolates were resistant to penicillin G, but susceptible to vancomycin, erythromycin and clindamycin. Conclusions: The results of this study confirm the importance of including B. cereus in disease control and prevention programs, as well as in routine clinical and food quality control laboratories in both Saudi Arabia and Egypt.

  6. Occurrence and characterization of toxigenic Bacillus cereus in food and infant feces

    Institute of Scientific and Technical Information of China (English)

    Sameer Rushdi Organji; Hussein Hasan Abulreesh; Khaled Elbanna; Gamal Ebrahim Haridy Osman; Manal Khider

    2015-01-01

    Objective:To investigate the true incidence of Bacillus cereus (B. cereus) in food and children diarrhea cases. Methods:A total of 110 samples of various dairy products such as raw milk, long life pasteurized milk, yoghurt and infant powdered milk formulas, raw rice, and feces were examined for the presence of B. cereus by selective plating on mannitol-egg-yolk-polymyxin agar. Confirmation of B. cereus was carried out by biochemical tests and PCR. Identification of non-B. cereus isolates was carried out by 16S rDNA sequencing. Antimicrobial susceptibility was done by disk diffusion method. Results:Overall 35 samples (31.8%, n=110) yielded Bacillus-like growth. Of which 19 samples (54.28%) were positive for B. cereus. All isolates were positive for enterotoxin production. No psychrotolerant B. cereus strains were detected in all samples. All B. cereus isolates were resistant to penicillin G, but susceptible to vancomycin, erythromycin and clindamycin. Conclusions:The results of this study confirm the importance of including B. cereus in disease control and prevention programs, as well as in routine clinical and food quality control laboratories in both Saudi Arabia and Egypt.

  7. Summoning the wind: Hydrodynamic cooperation of forcibly ejected fungal spores

    CERN Document Server

    Roper, Marcus; Cobb, Ann; Dillard, Helene R; Pringle, Anne

    2009-01-01

    The forcibly launched spores of the crop pathogen \\emph{Sclerotinia sclerotiorum} must eject through many centimeters of nearly still air to reach the flowers of the plants that the fungus infects. Because of their microscopic size, individually ejected spores are quickly brought to rest by drag. In the accompanying fluid dynamics video we show experimental and numerical simulations that demonstrate how, by coordinating the nearly simultaneous ejection of hundreds of thousands of spores,\\emph{Sclerotinia} and other species of apothecial fungus are able to sculpt a flow of air that carries spores across the boundary layer and around intervening obstacles. Many spores are sacrificed to create this flow of air. Although high speed imaging of spore launch in a wild isolate of the dung fungus \\emph{Ascobolus} shows that the synchronization of spore ejections is self-organized, which could lead to spores delaying their ejection to avoid being sacrificed, simulations and asymptotic analysis show that, close the frui...

  8. Production of Alpha Amylase by Bacillus cereus in Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Helen H. Raplong

    2014-09-01

    Full Text Available Microorganisms have the ability to secrete enzymes when they are grown in the presence of certain substrates. Amylases are among the most important industrial enzymes and are of great significance in biotechnological studies. Bacteria belonging to the genus Bacillus were isolated using mannitol egg yolk polymyxin B (MYP agar a highly selective media for Bacillus cereus isolation. The isolates were tested for α-amylase production on nutrient agar supplemented with starch and in submerged fermentation. The bacteria isolated and identified (using the Microgen Bacillus identification kit were all Bacillus cereus and SB2 had the largest zone of hydrolysis of 12mm on nutrient agar supplemented with starch as well as the highest enzyme activity of 1.62U/ml. Amylase activity of 2.56U/ml was obtained after 24 hours incubation in submerged fermentation. When amylase enzyme production parameters where optimized, maximum amylase activity was obtained at a pH of 6.5, temperature of 350C, incubation time of 24 hours and 4% inoculums concentration. Bacillus cereus SB2 is a potential isolate for alpha-amylase production with soluble starch as the sole carbon source in submerged fermentation.

  9. Cr(VI) uptake mechanism of Bacillus cereus.

    Science.gov (United States)

    Chen, Zhi; Huang, Zhipeng; Cheng, Yangjian; Pan, Danmei; Pan, Xiaohong; Yu, Meijuan; Pan, Zhiyun; Lin, Zhang; Guan, Xiong; Wu, Ziyu

    2012-04-01

    In this study, we investigated the Cr(VI) uptake mechanism in an indigenous Cr(VI)-tolerant bacterial strain -Bacillus cereus through batch and microscopic experiments. We found that both the cells and the supernatant collected from B. cereus cultivation could reduce Cr(VI). The valence state analysis revealed the complete transformation from Cr(VI) into Cr(III) by living B. cereus. Further X-ray absorption fine structure and Fourier transform infrared analyses showed that the reduced Cr(III) was coordinated with carboxyl and amido functional groups from either the cells or supernatant. Scanning electron microscopy and atomic force microscopy observation showed that noticeable Cr(III) precipitates were accumulated on bacterial surfaces. However, Cr(III) could also be detected in bacterial inner portions by using transmission electron microscopy thin section analysis coupled with energy dispersive X-ray spectroscopy. Through quantitative analysis of chromium distribution, we determined the binding ratio of Cr(III) in supernatant, cell debris and cytoplasm as 22%, 54% and 24%, respectively. Finally, we further discussed the role of bacterium-origin soluble organic molecules to the remediation of Cr(VI) pollutants.

  10. Chemodiversity of cereulide, the emetic toxin of Bacillus cereus.

    Science.gov (United States)

    Marxen, Sandra; Stark, Timo D; Frenzel, Elrike; Rütschle, Andrea; Lücking, Genia; Pürstinger, Gabriel; Pohl, Elena E; Scherer, Siegfried; Ehling-Schulz, Monika; Hofmann, Thomas

    2015-03-01

    Food-borne intoxications are increasingly caused by the dodecadepsipeptide cereulide, the emetic toxin produced by Bacillus cereus. As such intoxications pose a health risk to humans, a more detailed understanding on the chemodiversity of this toxin is mandatory for the reliable risk assessment of B. cereus toxins in foods. Mass spectrometric screening now shows a series of at least 18 cereulide variants, among which the previously unknown isocereulides A-G were determined for the first time by means of UPLC-TOF MS and ion-trap MS(n) sequencing, (13)C-labeling experiments, and post-hydrolytic dipeptide and enantioselective amino acid analysis. The data demonstrate a high microheterogeneity in cereulide and show evidence for a relaxed proof reading function of the non-ribosomal cereulide peptide synthetase complex giving rise to an enhanced cereulide chemodiversity. Most intriguingly, the isocereulides were found to differ widely in their cell toxicity correlating with their ionophoric properties (e.g., purified isocereulide A showed about 8-fold higher cytotoxicity than purified cereulide in the HEp-2 assay and induced an immediate breakdown of bilayer membranes). These findings provide a substantial contribution to the knowledge-based risk assessment of B. cereus toxins in foods, representing a still unsolved challenge in the field of food intoxications.

  11. Measurement of Metabolic Activity in Dormant Spores of Bacillus Species

    Science.gov (United States)

    2015-01-14

    SECURITY CLASSIFICATION OF: Spores of Bacillus megaterium and Bacillus subtilis were harvested shortly after release from sporangia, incubated under...Dec-2014 Approved for Public Release; Distribution Unlimited Final Report: Measurement of Metabolic Activity in Dormant Spores of Bacillus Species...Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 spores, Bacillus , spore dormancy, 3-phosphoglycerate REPORT DOCUMENTATION PAGE 11

  12. 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.

  13. Modeling Radiation Effectiveness for Inactivation of Bacillus Spores

    Science.gov (United States)

    2015-09-17

    will undergo germination which is the first step in the process by which bacteria transforms from a dormant spore into a vegetative cell [28]. The...order to survive a dose of radiation, a spore must repair its damaged DNA during germination . The DNA repair process is dependent on reactions catalyzed...in the next section. 2.2 Life Cycle of a Bacterial Spore A dormant spore is formed via a multi- step process called sporulation (refer to Figure 2.5

  14. Dendritic Cells Endocytose Bacillus Anthracis Spores: Implications for Anthrax Pathogenesis

    Science.gov (United States)

    2007-11-02

    Dendritic Cells Endocytose Bacillus anthracis Spores: Implications for Anthrax Pathogenesis1 Katherine C. Brittingham,* Gordon Ruthel,* Rekha G...germination and dissemination of spores. Found in high frequency throughout the respiratory track, dendritic cells (DCs) routinely take up foreign...COVERED - 4. TITLE AND SUBTITLE Dendritic cells endocytose Bacillus anthracis spores: implications for anthrax pathogenesis, The Journal of

  15. 9 CFR 113.66 - Anthrax Spore Vaccine-Nonencapsulated.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Anthrax Spore Vaccine-Nonencapsulated. 113.66 Section 113.66 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... REQUIREMENTS Live Bacterial Vaccines § 113.66 Anthrax Spore Vaccine—Nonencapsulated. Anthrax Spore...

  16. 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.

  17. Toxicity of terpenes to spores and mycelium of Penicillium digitatum

    NARCIS (Netherlands)

    Wolken, W.A.M.; Tramper, J.; Werf, M.J. van der

    2002-01-01

    Spores, although often considered metabolically inert, catalyze a variety of reactions. The use of spores instead of mycelium for bioconversions has several advantages. In this paper, we describe the difference in susceptibility of mycelium and spores against toxic substrates and products. A higher

  18. Toxicity of terpenes to spores and mycelium op Penicillium digitatum

    NARCIS (Netherlands)

    Wolken, W.A.M.; Tramper, J.; Werf, van der M.J.

    2002-01-01

    Spores, although often considered metabolically inert, catalyze a variety of reactions. The use of spores instead of mycelium for bioconversions has several advantages. In this paper, we describe the difference in susceptibility of mycelium and spores against toxic substrates and products. A higher

  19. Geraniol biotransformation-pathway in spores of Penicillium digitatum

    NARCIS (Netherlands)

    Wolken, W.A.M.; Werf, M.J. van der

    2001-01-01

    Spores of Penicillium digitatum ATCC 201167 transform geraniol, nerol, citral, and geranic acid into methylheptenone. Spore extracts of P. digitatum convert geraniol and nerol NAD+-dependently into citral. Spore extract also converts citral NAD+-dependently into geranic acid. Furthermore, a novel en

  20. Detection of Bacillus cereus on selected retail chicken products.

    Science.gov (United States)

    Smith, D P; Berrang, M E; Feldner, P W; Phillips, R W; Meinersmann, R J

    2004-08-01

    Samples from five chicken meat products, obtained at retail stores, were evaluated for the presence of Bacillus cereus. The products tested were as follows: breaded, fully cooked, frozen nuggets (NUGGETS); breaded, fully cooked, frozen tenders (TENDERS); fully cooked, frozen, white-meat fajita-style strips (STRIPS); raw, refrigerated, boneless, skinless, marinated breast fillets (FILLETS); and raw, refrigerated, cut-up, tray-pack bone-in parts (PARTS), either split breasts or thighs. Four packages of each item were obtained on three different days (n = 60). Frozen and refrigerated products were held overnight in their respective environments as appropriate; then packages were opened aseptically, and a total of 25 g of tissue was excised from multiple pieces within a package. The 25-g samples were enriched in 225 ml of Trypticase soy-polymixin broth for 18 to 24 h at 30 degrees C and then plated on mannitol-egg yolk-polymixin agar and incubated for 18 to 24 h at 30 degrees C. Colonies characteristic of B. cereus were chosen and replated for isolation on mannitol-egg yolk-polymixin agar. Suspect colonies were confirmed as Bacillus spp. by Gram stain, hemolysis on blood agar, and a biochemical test strip. Isolates were further confirmed as B. cereus using Bacteriological Analytical Manual procedures, including tests for motility, rhizoid growth, hemolysis, and protein toxin crystal production. B. cereus was detected in 27 of 60 total samples. By product, the prevalence levels were as follows: NUGGETS, 11 of 12 positive; TENDERS, 8 of 12 positive; STRIPS, 6 of 12 positive; FILLETS, 0 of 12 positive; and PARTS, 2 of 12 positive. Isolates were tested by PCR for presence of the toxin-encoding genes bceT, nheABC, hblACD, and cytK. Results indicate that B. cereus organisms were present on four of the five retail poultry products tested in this study, with the highest rates reported for the three fully cooked items, especially the two breaded products. All strains isolated

  1. Phospholipase Cδ regulates germination of Dictyostelium spores

    NARCIS (Netherlands)

    Dijken, Peter van; Haastert, Peter J.M. van

    2001-01-01

    Background: Many eukaryotes, including plants and fungi make spores that resist severe environmental stress. The micro-organism Dictyostelium contains a single phospholipase C gene (PLC); deletion of the gene has no effect on growth, cell movement and differentiation. In this report we show that PLC

  2. Mechanisms of Resistance in Microbial Spores

    Science.gov (United States)

    1990-12-20

    solids (and water) content by immersion refractometry . Heat-activated spores of Bacillus stearotherrnophilus were found to be separable into two...incrC· ment of bacterial cells, enabling determination of their solids content by immersion refractometry . The results agreed well with values for

  3. Analysis of Alicyclobacillus acidoterrestris spores from different sporulation media subjected to wet-heat

    Directory of Open Access Journals (Sweden)

    Celenk Molva

    2015-12-01

    Full Text Available The presence of Alicyclobacillus acidoterrestris endospores in fruit juices is a significant problem for the juice industry since they are able to survive pasteurization subsequently leading to the spoilage. To evaluate the mechanism of wet-heat, structural damage and the leakages of intracellular materials of A. acidoterrestris DSM 3922 spores from different sporulation media was studied at 90°C (15-45 min. For sporulation, Bacillus acidoterrestris agar, Bacillus acidocaldarius agar, potato dextrose agar and malt extract agar were used. Based on the scanning electron microscopy, loss of internal volume and structural integrity were observed following heating which were further confirmed by the leakages of intracellular components. The obtained results suggest that the inactivation of A. acidoterrestris DSM 3922 spores by wet-heat is associated with damage to the coat and inner membrane depending on the sporulation medium composition and heating time.

  4. Monitoring Rates and Heterogeneity of High-Pressure Germination of Bacillus Spores by Phase-Contrast Microscopy of Individual Spores

    Science.gov (United States)

    2014-01-01

    SECURITY CLASSIFICATION OF: The germination of multiple individual Bacillus subtilis spores by a high pressure (HP) of 140-150 (unless noted...otherwise) megaPascals (MPa) that activates spore germinant receptors (GRs) was monitored by phase contrast microscopy in a diamond anvil cell. Major...conclusions were that: i) >95% of spores germinated in 40 min; ii) individual spore’s HP germination kinetics were very similar to those for nutrient

  5. Bacterial spore detection and analysis using hyperpolarized (129)Xe chemical exchange saturation transfer (Hyper-CEST) NMR.

    Science.gov (United States)

    Bai, Yubin; Wang, Yanfei; Goulian, Mark; Driks, Adam; Dmochowski, Ivan J

    2014-08-01

    Previously, we reported hyperpolarized (129)Xe chemical exchange saturation transfer (Hyper-CEST) NMR techniques for the ultrasensitive (i.e., 1 picomolar) detection of xenon host molecules known as cryptophane. Here, we demonstrate a more general role for Hyper-CEST NMR as a spectroscopic method for probing nanoporous structures, without the requirement for cryptophane or engineered xenon-binding sites. Hyper-CEST (129)Xe NMR spectroscopy was employed to detect Bacillus anthracis and Bacillus subtilis spores in solution, and interrogate the layers that comprise their structures. (129)Xe-spore samples were selectively irradiated with radiofrequency pulses; the depolarized (129)Xe returned to aqueous solution and depleted the (129)Xe-water signal, providing measurable contrast. Removal of the outermost spore layers in B. anthracis and B. subtilis (the exosporium and coat, respectively) enhanced (129)Xe exchange with the spore interior. Notably, the spores were invisible to hyperpolarized (129)Xe NMR direct detection methods, highlighting the lack of high-affinity xenon-binding sites, and the potential for extending Hyper-CEST NMR structural analysis to other biological and synthetic nanoporous structures.

  6. Modelling the influence of the sporulation temperature upon the bacterial spore heat resistance, application to heating process calculation.

    Science.gov (United States)

    Leguérinel, I; Couvert, O; Mafart, P

    2007-02-28

    Environmental conditions of sporulation influence bacterial heat resistance. For different Bacillus species a linear Bigelow type relationship between the logarithm of D values determined at constant heating temperature and the temperature of sporulation was observed. The absence of interaction between sporulation and heating temperatures allows the combination of this new relationship with the classical Bigelow model. The parameters zT and zT(spo) of this global model were fitted to different sets of data regarding different Bacillus species: B. cereus, B. subtilis, B. licheniformis, B. coagulans and B. stearothermophilus. The origin of raw products or food process conditions before a heat treatment can lead to warm temperature conditions of sporulation and to a dramatic increase of the heat resistance of the generated spores. In this case, provided that the temperature of sporulation can be assessed, this model can be easily implemented to rectify F values on account of possible increase of thermal resistance of spores and to ensure the sterilisation efficacy.

  7. Detection of chlorophylls in spores of seven ferns.

    Science.gov (United States)

    Tseng, Mei-Hwei; Lin, Kuei-Huei; Huang, Yi-Jia; Chang, Ya-Lan; Huang, Sheng-Cih; Kuo, Li-Yaung; Huang, Yao-Moan

    2017-03-01

    Fern spores were traditionally classified into chlorophyllous (green) and nonchlorophyllous (nongreen) types based on the color visible to the naked eye. Recently, a third type, "cryptochlorophyllous spores", is recognized, and these spores are nongreen under white light but contain chlorophylls. Epifluorescence microscopy was previously used to detect chlorophylls in cryptochlorophyllous spores. In addition to epifluorescence microscopy, current study performed some other approaches, including spore-squash epifluorescence, absorption spectra, laser-induced fluorescence emission spectra, thin layer chromatography (TLC), and ultra-high performance liquid chromatography with ultraviolet and mass spectrometric detection (UHPLC-UV-MS) in order to detect chlorophylls of spores of seven ferns (Sphaeropteris lepifera, Ceratopteris thalictroides, Leptochilus wrightii, Leptochilus pothifolius, Lepidomicrosorum buergerianum, Osmunda banksiifolia, and Platycerium grande). Destructive methods, such as TLC and UHPLC-UV-MS, successfully detected chlorophylls inside the spores when their signals of red fluorescence under epifluorescence microscope were masked by spore wall. Although UHPLC-UV-MS analysis was the most sensitive and reliable for determining the chlorophylls of spores, spore-squash epifluorescence is not only reliable but also cost- and time-effective one among our study methods. In addition, we first confirmed that Lepidomicrosorium buergerianum, Leptochilus pothifolius, Leptochilus wrightii, and Platycerium grande, produce cryptochlorophyllous spores.

  8. Source strength of fungal spore aerosolization from moldy building material

    Energy Technology Data Exchange (ETDEWEB)

    Gorny, Rafa L.; Reponen, Tiina; Grinshpun, Sergey A.; Willeke, Klaus [Cincinnati Univ., Dept. of Environmental Health, Cincinnati, OH (United States)

    2001-07-01

    The release of Aspergillus versicolor, Cladosporium cladosporioides, and Penicillium melinii spores from agar and ceiling tile surfaces was tested under different controlled environmental conditions using a newly designed and constructed aerosolization chamber. This study revealed that all the investigated parameters, such as fungal species, air velocity above the surface, texture of the surface, and vibration of contaminated material, affected the fungal spore release. It was found that typical indoor air currents can release up to 200 spores cm {sup -2} from surface with fungal spores during 30-min experiments. The release of fungal spores from smooth agar surfaces was found to be inadequate for accurately predicting the emission from rough ceiling tile surfaces because the air turbulence increases the spore release from a rough surface. A vibration of a frequency of 1Hz at a power level of 14W resulted in a significant increase in the spore release rate. The release appears to depend on the morphology of the fungal colonies grown on ceiling tile surfaces including the thickness of conidiophores, the length of spore chains, and the shape of spores. The spores were found to be released continuously during each 30-min experiment. However, the release rate was usually highest during the first few minutes of exposure to air currents and mechanical vibration. About 71-88% of the spores released during a 30-min interval became airborne during the first 10min. (Author)

  9. Polymerase chain reaction assay for the detection of Bacillus cereus group cells

    DEFF Research Database (Denmark)

    Hansen, Bjarne Munk; Leser, Thomas D.; Hendriksen, Niels Bohse

    2001-01-01

    Recent investigations have shown that members of the Bacillus cereus group carry genes which have the potential to cause gastrointestinal and somatic diseases. Although most cases of diseases caused by the B. cereus group bacteria are relatively mild, it is desirable to be able to detect members ...

  10. Association between tea ingestion and invasive Bacillus cereus infection among children with cancer.

    Science.gov (United States)

    El Saleeby, C M; Howard, S C; Hayden, R T; McCullers, J A

    2004-11-15

    Bacillus cereus is an emerging pathogen that causes invasive disease in immunocompromised hosts. A case-control study, prompted by a clinical case, demonstrated an association between dietary tea ingestion and B. cereus bacteremia. Policies designed to interrupt transmission of this pathogen to susceptible patients should be considered.

  11. Persistent Bacillus cereus Bacteremia in 3 Persons Who Inject Drugs, San Diego, California, USA.

    Science.gov (United States)

    Schaefer, Gabrielle; Campbell, Wesley; Jenks, Jeffrey; Beesley, Cari; Katsivas, Theodoros; Hoffmaster, Alex; Mehta, Sanjay R; Reed, Sharon

    2016-09-01

    Bacillus cereus is typically considered a blood culture contaminant; however, its presence in blood cultures can indicate true bacteremia. We report 4 episodes of B. cereus bacteremia in 3 persons who inject drugs. Multilocus sequence typing showed that the temporally associated infections were caused by unrelated clones.

  12. Persistent Bacillus cereus Bacteremia in 3 Persons Who Inject Drugs, San Diego, California, USA

    OpenAIRE

    Schaefer, Gabrielle; Campbell, Wesley; Jenks, Jeffrey; Beesley, Cari; Katsivas, Theodoros; Hoffmaster, Alex; Mehta, Sanjay R.; Reed, Sharon

    2016-01-01

    Bacillus cereus is typically considered a blood culture contaminant; however, its presence in blood cultures can indicate true bacteremia. We report 4 episodes of B. cereus bacteremia in 3 persons who inject drugs. Multilocus sequence typing showed that the temporally associated infections were caused by unrelated clones.

  13. Metabolic capacity of Bacillus cereus strains ATCC 14579 and ATCC 10987 interlinked with comparative genomics.

    NARCIS (Netherlands)

    Mols, M.; Been, M.W.H.J. de; Zwietering, M.H.; Moezelaar, R.; Abee, T.

    2007-01-01

    Bacillus cereus is an important food-borne pathogen and spoilage organism. In this study, numerous phenotypes and the genomes of B.?cereus strains ATCC 14579 and ATCC 10987 were analysed to compare their metabolic capacity and stress resistance potential. The growth performance of the two strains wa

  14. Identification of proteins involved in the heat stress response of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Periago, P.M.; Schaik, van W.; Abee, T.; Wouters, J.A.

    2002-01-01

    To monitor the ability of the food-borne opportunistic pathogen Bacillus cereus to survive during minimal processing of food products, we determined its heat-adaptive response. During pre-exposure to 42°C, B. cereus ATCC 14579 adapts to heat exposure at the lethal temperature of 50°C (maximum protec

  15. Adaptation of the food-borne pathogen Bacillus cereus to carvacrol

    NARCIS (Netherlands)

    Ultee, A.; Kets, E.P.W.; Alberda, M.; Hoekstra, F.A.; Smid, E.J.

    2000-01-01

    Carvacrol, a natural antimicrobial compound present in the essential oil fraction of oregano and thyme, is bactericidal towards Bacillus cereus. A decrease of the sensitivity of B. cereus towards carvacrol was observed after growth in the presence of non-lethal carvacrol concentrations. A decrease o

  16. Prevalence, genetic diversity, and antibiotic resistance of Bacillus cereus isolated from Korean fermented soybean products.

    Science.gov (United States)

    Kim, Cheol-Woo; Cho, Seung-Hak; Kang, Suk-Ho; Park, Yong-Bae; Yoon, Mi-Hye; Lee, Jong-Bok; No, Wan-Seob; Kim, Jung-Beom

    2015-01-01

    Bacillus cereus contamination is a major food safety problem for Korean fermented soybean products, but few studies have assessed its potential to cause foodborne illness. The objectives of this study were to investigate the prevalence and characteristics of B. cereus isolated from Korean fermented soybean products. B. cereus was detected in 110 of 162 (67.9%) samples. The highest B. cereus frequency was observed in deonjang (68 of 93 samples, 73.1%) and cheonggukjang (18 of 25, 72.0%); however, nonhemolytic enterotoxin was detected only in 22 of 162 samples (13.6%). Although the tested B. cereus isolates showed diverse pulsotypes according to repetitive sequence-PCR banding patterns, they displayed similar antibiotic sensitivity spectra. The low frequency of enterotoxin detection suggests that the potential risk of B. cereus foodborne illness associated with Korean fermented soybean products is lower than generally presumed. However, considering the prevalence of B. cereus and the high content of fermented soybean products in the Korean diet, it is necessary to constantly monitor the level of contamination with B. cereus and its toxins in such Korean food products.

  17. Plant compounds enhance assay sensitivity for detection of active bacillus cereus toxin

    Science.gov (United States)

    Bacillus cereus is an important food pathogen, producing emetic and diarrheal syndromes, the latter mediated by enterotoxins. It has been estimated that there are 84,000 cases of B. cereus food poisoning in the US each year, with an annual cost of USD 36 million. The ability to sensitively trace and...

  18. Influence of carvacrol on growth and toxin production by Bacillus cereus. International

    NARCIS (Netherlands)

    Ultee, A.; Smid, E.J.

    2001-01-01

    The natural antimicrobial compound carvacrol was investigated for its effect on diarrheal toxin production by Bacillus cereus. Carvacrol (0-0.06 mg/ml) reduced the viable count and the maximal specific growth rate (μmax) of B. cereus in BHI broth. The total amount of protein was not affected by carv

  19. Detection and expression of enterotoxin genes in plant-associated strains of Bacillus cereus

    Science.gov (United States)

    Bacillus cereus is an environmental microbe that commonly inhabits plants and soil. Twenty five plant-associated B. cereus isolates were obtained from apple, cacao, tomato, and potato. The isolates were screened for the presence and expression of enterotoxin B (BcET) components of the nonhemolytic e...

  20. Concerted action of sphingomyelinase and non-hemolytic enterotoxin in pathogenic Bacillus cereus.

    Directory of Open Access Journals (Sweden)

    Viktoria M Doll

    Full Text Available Bacillus cereus causes food poisoning and serious non-gastrointestinal-tract infections. Non-hemolytic enterotoxin (Nhe, which is present in most B. cereus strains, is considered to be one of the main virulence factors. However, a B. cereus ΔnheBC mutant strain lacking Nhe is still cytotoxic to intestinal epithelial cells. In a screen for additional cytotoxic factors using an in vitro model for polarized colon epithelial cells we identified B. cereus sphingomyelinase (SMase as a strong inducer of epithelial cell death. Using single and double deletion mutants of sph, the gene encoding for SMase, and nheBC in B. cereus we demonstrated that SMase is an important factor for B. cereus cytotoxicity in vitro and pathogenicity in vivo. SMase substantially complemented Nhe induced cytotoxicity in vitro. In addition, SMase but not Nhe contributed significantly to the mortality rate of larvae in vivo in the insect model Galleria mellonella. Our study suggests that the role of B. cereus SMase as a secreted virulence factor for in vivo pathogenesis has been underestimated and that Nhe and SMase complement each other significantly to cause full B. cereus virulence hence disease formation.

  1. Analysis of a Novel Spore Antigen in Bacillus anthracis That Contributes to Spore Opsonization

    Science.gov (United States)

    2008-01-01

    identity with homologues in B. cereus and Bacillus thuringiensis (99 and 94 %, respectively). In addition, a small ORF (BA5270) was located immediately...N. R. (1962). Field evaluation of a human anthrax vaccine. Am J Public Health 52, 632–645. Brossier, F. & Mock, M. (2001). Toxins of Bacillus ...authors (2007). The complete genome sequence of Bacillus thuringiensis Al Hakam. J Bacteriol 189, 3680–3681. Clements, M. O. & Moir, A. (1998). Role of

  2. Mushroom spore dispersal by convectively-driven winds

    CERN Document Server

    Dressaire, Emilie; Song, Boya; Roper, Marcus

    2015-01-01

    Thousands of fungal species rely on mushroom spores to spread across landscapes. It has long been thought that spores depend on favorable airflows for dispersal -- that active control of spore dispersal by the parent fungus is limited to an impulse delivered to the spores to carry them clear of the gill surface. Here we show that evaporative cooling of the air surrounding the mushroom pileus creates convective airflows capable of carrying spores at speeds of centimeters per second. Convective cells can transport spores from gaps that may be only a centimeter high, and lift spores ten centimeters or more into the air. The work reveals how mushrooms tolerate and even benefit from crowding, and provides a new explanation for their high water needs.

  3. Bacterial spore structures and their protective role in biocide resistance.

    Science.gov (United States)

    Leggett, M J; McDonnell, G; Denyer, S P; Setlow, P; Maillard, J-Y

    2012-09-01

    The structure and chemical composition of bacterial spores differ considerably from those of vegetative cells. These differences largely account for the unique resistance properties of the spore to environmental stresses, including disinfectants and sterilants, resulting in the emergence of spore-forming bacteria such as Clostridium difficile as major hospital pathogens. Although there has been considerable work investigating the mechanisms of action of many sporicidal biocides against Bacillus subtilis spores, there is far less information available for other species and particularly for various Clostridia. This paucity of information represents a major gap in our knowledge given the importance of Clostridia as human pathogens. This review considers the main spore structures, highlighting their relevance to spore resistance properties and detailing their chemical composition, with a particular emphasis on the differences between various spore formers. Such information will be vital for the rational design and development of novel sporicidal chemistries with enhanced activity in the future.

  4. [Bacterial spore--a new vaccine vehicle--a review].

    Science.gov (United States)

    Wang, Yanchun; Zhang, Zhaoshan

    2008-03-01

    Bacterial spores are robust and dormant life forms with formidable resistance properties. Spores of the genus Bacillus have been used for a long time as probiotics for oral bacteriotherapy both in humans and animals. Recently, genetically modified B. subtilis spores and B. anthracis spores have been used as indestructible delivery vehicles for vaccine antigens. They were used as vaccine vehicles or spore vaccine for oral immunization against tetanus and anthrax, and the results were very exciting. Unlike many second generation vaccine systems currently under development, bacterial spores offer heat stability and the flexibility for genetic manipulation. At the same time, they can elicit mucosal immune response by oral and nasal administration. This review focuses on the use of recombinant spores as vaccine delivery vehicles.

  5. Determination of fungal spore release from wet building materials

    DEFF Research Database (Denmark)

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

    2003-01-01

    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...

  6. INCORPORATION OF BACTERIOPHAGE GENOME BY SPORES OF BACILLUS SUBTILIS.

    Science.gov (United States)

    TAKAHASHI, I

    1964-06-01

    Takahashi, I. (Microbiology Research Institute, Ottawa, Ontario, Canada). Incorporation of bacteriophage genome by spores of Bacillus subtilis. J. Bacteriol. 87:1499-1502. 1964-The buoyant density in a CsCl gradient of deoxyribonucleic acid (DNA) extracted from spores of Bacillus subtilis was found to be identical to that of DNA from vegetative cells. Density-gradient centrifugation of DNA of spores derived from cultures infected with phage PBS 1 revealed the presence of a minor band whose density corresponded to that of the phage DNA in addition to the spore DNA. No intermediate bands were present. The relative amount of the phage DNA present in the spores was estimated to be 11%, suggesting that spores of this organism may incorporate several copies of the phage genome. Although the possibility that true lysogeny may occur cannot be entirely eliminated, the results seem to indicate that the phage genomes incorporated into spores are not attached to the host chromosome in this system.

  7. Inactivation of Clostridium difficile spores by microwave irradiation.

    Science.gov (United States)

    Ojha, Suvash Chandra; Chankhamhaengdecha, Surang; Singhakaew, Sombat; Ounjai, Puey; Janvilisri, Tavan

    2016-04-01

    Spores are a potent agent for Clostridium difficile transmission. Therefore, factors inhibiting spores have been of continued interest. In the present study, we investigated the influence of microwave irradiation in addition to conductive heating for C. difficile spore inactivation in aqueous suspension. The spores of 15 C. difficile isolates from different host origins were exposed to conductive heating and microwave irradiation. The complete inhibition of spore viability at 10(7) CFU/ml was encountered following microwave treatment at 800 W for 60 s, but was not observed in the conductive-heated spores at the same time-temperature exposure. The distinct patterns of ultrastructural alterations following microwave and conductive heat treatment were observed and the degree of damages by microwave was in the exposure time-dependent manner. Microwave would therefore be a simple and time-efficient tool to inactivate C. difficile spores, thus reducing the risk of C. difficile transmission.

  8. Scanning electron microscopy of food-poisoning bacterium Bacillus cereus using a variable-pressure SEM.

    Science.gov (United States)

    Nishimura, Masako; Wada, Masao; Akiba, Tsuneo; Yamada, Mitsuhiko

    2003-01-01

    A variable-pressure scanning electron microscopy (VP-SEM) with a cooling stage permits long hours of observation of water-containing specimens in their natural or close to natural state, without the conventional specimen preparations of fixation, dehydration, drying and metal coating. It reduces water vaporization and beam damage by keeping the specimens at a low temperature. We observed Bacillus cereus colonies on nutrient agar, which would shrink significantly if any conventional specimen preparation technique were used. We also studied the growing process of the bacteria on raw and steamed rice using the VP-SEM without conventional preparation techniques. Original specimens were directly mounted onto specimen holders and their backscattered electron images observed under the following conditions: specimen stage temperature, -10 degrees C; specimen chamber vacuum level, 30-70 Pa; and accelerating voltage, 15-20 kV. We recognized that the VP-SEM minimized deformation of the colonies due to shrinkage of the nutrient agar, and successfully imaged the morphology of the colonies and bacteria without a decline in bacteria number, which is apt to occur during fixation and dehydration. Also, the growth process of the bacteria on raw or steamed rice could be observed promptly, since there is no specimen preparation step.

  9. Spore development and nuclear inheritance in arbuscular mycorrhizal fungi

    Directory of Open Access Journals (Sweden)

    Hijri Mohamed

    2011-02-01

    Full Text Available Abstract Background A conventional tenet of classical genetics is that progeny inherit half their genome from each parent in sexual reproduction instead of the complete genome transferred to each daughter during asexual reproduction. The transmission of hereditary characteristics from parents to their offspring is therefore predictable, although several exceptions are known. Heredity in microorganisms, however, can be very complex, and even unknown as is the case for coenocytic organisms such as Arbuscular Mycorrhizal Fungi (AMF. This group of fungi are plant-root symbionts, ubiquitous in most ecosystems, which reproduce asexually via multinucleate spores for which sexuality has not yet been observed. Results We examined the number of nuclei per spore of four AMF taxa using high Z-resolution live confocal microscopy and found that the number of nuclei was correlated with spore diameter. We show that AMF have the ability, through the establishment of new symbioses, to pass hundreds of nuclei to subsequent generations of multinucleated spores. More importantly, we observed surprising heterogeneity in the number of nuclei among sister spores and show that massive nuclear migration and mitosis are the mechanisms by which AMF spores are formed. We followed spore development of Glomus irregulare from hyphal swelling to spore maturity and found that the spores reached mature size within 30 to 60 days, and that the number of nuclei per spores increased over time. Conclusions We conclude that the spores used for dispersal of AMF contain nuclei with two origins, those that migrate into the spore and those that arise by mitosis in the spore. Therefore, these spores do not represent a stage in the life cycle with a single nucleus, raising the possibility that AMF, unlike all other known eukaryotic organisms, lack the genetic bottleneck of a single-nucleus stage.

  10. The Pathogenomic Sequence Analysis of B. cereus and B. Thuringiensis isolates closely related to Bacillus anthracis

    Energy Technology Data Exchange (ETDEWEB)

    Han, C S; Xie, G; Challacombe, J F; Altherr, M R; Bhotika, S S; Bruce, D; Campbell, C S; Campbell, M L; Chen, J; Chertkov, O; Cleland, C; Dimitrijevic-Bussod, M; Doggett, N A; Fawcett, J J; Glavina, T; Goodwin, L A; Hill, K K; Hitchcock, P; Jackson, P J; Keim, P; Kewalramani, A R; Longmire, J; Lucas, S; Malfatti, S; McMurry, K; Meincke, L J; Misra, M; Moseman, B L; Mundt, M; Munk, A C; Okinaka, R T; Parson-Quintana, B; Reilly, L P; Richardson, P; Robinson, D L; Rubin, E; Saunders, E; Tapia, R; Tesmer, J G; Thayer, N; Thompson, L S; Tice, H; Ticknor, L O; Wills, P L; Gilna, P; Brettin, T S

    2005-10-12

    The sequencing and analysis of two close relatives of Bacillus anthracis are reported. AFLP analysis of over 300 isolates of B. cereus, B. thuringiensis and B. anthracis identified two isolates as being very closely related to B. anthracis. One, a B. cereus, BcE33L, was isolated from a zebra carcass in Nambia; the second, a B. thuringiensis, 97-27, was isolated from a necrotic human wound. The B. cereus appears to be the closest anthracis relative sequenced to date. A core genome of over 3,900 genes was compiled for the Bacillus cereus group, including B anthracis. Comparative analysis of these two genomes with other members of the B. cereus group provides insight into the evolutionary relationships among these organisms. Evidence is presented that differential regulation modulates virulence, rather than simple acquisition of virulence factors. These genome sequences provide insight into the molecular mechanisms contributing to the host range and virulence of this group of organisms.

  11. Fern spore longevity in saline water: can sea bottom sediments maintain a viable spore bank?

    Science.gov (United States)

    de Groot, G Arjen; During, Heinjo

    2013-01-01

    Freshwater and marine sediments often harbor reservoirs of plant diaspores, from which germination and establishment may occur whenever the sediment falls dry. Therewith, they form valuable records of historical inter- and intraspecific diversity, and are increasingly exploited to facilitate diversity establishment in new or restored nature areas. Yet, while ferns may constitute a considerable part of a vegetation's diversity and sediments are known to contain fern spores, little is known about their longevity, which may suffer from inundation and--in sea bottoms--salt stress. We tested the potential of ferns to establish from a sea or lake bottom, using experimental studies on spore survival and gametophyte formation, as well as a spore bank analysis on sediments from a former Dutch inland sea. Our experimental results revealed clear differences among species. For Asplenium scolopendrium and Gymnocarpium dryopteris, spore germination was not affected by inundated storage alone, but decreased with rising salt concentrations. In contrast, for Asplenium trichomanes subsp. quadrivalens germination decreased following inundation, but not in response to salt. Germination rates decreased with time of storage in saline water. Smaller and less viable gametophytes were produced when saline storage lasted for a year. Effects on germination and gametophyte development clearly differed among genotypes of A. scolopendrium. Spore bank analyses detected no viable spores in marine sediment layers. Only two very small gametophytes (identified as Thelypteris palustris via DNA barcoding) emerged from freshwater sediments. Both died before maturation. We conclude that marine, and likely even freshwater sediments, will generally be of little value for long-term storage of fern diversity. The development of any fern vegetation on a former sea floor will depend heavily on the deposition of spores onto the drained land by natural or artificial means of dispersal.

  12. Bacillus cereus un patógeno importante en el control microbiológico de los alimentos / Bacillus cereus an important pathogen the microbiological control of food

    Directory of Open Access Journals (Sweden)

    Jennifer Sánchez

    2016-05-01

    Full Text Available Resumen Bacillus cereus es una bacteria genéticamente diversa que se encuentra comúnmente en el ambiente. Contamina los alimentos afectando la salud humana, al ingerir el microorganismo y/o sus toxinas, la emética o las enterotoxinas. En Colombia son escasos los reportes de intoxicación por B. cereus y se estima que hay un gran subregistro. Por lo anterior, se recomienda aumentar la vigilancia de este patógeno y realizar estudios sobre aspectos relevantes que permitan aplicar medidas de control para disminuir las intoxicaciones por B. cereus. El objetivo de esta revisión bibliográfica es presentar información actualizada sobre B. cereus, que incluye aspectos de su biología, taxonomía, toxinas, alimentos que contamina y metodologías para detectar, prevenir y controlar este microorganismo. La información presentada es de utilidad para el público en general, especialmente personas vinculadas al sector de alimentos, inocuidad alimentaria y control de procesos. / Abstract Bacillus cereus is a genetically diverse bacterium commonly found in the environment. It contaminates food, thus affecting human health upon ingestion of the microorganism and/or its toxins, the emetic or enterotoxins. In Colombia, reports of intoxication by B. cereus are scarce and under-registration is presumed. Because of this, it is recommended to increase surveillance of this pathogen and to develop studies on relevant aspects that allow the application of control measures to reduce intoxications by B. cereus. The aim of this review is to present current information on B. cereus, including aspects of its biology, taxonomy, toxins, food that it contaminates and methodologies for the detection, prevention and control of this microorganism. This information is useful for the general public, especially people involved with the food sector, food safety and process control.

  13. Effects of two probiotic additives containing Bacillus spores on carcass characteristics, blood lipids and cecal volatile fatty acids in meat type chickens.

    Science.gov (United States)

    Novak, R; Bogovič Matijašić, B; Terčič, D; Cervek, M; Gorjanc, G; Holcman, A; Levart, A; Rogelj, I

    2011-08-01

    The objective of this study was to evaluate effects of two commercially available probiotic additives, containing Bacillus spores, on carcass and meat characteristics, serum lipids and concentration of cecal volatile fatty acids of meat type chickens. Birds were fed regular corn-soy meal based feed (control), supplemented with additive A, containing 1.6 × 10(6) spores per gram of feed of Bacillus subtilis and Bacillus licheniformis (group A) or additive B, containing the same concentration of Bacillus cereus var. toyoi spores (group B). One hundred and twenty birds (20 per replicate) were slaughtered at the age of 55 days. Results showed that birds in group B had higher (p blood serum cholesterol profile. Both probiotics influenced the cecal fermentation, which was observed as decrease in cecal concentrations of propionic, butyric, n-butyric and n-valeric acids, but the differences compared to control group were statistically significant for group A only. It was established that probiotic additive B was more effective regarding carcass and meat part weights than additive A, however the animals from group B also had more abdominal fat and their meat had significantly higher conductivity than control group, which is not considered as beneficial.

  14. Is Cytotoxin K from Bacillus cereus a bona fide enterotoxin?

    Science.gov (United States)

    Castiaux, Virginie; Liu, Xiaojin; Delbrassinne, Laurence; Mahillon, Jacques

    2015-10-15

    Cytotoxin K (CytK) produced by Bacillus cereus s.l. has generally been considered to be associated with the foodborne diarrhoeal syndrome. Two distinct variants of CytK have been reported: CytK-1 from Bacillus cytotoxicus and CytK-2 from B. cereus. In order to determine whether CytK plays a significant role in the diarrhoeal disease, the occurrence of cytK genes was assessed among 390 B. cereus isolates with different origins including clinical and food poisoning samples and was found to be 46%. Interestingly, the cytK occurrence was slightly lower in food poisoning and clinical isolates than in environmental samples. Seventy cytK-2 positive strains (including 28 isolates from foodborne outbreaks) were then selected in order to assess their genetic diversity. A genetic dendrogram based on the cytK-2 sequences of these 70 strains and on two cytK-1 sequences from strains NVH 391-98 and 883-00 showed an important diversity. However, no strain clustering according to the origin or source of isolation was observed. These observations were confirmed by Multi-Locus Sequences Typing (MLST) based on five different loci of housekeeping genes (ccpA, recF, sucC, purF and gdpD) for which no grouping of foodborne outbreak strains could be identified. Therefore, the choice of cytK as virulence factor for the diarrhoeal pathotype does not seem to be relevant per se, even though the involvement of CytK in the diarrhoeal syndrome cannot be fully excluded. Potential synergistic effects between CytK and other virulence factors, together with their potential variable expression levels should be further investigated.

  15. The PlcR virulence regulon of Bacillus cereus.

    Directory of Open Access Journals (Sweden)

    Michel Gohar

    Full Text Available PlcR is a Bacillus cereus transcriptional regulator, which activates gene expression by binding to a nucleotidic sequence called the 'PlcR box'. To build a list of all genes included in the PlcR regulon, a consensus sequence was identified by directed mutagenesis. The reference strain ATCC14579 sequenced genome was searched for occurrences of this consensus sequence to produce a virtual regulon. PlcR control of these genes was confirmed by comparing gene expression in the reference strain and its isogenic Delta-plcR strain using DNA microarrays, lacZ fusions and proteomics methods. The resulting list included 45 genes controlled by 28 PlcR boxes. Forty of the PlcR controlled proteins were exported, of which 22 were secreted in the extracellular medium and 18 were bound or attached to cell wall structures (membrane or peptidoglycan layer. The functions of these proteins were related to food supply (phospholipases, proteases, toxins, cell protection (bacteriocins, toxins, transporters, cell wall biogenesis and environment-sensing (two-component sensors, chemotaxis proteins, GGDEF family regulators. Four genes coded for cytoplasmic regulators. The PlcR regulon appears to integrate a large range of environmental signals, including food deprivation and self cell-density, and regulate the transcription of genes designed to overcome obstacles that hinder B. cereus growth within the host: food supply, host barriers, host immune defenses, and competition with other bacterial species. PlcR appears to be a key component in the efficient adaptation of B. cereus to its host environment.

  16. 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

    , 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....... 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......., Hall, & Kowbel 2011). Precise genetic distances between mating pairs were calculated to control for the effects of crossing distance on offspring production. We performed reciprocal crosses of all 121 strain pairings and collected data on perithecial production, ascospore (sexual spore) production...

  17. Detection of presumptive Bacillus cereus in the Irish dairy farm environment

    Directory of Open Access Journals (Sweden)

    O’Connell A.

    2016-12-01

    Full Text Available The objective of the study was to isolate potential Bacillus cereus sensu lato (B. cereus s.l. from a range of farm environments. Samples of tap water, milking equipment rinse water, milk sediment filter, grass, soil and bulk tank milk were collected from 63 farms. In addition, milk liners were swabbed at the start and the end of milking, and swabs were taken from cows’ teats prior to milking. The samples were plated on mannitol egg yolk polymyxin agar (MYP and presumptive B. cereus s.l. colonies were isolated and stored in nutrient broth with 20% glycerol and frozen at -80 °C. These isolates were then plated on chromogenic medium (BACARA and colonies identified as presumptive B. cereus s.l. on this medium were subjected to 16S ribosomal RNA (rRNA sequencing. Of the 507 isolates presumed to be B. cereus s.l. on the basis of growth on MYP, only 177 showed growth typical of B. cereus s.l. on BACARA agar. The use of 16S rRNA sequencing to identify isolates that grew on BACARA confirmed that the majority of isolates belonged to B. cereus s.l. A total of 81 of the 98 isolates sequenced were tentatively identified as presumptive B. cereus s.l. Pulsed-field gel electrophoresis was carried out on milk and soil isolates from seven farms that were identified as having presumptive B. cereus s.l. No pulsotype was shared by isolates from soil and milk on the same farm. Presumptive B. cereus s.l. was widely distributed within the dairy farm environment.

  18. Isolation of Bacillus Cereus from wounds and burns

    Directory of Open Access Journals (Sweden)

    Behzadiannejhad Gh

    1997-09-01

    Full Text Available The culture results of 203 cases with different wounds were studies; 150 of the latter were burn cases (mainly second and third degree burns, and 53 were of other types (surgical, traumatic, ect. Four subtypes of Bacillus cereus were isolated upon culture, and the different toxins produced in DHT broth with 0.1% glucose were assessed. The lethal toxin was injected intravenously to Syrian rats, none of whom died. VPR factor was assessed in the 4 subtypes. Three subtypes produced VPR in significant amounts.

  19. CLOSTRIDIUM SPORE ATTACHMENT TO HUMAN CELLS

    Energy Technology Data Exchange (ETDEWEB)

    PANESSA-WARREN,B.; TORTORA,G.; WARREN,J.

    1997-08-10

    This paper uses high resolution scanning electron microscopy (SEM) with a LaB6 gun and the newest commercial field emission guns, to obtain high magnification images of intact clostridial spores throughout the activation/germination/outgrowth process. By high resolution SEM, the clostridial exosporial membrane can be seen to produce numerous delicate projections (following activation), that extend from the exosporial surface to a nutritive substrate (agar), or cell surface when anaerobically incubated in the presence of human cells (embryonic fibroblasts and colon carcinoma cells). Magnifications of 20,000 to 200,000Xs at accelerating voltages low enough to minimize or eliminate specimen damage (1--5 kV) have permitted the entire surface of C.sporogenes and C.difficile endospores to be examined during all stages of germination. The relationships between the spore and the agar or human cell surface were also clearly visible.

  20. Phosphoproteome dynamics mediate revival of bacterial spores

    OpenAIRE

    2015-01-01

    Background Bacterial spores can remain dormant for decades, yet harbor the exceptional capacity to rapidly resume metabolic activity and recommence life. Although germinants and their corresponding receptors have been known for more than 30 years, the molecular events underlying this remarkable cellular transition from dormancy to full metabolic activity are only partially defined. Results Here, we examined whether protein phospho-modifications occur during germination, the first step of exit...

  1. 3种细菌芽胞遗传同源性及抗力对比研究%Comparison of Genetic Homology and Resistance of Three Bacillus Spores

    Institute of Scientific and Technical Information of China (English)

    许斌; 程立庆; 黄国荣; 张林; 饶中林; 张廷惠; 熊鸿燕

    2009-01-01

    比较细菌芽胞的遗传同源性、结构和抗力差异,为炭疽芽胞的替代试验菌的可靠评价提供依据.采用资料检索、生物信息分析、显微镜观察和微生物学技术分析不同芽胞的遗传同源性、超微结构和抗力差异.炭疽芽胞与腊样芽胞的结构和大小相似,生物遗传同源性最近,对热力、UVC和有效氯的抗力相近.腊样芽胞对炭疽芽胞的代表性最好,可以倾向性选用其替代炭疽芽胞进行试验研究.%Genetic homology, structure, and resistance difference of bacillus spores were compared to provide founda-tion to evaluate the reliability of experimental substitute for Bacillus anthracis spore using data-search, bio-information analysis, microscopic observation, and microbiological technology to analyze the genetic homology, ultrastructure, and resistance difference of different spores. The results showed that the structure and the size of B. anthracis was similar to those of B. cereus, very close in genetic homology, close in the resistance against heat, UVC, and effective chlo-rine. Therefore, B. cereus had the best representativeness of B. anthracis, and could be used tendentiously and selec-tively to study their spores.

  2. Methyl Iodide Fumigation of Bacillus anthracis Spores.

    Science.gov (United States)

    Sutton, Mark; Kane, Staci R; Wollard, Jessica R

    2015-09-01

    Fumigation techniques such as chlorine dioxide, vaporous hydrogen peroxide, and paraformaldehyde previously used to decontaminate items, rooms, and buildings following contamination with Bacillus anthracis spores are often incompatible with materials (e.g., porous surfaces, organics, and metals), causing damage or residue. Alternative fumigation with methyl bromide is subject to U.S. and international restrictions due to its ozone-depleting properties. Methyl iodide, however, does not pose a risk to the ozone layer and has previously been demonstrated as a fumigant for fungi, insects, and nematodes. Until now, methyl iodide has not been evaluated against Bacillus anthracis. Sterne strain Bacillus anthracis spores were subjected to methyl iodide fumigation at room temperature and at 550C. Efficacy was measured on a log-scale with a 6-log reduction in CFUs being considered successful compared to the U.S. Environmental Protection Agency biocide standard. Such efficacies were obtained after just one hour at 55 °C and after 12 hours at room temperature. No detrimental effects were observed on glassware, PTFE O-rings, or stainless steel. This is the first reported efficacy of methyl iodide in the reduction of Bacillus anthracis spore contamination at ambient and elevated temperatures.

  3. Role of YpeB in Cortex Hydrolysis during Germination of Bacillus anthracis Spores

    OpenAIRE

    2014-01-01

    The infectious agent of the disease anthrax is the spore of Bacillus anthracis. Bacterial spores are extremely resistant to environmental stresses, which greatly hinders spore decontamination efforts. The spore cortex, a thick layer of modified peptidoglycan, contributes to spore dormancy and resistance by maintaining the low water content of the spore core. The cortex is degraded by germination-specific lytic enzymes (GSLEs) during spore germination, rendering the cells vulnerable to common ...

  4. Airborne Spore Analysis of Karabük Atmosphere

    Directory of Open Access Journals (Sweden)

    Ayşe Kaplan

    2014-05-01

    Full Text Available In order to identify types and amounts of airborne allergenic spore dispersal in the atmosphere of Karabük by gravimetric method in 2006 and 2007, two Durham samplers were situated on roof and garden of Technical Education Faculty of Karabük University between the dates January 1, 2006 and December 31, 2007. As a result of the analysis a total of 2822.2±625.01 spore/cm2 spore quantity belonging to 21 types was identified. Of this total, 1106±250.33 spore/cm² was observed in 2006 and 1716±374.68 spore/cm² was observed in 2007. Spore concentrations revealed no statistically differences between two samplers (t=0.1527-1.1355, p>0.05. The relationship between spore concentrations and meteorological factors was displayed by Spearman Correlation analysis. The highest quantity of fungal spores and Myxomycetes were determined in June and July. Cladosporium, Alternaria, Ustilago, Myxomycetes and unidentified Ascomycetes spores were recorded as dominant. In the end of this study, a two-year spore calendar was prepared.

  5. Thirty-four identifiable airborne fungal spores in Havana, Cuba.

    Science.gov (United States)

    Almaguer, Michel; Aira, María-Jesús; Rodríguez-Rajo, F Javier; Fernandez-Gonzalez, Maria; Rojas-Flores, Teresa I

    2015-01-01

    The airborne fungal spore content in Havana, Cuba, collected by means a non-viable volumetric methodology, was studied from November 2010 - October 2011. The study, from a qualitative point of view, allowed the characterization of 29 genera and 5 fungal types, described following the Saccardo´s morphotypes, as well as their morphobiometrical characteristics. In the amerospores morphotype, the conidia of 7 genera (with ascospores, basidiospores and uredospores) and 5 fungal types were included. Among phragmospores morphotype, the ascospores and conidia of 12 different genera were identified. The dictyospores morphotype only included conidial forms from 6 genera. Finally, the less frequent morphotypes were staurospores, didymospores and distosepted spores. In general, the main worldwide spread mitosporic fungi also predominated in the Havana atmosphere, accompanied by some ascospores and basidiospores. Cladosporium cladosporioides type was the most abundant with a total of 148,717 spores, followed by Leptosphaeria, Coprinus and the Aspergillus-Penicillium type spores, all of them with total values ranging from 20,591 - 16,392 spores. The higher monthly concentrations were registered in January (31,663 spores) and the lowest in December (7,314 spores). Generally, the average quantity of spores recorded during the months of the dry season (20,599 spores) was higher compared with that observed during the rainy season (17,460 spores).

  6. Thirty-four identifiable airborne fungal spores in Havana, Cuba

    Directory of Open Access Journals (Sweden)

    Michel Almaguer

    2015-05-01

    Full Text Available The airborne fungal spore content in Havana, Cuba, collected by means a non-viable volumetric methodology, was studied from November 2010 – October 2011. The study, from a qualitative point of view, allowed the characterization of 29 genera and 5 fungal types, described following the Saccardo´s morphotypes, as well as their morphobiometrical characteristics. In the amerospores morphotype, the conidia of 7 genera (with ascospores, basidiospores and uredospores and 5 fungal types were included. Among phragmospores morphotype, the ascospores and conidia of 12 different genera were identified. The dictyospores morphotype only included conidial forms from 6 genera. Finally, the less frequent morphotypes were staurospores, didymospores and distosepted spores. In general, the main worldwide spread mitosporic fungi also predominated in the Havana atmosphere, accompanied by some ascospores and basidiospores. [i]Cladosporium[/i] cladosporioides type was the most abundant with a total of 148,717 spores, followed by [i]Leptosphaeria, Coprinus[/i] and the [i]Aspergillus-Penicillium [/i]type spores, all of them with total values ranging from 20,591 – 16,392 spores. The higher monthly concentrations were registered in January (31,663 spores and the lowest in December (7,314 spores. Generally, the average quantity of spores recorded during the months of the dry season (20,599 spores was higher compared with that observed during the rainy season (17,460 spores.

  7. Bryophyte spore germinability is inhibited by peatland substrates

    Science.gov (United States)

    Bu, Zhao-Jun; Li, Zhi; Liu, Li-Jie; Sundberg, Sebastian; Feng, Ya-Min; Yang, Yun-He; Liu, Shuang; Song, Xue; Zhang, Xing-Lin

    2017-01-01

    Bryophyte substrates and species may affect spore germination through allelopathy. Polytrichum strictum is currently expanding in peatlands in north-eastern China - is this an effect of its superior spore germinability or do its gametophytes have a stronger allelopathic effect than do Sphagnum? We conducted a spore burial experiment to test the effect of species identity, substrate and water table depth (WTD) on spore germinability and bryophyte allelopathic effect with P. strictum and two Sphagnum species (S. palustre and S. magellanicum). After 5 months of burial during a growing season, the spores were tested for germinability. Allelopathic effect of bryophyte substrates was assessed by the difference between spore germinability after being stored inside or outside the substrates. After burial, more than 90% of the spores lost their germinability across all three species due to ageing and allelopathy. Spore germinability differed among species, where the spores in S. palustre had a higher germination frequency than those in P. strictum. The three bryophytes maintained a higher germinability in Sphagnum than in Polytrichum hummocks, probably due to a stronger allelopathic effect of P. strictum. Water table drawdown by 10 cm increased germinability by more than 60% across the three species. The study indicates that P. strictum does not possess an advantage regarding spore germination but rather its gametophytes have a stronger allelopathic effect. Due to the weaker inhibitive effect of Sphagnum gametophytes, P. strictum may have a potential establishment superiority over Sphagnum in peatlands, in addition to a better drought tolerance, which may explain its current expansion.

  8. Cash management system 'CEREUS-SM'; Supermarket muke kinsen shoriki 'CEREUS-SM'

    Energy Technology Data Exchange (ETDEWEB)

    Niizuma, N.; Fukushima, Y.; Kinoshita, S. [Fuji Electric Co. Ltd., Tokyo (Japan)

    2000-03-10

    The adjustment system with change in coin has reduced cashier work in supermarkets, and has spread as the standard peripheral equipment of the point of sales (POS) system. Recently, there have been customer needs for a type with change in bill and coin. Fuji Electric has developed cash management system 'CEREUS-SM' which is as small as the current system only with change in coin and can be handled by anyone, including a part-timer or a student worker. (author)

  9. Erratum to: Seasonal trend and clinical presentation of Bacillus cereus bloodstream infection: association with summer and indwelling catheter.

    Science.gov (United States)

    Kato, K; Matsumura, Y; Yamamoto, M; Nagao, M; Ito, Y; Takakura, S; Ichiyama, S

    2016-05-01

    Bacillus cereus, an opportunistic pathogen, can cause fatal infection. However, B. cereus bloodstream infections (BSIs) have not been well characterised. From 2008 to 2013, B. cereus isolates from all of the specimens and patients with B. cereus BSIs were identified. Environmental samples were collected to detect B. cereus contamination. We also characterised the clinical presentation of B. cereus BSI through analyses of risk factors for BSI and mortality. A total of 143 clinical B. cereus isolates was detected. Fifty-one patients with nosocomial infections were diagnosed as B. cereus BSI, and 37 had contaminated blood cultures. The number of B. cereus isolates and BSI patients was significantly greater from June to September than from January to April (3.4 vs. 1.0 per month and 1.4 vs. 0.2, respectively). All BSIs were nosocomial and related to central or peripheral vascular catheter. Urinary catheter [odds ratio (OR) 6.93, 95 % confidence interval (CI) 2.40-20.0] was the independent risk factor associated with BSI patients when compared to patients regarded as contaminated. In-hospital mortality among BSI patients was 20 % and was associated with urinary catheter (OR 12.3, 95 % CI 0.67-225, p=0.045) and higher Charlson index (OR 1.99, 95 % CI 1.26-3.12). The number of B. cereus isolates and BSI increased during summer. Inpatients with indwelling vascular or urinary catheters should be carefully monitored for potential B. cereus BSIs.

  10. Bacillus subtilis HJ18-4 from traditional fermented soybean food inhibits Bacillus cereus growth and toxin-related genes.

    Science.gov (United States)

    Eom, Jeong Seon; Lee, Sun Young; Choi, Hye Sun

    2014-11-01

    Bacillus subtilis HJ18-4 isolated from buckwheat sokseongjang, a traditional Korean fermented soybean food, exhibits broad-spectrum antimicrobial activity against foodborne pathogens, including Bacillus cereus. In this study, we investigated the antibacterial efficacy and regulation of toxin gene expression in B. cereus by B. subtilis HJ18-4. Expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM) was downregulated by B. subtilis HJ18-4, which also exhibited strong antibacterial activity against B. cereus. We also found that water extracts of soy product fermented with B. subtilis HJ18-4 significantly inhibited the growth of B. cereus and toxin expression. These results indicate that B. subtilis HJ18-4 could be used as an antimicrobial agent to control B. cereus in the fermented soybean food industry. Our findings also provide an opportunity to develop an efficient biological control agent against B. cereus.

  11. Genetic relationships between sympatric populations of Bacillus cereus and Bacillus thuringiensis, as revealed by rep-PCR genomic fingerprinting

    Directory of Open Access Journals (Sweden)

    Ana Paula S Peruca

    2008-08-01

    Full Text Available The bacterial strain Bacillus cereus is closely related to Bacillus thuringiensis, although any genetic relationship between the two strains is still in debate. Using rep-PCR genomic fingerprinting, we established the genetic relationships between Brazilian sympatric populations of B. cereus and B. thuringiensis simultaneously collected from two geographically separate sites. We observed the formation of both B. thuringiensis and B. cereus clusters, as well as strains of B. cereus that are more closely related to B. thuringiensis than to other B. cereus strains. In addition, lower genetic variability was observed among B. thuringiensis clusters compared to B. cereus clusters, indicating that either the two species should be categorized as separate or that B. thuringiensis may represent a clone from a B. cereus background.

  12. Assessment of hydrophobicity and roughness of stainless steel adhered by an isolate of Bacillus cereus from a dairy plant

    Directory of Open Access Journals (Sweden)

    Patrícia Campos Bernardes

    2010-12-01

    Full Text Available The interaction between the surface of stainless steel and Bacillus cereus was studied in terms of the characteristics of interfacial interaction determined from the measurement of the contact angle of the surface of B. cereus and stainless steel in the presence or absence of B. cereus adherence. The microtopographies and the roughness of the surface of stainless steel and stainless steel adhered by B. cereus were evaluated with the help of atomic force microscopy and perfilometry. The strain of B. cereus studied was considered hydrophilic, whereas the stainless steel was considered hydrophobic. The adhesion was not thermodynamically favorable (ΔGadhesion > 0 between the stainless steel and the strain of B. cereus studied. Thus, the interaction between them was not favored by the thermodynamic aspect of adhesion. There was no difference (p > 0.05 in the roughness of the surfaces of stainless steel adhered by B. cereus when analyzed by atomic force microscope and perfilometry.

  13. Assessment of hydrophobicity and roughness of stainless steel adhered by an isolate of Bacillus cereus from a dairy plant

    Science.gov (United States)

    Bernardes, Patrícia Campos; de Andrade, Nélio José; Ferreira, Sukarno Olavo; de Sá, João Paulo Natalino; Araújo, Emiliane Andrade; Delatorre, Deyse Maria Zanom; Luiz, Lívia Maria Pinheiro

    2010-01-01

    The interaction between the surface of stainless steel and Bacillus cereus was studied in terms of the characteristics of interfacial interaction determined from the measurement of the contact angle of the surface of B. cereus and stainless steel in the presence or absence of B. cereus adherence. The microtopographies and the roughness of the surface of stainless steel and stainless steel adhered by B. cereus were evaluated with the help of atomic force microscopy and perfilometry. The strain of B. cereus studied was considered hydrophilic, whereas the stainless steel was considered hydrophobic. The adhesion was not thermodynamically favorable (ΔGadhesion > 0) between the stainless steel and the strain of B. cereus studied. Thus, the interaction between them was not favored by the thermodynamic aspect of adhesion. There was no difference (p > 0.05) in the roughness of the surfaces of stainless steel adhered by B. cereus when analyzed by atomic force microscope and perfilometry. PMID:24031578

  14. [Morphologic detection of Bacillus cereus in blank cartridges].

    Science.gov (United States)

    Rothschild, M A; Mülling, C

    1998-01-01

    Wound infections after gunshot wounds from live ammunition can produce serious complications. It is well known that projectiles per se are neither sterile nor does their firing cause sterilization. The germs on the surface of a projectile enter the body together with the projectile and are thus introduced into the wound together with skin bacteria. However it is less known that wound infections can occur in wounds caused by the gas jet from blank ammunition (mainly from shots at very close range). In such ammunition without a projectile, the propellant particles are usually contaminated with bacteria which find their way into the wound together with skin germs. In previous investigations, we have microbiologically detected the species Bacillus cereus in the propellant of blank cartridges. In the present study, we have applied scanning electron microscopic methods to find out which areas of the blank cartridges are colonized by these bacteria. For this purpose 20 blank cartridges, each from 4 different manufacturers, were electronmicroscopically examined. B. cereus only found on the surface of intact nitrocellulose particles but not in the interior of broken prepared propellant particles. Bacterial structures were not morphologically identified on black powder particles.

  15. Bacillus cereus var. toyoi enhanced systemic immune response in piglets.

    Science.gov (United States)

    Schierack, Peter; Wieler, Lothar H; Taras, David; Herwig, Volker; Tachu, Babila; Hlinak, Andreas; Schmidt, Michael F G; Scharek, Lydia

    2007-07-15

    Probiotic bacteria have been suggested to stimulate the host immune system. In this study we evaluated the immunomodulatory effects of probiotic Bacillus cereus var. toyoi on the systemic immunity of piglets. A pool of 70 piglets was divided into a probiotic or control group. We determined the ratios of peripheral blood mononuclear cell (PBMC) subsets and measured proliferative responses and cytokine production of PBMCs and effects on vaccination responses. Blood samples of probiotic-treated piglets showed a significantly lower frequency of CD8(high)/CD3+ T cells and CD8(low)/CD3+ T cells and a significant higher CD4+/CD8+ ratio. IL-4 and IFN-gamma production of polyclonally stimulated PBMCs was on average higher in the probiotic group. Specific proliferative responses of PBMCs to Influenza vaccination antigens were significantly higher and antibody titers against H3N2 Influenza and Mycoplasma vaccination antigens were on average higher in the probiotic group. In conclusion, B. cereus var. toyoi therefore alters the immune status of piglets as indicated by changes in the ratios as well as functionalities of systemic immune cell populations.

  16. Environment driven cereulide production by emetic strains of Bacillus cereus.

    Science.gov (United States)

    Apetroaie-Constantin, Camelia; Shaheen, Ranad; Andrup, Lars; Smidt, Lasse; Rita, Hannu; Salkinoja-Salonen, Mirja

    2008-09-30

    The impacts of growth media and temperature on production of cereulide, the emetic toxin of Bacillus cereus, were measured for seven well characterised strains selected for diversity of biochemical and genetic properties and sources of origin. All strains carried cereulide synthase gene, ces, on a megaplasmid of ca. 200 kb and all grew up to 48-50 degrees C, but produced cereulide only up to 39 degrees C. On tryptic soy agar five strains, originating from foods, food poisonings and environment, produced highest amounts of cereulide at 23 to 28 degrees C, whereas two strains, from human faeces, produced cereulide similarly from 23 to 39 degrees C, with no clear temperature trend. These two strains differed from the others also by producing more cereulide on tryptic soy agar if supplemented with 5 vol.% of blood, whereas the other five strains produced similarly, independent on the presence of blood. On oatmeal agar only one strain produced major amounts of cereulide. On skim milk agar, raw milk agar, and MacConkey agar most strains grew well but produced only low amounts of cereulide. Three media components, the ratio [K+]:[Na+], contents of glycine and [Na+], appeared of significance for predicting cereulide production. Increase of [K+]:[Na+] (focal variable) predicted (P cereus in a complex manner. The relevance of the findings to production of cereulide in the gut and to the safety of amino acids as additives in foods containing live toxinogenic organisms is discussed.

  17. Investigating synergism during sequential inactivation of Bacillus subtilis spores with several disinfectants.

    Science.gov (United States)

    Cho, Min; Kim, Jae-Hong; Yoon, Jeyong

    2006-08-01

    The sequential application of ozone, chlorine dioxide, or UV followed by free chlorine was performed to investigate the synergistic inactivation of Bacillus subtilis spores. The greatest synergism was observed when chlorine dioxide was used as a primary disinfectant followed by secondary disinfection with free chlorine. A lesser synergistic effect was observed when ozone was used as the primary disinfectant, but no synergism was observed when UV was used as the primary disinfectant. When free chlorine was used as the primary disinfectant (i.e., sequential application in the reverse order), the synergistic effect was shown only when chlorine dioxide was applied as the secondary disinfectant. The synergistic effect observed could be related to damage to the spore coat during primary disinfection, suggested by the loss of proteins from spores during disinfectant treatment. The greatest synergism observed by the chlorine dioxide/free chlorine pair suggested that common reaction sites might exist for these disinfectants. The concept of percent synergistic effect was introduced to quantitatively compare the extent of synergistic effects in the sequential disinfection processes.

  18. Development of a heat-stable and orally delivered recombinant M2e-expressing B. subtilis spore-based influenza vaccine.

    Science.gov (United States)

    Zhao, Guangyu; Miao, Yu; Guo, Yan; Qiu, Hongjie; Sun, Shihui; Kou, Zhihua; Yu, Hong; Li, Junfeng; Chen, Yue; Jiang, Shibo; Du, Lanying; Zhou, Yusen

    2014-01-01

    Highly conserved ectodomain of influenza virus M2 protein (M2e) is an important target for the development of universal influenza vaccines. Today, the use of chemical or genetic fusion constructs have been undertaken to overcome the low immunogenicity of M2e in vaccine formulation. However, current M2e vaccines are neither orally delivered nor heat-stable. In this study, we evaluated the immune efficacy of an orally delivered recombinant M2e vaccine containing 3 molcules of M2e consensus sequence of influenza A viruses, termed RSM2e3. To accomplish this, CotB, a spore coat of Bacillus subtilis (B. subtilis), was used as a fusion partner, and heat-stable nonpathogenic B. subtilis spores were used as the carrier. Our results showed that CotB-M2e3 fusion had no effect on spore structure or function in the resultant recombinant RSM2e3 strain and that heterologous influenza virus M2e protein was successfully displayed on the surface of the recombinant RSM2e3 spore. Importantly, recombinant RSM2e3 spores elicited strong and long-term M2e-specific systemic and mucosal immune responses, completely protecting immunized mice from lethal challenge of A/PR/8/34(H1N1) influenza virus. Taken together, our study forms a solid basis for the development of a novel orally delivered and heat-stable influenza vaccine based on B. subtilis spore surface display.

  19. Occurrence of Toxigenic Bacillus cereus and Bacillus thuringiensis in Doenjang, a Korean Fermented Soybean Paste.

    Science.gov (United States)

    Park, Kyung Min; Kim, Hyun Jung; Jeong, Moon Cheol; Koo, Minseon

    2016-04-01

    This study determined the prevalence and toxin profile of Bacillus cereus and Bacillus thuringiensis in doenjang, a fermented soybean food, made using both traditional and commercial methods. The 51 doenjang samples tested were broadly contaminated with B. cereus; in contrast, only one sample was positive for B. thuringiensis. All B. cereus isolates from doenjang were positive for diarrheal toxin genes. The frequencies of nheABC and hblACD in traditional samples were 22.7 and 0%, respectively, whereas 5.1 and 5.1% of B. cereus isolates from commercial samples possessed nheABC and hblACD, respectively. The detection rate of ces gene was 10.8%. The predominant toxin profile among isolates from enterotoxigenic B. cereus in doenjang was profile 4 (entFM-bceT-cytK). The major enterotoxin genes in emetic B. cereus were cytK, entFM, and nheA genes. The B. thuringiensis isolate was of the diarrheagenic type. These results provide a better understanding of the epidemiology of the enterotoxigenic and emetic B. cereus groups in Korean fermented soybean products.

  20. Bacillus cereus induces permeability of an in vitro blood-retina barrier.

    Science.gov (United States)

    Moyer, A L; Ramadan, R T; Thurman, J; Burroughs, A; Callegan, M C

    2008-04-01

    Most Bacillus cereus toxin production is controlled by the quorum-sensing-dependent, pleiotropic global regulator plcR, which contributes to the organism's virulence in the eye. The purpose of this study was to analyze the effects of B. cereus infection and plcR-regulated toxins on the barrier function of retinal pigment epithelium (RPE) cells, the primary cells of the blood-retina barrier. Human ARPE-19 cells were apically inoculated with wild-type or quorum-sensing-deficient B. cereus, and cytotoxicity was analyzed. plcR-regulated toxins were not required for B. cereus-induced RPE cytotoxicity, but these toxins did increase the rate of cell death, primarily by necrosis. B. cereus infection of polarized RPE cell monolayers resulted in increased barrier permeability, independent of plcR-regulated toxins. Loss of both occludin and ZO-1 expression occurred by 8 h postinfection, but alterations in tight junctions appeared to precede cytotoxicity. Of the several proinflammatory cytokines analyzed, only interleukin-6 was produced in response to B. cereus infection. These results demonstrate the deleterious effects of B. cereus infection on RPE barrier function and suggest that plcR-regulated toxins may not contribute significantly to RPE barrier permeability during infection.

  1. Probiotic Bacillus cereus Strains, a Potential Risk for Public Health in China.

    Science.gov (United States)

    Zhu, Kui; Hölzel, Christina S; Cui, Yifang; Mayer, Ricarda; Wang, Yang; Dietrich, Richard; Didier, Andrea; Bassitta, Rupert; Märtlbauer, Erwin; Ding, Shuangyang

    2016-01-01

    Bacillus cereus is an important cause of foodborne infectious disease and food poisoning. However, B. cereus has also been used as a probiotic in human medicine and livestock production, with low standards of safety assessment. In this study, we evaluated the safety of 15 commercial probiotic B. cereus preparations from China in terms of mislabeling, toxin production, and transferable antimicrobial resistance. Most preparations were incorrectly labeled, as they contained additional bacterial species; one product did not contain viable B. cereus at all. In total, 18 B. cereus group strains-specifically B. cereus and Bacillus thuringiensis-were isolated. Enterotoxin genes nhe, hbl, and cytK1, as well as the ces-gene were assessed by PCR. Enterotoxin production and cytotoxicity were confirmed by ELISA and cell culture assays, respectively. All isolated B. cereus group strains produced the enterotoxin Nhe; 15 strains additionally produced Hbl. Antimicrobial resistance was assessed by microdilution; resistance genes were detected by PCR and further characterized by sequencing, transformation and conjugation assays. Nearly half of the strains harbored the antimicrobial resistance gene tet(45). In one strain, tet(45) was situated on a mobile genetic element-encoding a site-specific recombination mechanism-and was transferable to Staphylococcus aureus and Bacillus subtilis by electro-transformation. In view of the wide and uncontrolled use of these products, stricter regulations for safety assessment, including determination of virulence factors and transferable antimicrobial resistance genes, are urgently needed.

  2. Probiotic Bacillus cereus strains, a potential risk for public health in China

    Directory of Open Access Journals (Sweden)

    Kui eZhu

    2016-05-01

    Full Text Available Bacillus cereus is an important cause of foodborne infectious disease and food poisoning. However, B. cereus has also been used as a probiotic in human medicine and livestock production, with low standards of safety assessment. In this study, we evaluated the safety of 15 commercial probiotic B. cereus preparations from China in terms of mislabeling, toxin production, and transferable antimicrobial resistance. Most preparations were incorrectly labeled, as they contained additional bacterial species; one product did not contain viable B. cereus at all. In total, 18 B. cereus group strains – specifically B. cereus and B. thuringiensis – were isolated. Enterotoxin genes nhe, hbl, and cytK1, as well as the ces-gene were assessed by PCR. Enterotoxin production and cytotoxicity were confirmed by ELISA and cell culture assays, respectively. All isolated B. cereus group strains produced the enterotoxin Nhe; 15 strains additionally produced Hbl. Antimicrobial resistance was assessed by microdilution; resistance genes were detected by PCR and further characterized by sequencing, transformation and conjugation assays. Nearly half of the strains harbored the antimicrobial resistance gene tet(45. In one strain, tet(45 was situated on a mobile genetic element – encoding a site specific recombination mechanism – and was transferable to Staphylococcus aureus and B. subtilis by electro-transformation. In view of the wide and uncontrolled use of these products, stricter regulations for safety assessment, including determination of virulence factors and transferable antimicrobial resistance genes, are urgently needed.

  3. 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.

  4. Protection of Penaeus monodon against white spot syndrome by continuous oral administration of a low concentration of Bacillus subtilis spores expressing the VP28 antigen.

    Science.gov (United States)

    Pham, K-C; Tran, H T T; Van Doan, C; Le, P H; Van Nguyen, A T; Nguyen, H A; Hong, H A; Cutting, S M; Phan, T-N

    2017-03-01

    In this study, Bacillus subtilis spores expressing a chimeric protein, CotB-VP28, were used as a probiotic vaccine to protect black tiger shrimps (Penaeus monodon) against white spot syndrome virus (WSSV) infection. Oral administration of pellets coated with CotB-VP28 spores (at ≥1 × 10(9 ) CFU per g pellet) to shrimps induced immune-relating phenoloxydase activity (PO) in shrimps after 14 days of feeding (prior challenge) and at day 3 post challenge (1·26 and 1·70 fold increase respectively). A 75% protection rate was obtained by continuous feeding of the spore-coated pellets at ≥1 × 10(9 ) CFU per g for 14 days prior to WSSV challenge and during all the postchallenge period. Even when the amount of CotB-VP28 spores in feed pellets was reduced down to ≥5 × 10(7)  CFU per g and ≥1 × 10(6)  CFU per g, relatively high protection rates of 70 and 67·5%, respectively, were still obtained. By contrast, feeding pellets without spores (untreated group) and with naked spores (PY79 group) at ≥1 × 10(9)  CFU per g could not protect shrimps against WSSV. These data suggest that supplementation of CotB-VP28 spores at low dose of ≥1 × 10(6)  CFU per g could be effective as a prophylactic treatment of WSS for black tiger shrimps.

  5. Genetic diversity of clinical isolates of Bacillus cereus using multilocus sequence typing

    Directory of Open Access Journals (Sweden)

    Pruckler James M

    2008-11-01

    Full Text Available Abstract Background Bacillus cereus is most commonly associated with foodborne illness (diarrheal and emetic but is also an opportunistic pathogen that can cause severe and fatal infections. Several multilocus sequence typing (MLST schemes have recently been developed to genotype B. cereus and analysis has suggested a clonal or weakly clonal population structure for B. cereus and its close relatives B. anthracis and B. thuringiensis. In this study we used MLST to determine if B. cereus isolates associated with illnesses of varying severity (e.g., severe, systemic vs. gastrointestinal (GI illness were clonal or formed clonal complexes. Results A retrospective analysis of 55 clinical B. cereus isolates submitted to the Centers for Disease Control and Prevention between 1954 and 2004 was conducted. Clinical isolates from severe infections (n = 27, gastrointestinal (GI illness (n = 18, and associated isolates from food (n = 10 were selected for analysis using MLST. The 55 isolates were diverse and comprised 38 sequence types (ST in two distinct clades. Of the 27 isolates associated with serious illness, 13 clustered in clade 1 while 14 were in clade 2. Isolates associated with GI illness were also found throughout clades 1 and 2, while no isolates in this study belonged to clade 3. All the isolates from this study belonging to the clade 1/cereus III lineage were associated with severe disease while isolates belonging to clade1/cereus II contained isolates primarily associated with severe disease and emetic illness. Only three STs were observed more than once for epidemiologically distinct isolates. Conclusion STs of clinical B. cereus isolates were phylogenetically diverse and distributed among two of three previously described clades. Greater numbers of strains will need to be analyzed to confirm if specific lineages or clonal complexes are more likely to contain clinical isolates or be associated with specific illness, similar to B. anthracis and

  6. Bacteriophage PBC1 and its endolysin as an antimicrobial agent against Bacillus cereus.

    Science.gov (United States)

    Kong, Minsuk; Ryu, Sangryeol

    2015-04-01

    Bacillus cereus is an opportunistic human pathogen responsible for food poisoning and other, nongastrointestinal infections. Due to the emergence of multidrug-resistant B. cereus strains, the demand for alternative therapeutic options is increasing. To address these problems, we isolated and characterized a Siphoviridae virulent phage, PBC1, and its lytic enzymes. PBC1 showed a very narrow host range, infecting only 1 of 22 B. cereus strains. Phylogenetic analysis based on the major capsid protein revealed that PBC1 is more closely related to the Bacillus clarkii phage BCJA1c and phages of lactic acid bacteria than to the phages infecting B. cereus. Whole-genome comparison showed that the late-gene region, including the terminase gene, structural genes, and holin gene of PBC1, is similar to that from B. cereus temperate phage 250, whereas their endolysins are different. Compared to the extreme host specificity of PBC1, its endolysin, LysPBC1, showed a much broader lytic spectrum, albeit limited to the genus Bacillus. The catalytic domain of LysPBC1 when expressed alone also showed Bacillus-specific lytic activity, which was lower against the B. cereus group but higher against the Bacillus subtilis group than the full-length protein. Taken together, these results suggest that the virulent phage PBC1 is a useful component of a phage cocktail to control B. cereus, even with its exceptionally narrow host range, as it can kill a strain of B. cereus that is not killed by other phages, and that LysPBC1 is an alternative biocontrol agent against B. cereus.

  7. The Molecular Timeline of a Reviving Bacterial Spore

    OpenAIRE

    2015-01-01

    Summary The bacterial spore can rapidly convert from a dormant to a fully active cell. Here we study this remarkable cellular transition in Bacillus subtilis and reveal the identity of the newly synthesized proteins throughout spore revival. Our analysis uncovers a highly ordered developmental program that correlates with the spore morphological changes and reveals the spatial and temporal molecular events fundamental to reconstruct a cell. As opposed to current knowledge, we found that trans...

  8. Biocidal Energetic Materials for the Destruction of Spore Forming Bacteria

    Science.gov (United States)

    2015-07-01

    L R E P O R T DTRA-TR-13-52 Biocidal Energetic Materials for the Destruction of Spore Forming Bacteria Distribution Statement A...Z39.18 00-07-2015 Technical N/A Biocidal Energetic Materials for the Destruction of Spore Forming Bacteria HDTRA1-10-1-0108 Emily M. Hunt, Ph.D. West...understand the interaction between spore forming bacteria and thermite reactions and products and to exploit energetic material reactions with

  9. Bacillus cereus catheter related bloodstream infection in a patient with acute lymphoblastic leukemia.

    Science.gov (United States)

    Gurler, N; Oksuz, L; Muftuoglu, M; Sargin, Fd; Besisik, Sk

    2012-01-01

    Bacillus cereus infection is rarely associated with actual infection and for this reason single positive blood culture is usually regarded as contamination . However it may cause a number of infections, such catheter-related bloodstream infections. Significant catheter-related bloodstream infections (CRBSI) caused by Bacillus spp. are mainly due to B. cereus and have been predominantly reported in immunocompromised hosts. Catheter removal is generally advised for management of infection. In this report, catheter-related bacteremia caused by B. cereus in a patient with acute lymphoblast c leukemia (ALL) in Istanbul Medical Faculty was presented.

  10. Bacillus cereus bacteremia and hemolytic anemia in a patient with hemoglobin SC disease.

    Science.gov (United States)

    Rodgers, G M; Barrera, E; Martin, R R

    1980-08-01

    A patient with hemoglobin SC disease and cholelithiasis was found to have Bacillus cereus bacteremia. Hemolytic anemia developed, for which common causes of hemolysis were excluded, suggesting a relationship with the bacteremia. Following in vitro incubation, type O erythrocytes were hemolyzed by the culture, but not by a bacteria-free filtrate. This case confirms the association between sickle cell disorders and cholelithiasis with B cereus infections. In addition, it provides evidence for in vivo hemolysis with B cereus bacteremia, an organism not previously associated with hemolytic anemia.

  11. Successful surgical drainage and aggressive medical therapy in a preterm neonate with Bacillus cereus meningitis.

    Science.gov (United States)

    Drazin, Doniel; Lehman, Deborah; Danielpour, Moise

    2010-01-01

    Bacillus cereus meningitis is a rare disease with a very high mortality rate in neonates. The authors present the rare case of a premature infant with B. cereus bacteremia and subsequent intracranial abscesses. In addition to aggressive medical therapy, surgical drainage was performed via a left frontal mini-craniotomy. At 15 months of age, the patient had mild developmental delay, cortical blindness, and sensorineural hearing loss. The clinical case is described and difficulties in the management of B. cereus meningoencephalitis in infants are discussed.

  12. Occurrence and significance of Bacillus cereus and Bacillus thuringiensis in ready-to-eat food

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Ørum-Smidt, Lasse; Andersen, Sigrid R

    2005-01-01

    Among 48,901 samples of ready-to-eat food products at the Danish retail market, 0.5% had counts of Bacillus cereus-like bacteria above 10(4) cfu g(-1). The high counts were most frequently found in starchy, cooked products, but also in fresh cucumbers and tomatoes. Forty randomly selected strains...... had at least one gene or component involved in human diarrhoeal disease, while emetic toxin was related to only one B. cereus strain. A new observation was that 31 out of the 40 randomly selected B. cereus-like strains could be classified as Bacillus thuringiensis due to crystal production and...

  13. Bacillus cereus Induces Permeability of an In Vitro Blood-Retina Barrier▿

    OpenAIRE

    Moyer, A. L.; Ramadan, R.T.; Thurman, J.; Burroughs, A; Callegan, M.C.

    2008-01-01

    Most Bacillus cereus toxin production is controlled by the quorum-sensing-dependent, pleiotropic global regulator plcR, which contributes to the organism's virulence in the eye. The purpose of this study was to analyze the effects of B. cereus infection and plcR-regulated toxins on the barrier function of retinal pigment epithelium (RPE) cells, the primary cells of the blood-retina barrier. Human ARPE-19 cells were apically inoculated with wild-type or quorum-sensing-deficient B. cereus, and ...

  14. Bacillus cereus Bloodstream Infection in a Preterm Neonate Complicated by Late Meningitis

    Directory of Open Access Journals (Sweden)

    Toshinobu Horii

    2012-01-01

    Full Text Available Central nervous system infections caused by Bacillus cereus have rarely been reported in infants. In this paper, the case of a 2-month-old low-birth-weight female who developed meningitis 45 days after resolution of a bloodstream infection (BSI is described. The pulsed-field gel electrophoresis results revealed that the patterns of both B. cereus isolates responsible for the acute meningitis and for the prior bacteraemic episode were closely related. Although the source of the infection from within the patient was not clear, it is suggested that the B. cereus BSI developed in the neonate was complicated by acute meningitis.

  15. Characteristics and phylogeny of Bacillus cereus strains isolated from Maari, a traditional West African food condiment

    DEFF Research Database (Denmark)

    Thorsen, Line; Kando, Christine Kere; Sawadogo, Hagrétou

    2015-01-01

    cereus occasionally occurs in Maari. This study characterizes succession patterns and pathogenic potential of B. cereus isolated from the raw materials (ash, water from a drilled well (DW) and potash), seed mash throughout fermentation (0-96h), after steam cooking and sun drying (final product) from two...... found in potash, DW, cooking water and at 8h fermentation. The "emetic" type B. cereus were present in DW, the seed mash at 48-72h of fermentation and in the final product, while the remaining isolates (PanC type IV) were detected in ash, at 48-72h fermentation and in the final product. This work sheds...

  16. Surface tension propulsion of fungal spores by use of microdroplets

    CERN Document Server

    Noblin, Xavier; 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 of Fluid Motion 2010 which is showcase of fluid dynamics videos.

  17. Quantitative and Sensitive RNA Based Detection of Bacillus Spores

    Directory of Open Access Journals (Sweden)

    Ekaterina eOsmekhina

    2014-03-01

    Full Text Available The fast and reliable detection of bacterial spores is of great importance and still remains a challenge. Here we describe a direct RNA based diagnostic method for the specific detection of viable bacterial spores which does not depends on an enzymatic amplification step and therefore is directly appropriate for quantification. The procedure includes the following steps: (i heat activation of spores, (ii germination and enrichment cultivation, (iii cell lysis, and (iv analysis of 16S rRNA in crude cell lysates using a sandwich hybridization assay. The sensitivity of the method is dependent on the cultivation time and the detection limit; it is possible to detect 10 spores per ml when the RNA analysis is performed after 6 h of enrichment cultivation. At spore concentrations above 106 spores per ml the cultivation time can be shortened to 30 min. Total analysis times are in the range of 2 to 8 hours depending on the spore concentration in samples. The developed procedure is optimized at the example of Bacillus subtilis spores but should be applicable to other organisms. The new method can easily be modified for other target RNAs and is suitable for specific detection of spores from known groups of organisms.

  18. Model simulations of fungal spore distribution over the Indian region

    Science.gov (United States)

    Ansari, Tabish U.; Valsan, Aswathy E.; Ojha, N.; Ravikrishna, R.; Narasimhan, Balaji; Gunthe, Sachin S.

    2015-12-01

    Fungal spores play important role in the health of humans, animals, and plants by constituting a class of the primary biological aerosol particles (PBAPs). Additionally, these could mediate the hydrological cycle by acting as nuclei for ice and cloud formation (IN and CCN respectively). Various processes in the biosphere and the variations in the meteorological conditions control the releasing mechanism of spores through active wet and dry discharge. In the present paper, we simulate the concentration of fungal spores over the Indian region during three distinct meteorological seasons by combining a numerical model (WRF-Chem) with the fungal spore emissions based on land-use type. Maiden high-resolution regional simulations revealed large spatial gradient and strong seasonal dependence in the concentration of fungal spores over the Indian region. The fungal spore concentrations are found to be the highest during winter (0-70 μg m-3 in December), moderately higher during summer (0-35 μg m-3 in May) and lowest during the monsoon (0-25 μg m-3 in July). The elevated concentrations during winter are attributed to the shallower boundary layer trapping the emitted fungal spores in smaller volume. In contrast, the deeper boundary layer mixing in May and stronger monsoonal-convection in July distribute the fungal spores throughout the lower troposphere (∼5 km). We suggest that the higher fungal spore concentrations during winter could have potential health impacts. While, stronger vertical mixing could enable fungal spores to influence the cloud formation during summer and monsoon. Our study provides the first information about the distribution and seasonal variation of fungal spores over the densely populated and observationally sparse Indian region.

  19. Synergistic Activity Between S-Layer Protein and Spore-Crystal Preparations from Lysinibacillus sphaericus Against Culex quinquefasciatus Larvae.

    Science.gov (United States)

    Lozano, Lucía C; Dussán, Jenny

    2017-03-01

    Lysinibacillus sphaericus is used for the biological control of mosquitoes. The main toxicity mechanism of pathogenic strains is a binary toxin produced during sporulation. S-layer is a proteinaceous structure on the surface of bacteria; its functions have been involved in the interaction between bacterial cells and the environment, for example, as protective coats, surface recognition, and biological control. In L. sphaericus, S-layer protein (SlpC) is expressed in vegetative cells, and is also found in spore-crystal preparations; it has larvicidal activity in Culex spp. In this study, partial and completed sporulated culture toxicities were compared; also, S-layer protein and spore-crystal proteins were tested against Culex quinquefasciatus larvae for possible interactions. Larvicidal activity obtained with a combination of SlpC and spore-crystal proteins from strain III(3)7 showed no significant interaction, whereas, combinations of both preparations from strain 2362 showed synergistic effect. The highest synergistic activity observed was between spore protein complex from strain 2362 and SlpC from III(3)7. S-layer protein could be considered a good alternative in formulation improvement, for biological control of mosquitoes.

  20. Characterization of arabinogalactan-rich mucilage from Cereus triangularis cladodes.

    Science.gov (United States)

    Petera, B; Delattre, C; Pierre, G; Wadouachi, A; Elboutachfaiti, R; Engel, E; Poughon, L; Michaud, P; Fenoradosoa, T A

    2015-01-01

    Cereus triangularis (Cactaceae) is a cactus used in food decoction as a traditional medicine in the North region of Madagascar to reduce stomach ache and intestinal diseases. Hydrocolloids were sequentially extracted from its cladodes with a yield of 24% (240 mg/g based on dried cladodes powder). Structural analyses has revealed that this polysaccharide with a molecular mass of 8430,000g/mol was mainly composed of a galactan backbone of a (1 → 4) linked β-d-Galp residues probably substituted at position 3 by L-arabinofuranosyl residues. In vitro antioxidant activity of this arabinogalactan-rich fraction was detected and quantified by radical DPPH scavenging, hydroxyl radical scavenging, radical anion superoxide scavenging and reducing power method.

  1. Alkaloid production by callous tissue cultures of Cereus peruvianus (Cactaceae).

    Science.gov (United States)

    de Oliveira, Arildo José Braz; Machado, Maria Fátima Pires da Silva

    2003-02-01

    The morphologically undifferentiated cells of nonregenerant callous tissue of Cereus peruvianus cultured in the original medium and in medium supplemented with tyrosine were used as an alkaloid source. Comparison of alkaloid production by C. peruvianus plants and by callous tissues indicated that alkaloid levels were almost twice as high in callous tissues as in shoots of C. peruvianus plants. The ratio of alkaloid concentration between mature plant and morphologically undifferentiated cells of callous tissue was 1:1.7. A relationship between culture medium containing tyrosine and alkaloid production was also observed in the callous tissues of C. peruvianus. Since increased alkaloid production may be induced by additional factors such as tyrosine, increasing levels of tyrosine or other conditions of the culture medium may be considered factors for inducing higher alkaloid production by C. peruvianus callous tissues.

  2. Cereus peruvianus (Koubo new cactus fruit for the world

    Directory of Open Access Journals (Sweden)

    Yosef Mizrahi

    2014-03-01

    Full Text Available Several different species of the columnar cacti of the genera Stenocereus and Pachycereus, were introduced into different semi-arid ecozones in Israel and most of these efforts were of disappointing outcomes, the only exception being the Cereus peruvianus (L. Miller,which bore plenty of fruits, some of them of good taste. The original seeds of this plant were obtained from the late Mr. Amram (Ron Kodish, who collected seeds from various private gardens in Southern California which bore fruits of reasonable qualities. The initial success of this species led us to initiate an intensive research study, and today it is already fruit-crop, marketed mainly in Israel under the name " Koubo" . This paper will describe our work of domestication of this new cactus fruit crop in Israel.

  3. Nosocomial bacteremia and catheter infection by Bacillus cereus in an immunocompetent patient.

    Science.gov (United States)

    Hernaiz, C; Picardo, A; Alos, J I; Gomez-Garces, J L

    2003-09-01

    We present a case of Bacillus cereus bacteremia and catheter infection in an immunocompetent patient subjected to abdominal surgery, who recovered following central catheter removal and treatment with piperacillin/tazobactam.

  4. [Two cases of acute myelogenous leukemia with Bacillus cereus bacteremia resulting in fatal intracranial hemorrhage].

    Science.gov (United States)

    Yoshida, H; Moriyama, Y; Tatekawa, T; Tominaga, N; Teshima, H; Hiraoka, A; Masaoka, T; Yoshinaga, T

    1993-12-01

    This manuscript reports Bacillus cereus sepsis in two cases with acute myelogenous leukemia (AML) who suffered complications of fatal intracranial hemorrhage during remission induction therapy. The first case was 43-year-old male with AML (M0) receiving first consolidation chemotherapy who developed sudden diarrhea, abdominal pain and spiking fever. Two days later, he died of intracranial hemorrhage. The second case was 15-year-old male with AML (M5b) who was receiving first induction chemotherapy. He developed headache and vomiting following spiking fever and diarrhea. He died of subarachnoid hemorrhage the next day. In both cases, Bacillus cereus was isolated from blood culture. Fatal intracranial hemorrhage due to severe bleeding tendency caused rapid to death in both cases. These bleeding tendencies might have been induced by B. cereus sepsis. In addition, we should not overlook B. cereus as contamination, but rather consider it as a potential pathogen, when isolated from blood culture.

  5. Relapsing peritonitis with Bacillus cereus in a patient on continuous ambulatory peritoneal dialysis.

    Science.gov (United States)

    Magnussen, Eyð Tausen; Vang, Amanda Gratton; á Steig, Torkil; Gaini, Shahin

    2016-04-26

    We present a case where Bacillus cereus was determined to be the causative agent of relapsing peritonitis in a patient on continuous ambulatory peritoneal dialysis (CAPD). The patient, a 70-year-old man from the Faroe Islands, was admitted with relapsing peritonitis four times over a 3-month period. Peritoneal cultures were positive for growth of B. cereus, a rare bacterial cause of peritonitis. The cultures demonstrated susceptibility to vancomycin, and therefore the patient was treated with intraperitoneal vancomycin, intraperitoneal gentamycin and oral ciprofloxacin. As a result of the relapsing B. cereus peritonitis diagnosis and a CT scan showing contraction of the peritoneum after longstanding inflammation, the peritoneal catheter was removed and the patient converted to haemodialysis. To date, the patient has not been readmitted due to peritonitis. A lack of proper hygiene when changing the dialysis bag was the suspected source of infection with B. cereus.

  6. A case of intoxication due to a highly cytotoxic Bacillus cereus strain isolated from cooked chicken.

    Science.gov (United States)

    López, Ana C; Minnaard, Jessica; Pérez, Pablo F; Alippi, Adriana M

    2015-04-01

    Outbreaks of Bacillus cereus infection/intoxication are not commonly reported because symptoms are often mild, and the disease is self-limiting. However, hypervirulent strains increase health risks. We report a case, which occurred in Argentina, of severe food poisoning illness on a healthy adult woman associated to B. cereus strain MVL2011. The studied strain was highly cytotoxic, showed high ability to detach Caco-2 cells and was positive for the hblA, hblB, and hblC genes of the hbl complex, bceT, entS and ces. As it is considered that B. cereus emetic cluster evolved from a panmictic population of diarrheal strains, B. cereus MVL2011 could constitute an intermediate strain between diarrheal and emetic strains.

  7. Bacillus Cereus Catheter Related Bloodstream Infection in a Patient with Acute Lymphoblastic Leukemia

    OpenAIRE

    N Gurler; Oksuz, L; M Muftuoglu; Sargin, FD; Besisik, SK

    2012-01-01

    Bacillus cereus infection is rarely associated with actual infection and for this reason single positive blood culture is usually regarded as contamination . However it may cause a number of infections, such catheter-related bloodstream infections. Significant catheter-related bloodstream infections (CRBSI) caused by Bacillus spp. are mainly due to B. cereus and have been predominantly reported in immunocompromised hosts. Catheter removal is generally advised for management of infection. In t...

  8. Association of Genotyping of Bacillus cereus with Clinical Features of Post-Traumatic Endophthalmitis.

    Science.gov (United States)

    Hong, Meng; Wang, Qian; Tang, Zhide; Wang, Youpei; Gu, Yunfeng; Lou, Yongliang; Zheng, Meiqin

    2016-01-01

    Bacillus cereus is the second most frequent cause of post-traumatic bacterial endophthalmitis. Although genotyping of B. cereus associated with gastrointestinal infections has been reported, little is known about the B. cereus clinical isolates associated with post-traumatic endophthalmitis. This is largely due to the limited number of clinical strains available isolated from infected tissues of patients with post-traumatic endophthalmitis. In this study, we report successful isolation of twenty-four B. cereus strains from individual patients with different disease severity of post-traumatic endophthalmitis. Phylogenetic analysis showed that all strains could be categorized into three genotypes (GTI, GTII and GTIII) and the clinical score showed significant differences among these groups. We then further performed genotyping using the vrrA gene, and evaluated possible correlation of genotype with the clinical features of B. cereus-caused post-traumatic endophthalmitis, and with the prognosis of infection by conducting follow-up with patients for up to 2 months. We found that the disease of onset and final vision acuity were significantly different among the three groups. These results suggested that the vrrA gene may play a significant role in the pathogenesis of endophthalmitis, and genotyping of B. cereus has the potential for predicting clinical manifestation and prognosis of endophthalmitis. To the best of our knowledge, this is the first report of isolation of large numbers of clinical isolates of B. cereus from patients with endophthalmitis. This work sets the foundation for future investigation of the pathogenesis endophthalmitis caused by B. cereus infection.

  9. Association of Genotyping of Bacillus cereus with Clinical Features of Post-Traumatic Endophthalmitis.

    Directory of Open Access Journals (Sweden)

    Meng Hong

    Full Text Available Bacillus cereus is the second most frequent cause of post-traumatic bacterial endophthalmitis. Although genotyping of B. cereus associated with gastrointestinal infections has been reported, little is known about the B. cereus clinical isolates associated with post-traumatic endophthalmitis. This is largely due to the limited number of clinical strains available isolated from infected tissues of patients with post-traumatic endophthalmitis. In this study, we report successful isolation of twenty-four B. cereus strains from individual patients with different disease severity of post-traumatic endophthalmitis. Phylogenetic analysis showed that all strains could be categorized into three genotypes (GTI, GTII and GTIII and the clinical score showed significant differences among these groups. We then further performed genotyping using the vrrA gene, and evaluated possible correlation of genotype with the clinical features of B. cereus-caused post-traumatic endophthalmitis, and with the prognosis of infection by conducting follow-up with patients for up to 2 months. We found that the disease of onset and final vision acuity were significantly different among the three groups. These results suggested that the vrrA gene may play a significant role in the pathogenesis of endophthalmitis, and genotyping of B. cereus has the potential for predicting clinical manifestation and prognosis of endophthalmitis. To the best of our knowledge, this is the first report of isolation of large numbers of clinical isolates of B. cereus from patients with endophthalmitis. This work sets the foundation for future investigation of the pathogenesis endophthalmitis caused by B. cereus infection.

  10. Exoproteome analysis of a novel strain of Bacillus cereus implicated in disease resembling cutaneous anthrax.

    Science.gov (United States)

    Ghosh, Neha; Goel, Ajay Kumar; Alam, Syed Imteyaz

    2014-03-01

    Bacillus cereus belongs to B. cereus sensu lato group, shared by six other related species including Bacillus anthracis. B. anthracis is the causative agent for serious illness affecting a wide range of animals as well as humans and is a category A Biological and Toxin Warfare (BTW) agent. Recent studies indicate that a Bacillus species other than B. anthracis can cause anthrax-like disease and role of anthrax virulence plasmids (pXO1 and pXO2) on the pathogenicity of B. cereus has been documented. B. cereus strain TF5 was isolated from the tissue fluid of cutaneous anthrax-like skin lesions of a human patient from an anthrax endemic area in India. The strain harboured a PA gene, however, presence of pXO1 or pXO2-like plasmids could not be ascertained using reported primers. Abundant exoproteome of the strain in the early stationary phase was elucidated using a 2-DE MS approach and compared with that from a reference B. cereus strain. Analysis of proteins showing qualitative and quantitative differences between the two strains indicated an altered regulatory mechanism and putative role of S-layer protein and sphingomyelinase in the pathogenesis of strain TF5. Phylogenetic analysis of the S-layer protein indicated close affiliation of the strain with anthracis-like B. cereus strains such as B. cereus var. anthracis strain CI; whereas sphingomyelinase exhibited specific relationship with all the strains of B. anthracis apart from that with anthracis-like B. cereus strains.

  11. Enterotoxin Production in Natural Isolates of Bacillaceae outside the Bacillus cereus Group

    OpenAIRE

    Phelps, Rebecca J.; McKillip, John L.

    2002-01-01

    Thirty-nine Bacillus strains obtained from a variety of environmental and food sources were screened by PCR for the presence of five gene targets (hblC, hblD, hblA, nheA, and nheB) in two enterotoxin operons (HBL and NHE) traditionally harbored by Bacillus cereus. Seven isolates exhibited a positive signal for at least three of the five possible targets, including Bacillus amyloliquefaciens, B. cereus, Bacillus circulans, Bacillus lentimorbis, Bacillus pasteurii, and Bacillus thuringiensis su...

  12. Infective endocarditis due to Bacillus cereus in a pregnant female: A case report and literature review

    OpenAIRE

    Mahek Shah; Soumya Patnaik; Supakanya Wongrakpanich; Yaser Alhamshari; Talal Alnabelsi

    2015-01-01

    Incidence of infective endocarditis during pregnancy is around 0.006% with high maternal and fetal mortality. Bacillus cereus is an extremely rare cause for endocarditis in intravenous drug abusers (IVDA) or those with valvular disease or devices such as pacemakers. We report a case of B. cereus endocarditis, which, to the best of our knowledge, has never been reported in pregnancy. A 30-year-old, 25-week pregnant female presented with right shoulder pain, swelling and erythema on the lateral...

  13. Group II introns in the Bacillus cereus group with unusual splicing properties

    OpenAIRE

    Stabell, Fredrik Bernhard

    2009-01-01

    Mobile genetic elements have had, and still have an impact on the evolution of the genomes providing means for adaptation and structural organization. These elements are one of the major driving forces for the general evolution of all life forms. For the organisms and their genomes these elements are essential for development and adaptation to different environments. The Bacillus cereus group of bacteria includes the related species B. cereus (sensu stricto), B. thuringiensis, B. weihenst...

  14. Comparison of enterotoxin production and phenotypic characteristics between emetic and enterotoxic Bacillus cereus.

    Science.gov (United States)

    Kim, Jung-Beom; Kim, Jai-Moung; Kim, So-Yeong; Kim, Jong-Hyun; Park, Yong-Bae; Choi, Na-Jung; Oh, Deog-Hwan

    2010-07-01

    Bacillus cereus was divided into emetic toxin (cereulide)- and enterotoxin-producing strains, but emetic toxin-producing B. cereus is difficult to detect immunochemically. Screening methods for emetic toxin-producing B. cereus are needed. The objectives of this study were to identify and detect emetic toxin-producing B. cereus among 160 B. cereus strains, and to compare enterotoxin production and phenotypic characteristics between the emetic toxin-producing and enterotoxin-producing strains. Forty emetic toxin-producing B. cereus strains were determined with high-pressure liquid chromatography-mass spectrometry analysis. Among the emetic toxin-producing strains (n = 40), 31 (77.5%) and 3 (7.5%) strains produced nonhemolytic enterotoxin (NHE) and hemolysin BL (HBL) enterotoxins, respectively. In addition, 107 (89.2%) and 100 (83.3%) strains produced NHE and HBL enterotoxins among the enterotoxin-producing strains (n = 120). The number of strains positive for starch hydrolysis, salicin fermentation, and hemolysis among the emetic toxin-producing strains were 3 (7.5%), 3 (7.5%), and 26 (65.0%), respectively, and among enterotoxin-producing strains, these numbers were 101 (84.2%), 100 (83.3%), and 111 (92.5%), respectively. In particular, the three emetic toxin-producing B. cereus strains (JNHE 6, JNHE 36, and KNIH 28) produced the HBL and NHE enterotoxins and were capable of starch hydrolysis and salicin fermentation. The absence of HBL enterotoxin and certain phenotypic properties, such as starch hydrolysis and salicin fermentation, indicates that these properties were not critical characteristics of the emetic toxin-producing B. cereus tested in this study.

  15. PENGARUH EKSTRAK ANDALIMAN (Zanthoxyium acanthopodium DC TERHADAP PERMEABILITAS DAN HIDROFOBISITAS Bacillus cereus [Effect of Andaliman (Zanthoxylum acanthopodium DC Extracts upon Permeability and Hidrophobicity of Bacillus cereus

    Directory of Open Access Journals (Sweden)

    Sedarnawati Yasni2

    2005-04-01

    Full Text Available Andaliman spice is usually added as one of main spices in cooked fish and meat. Andaliman seeds were extracted using maceration method with nonpolar, semipolar and polar solvents. The result showed that the three kinds of andaliman extract had antibacterial activity on Bacillus cereus, especially during exponential phase (8 hour incubation period. Ethyl-acetate extract of Andaliman showed the highest antibacterial activity toward B. cereus with MIC and MBC values being 0.2% and 0.8% respectively. The permeability of B. cereus was observed at the dose of 2.5 MIC and 60.30% hydrophobicity leakage was obtained at 6% andaliman extracted by ethyl-acetate.

  16. Improvement of immunodetection of bacterial spore antigen by ultrasonic cavitation.

    Science.gov (United States)

    Borthwick, Kathryn A J; Love, Tracey E; McDonnell, Martin B; Coakley, W Terence

    2005-11-15

    Ultrasonic cavitation was employed to enhance sensitivity of bacterial spore immunoassay detection, specifically, enzyme-linked immunosorbent assay (ELISA) and resonant mirror (RM) sensing. Bacillus spore suspensions were exposed to high-power ultrasound in a tubular sonicator operated at 267 kHz in both batch and flow modes. The sonicator was designed to deliver high output power and is in a form that can be cooled efficiently to avoid thermal denaturation of antigen. The 30-s batch and cooled flow (0.3 mL/min) sonication achieved an approximately 20-fold increase in ELISA sensitivity compared to unsonicated spores by ELISA. RM sensing of sonicated spores achieved detection sensitivity of approximately 10(6) spores/mL, whereas unsonicated spores were undetectable at the highest concentration tested. Improvements in detection were associated with antigen released from the spores. Equilibrium temperature increase in the tubular sonicator was limited to 14 K after 30 min and was maintained for 6 h with cooling and flow (0.3 mL/min). The work described here demonstrates the utility of the tubular sonicator for the improvement in the sensitivity of the detection of spores and its suitability as an in-line component of a rapid detection system.

  17. 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 elimina

  18. 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.

  19. 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.

  20. Mushrooms as Rainmakers: How Spores Act as Nuclei for Raindrops.

    Science.gov (United States)

    Hassett, Maribeth O; Fischer, Mark W F; Money, Nicholas P

    2015-01-01

    Millions of tons of fungal spores are dispersed in the atmosphere every year. These living cells, along with plant spores and pollen grains, may act as nuclei for condensation of water in clouds. Basidiospores released by mushrooms form a significant proportion of these aerosols, particularly above tropical forests. Mushroom spores are discharged from gills by the rapid displacement of a droplet of fluid on the cell surface. This droplet is formed by the condensation of water on the spore surface stimulated by the secretion of mannitol and other hygroscopic sugars. This fluid is carried with the spore during discharge, but evaporates once the spore is airborne. Using environmental electron microscopy, we have demonstrated that droplets reform on spores in humid air. The kinetics of this process suggest that basidiospores are especially effective as nuclei for the formation of large water drops in clouds. Through this mechanism, mushroom spores may promote rainfall in ecosystems that support large populations of ectomycorrhizal and saprotrophic basidiomycetes. Our research heightens interest in the global significance of the fungi and raises additional concerns about the sustainability of forests that depend on heavy precipitation.

  1. Differentiation of strains from the Bacillus cereus group by RFLP-PFGE genomic fingerprinting.

    Science.gov (United States)

    Otlewska, Anna; Oltuszak-Walczak, Elzbieta; Walczak, Piotr

    2013-11-01

    Bacillus mycoides, Bacillus pseudomycoides, Bacillus weihenstephanensis, Bacillus anthracis, Bacillus thuringiensis, and Bacillus cereus belong to the B. cereus group. The last three species are characterized by different phenotype features and pathogenicity spectrum, but it has been shown that these species are genetically closely related. The macrorestriction analysis of the genomic DNA with the NotI enzyme was used to generate polymorphism of restriction profiles for 39 food-borne isolates (B. cereus, B. mycoides) and seven reference strains (B. mycoides, B. thuringiensis, B. weihenstephanensis, and B. cereus). The PFGE method was applied to differentiate the examined strains of the B. cereus group. On the basis of the unweighted pair group method with the arithmetic mean method and Dice coefficient, the strains were divided into five clusters (types A-E), and the most numerous group was group A (25 strains). A total of 21 distinct pulsotypes were observed. The RFLP-PFGE analysis was successfully used for the differentiation and characterization of B. cereus and B. mycoides strains isolated from different food products.

  2. Fulminant septicemia of Bacillus cereus resistant to carbapenem in a patient with biphenotypic acute leukemia.

    Science.gov (United States)

    Kiyomizu, Kazunobu; Yagi, Toshinari; Yoshida, Hitoshi; Minami, Ryota; Tanimura, Akira; Karasuno, Takahiro; Hiraoka, Akira

    2008-10-01

    We report a case of fulminant septicemia with Bacillus cereus resistant to carbapenem. A 33-year-old man was suffering from febrile neutropenia (FN) on day 15 after the start of remission-induction therapy for biphenotypic acute leukemia under gut decontamination with polymyxin B and nystatin. Meropenem, a carbapenem, was administered according to the guideline for FN. Two days later (on day 17), he complained of severe abdominal pain, lost consciousness, went into sudden cardiopulmonary arrest, and died. Autopsy showed multiple spots of hemorrhage and necrosis caused by bacterial plaque in the brain, lungs, and liver. B. cereus was isolated from a blood sample obtained in the morning on day 17 and it was after his death that the isolated B. cereus was revealed to be resistant to carbapenem. B. cereus obtained from blood samples has been reported to be usually sensitive to carbapenem and also to vancomycin, new quinolones, and clindamycin. If B. cereus resistant to carbapem increases, our method of gut decontamination with polymyxin B and nystatin may have to be changed to one containing a new quinolone for the prevention of septicemia. Careful watching to determine whether B. cereus resistant to carbapem increases may be also important for empiric therapy, because carbapenem is often selected as the initial therapy for FN in patients with severe neutropenia.

  3. Proteomic evidences for rex regulation of metabolism in toxin-producing Bacillus cereus ATCC 14579.

    Science.gov (United States)

    Laouami, Sabrina; Clair, Géremy; Armengaud, Jean; Duport, Catherine

    2014-01-01

    The facultative anaerobe, Bacillus cereus, causes diarrheal diseases in humans. Its ability to deal with oxygen availability is recognized to be critical for pathogenesis. The B. cereus genome comprises a gene encoding a protein with high similarities to the redox regulator, Rex, which is a central regulator of anaerobic metabolism in Bacillus subtilis and other Gram-positive bacteria. Here, we showed that B. cereus rex is monocistronic and down-regulated in the absence of oxygen. The protein encoded by rex is an authentic Rex transcriptional factor since its DNA binding activity depends on the NADH/NAD+ ratio. Rex deletion compromised the ability of B. cereus to cope with external oxidative stress under anaerobiosis while increasing B. cereus resistance against such stress under aerobiosis. The deletion of rex affects anaerobic fermentative and aerobic respiratory metabolism of B. cereus by decreasing and increasing, respectively, the carbon flux through the NADH-recycling lactate pathway. We compared both the cellular proteome and exoproteome of the wild-type and Δrex cells using a high throughput shotgun label-free quantitation approach and identified proteins that are under control of Rex-mediated regulation. Proteomics data have been deposited to the ProteomeXchange with identifier PXD000886. The data suggest that Rex regulates both the cross-talk between metabolic pathways that produce NADH and NADPH and toxinogenesis, especially in oxic conditions.

  4. Proteomic evidences for rex regulation of metabolism in toxin-producing Bacillus cereus ATCC 14579.

    Directory of Open Access Journals (Sweden)

    Sabrina Laouami

    Full Text Available The facultative anaerobe, Bacillus cereus, causes diarrheal diseases in humans. Its ability to deal with oxygen availability is recognized to be critical for pathogenesis. The B. cereus genome comprises a gene encoding a protein with high similarities to the redox regulator, Rex, which is a central regulator of anaerobic metabolism in Bacillus subtilis and other Gram-positive bacteria. Here, we showed that B. cereus rex is monocistronic and down-regulated in the absence of oxygen. The protein encoded by rex is an authentic Rex transcriptional factor since its DNA binding activity depends on the NADH/NAD+ ratio. Rex deletion compromised the ability of B. cereus to cope with external oxidative stress under anaerobiosis while increasing B. cereus resistance against such stress under aerobiosis. The deletion of rex affects anaerobic fermentative and aerobic respiratory metabolism of B. cereus by decreasing and increasing, respectively, the carbon flux through the NADH-recycling lactate pathway. We compared both the cellular proteome and exoproteome of the wild-type and Δrex cells using a high throughput shotgun label-free quantitation approach and identified proteins that are under control of Rex-mediated regulation. Proteomics data have been deposited to the ProteomeXchange with identifier PXD000886. The data suggest that Rex regulates both the cross-talk between metabolic pathways that produce NADH and NADPH and toxinogenesis, especially in oxic conditions.

  5. Direct detection of toxigenic Bacillus cereus in dietary complement for children and cassava starch

    Directory of Open Access Journals (Sweden)

    Jnnifer A. Sánchez

    2015-09-01

    Full Text Available Bacillus cereus is a food contaminant anda known human pathogen that can causeemetic and diarrheal syndromes. In this studywe evaluated the presence of toxigenic B.cereus by multiplex PCR directly in dietarycomplement for children and cassava starchsamples collected on Medellin, Colombia.Of 75 dietary complement for childrensamples evaluated, 70.7% were contaminatedwith toxigenic B. cereus and four differenttoxigenic consortia were detected: I: nheA,hblC, cytK (9.8%, II: nheA, hblC (2%,III: hblC, cytK (41.2%, IV: hblC (47%.Of 75 cassava starch samples, 44% werecontaminated with toxigenic B. cereus andfour different toxigenic consortia weredetermined: I: nheA, hblC, cytK (48.5%,II: nheA, hblC, cytK, cesB (3%, III: hblC,cytK (30.3%, IV: hblC (18.2%. In general,in dietary complement for children onlyenterotoxigenic consortia were detectedwhile in cassava starch the enterotoxigenicconsortia predominated over the emetic.Multiplex PCR was useful to detect toxigenicB. cereus contamination allowing directand simultaneous detection of all toxingenes in foods. This study is the first inColombia to evaluate toxigenic B. cereus,providing information of importance formicrobiological risk evaluation in driedfoods.

  6. Bacillus cereus strain S2 shows high nematicidal activity against Meloidogyne incognita by producing sphingosine

    Science.gov (United States)

    Gao, Huijuan; Qi, Gaofu; Yin, Rong; Zhang, Hongchun; Li, Chenggang; Zhao, Xiuyun

    2016-01-01

    Plant-parasitic nematodes cause serious crop losses worldwidely. This study intended to discover the antagonistic mechanism of Bacillus cereus strain S2 against Meloidogyne incognita. Treatment with B. cereus strain S2 resulted in a mortality of 77.89% to Caenorhabditis elegans (a model organism) and 90.96% to M. incognita. In pot experiment, control efficiency of B. cereus S2 culture or supernatants were 81.36% and 67.42% towards M. incognita, respectively. In field experiment, control efficiency was 58.97% towards M. incognita. Nematicidal substances were isolated from culture supernatant of B. cereus S2 by polarity gradient extraction, silica gel column chromatography and HPLC. Two nematicidal compounds were identified as C16 sphingosine and phytosphingosine by LC-MS. The median lethal concentration of sphingosine was determined as 0.64 μg/ml. Sphingosine could obviously inhibit reproduction of C. elegans, with an inhibition rate of 42.72% for 24 h. After treatment with sphingosine, ROS was induced in intestinal tract, and genital area disappeared in nematode. Furthermore, B. cereus S2 could induce systemic resistance in tomato, and enhance activity of defense-related enzymes for biocontrol of M. incognita. This study demonstrates the nematicidal activity of B. cereus and its product sphingosine, as well provides a possibility for biocontrol of M. incognita. PMID:27338781

  7. The molecular timeline of a reviving bacterial spore.

    Science.gov (United States)

    Sinai, Lior; Rosenberg, Alex; Smith, Yoav; Segev, Einat; Ben-Yehuda, Sigal

    2015-02-19

    The bacterial spore can rapidly convert from a dormant to a fully active cell. Here we study this remarkable cellular transition in Bacillus subtilis and reveal the identity of the newly synthesized proteins throughout spore revival. Our analysis uncovers a highly ordered developmental program that correlates with the spore morphological changes and reveals the spatial and temporal molecular events fundamental to reconstruct a cell. As opposed to current knowledge, we found that translation takes place during the earliest revival event, termed germination, a process hitherto considered to occur without the need for any macromolecule synthesis. Furthermore, we demonstrate that translation is required for execution of germination and relies on the bona fide translational factors RpmE and Tig. Our study sheds light on the spore revival process and on the vital building blocks underlying cellular awakening, thereby paving the way for designing new antimicrobial agents to eradicate spore-forming pathogens.

  8. 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.

  9. Unlocking the Sporicidal Potential of Ethanol: Induced Sporicidal Activity of Ethanol against Clostridium difficile and Bacillus Spores under Altered Physical and Chemical Conditions

    Science.gov (United States)

    Nerandzic, Michelle M.; Sunkesula, Venkata C. K.; C., Thriveen Sankar; Setlow, Peter; Donskey, Curtis J.

    2015-01-01

    Background Due to their efficacy and convenience, alcohol-based hand sanitizers have been widely adopted as the primary method of hand hygiene in healthcare settings. However, alcohols lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We hypothesized that sporicidal activity could be induced in alcohols through alteration of physical or chemical conditions that have been shown to degrade or allow penetration of spore coats. Principal Findings Acidification, alkalinization, and heating of ethanol induced rapid sporicidal activity against C. difficile, and to a lesser extent Bacillus thuringiensis and Bacillus subtilis. The sporicidal activity of acidified ethanol was enhanced by increasing ionic strength and mild elevations in temperature. On skin, sporicidal ethanol formulations were as effective as soap and water hand washing in reducing levels of C. difficile spores. Conclusions These findings demonstrate that novel ethanol-based sporicidal hand hygiene formulations can be developed through alteration of physical and chemical conditions. PMID:26177038

  10. 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.

  11. Determination of fungal spore release from wet building materials.

    Science.gov (United States)

    Kildesø, J; Würtz, H; Nielsen, K F; Kruse, P; Wilkins, K; Thrane, U; Gravesen, S; Nielsen, P A; Schneider, T

    2003-06-01

    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 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.

  12. 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.

  13. Methods for neutralizing anthrax or anthrax spores

    Science.gov (United States)

    Sloan, Mark A; Vivekandanda, Jeevalatha; Holwitt, Eric A; Kiel, Johnathan L

    2013-02-26

    The present invention concerns methods, compositions and apparatus for neutralizing bioagents, wherein bioagents comprise biowarfare agents, biohazardous agents, biological agents and/or infectious agents. The methods comprise exposing the bioagent to an organic semiconductor and exposing the bioagent and organic semiconductor to a source of energy. Although any source of energy is contemplated, in some embodiments the energy comprises visible light, ultraviolet, infrared, radiofrequency, microwave, laser radiation, pulsed corona discharge or electron beam radiation. Exemplary organic semiconductors include DAT and DALM. In certain embodiments, the organic semiconductor may be attached to one or more binding moieties, such as an antibody, antibody fragment, or nucleic acid ligand. Preferably, the binding moiety has a binding affinity for one or more bioagents to be neutralized. Other embodiments concern an apparatus comprising an organic semiconductor and an energy source. In preferred embodiments, the methods, compositions and apparatus are used for neutralizing anthrax spores.

  14. DnaJ sequences of Bacillus cereus strains isolated from outbreaks of hospital infection are highly similar to Bacillus anthracis.

    Science.gov (United States)

    Zhang, Jiwei; van Hung, Pham; Hayashi, Masahiro; Yoshida, Shigeru; Ohkusu, Kiyofumi; Ezaki, Takayuki

    2011-07-01

    Bacillus cereus is becoming an important nomosomial pathogen because of frequent isolation from blood cultures and from severe systemic infections. To differentiate highly pathogenic outbreak strain of B. cereus from other sources of the Bacillus cereus, we attempted to analyze their dnaJ sequences. Assays indicated that dnaJ sequence similarity of all of 52 blood culture isolates of B. cereus ranged from 92.8% to 100%. The distance between B. anthracis and B. cereus except six outbreak isolates ranged from 3.8% to 6.4%. The dnaJ sequences of six outbreak strains of B. cereus (GTC 02891, GTC 02896, GTC 02916, GTC 02917, GTC 03221, and GTC 03222) were closely related to those of B. anthracis (99.2%-99.5% sequence similarity). Ba813 sequences were only found in the six outbreak strains of B. cereus. The other pathogenic factors of B. anthracis were not found in these six outbreak strains, with the exception of GTC 02891 (cap-positive). The six outbreak strains formed clear β-hemolytic colonies on a sheep blood agar plate. Our findings suggest that outbreak strains of B. cereus isolated from blood cultures are likely to have the risk of causing serious infection, and dnaJ and Ba813 are important markers to identify such strains. Phylogenetic analysis of dnaJ and MLST revealed that the six outbreak strains of B. cereus are closely related to B. anthracis.

  15. Recombinant Bacillus subtilis spores expressing cholera toxin B subunit and Helicobacter pylori urease B confer protection against H. pylori in mice.

    Science.gov (United States)

    Zhou, Zhenwen; Dong, Hui; Huang, Yanmei; Yao, Shuwen; Liang, Bingshao; Xie, Yongqiang; Long, Yan; Mai, Jialiang; Gong, Sitang

    2017-01-01

    Helicobacter pylori infection is associated with chronic gastritis, peptic ulcers, gastric cancer and mucosa-associated lymphoid tissue lymphoma. The limitations of current therapies for H. pylori infection include poor compliance and antibiotic resistance. Therefore, an effective anti-H. pylori vaccine would be an alternative or complement to antibiotic treatment. Urease B (UreB) is considered an ideal vaccine antigen against H. pylori infection. In this study, cholera toxin B subunit (CTB), a mucosal adjuvant, was used to enhance the immunogenicity of a novel Bacillus subtilis spore vaccine expressing CTB-UreB, along with the B. subtilis spore coat protein CotC as a fusion protein. Oral administration of B. subtilis spores expressing CotC-UreB or CotC-CTB-UreB led to increased levels of UreB-specific IgG in serum and UreB-specific IgA in faeces, as well as elevated levels of IL-10 and IFN-γ in splenocytes. In addition, oral administration of CotC-UreB or CotC-CTB-UreB spores induced significant reductions (80.0 and 90.5 %, respectively) in gastric H. pylori bacterial load (1.11±0.36×105 and 0.53±0.21×105 c.f.u., respectively) compared to that of the CotC control group (5.56±1.64×105 c.f.u., P<0.01). Moreover, CotC-CTB-UreB spores were significantly more effective at reducing the bacterial load than CotC-UreB spores (P<0.05). These results indicate that CotC-CTB-UreB-expressing B. subtilis spores are a potential vaccine candidate for the control of H. pylori infection.

  16. High avidity binding of engineered papaya mosaic virus virus-like particles to resting spores of Plasmodiophora brassicae.

    Science.gov (United States)

    Morin, Hélène; Tremblay, Marie-Hélène; Plante, Edith; Paré, Christine; Majeau, Nathalie; Hogue, Richard; Leclerc, Denis

    2007-02-01

    Papaya mosaic virus (PapMV) like particles (VLPs) were used as a platform for fusion of affinity peptides binding to resting spores of Plasmodiophora brassicae-a major pathogen of crucifers. Three peptides with specific affinity to the target were isolated and cloned at the C-terminus of the PapMV coat protein (CP), generating three different high avidity VLPs. The peptides were exposed at the surface of the VLPs and their avidity to resting spores of P. brassicae was measured by flow cytometry. NLP-A, with the peptide DPAPRPR, showed the highest avidity. The binding avidity of NLP-A to P. brassicae spores was comparable to that of a polyclonal antibody. NLP-A was also shown to be more specific than the antibody. Fusion of the affinity peptide to a monomeric form (mCP) of the CP [Lecours, K., Tremblay, M.-H., Laliberté Gagné, M.-E., Gagné, S.M., Leclerc, D., 2006. Purification and biochemical characterization of a monomeric form of papaya mosaic potexvirus coat protein. Protein Express. Purific. 47, 273-280] generated a fusion protein that was unable to assemble into VLPs, and mCP-A fusions failed to bind resting spores. The avidity of VLP-A was increased by adding a glycine spacer between the C-terminus of the PapMV CP and the peptide, and improved even further by using a duplicated A peptide in the fusion protein. The use of high avidity VLPs has advantages over polyclonal antibodies because of target specificity. VLPs offers the specificity of monoclonal antibodies but can be more easily generated using the powerful selection of phage display.

  17. 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

  18. 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

    2015-11-01

    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.

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

    Directory of Open Access Journals (Sweden)

    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

  20. Elucidation of enterotoxigenic Bacillus cereus outbreaks in Austria by complementary epidemiological and microbiological investigations, 2013.

    Science.gov (United States)

    Schmid, Daniela; Rademacher, Corinna; Kanitz, Elisabeth Eva; Frenzel, Elrike; Simons, Erica; Allerberger, Franz; Ehling-Schulz, Monika

    2016-09-01

    Identifying Bacillus cereus as the causative agent of a foodborne outbreak still poses a challenge. We report on the epidemiological and microbiological investigation of three outbreaks of food poisoning (A, B, and C) in Austria in 2013. A total of 44% among 32 hotel guests (A), 22% among 63 employees (B) and 29% among 362 residents of a rehab clinic (C) fell sick immediately after meal consumption. B. cereus isolated from left overs or retained samples from related foods were characterized by toxin gene profiling, and molecular typing using panC sequencing and M13-PCR typing (in outbreak A and C). We identified two B. cereus strains in outbreak A, and six B. cereus strains, each in outbreak B and C; we also found Staphylococcus aureus and staphylococcal enterotoxins in outbreak A. The panC sequence based phylogenetic affiliation of the B. cereus strains, together with findings of the retrospective cohort analyses, helped determining their etiological role. Consumption of a mashed potatoes dish in outbreak A (RR: ∞), a pancake strips soup in outbreak B (RR 13.0; 95% CI 1.8-93.0) and for outbreak C of a fruit salad (RR 1.50; 95% CI 1.09-2.00), deer ragout (RR: 1.99; 95% CI 1.23-3.22) and a cranberry/pear (RR 2.46; 95% CI 1.50-4.03)were associated with increased risk of falling sick. An enterotoxigenic strain affiliated to the phylogenetic group with the highest risk of food poisoning was isolated from the crème spinach and the strawberry buttermilk, and also from the stool samples of the one B. cereus positive outbreak case-patient, who ate both. Our investigation of three food poisoning outbreaks illustrates the added value of a combined approach by using epidemiological, microbiological and genotyping methods in identifying the likely outbreak sources and the etiological B. cereus strains.

  1. 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

  2. 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

  3. Vacuum Distillation Residue Upgrading by an Indigenous Bacillus Cereus

    Directory of Open Access Journals (Sweden)

    Mitra Sadat Tabatabaee

    2013-07-01

    Full Text Available Background:Biological processing of heavy fractions of crude oils offers less severe process conditions and higher selectivity for refining. Biochemical Processes are expected to be low demand energy processes and certainly ecofriendly.Results:A strain of biosurfactant producing bacterium was isolated from an oil contaminated soil at Tehran refinery distillation unit. Based on selected phenotypic and genotypic characteristic including morphology, biochemical proprety, and 16 SrRNA sequencing identified as a novel strain of Bacillus cereus (JQ178332. This bacterium endures a wide range of pH, salinity and temperature. This specific strain utilizes both paraffin and anthracene as samples of aliphatic and polycyclic aromatic hydrocarbons. The ability of this bacterium to acquire all its energy and chemical requirements from Vacuum Distillation Residue (VR, as a net sample of problematic hydrocarbons in refineries, was studied. SARA test ASTM D4124-01 revealed 65.5% decrease in asphaltenic, 22.1% in aliphatics and 30.3% in Aromatics content of the VR in MSM medium. Further results with 0.9% saline showed 55% decrease in asphaltene content and 2.1% Aromatics respectively.Conclusion:Remarkable abilities of this microorganism propose its application in an ecofriendly technology to upgrade heavy crude oils.

  4. Compensatory Evolution of Intrinsic Transcription Terminators in Bacillus Cereus

    Science.gov (United States)

    Safina, Ksenia R.; Mironov, Andrey A.

    2017-01-01

    Many RNA molecules possess complicated secondary structure critical to their function. Mutations in double-helical regions of RNA may disrupt Watson–Crick (WC) interactions causing structure destabilization or even complete loss of function. Such disruption can be compensated by another mutation restoring base pairing, as has been shown for mRNA, rRNA and tRNA. Here, we investigate the evolution of intrinsic transcription terminators between closely related strains of Bacillus cereus. While the terminator structure is maintained by strong natural selection, as evidenced by the low frequency of disrupting mutations, we observe multiple instances of pairs of disrupting-compensating mutations in RNA structure stems. Such two-step switches between different WC pairs occur very fast, consistent with the low fitness conferred by the intermediate non-WC variant. Still, they are not instantaneous, and probably involve transient fixation of the intermediate variant. The GU wobble pair is the most frequent intermediate, and remains fixed longer than other intermediates, consistent with its less disruptive effect on the RNA structure. Double switches involving non-GU intermediates are more frequent at the ends of RNA stems, probably because they are associated with smaller fitness loss. Together, these results show that the fitness landscape of bacterial transcription terminators is rather rugged, but that the fitness valleys associated with unpaired stem nucleotides are rather shallow, facilitating evolution. PMID:28201729

  5. A biochemically active MCM-like helicase in Bacillus cereus

    Science.gov (United States)

    Samuels, Martin; Gulati, Gaurav; Shin, Jae-Ho; Opara, Rejoice; McSweeney, Elizabeth; Sekedat, Matt; Long, Stephen; Kelman, Zvi; Jeruzalmi, David

    2009-01-01

    The mini-chromosome maintenance (MCM) proteins serve as the replicative helicases in archaea and eukaryotes. Interestingly, an MCM homolog was identified, by BLAST analysis, within a phage integrated in the bacterium Bacillus cereus (Bc). BcMCM is only related to the AAA region of MCM-helicases; the typical amino-terminus is missing and is replaced by a segment with weak homology to primases. We show that BcMCM displays 3′→5′ helicase and ssDNA-stimulated ATPase activity, properties that arise from its conserved AAA domain. Isolated BcMCM is a monomer in solution but likely forms the functional oligomer in vivo. We found that the BcMCM amino-terminus can bind ssDNA and harbors a zinc atom, both hallmarks of the typical MCM amino-terminus. No BcMCM-catalyzed primase activity could be detected. We propose that the divergent amino-terminus of BcMCM is a paralog of the corresponding region of MCM-helicases. A divergent amino terminus makes BcMCM a useful model for typical MCM-helicases since it accomplishes the same function using an apparently unrelated structure. PMID:19474351

  6. Purification and characterization of two polyhydroxyalcanoates from Bacillus cereus.

    Science.gov (United States)

    Zribi-Maaloul, Emna; Trabelsi, Imen; Elleuch, Lobna; Chouayekh, Hichem; Ben Salah, Riadh

    2013-10-01

    This work aimed to study the potential of 155 strains of Bacillus sp., isolated from a collection of Tunisian microorganisms, for polyhydroxyalcanoates production. The strains were submitted to a battery of standard tests commonly used for determining bioplastic properties. The findings revealed that two of the isolates, namely Bacillus US 163 and US 177, provided red excitations at a wavelength of approximately 543 nm. The polyhydroxyalcanoates produced by the two strains were purified. Gas chromatography-mass spectroscopy (GC-MS), Fourier transformed infrared spectroscopy (FTIR), and gel permeation chromatography (GPC) were used to characterize the two biopolymers. Bacillus US 163 was noted to produce a poly methyl-3-hydroxy tetradecanoic acid (P-3HTD) with an average molecular weight of 455 kDa, a completely amorphous homopolymer without crystallinity. The US 177 strain produced a homopolymer of methyl-3-hydroxy octadecanoic acid (P3-HOD) with an average molecular weight of 555 kDa. Exhibiting the highest performance, US 163 and US 177 were submitted to 16S rRNA gene sequencing, and the results revealed that they belonged to the Bacillus cereus species. Overall, the findings indicated that the Bacilli from petroleum soil have a number of promising properties that make them promising candidates for bioplastic production.

  7. On the neutralization of bacterial spores in post-detonation flows

    Science.gov (United States)

    Gottiparthi, K. C.; Schulz, J. C.; Menon, S.

    2014-09-01

    In multiple operational scenarios, explosive charges are used to neutralize confined or unconfined stores of bacterial spores. The spore destruction is achieved by post-detonation combustion and mixing of hot detonation product gases with the ambient flow and spore clouds. In this work, blast wave interaction with bacterial spore clouds and the effect of post-detonation combustion on spore neutralization are investigated using numerical simulations. Spherical explosive charges (radius, = 5.9 cm) comprising of nitromethane are modeled in the vicinity of a spore cloud, and the spore kill in the post-detonation flow is quantified. The effect of the mass of the spores and the initial distance, , of the spore cloud from the explosive charge on the percentage of spores neutralized is investigated. When the spores are initially placed within a distance of 3.0, within 0.1 ms after detonation of the charge, all the spores are neutralized by the blast wave and the hot detonation product gases. In contrast, almost all the spores survived the explosion when is greater than 8.0. The percentage of intact spores varied from 0 to 100 for 3.0 8.0 with spore neutralization dependent on time spent by the spores in the post-detonation mixing/combustion zone.

  8. Does proximity to neighbours affect germination of spores of non-proteolytic Clostridium botulinum?

    Science.gov (United States)

    Webb, Martin D; Stringer, Sandra C; Le Marc, Yvan; Baranyi, József; Peck, Michael W

    2012-10-01

    It is recognised that inoculum size affects the rate and extent of bacterial spore germination. It has been proposed that this is due to spores interacting: molecules released from germinated spores trigger germination of dormant neighbours. This study investigated whether changes to the total number of spores in a system or proximity to other spores (local spore density) had a more significant effect on interaction between spores of non-proteolytic Clostridium botulinum strain Eklund 17B attached to defined areas of microscope slides. Both the number of spores attached to the slides and local spore density (number of spores per mm(2)) were varied by a factor of nine. Germination was observed microscopically at 15 °C for 8 h and the probability of, and time to, germination calculated from image analysis measurements. Statistical analysis revealed that the effect of total spore number on the probability of germination within 8 h was more significant than that of proximity to neighbours (local spore density); its influence on germination probability was approximately four-times greater. Total spore number had an even more significant affect on time to germination; it had a nine-fold greater influence than proximity to neighbours. The applied models provide a means to characterise, quantitatively, the effect of the total spore number on spore germination relative to the effect of proximity to neighbouring spores.

  9. Effects of temperature and desiccation on ex situ conservation of nongreen fern spores

    Science.gov (United States)

    Conservation of the genetic diversity of ferns is limited by the paucity of ex situ spore banks. Conflicting reports of fern spore response to low temperature and moisture impedes establishment of fern spore banks. There is little information available to evaluate longevity of fern spores under dif...

  10. Sperm bioassay for rapid detection of cereulide-producing Bacillus cereus in food and related environments.

    Science.gov (United States)

    Andersson, Maria A; Jääskeläinen, Elina L; Shaheen, Ranad; Pirhonen, Tuula; Wijnands, Luc M; Salkinoja-Salonen, Mirja S

    2004-07-15

    A novel in vitro method, sperm micro assay for rapidly distinguishing cereulide, the emetic toxin producing Bacillus cereus from non-producers is described and its use for quantitating cereulide and screening large numbers of B. cereus strains/colonies evaluated. The assay is non-laborious and can be executed with equipment present in most laboratories. Boar spermatozoa, purchased as standard semen from artificial insemination suppliers, are used to detect toxicity. Boar sperms respond within 5 min by cessation of motility when exposed at 37 degrees C to heat-treated (100 degrees C) extract prepared from a cereulide containing B. cereus. The assay can be done on individual colonies on the primary plate, with no need for pure culture and the qualitative result is obtained within 30 min. The assay is robust, not sensitive to age or storage of the culture plates. The use of the sperm micro assay for semiquantitative estimation of cereulide in B. cereus was validated with 14 different B. cereus strains using as reference the specific chemical assay for cereulide, based on liquid chromatography-ion trap mass spectrometry (LC-ion trap MS). The cereulide contents calculated from endpoint dilutions of the sperm micro assay matched the result of the chemical analysis closely. The detection threshold of the sperm micro assay was measured as 0.3 +/- 0.1 ng of cereulide per 5.4 x 10(6) sperm cells in 0.2 ml or 0.9 ng of cereulide per mg of B. cereus biomass (wet wt.). Food-related B. cereus strains contained 4-400 ng of cereulide per mg (wet wt.). When a large number of B. cereus of food, non-food, clinical and environmental origins were screened and 107 independent strains/isolates were identified as cereulide producers, it was observed that all of these had low or no haemolytic activity when cultivated on bovine blood agar. None of the strains/isolates with wide, clear zones of haemolysis, considered typical of B. cereus, produced cereulide.

  11. Quantification of [i]C. globosum [/i]spores in house dust samples

    Directory of Open Access Journals (Sweden)

    Chunhua Shi

    2014-09-01

    Full Text Available [i]Chaetomium globosum [/i]is one of the most common fungi that grows in damp buildings and occurs in agricultural and forestry workplaces. Using sera from atopic patients, we characterized and purified an extracellular chitosanase (Chg47 from [i]C. globosum[/i] that is antigenic to humans. The study reports the production of monoclonal antibodies to the protein. Three capture ELISAs were developed for Chg47 for detection of spores and spore and mycelial fragments in dust samples using different mono- and polyclonal antibody combinations. One method is based on an enhanced biotinylated polyclonal antibody as the secondary antibody and coating anti-IgM to capture one of two clones of IgM monoclonal antibodies as the capture antibody. The other method makes use of an enhanced rabbit polyclonal antibody as both the primary and capture antibody. The detection limit of the double PAb method for the Chg47 antigen was 7.6 pg/ml. When the anti-IgM+10B3 clone was used, the detection limit was 61 pg/ml and for anti-IgM+5F12, 122 pg/ml. The detection limit of double PAb method is comparable to methods for the allergen and spores of [i]Aspergillus versicolor[/i] in house dust and is more sensitive than other immunoassays for allergens in house including for [i]Stachybotrys chartarum[/i], [i]Aspergillus fumigatus[/i] and [i]Alternaria alternata[/i]. All three methods had limited cross-reactivity to fungi common in house dust representing a diverse array of taxa.

  12. Bacillus Cereus catheter related bloodstream infection in a patient in a patient with acute lymphblastic leukemia

    Directory of Open Access Journals (Sweden)

    Lütfiye Öksüz

    2012-01-01

    Full Text Available Bacillus cereus infection is rarely associated with actual infection and for this reason single positive blood culture is usually regarded as contamination . However it may cause a number of infections, such catheter-related blood stream infections. Significant catheter-related bloodstream infections (CRBSI caused by Bacillus spp. are mainly due to B.cereus and have been predominantly reported in immunocompromised hosts1 . Catheter removal is generally advised for management of infection. In this report, catheter-related bacteremia caused by B.cereus in a patient with acute lymphoblastıc leukemia (ALL in Istanbul Medical Faculty was presented.A 44-year old man presented with fatigue, weight loss, epistaxis and high fever. A double-lumen Hickman–catheter (Bard 12.0 Fr, Round Dual Lumen was inserted by surgical cut-down to access the right subclavian vein which would be necessary for allogeneic stem cell transplantation. Three weeks later the patient presented with high fever and headache. Bacillus spp. was isolated from the cathether while blood culture obtained from the peripheral vein remained negative. The bacterial identification was confirmed as B.cereus using VITEK identification system It has been reported Bacillus cereus septicemia may be fatal in immunocompromised hosts despite broad-spectrum appropriate treatment10. Catheter removal is essential for prevention of recurrent bacteremia. Long-term cathater salvage should be reserved for appropriate patient group.

  13. Infective endocarditis due to Bacillus cereus in a pregnant female: A case report and literature review.

    Science.gov (United States)

    Shah, Mahek; Patnaik, Soumya; Wongrakpanich, Supakanya; Alhamshari, Yaser; Alnabelsi, Talal

    2015-01-01

    Incidence of infective endocarditis during pregnancy is around 0.006% with high maternal and fetal mortality. Bacillus cereus is an extremely rare cause for endocarditis in intravenous drug abusers (IVDA) or those with valvular disease or devices such as pacemakers. We report a case of B. cereus endocarditis, which, to the best of our knowledge, has never been reported in pregnancy. A 30-year-old, 25-week pregnant female presented with right shoulder pain, swelling and erythema on the lateral aspect of deltoid muscle from large abscess over her deltoid muscle. She was found to have a vegetation on the native tricuspid valve. Cultures from abscess fluid and blood cultures grew B. cereus, she was appropriately treated with antimicrobials and had favorable outcomes. There are cereus endocarditis reported but none during pregnancy. When cultures grow unusual organisms the case must be thoroughly investigated. This case illustrates a rare situation (endocarditis in pregnancy) with an unusual outcome (B. cereus) on an uncommon valve (tricuspid valve).

  14. Infective endocarditis due to Bacillus cereus in a pregnant female: A case report and literature review

    Directory of Open Access Journals (Sweden)

    Mahek Shah

    2015-01-01

    Full Text Available Incidence of infective endocarditis during pregnancy is around 0.006% with high maternal and fetal mortality. Bacillus cereus is an extremely rare cause for endocarditis in intravenous drug abusers (IVDA or those with valvular disease or devices such as pacemakers. We report a case of B. cereus endocarditis, which, to the best of our knowledge, has never been reported in pregnancy. A 30-year-old, 25-week pregnant female presented with right shoulder pain, swelling and erythema on the lateral aspect of deltoid muscle from large abscess over her deltoid muscle. She was found to have a vegetation on the native tricuspid valve. Cultures from abscess fluid and blood cultures grew B. cereus, she was appropriately treated with antimicrobials and had favorable outcomes. There are <20 cases of B. cereus endocarditis reported but none during pregnancy. When cultures grow unusual organisms the case must be thoroughly investigated. This case illustrates a rare situation (endocarditis in pregnancy with an unusual outcome (B. cereus on an uncommon valve (tricuspid valve.

  15. [Bacillus cereus sepsis and subarachnoid hemorrhage following consolidation chemotherapy for acute myelogenous leukemia].

    Science.gov (United States)

    Kawatani, Eri; Kishikawa, Yuki; Sankoda, Chikahiro; Kuwahara, Nobuo; Mori, Daisuke; Osoegawa, Kouichi; Matsuishi, Eijo; Gondo, Hisashi

    2009-04-01

    A 64-year-old man with acute myelogenous leukemia (FAB classification, M7) in remission received consolidation chemotherapy with mitoxantrone/cytosine arabinoside. WBC counts decreased to 0/microl on day 14, and fever (39.3 degrees C) and epigastralgia developed on day 15. Cefozopran was instituted for febrile neutropenia; however, on day 16, he was found to be in cardiac arrest. CT scan on day 16 revealed subarachnoid hemorrhage. Gram-positive rods were isolated from blood cultures on day 15, and were later identified as B.cereus. He recovered transiently, but eventually died on day 19. Postmortem examination demonstrated many colonies of B. cereus in the cerebrum, cerebellum, lung, and liver. Hepatocyte necrosis was also observed in the liver. Bacterial aneurysms or septic emboli were not identified in the arachnoid vessels, but necrosis of cerebral vessels was prominent, which was considered to be the cause of subarachnoid hemorrhage. Fatal subarachnoid hemorrhage has been reported to be associated with B. cereus sepsis, which developed at nadir following chemotherapy for leukemia patients. Because of the aggressive clinical course of B. cereus sepsis, including the risk for subarachnoid hemorrhage, early treatment with effective antibiotics for B. cereus sepsis would be important in the management of leukemia patients after chemotherapy.

  16. Comparative proteomic analysis of experimental evolution of the Bacillus cereus-Ketogulonicigenium vulgare co-culture.

    Directory of Open Access Journals (Sweden)

    Qian Ma

    Full Text Available The microbial co-culture system composing of Ketogulonicigenium vulgare and Bacillus cereus was widely adopted in industry for the production of 2-keto-gulonic acid (2-KGA, the precursor of vitamin C. We found serial subcultivation of the co-culture could enhance the yield of 2-KGA by 16% in comparison to that of the ancestral co-culture. To elucidate the evolutionary dynamics and interaction mechanisms of the two microbes, we performed iTRAQ-based quantitative proteomic analyses of the pure cultures of K. vulgare, B. cereus and their co-culture during serial subcultivation. Hierarchy cluster analyses of the proteomic data showed that the expression level of a number of crucial proteins associated with sorbose conversion and oligopeptide transport was significantly enhanced by the experimental evolution. In particular, the expression level of sorbose/sorbosone dehydrogenase was enhanced in the evolved K. vulgare, while the expression level of InhA and the transport efficiency of oligopeptides were increased in the evolved B. cereus. The decreased sporulating protein expression and increased peptide transporter expression observed in evolved B. cereus, together with the increased amino acids synthesis in evolved K. vulgare suggested that serial subcultivation result in enhanced synergistic cooperation between K. vulgare and B. cereus, enabling an increased production of 2-KGA.

  17. BACILLUS CEREUS EM PRODUTOS LÁCTEOS - UMA REVISÃO

    Directory of Open Access Journals (Sweden)

    Maike Taís Maziero

    2011-06-01

    Full Text Available Bacillus cereus é uma bactéria termodúrica, formadora de esporos, capaz de se multiplicar em temperatura de refrigeração. Pode causar dois tipos de doenças de origem alimentar: a síndrome emética e a síndrome diarréica. Além do aspecto epidemiológico, B. cereus causa defeitos tecnológicos em produtos lácteos, relacionados com a produção de lipases e proteases. A presença de B. cereus em produtos lácteos vem sendo relatada por pesquisas feitas em vários países, bem como os defeitos tecnológicos associados à contaminação dos produtos por essa bactéria. Destacam-se, na presente revisão, os principais aspectos relacionados à presença de B. cereus em produtos lácteos. Para tanto, foram selecionados trabalhos relevantes e inovadores sobre B. cereus em várias bases de dados, entre elas, Science Direct, SciELO, Scirus e Google Acadêmico.

  18. Detection of toxin genes and RAPD analysis of bacillus cereus isolates from different soil types

    Directory of Open Access Journals (Sweden)

    Savic Dejana

    2015-01-01

    Full Text Available The aim of this study was to detect genes for enterotoxins (hbla, entFM and bceT and for emetic toxin (cer, to determine antibiotic resistance, and to estimate intraspecies diversity in B. cereus isolates by RAPD analysis. B. cereus was identified in 12 out of 117 indigenous Bacillus spp. using the classical microbiological methods and PCR. All isolates were resistant to penicillin and ampicillin, two to tetracyclin and four to trimethoprim-sulphamethoxazole. Also, all isolates produced inducible penicillinases and β-lactamase. Toxin genes were detected with PCR. EntFM and cer genes were present in all isolates, hbla in all, but two, and bceT in none. RAPD analysis was performed with four different primers, two of them designed for this study. The intraspecies diversity revealed 10 different patterns at the 90% similarity level. Two separate clusters were formed regardless of a soil type or utilization. The detection of genes encoding toxins in all B. cereus isolates indicated these bacteria as potentially pathogenic and seriously for human health. Regardless of a soil type or utilization, the RAPD analysis showed high intraspecies heterogeneity in B. cereus isolates. To the best of our knowledge, this is the first study to analyse the presence of entero- and emetic toxin genes and genetic heterogeneity in B. cereus isolates from different soil types and different soil utilization in Serbia. [Projekat Ministarstva nauke Republike Srbije, br. TR37006

  19. Enterotoxin production by Bacillus cereus under gastrointestinal conditions and their immunological detection by commercially available kits.

    Science.gov (United States)

    Ceuppens, Siele; Rajkovic, Andreja; Hamelink, Stefanie; Van de Wiele, Tom; Boon, Nico; Uyttendaele, Mieke

    2012-12-01

    Currently, three commercial kits for Bacillus cereus enterotoxins Nhe and/or Hbl detection are available, namely, the Bacillus diarrheal enterotoxin visual immunoassay (BDE VIA™) kit (3M Tecra), B. cereus enterotoxin reversed passive latex agglutination (BCET-RPLA) kit (Oxoid), and the Duopath(®) Cereus Enterotoxins (Merck). The performance of the kits and their applicability to gastrointestinal simulation samples were evaluated. Then, the stability and production of enterotoxins Hbl and Nhe under gastrointestinal conditions were investigated. Enterotoxin production was absent or impaired at acidic pH, i.e., in gastric medium with pH 5.0 and lasagne verde with pH 5.5. B. cereus did produce enterotoxins Nhe and Hbl during anaerobic growth in intestinal medium at pH 7.0, but the toxins were instantly degraded by the enzymes in the host's digestive secretions. Preformed enterotoxins did not withstand gastrointestinal passage under the simulated conditions, which suggests that preformed enterotoxins in food do not contribute to the diarrheal food poisoning syndrome. In conclusion, diarrhea is probably caused by de novo enterotoxin production by B. cereus cells located closely to the host's intestinal epithelium.

  20. Proteome data to explore the impact of pBClin15 on Bacillus cereus ATCC 14579

    Directory of Open Access Journals (Sweden)

    Jean-Paul Madeira

    2016-09-01

    Full Text Available This data article reports changes in the cellular and exoproteome of B. cereus cured from pBClin15.Time-course changes of proteins were assessed by high-throughput nanoLC-MS/MS. We report all the peptides and proteins identified and quantified in B. cereus with and without pBClin15. Proteins were classified into functional groups using the information available in the KEGG classification and we reported their abundance in term of normalized spectral abundance factor. The repertoire of experimentally confirmed proteins of B. cereus presented here is the largest ever reported, and provides new insights into the interplay between pBClin15 and its host B. cereus ATCC 14579. The data reported here is related to a published shotgun proteomics analysis regarding the role of pBClin15, “Deciphering the interactions between the Bacillus cereus linear plasmid, pBClin15, and its host by high-throughput comparative proteomics” Madeira et al. [1]. All the associated mass spectrometry data have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org via the PRIDE partner repository (http://www.ebi.ac.uk/pride/, with the dataset identifier PRIDE: PXD001568, PRIDE: PXD002788 and PRIDE: PXD002789.

  1. Alicyclobacillus acidoterrestris: new methods for inhibiting spore germination.

    Science.gov (United States)

    Bevilacqua, A; Sinigaglia, M; Corbo, M R

    2008-07-15

    For a long period the thermal processing has been considered as the only way to reduce the initial spore number of Alicyclobacillus acidoterrestris and prevent the spoilage of acidic beverage. New methods, however, were proposed by the literature to control spore germination both in laboratory media and in real systems. After a brief introduction on the impact of A. acidoterrestris in food microbiology and a description of enumeration methods and heat processing applied by the juices manufactures, a review of innovative approaches to inhibit and/or control spore germination is proposed. In particular, this paper focuses on two different topics; the 1st is the use of some natural compounds (monolaurin, lysozyme, nisin and essential oils) or some chemicals, conventional (like sodium-benzoate, organic acids, surfactants and chlorine dioxide) or not conventional (chlorine dioxide as gas). The 2nd topic is a description of some innovative methods to reduce the initial spore number (high hydrostatic and homogenisation pressures, radiation and microwaves).

  2. Pharmacologic and toxicologic evaluation of C. novyi-NT spores.

    Science.gov (United States)

    Diaz, Luis A; Cheong, Ian; Foss, Catherine A; Zhang, Xiaosong; Peters, Brock A; Agrawal, Nishant; Bettegowda, Chetan; Karim, Baktiar; Liu, Guosheng; Khan, Khalid; Huang, Xin; Kohli, Manu; Dang, Long H; Hwang, Paul; Vogelstein, Ahava; Garrett-Mayer, Elizabeth; Kobrin, Barry; Pomper, Martin; Zhou, Shibin; Kinzler, Kenneth W; Vogelstein, Bert; Huso, David L

    2005-12-01

    Clostridium novyi-NT (C. novyi-NT) spores have been shown to be potent therapeutic agents in experimental tumors of mice and rabbits. In the present study, pharmacologic and toxicologic studies were performed to better understand the factors influencing the efficacy and toxicity of this form of therapy. We found that spores were rapidly cleared from the circulation by the reticuloendothelial system. Even after large doses were administered, no clinical toxicity was observed in healthy mice or rabbits. The spores were also not toxic in mice harboring poorly vascularized non-neoplastic lesions, including myocardial infarcts. In tumor-bearing mice, toxicity appeared related to tumor size and spore dose, as expected with any bacterial infection. However, there was no laboratory or histopathologic evidence of sepsis, and the toxicity could be effectively controlled by simple hydration.

  3. Waterline ATS B. globigii spore water disinfection data

    Data.gov (United States)

    U.S. Environmental Protection Agency — Disinfection of B. globigii spores (a non-pathogenic surrogate for B. anthracis) in clean and dirty water using the ATS-Waterline system, which uses ultraviolet...

  4. Spore-forming bacteria and their utilisation as probiotics.

    Science.gov (United States)

    Bader, J; Albin, A; Stahl, U

    2012-03-01

    In this review article, the beneficial application of bacterial spore formers as probiotics in the food industry is discussed based on the knowledge gleaned from current publications. The summary of new scientific results provides evidence of the advantages of the utilisation of Bacillus or Clostridium strains in the food industry. Both bacteria are able to produce a very stable duration form: the endospore. Compared to the widely used lactic acid bacteria, bacterial spores offer the advantage of a higher survival rate during the acidic stomach passage and better stability during the processing and storage of the food product. In many food products, germination of the spores does not occur. Hence the product quality of the food is not affected because of their inactive metabolism. Besides the possible utilisation and functional properties, an overview of the fast-developing knowledge about the mechanisms of the beneficial health effects of spore-forming bacteria is provided.

  5. Small Probes for Orbital Return of Experiments (SPORE) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Analogous to the CubeSat standardization of micro-satellites, the SPORE flight system architecture will utilize a modular design approach to provide low-cost...

  6. Oxidation mechanism of Penicillium digitatum spores through neutral oxygen radicals

    Science.gov (United States)

    Hashizume, Hiroshi; Ohta, Takayuki; Takeda, Keigo; Ishikawa, Kenji; Hori, Masaru; Ito, Masafumi

    2014-01-01

    To investigate the inactivation process of Penicillium digitatum spores through neutral oxygen species, the spores were treated with an atmospheric-pressure oxygen radical source and observed in-situ using a fluorescent confocal-laser microscope. The treated spores were stained with two fluorescent dyes, 1,1‧-dioctadecyl-3,3,Y,3‧-tetramethylindocarbocyanine perchlorate (DiI) and diphenyl-1-pyrenylphosphine (DPPP). The intracellular organelles as well as the cell membranes in the spores treated with the oxygen radical source were stained with DiI without a major morphological change of the membranes. DPPP staining revealed that the organelles were oxidized by the oxygen radical treatment. These results suggest that neutral oxygen species, especially atomic oxygen, induce a minor structural change or functional inhibition of cell membranes, which leads to the oxidation of the intracellular organelles through the penetration of reactive oxygen species into the cell.

  7. Effect of Nanofibers on Spore Penetration and Lunar Dust Filtration

    OpenAIRE

    Phil Gibson, Ph.D.; Heidi Schreuder-Gibson, Ph.D.; Robert Stote; Margaret Roylance, Ph.D.; Cathy Capone; Masami Nakagawa, Ph.D.

    2008-01-01

    The results of two separate studies on biological spore penetration and simulated lunar dust filtration illustrate the use of nanofibers in some nonstandard filtration applications (nanofibers are generally defined as having diameters of less than a micron). In the first study, a variety of microporous liners containing microfibers and nanofibers were combined with cotton-based fabrics in order to filter aerosolized spores. The aerosol penetration resistance of the nanofiber-lined fabrics was...

  8. Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes

    Science.gov (United States)

    2014-10-30

    2010 31-May-2014 Approved for Public Release; Distribution Unlimited Final Report: (Life Science Division/Biochemistry) Inactivation of Bacillus ...S) AND ADDRESS (ES) U.S. Army Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 Bacillus Anthracis, Spores, Biofilm, Inhibition...Biochemistry) Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes Report Title The Specific Aims of the project were to investigate: 1) the

  9. Fate of ingested Clostridium difficile spores in mice.

    Directory of Open Access Journals (Sweden)

    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.

  10. Structural Analysis of Bacillus subtilis Spore Peptidoglycan during Sporulation

    OpenAIRE

    2000-01-01

    A major structural element of bacterial endospores is a peptidoglycan (PG) wall. This wall is produced between the two opposed membranes surrounding the developing forespore and is composed of two layers. The inner layer is the germ cell wall, which appears to have a structure similar to that of the vegetative cell wall and which serves as the initial cell wall following spore germination. The outer layer, the cortex, has a modified structure, is required for maintenance of spore dehydration,...

  11. Vacuum-induced Mutations In Bacillus Subtilis Spores

    Science.gov (United States)

    Munakata, N.; Maeda, M.; Hieda, K.

    During irradiation experiments with vacuum-UV radiation using synchrotron sources, we made unexpected observation that Bacillus subtilis spores of several recombination-deficient strains lost colony-forming ability by the exposure to high vacuum alone. Since this suggested the possible injury in spore DNA, we looked for mutation induction using the spores of strains HA101 (wild-type repair capability) and TKJ6312 (excision and spore repair deficient) that did not lose survivability. It was found that the frequency of nalidixic-acid resistant mutation increased several times in both of these strains by the exposure to high vacuum (10e-4 Pa after 24 hours). The analysis of sequence changes in gyrA gene showed that the majority of mutations carried a unique allele (gyrA12) of tandem double-base substitutions from CA to TT. The observation has been extended to rifampicin resistant mutations, the majority of that carried substitutions from CA to TT or AT in rpoB gene. On the other hand, when the spores of strains PS578 and PS2319 (obtained from P. Setlow) that are defective in a group of small acidic proteins (alpha/beta-type SASP) were similarly treated, none of the mutants analyzed carried such changes. This suggests that the unique mutations might be induced by the interaction of small acidic proteins with spore DNA under forced dehydration. The results indicate that extreme vacuum causes severe damage in spore DNA, and provide additional constraint to the long-term survival of bacterial spores in the space environment.

  12. 40 CFR 180.1181 - Bacillus cereus strain BPO1; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus cereus strain BPO1; exemption... FOOD Exemptions From Tolerances § 180.1181 Bacillus cereus strain BPO1; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance for residues of the Bacillus...

  13. The possibility of discriminating within the Bacillus cereus group using gyrB sequencing and PCR-RFLP

    DEFF Research Database (Denmark)

    Jensen, Gert B; Fisker, Niels; Sparsø, Thomas;

    2005-01-01

    Based on a combination of PCR and restriction endonuclease (RE) digestion (PCR-RE digestion), we have examined the possibility of differentiating members of the Bacillus cereus group. Fragments of the gyrB gene (362 bp) from pure cultures of 12 B. cereus, 25 B. thuringiensis, 25 B. mycoides and two...

  14. The worldwide distribution of genetically and phylogenetically diverse Bacillus cereus isolates harbouring Bacillus anthracis-like plasmids.

    Science.gov (United States)

    Kaminska, Paulina Sylwia; Yernazarova, Aliya; Drewnowska, Justyna Malgorzata; Zambrowski, Grzegorz; Swiecicka, Izabela

    2015-10-01

    Bacillus cereus is a close relative of B. anthracis, the causative agent of anthrax whose pathogenic determinants are located on pXO1 and pXO2 plasmids. Bacillus anthracis-like plasmids have been also noted among B. cereus, however, genetic features of B. cereus harbouring these elements remain largely undescribed, especially from the global perspective. Herein, we present the genetic polymorphism, population structure and phylogeny of B. cereus with pXO1-/pXO2-like plasmids originating from Argentina, Kazakhstan, Kenya and Poland. The plasmids were found in about 17% of the isolates, but their frequencies and expression of replicons differed within and between populations. In the multi-locus sequence typing, the bacteria exhibited high genetic polymorphism reflected by 116 sequencing types, including 84 singletons and 10 clonal complexes, which mainly consisted of isolates of the same origin. The phylogenetic analysis of pXO1-/pXO2-like positive B. cereus isolates revealed six independent clades; in certain clades individual populations predominated. Generally, B. cereus with pXO1-/pXO2-like plasmids did not indicate the genetic relationship with B. anthracis, and cannot be classified into an evolutionary independent anthrax line within the B. cereus group. Our report is of a crucial importance for discovering the genetic specificity and evolution of B. cereus bacilli.

  15. Toxin Profile, Biofilm Formation, and Molecular Characterization of Emetic Toxin-Producing Bacillus cereus Group Isolates from Human Stools.

    Science.gov (United States)

    Oh, Su Kyung; Chang, Hyun-Joo; Choi, Sung-Wook; Ok, Gyeongsik; Lee, Nari

    2015-11-01

    Emetic toxin-producing Bacillus cereus group species are an important problem, because the staple food for Korean is grains such as rice. In this study, we determined the prevalence (24 of 129 isolates) of emetic B. cereus in 36,745 stool samples from sporadic food-poisoning cases in Korea between 2007 and 2008. The toxin gene profile, toxin production, and biofilm-forming ability of the emetic B. cereus isolates were investigated. Repetitive element sequence polymorphism polymerase chain reaction fingerprints (rep-PCR) were also used to assess the intraspecific biodiversity of these isolates. Emetic B. cereus was present in 0.07% of the sporadic food-poisoning cases. The 24 emetic isolates identified all carried the nheABC and entFM genes and produced NHE enterotoxin. However, they did not have hemolysin BL toxin or related genes. A relationship between biofilm formation and toxin production was not observed in this study. The rep-PCR fingerprints of the B. cereus isolates were not influenced by the presence of toxin genes, or biofilm-forming ability. The rep-PCR assay discriminated emetic B. cereus isolates from nonemetic isolates, even if this assay did not perfectly discriminate these isolates. Further study on emetic isolates possessing a high degree of diversity may be necessary to evaluate the performance of the subtyping assay to discriminate emetic and nonemetic B. cereus isolates and could provide a more accurate indication of the risk from B. cereus strains.

  16. Characterization and exposure assessment of emetic bacillus cereus and cereulide production in food products on the Dutch market

    NARCIS (Netherlands)

    Biesta-Peters, Elisabeth G.; Dissel, Serge; Reij, Martine W.; Zwietering, Marcel H.; In't Veld, Paul H.

    2016-01-01

    The emetic toxin cereulide, which can be produced by Bacillus cereus, can be the cause of food poisoning upon ingestion by the consumer. The toxin causes vomiting and is mainly produced in farinaceous food products. This article includes the prevalence of B. cereus and of cereulide in food produc

  17. Identification of sigmaB-dependent genes in Bacillus cereus by proteome and in vitro transcription analysis

    NARCIS (Netherlands)

    Schaik, van W.; Zwietering, M.H.; Vos, de W.M.; Abee, T.

    2004-01-01

    The alternative sigma factor sigma(B) of the food pathogen Bacillus cereus is activated upon stress exposure and plays a role in the adaptive response of vegetative cells. This study describes the identification of sigma(B)-dependent genes in B. cereus. Two-dimensional gel electrophoresis was perfor

  18. The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host

    OpenAIRE

    2015-01-01

    Much of what we know regarding bacterial spore structure and function has been learned from studies of the genetically well-characterized bacterium Bacillus subtilis. Molecular aspects of spore structure, assembly, and function are well defined. However, certain bacteria produce spores with an outer spore layer, the exosporium, which is not present on B. subtilis spores. Our understanding of the composition and biological functions of the exosporium layer is much more limited than that of oth...

  19. Mechanisms of Induction of Germination of Bacillus subtilis Spores by High Pressure

    OpenAIRE

    Paidhungat, Madan; Setlow, Barbara; Daniels, William B.; Hoover, Dallas; Papafragkou, Efstathia; Setlow, Peter

    2002-01-01

    Spores of Bacillus subtilis lacking all germinant receptors germinate >500-fold slower than wild-type spores in nutrients and were not induced to germinate by a pressure of 100 MPa. However, a pressure of 550 MPa induced germination of spores lacking all germinant receptors as well as of receptorless spores lacking either of the two lytic enzymes essential for cortex hydrolysis during germination. Complete germination of spores either lacking both cortex-lytic enzymes or with a cortex not att...

  20. 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.

  1. 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.

  2. 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.

  3. Seed reserve composition and mobilization during germination and early seedling establishment of Cereus jamacaru D.C. ssp. jamacaru (Cactaceae).

    Science.gov (United States)

    Alencar, Nara L M; Innecco, Renato; Gomes-Filho, Enéas; Gallão, Maria Izabel; Alvarez-Pizarro, Juan C; Prisco, José T; Oliveira, Alexandre B De

    2012-09-01

    Cereus jamacaru, a Cactaceae found throughout northeast Brazil, is widely used as cattle food and as an ornamental and medicinal plant. However, there has been little information about the physiological and biochemical aspects involved in its germination. The aim of this study was to investigate its reserve mobilization during germination and early seedling growth. For this, C. jamacaru seeds were germinated in a growth chamber and collected at 0, 2, 4, 5, 6, 8 and 12 days after imbibition for morphological and biochemical analyses. Dry seeds had wrinkled seed coats and large, curved embryos. Lipids were the most abundant reserve, comprising approximately 55% and 65% of the dry mass for cotyledons and the hypocotylradicle axis, respectively. Soluble sugars and starch were the minor reserves, corresponding to approximately 2.2% of the cotyledons' dry mass, although their levels showed significant changes during germination. Soluble proteins corresponded to 40% of the cotyledons' dry mass, which was reduced by 81% at the final period of germination compared to dry seeds. C. jamacaru seed can be classified as an oil seed due to its high lipid content. Moreover, lipids were the main reserve mobilized during germination because their levels were strongly reduced after seed germination, while proteins were the second most utilized reserve in this process.

  4. COLONIZATION OF VIGNA RADIATA ROOTS BY CHROMIUM RESISTANT BACTERIAL STRAINS OF OCHROBACTRUM INTERMEDIUM, BACILLUS CEREUS AND BREVIBA CTERIUM SP.

    Institute of Scientific and Technical Information of China (English)

    MUHAMMAD Faisal; SHAHIDA Hasnain

    2005-01-01

    The present study deals with colonization potential of plant growth promoting bacterial strains ( Ochrobactrum intermedium, Bacillus cereus and Brevibacterium sp. ) on Vigna radiata roots. The roots were heavily colonized with O. intermedium and B. cereus as compared to Brevibacterium sp. O. intermedium mainly colonized rhizoplane while B. cereus occurred both on the rhizoplane and near root zone. O. intermedium and B. cereus were found to be present both on the rhizoplane and near root zone, while Brevibacterium only in the rhizosphere in the form of groups. The cells of B. cereus were found more in the sites where root exudates were existed. From the above results it was observed that the number of O. intermedium cells were large at root exudate site. Fig 2, Tab 1, Ref 15

  5. The Pathogenomic Sequence Analysis of B. cereus and B.thuringiensis Isolates Closely Related to Bacillus anthracis

    Energy Technology Data Exchange (ETDEWEB)

    Han, Cliff S.; Xie, Gary; Challacombe, Jean F.; Altherr, MichaelR.; Smriti, B.; Bruce, David; Campbell, Connie S.; Campbell, Mary L.; Chen, Jin; Chertkov, Olga; Cleland, Cathy; Dimitrijevic-Bussod, M.; Doggett, Norman A.; Fawcett, John J.; Glavina, Tijana; Goodwin, Lynne A.; Hill, Karen K.; Hitchcock, Penny; Jackson, Paul J.; Keim, Paul; Kewalramani, Avinash Ramesh; Longmire, Jon; Lucas, Susan; Malfatti,Stephanie; McMurry, Kim; Meincke, Linda J.; Misra, Monica; Moseman,Bernice L.; Mundt, Mark; Munk, A. Christine; Okinaka, Richard T.; Parson-Quintana, B.; Reilly, Lee P.; Richardson, Paul; Robinson, DonnaL.; Rubin, Eddy; Saunders, Elizabeth; Tapia, Roxanne; Tesmer, Judith G.; Thayer, Nina; Thompson, Linda S.; Tice, Hope; Ticknor, Lawrence O.; Wills, Patti L.; Gilna, Payl; Brettin, Thomas S.

    2005-08-18

    The sequencing and analysis of two close relatives of Bacillus anthracis are reported. AFLP analysis of over 300 isolates of B.cereus, B. thuringiensis and B. anthracis identified two isolates as being very closely related to B. anthracis. One, a B. cereus, BcE33L, was isolated from a zebra carcass in Nambia; the second, a B. thuringiensis, 97-27, was isolated from a necrotic human wound. The B. cereus appears to be the closest anthracis relative sequenced to date. A core genome of over 3,900 genes was compiled for the Bacillus cereus group, including Banthracis. Comparative analysis of these two genomes with other members of the B. cereus group provides insight into the evolutionary relationships among these organisms. Evidence is presented that differential regulation modulates virulence, rather than simple acquisition of virulence factors. These genome sequences provide insight into the molecular mechanisms contributing to the host range and virulence of this group of organisms.

  6. Multilocus sequence typing reveals that Bacillus cereus strains isolated from clinical infections have distinct phylogenetic origins.

    Science.gov (United States)

    Barker, Margaret; Thakker, Bishan; Priest, Fergus G

    2005-04-01

    Eight strains of Bacillus cereus isolated from bacteremia and soft tissue infections were assigned to seven sequence types (STs) by multilocus sequence typing (MLST). Two strains from different locations had identical STs. The concatenated sequences of the seven STs were aligned with 65 concatenated sequences from reference STs and a neighbor-joining tree was constructed. Two strains were distantly related to all reference STs. Three strains were recovered in a clade that included Bacillus anthracis, B. cereus and rare Bacillus thuringiensis strains while the other three strains were assigned to two STs that were more closely affiliated to most of the B. thuringiensis STs. We conclude that invasive B. cereus strains do not form a single clone or clonal complex of highly virulent strains.

  7. Relapsing peritonitis with Bacillus cereus in a patient on continuous ambulatory peritoneal dialysis

    DEFF Research Database (Denmark)

    Magnussen, Eyð Tausen; Vang, Amanda Gratton; á Steig, Torkil

    2016-01-01

    We present a case where Bacillus cereus was determined to be the causative agent of relapsing peritonitis in a patient on continuous ambulatory peritoneal dialysis (CAPD). The patient, a 70-year-old man from the Faroe Islands, was admitted with relapsing peritonitis four times over a 3-month period....... Peritoneal cultures were positive for growth of B. cereus, a rare bacterial cause of peritonitis. The cultures demonstrated susceptibility to vancomycin, and therefore the patient was treated with intraperitoneal vancomycin, intraperitoneal gentamycin and oral ciprofloxacin. As a result of the relapsing B....... cereus peritonitis diagnosis and a CT scan showing contraction of the peritoneum after longstanding inflammation, the peritoneal catheter was removed and the patient converted to haemodialysis. To date, the patient has not been readmitted due to peritonitis. A lack of proper hygiene when changing...

  8. Production of nanodrug for Bacillus cereus isolated from HIV positive patient using Mallotus philippensis

    Directory of Open Access Journals (Sweden)

    R. Bhuvaneswari

    2016-04-01

    Full Text Available The present investigation was aimed to synthesis of silver nanoparticles (AgNPs using Mallotus philippensis leaf extract and their antibacterial potential against Bacillus cereus isolated from HIV positive patient. In this, UV- Visible spectroscopy showed the high peak of absorption band at 450 nm. Based on XRD analysis, face centered cubic structure and average size of the AgNPs was around 16 nm. FTIR spectroscopy study revealed the seventeen functional groups of the AgNPs was observed. The morphology of AgNPs was spherical, oval shapes and diameter of the particle size ranges between 9 and 24 nm was measured using transmission electron microscopy (TEM. In addition to these green synthesized AgNPs were found to express the higher efficacy in inhibiting the growth of Bacillus cereus (B. cereus isolated from the HIV-positive patient.

  9. Phages Preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, Present and Future

    Directory of Open Access Journals (Sweden)

    Annika Gillis

    2014-07-01

    Full Text Available Many bacteriophages (phages have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here.

  10. Surface reaction of Bacillus cereus biomass and its biosorption for lead and copper ions

    Institute of Scientific and Technical Information of China (English)

    PAN Jian-hua; LIU Rui-xia; TANG Hong-xiao

    2007-01-01

    In this study, the Fourier transform infrared (FTIR) analytical technique identified the surface chemical functional groups of Bacillus cereus biomass. B. Cereus cells mainly contained carboxyl, hydroxyl, phosphate, amino, and amide functional groups. In order to explain the surface acid-base properties of aqueous B. Cereus biomass, the potentiometric titration was conducted . The computer program FITEQL 4.0 was used to perform the model calculations. The optimization results indicated that three sites-three pKas model, which assumed the cell surface to have three distinct types of surface organic functional groups based on IR analysis results, simulated the experimental results very well. Moreover, batch adsorption experiments were performed to investigate biosorption behavior of Cu (Ⅱ) and Pb (Ⅱ) ions onto the biomass. Obviously, the adsorption equilibrium data for the two ions were reasonably described by typical Langmuir isotherm.

  11. Optimal spore germination in Clostridium botulinum ATCC 3502 requires the presence of functional copies of SleB and YpeB, but not CwlJ.

    Science.gov (United States)

    Meaney, Carolyn A; Cartman, Stephen T; McClure, Peter J; Minton, Nigel P

    2015-08-01

    Germination, the process by which dormant endospores return to vegetative growth, is a critical process in the life cycle of the notorious pathogen Clostridium botulinum. Crucial is the degradation by hydrolytic enzymes of an inner peptidoglycan spore layer termed the cortex. Two mechanistically different systems of cortex lysis exist in spores of Clostridium species. C. botulinum ATCC 3502 harbours the Bacillus-like system of SleB, CwlJ and YpeB cortex lytic enzymes (CLEs). Through the construction of insertional gene knockout mutants in the sleB, cwlJ and ypeB genes of C. botulinum ATCC 3502 and the production of spores of each mutant strain, the effect on germination was assessed. This study demonstrates a reduced germination efficiency in spores carrying mutations in either sleB or ypeB with an approximate 2-fold reduction in heat resistant colony forming units (CFU/OD600) when plated on rich media. This reduction could be restored to wild-type levels by removing the spore coat and plating on media supplemented with lysozyme. It was observed that cwlJ spores displayed a similar germination efficiency as wild-type spores (P > 0.05). An optimal germinant commixture was identified to include a combination of l-alanine with sodium bicarbonate as it resulted in a 32% drop in OD600, while the additional incorporation of l-lactate resulted in a 57% decrease. Studies of the germination efficiency of spores prepared from all three CLE mutants was performed by monitoring the associated decrease in optical density but a germination defect was not observed in any of the CLE mutant strains. This was likely due to the lack of specificity of this particular assay. Taken together, these data indicate that functional copies of SleB and YpeB, but not CwlJ are required for the optimal germination of the spores of C. botulinum ATCC 3502.

  12. Phospholipase C from Pseudomonas aeruginosa and Bacillus cereus;characterization of catalytic activity

    Institute of Scientific and Technical Information of China (English)

    Nooran Sherif Elleboudy; Mohammad Mabrouk Aboulwafa; Nadia Abdel-Haleem Hassouna

    2014-01-01

    Objective:To study characteristics of phospholipases C (PLCs), their importance for producing microorganisms as well as the potential of their use for industrial purposes. Methods:PLC from Bacillus cereus (B. cereus) D101 was selected as an example of Gram-positive PLCs and PLC from Pseudomonas aeruginosa (P. aeruginosa) D183 of Gram-negative ones. Enzymes were partially purified by ammonium sulfate precipitation followed by membrane dialysis. Partially purified preparations were used to study effect of different factors on activities as well as in substrate specificity tests which were conducted using a turbidimetric assay method. Results: Maximum activity was at pH 7 and 8 and 40℃for P. aeruginosa PLC, and pH 8-10 and 37℃for B. cereus PLC. Both PLCs were inhibited by Pi at 5 mM or higher, whereas, PLC from B. cereus only was inhibited by EDTA. Activity of P. aeruginosa PLC was not affected by removing Zn2+ions from reaction mixture or their replacement with Ca2+, Ba2+, Mg2+or Mn2+ions. Vis-à-vis, activity of B. cereus PLC was found to be metal ion dependent. PLCs from both isolates were relatively thermostable and showed maximum affinity toward phosphatidylcholine. Sphingomyelin and phosphatidylethanolamine were not good substrates and phosphatidylinositol, phosphatidylserine, phosphatidylglycerol and cardiolipin could be considered non-substrates. Conclusions: Human body physiological conditions could favor activity of P. aeruginosa and B. cereus PLCs. These enzymes may participate in phosphate scavenging and virulence of producing isolates but not in autolysis. PLCs from both isolates are potential candidates for industrial use.

  13. Successful Treatment of Bacillus cereus Bacteremia in a Patient with Propionic Acidemia

    Directory of Open Access Journals (Sweden)

    Fatma Deniz Aygun

    2016-01-01

    Full Text Available Bacillus cereus can cause serious, life-threatening, systemic infections in immunocompromised patients. The ability of microorganism to form biofilm on biomedical devices can be responsible for catheter-related bloodstream infections. Other manifestations of severe disease are meningitis, endocarditis, osteomyelitis, and surgical and traumatic wound infections. The most common feature in true bacteremia caused by Bacillus is the presence of an intravascular catheter. Herein, we report a case of catheter-related bacteremia caused by B. cereus in a patient with propionic acidemia.

  14. Bacillus cereus Cerebral Abscess During Induction Chemotherapy for Childhood Acute Leukemia.

    Science.gov (United States)

    Dabscheck, Gabriel; Silverman, Lewis; Ullrich, Nicole J

    2015-10-01

    A 5-year-old boy with standard-risk B-cell acute lymphoblastic anemia developed fever during induction chemotherapy. The patient had no neurological symptoms. Blood cultures grew Bacillus cereus and neuroimaging studies demonstrated a cerebral abscess. Imaging changes resolved after completion of antibiotics. Bacillus cereus bacteremia is increasingly implicated as the cause of life-threatening infections, including cerebral abscesses, in compromised patients. Positive blood cultures for this organism should prompt neuroimaging and consideration of cerebrospinal fluid sampling, as well as catheter removal. Given the worse outcome with central nervous system involvement, there is a need for increased awareness and early diagnosis, particularly in immunocompromised individuals.

  15. Successful Treatment of Bacillus cereus Bacteremia in a Patient with Propionic Acidemia.

    Science.gov (United States)

    Aygun, Fatma Deniz; Aygun, Fatih; Cam, Halit

    2016-01-01

    Bacillus cereus can cause serious, life-threatening, systemic infections in immunocompromised patients. The ability of microorganism to form biofilm on biomedical devices can be responsible for catheter-related bloodstream infections. Other manifestations of severe disease are meningitis, endocarditis, osteomyelitis, and surgical and traumatic wound infections. The most common feature in true bacteremia caused by Bacillus is the presence of an intravascular catheter. Herein, we report a case of catheter-related bacteremia caused by B. cereus in a patient with propionic acidemia.

  16. Resistance and biosorption mechanism of silver ions by Bacillus cereus biomass

    Institute of Scientific and Technical Information of China (English)

    Li Li; Qing Hu; Jinghai Zeng; Hongyan Qi; Guoqiang Zhuang

    2011-01-01

    Biosorption of silver ions onto Bacillus cereus biomass was investigated.Overall kinetic experiments were performed for the determination of the necessary contact time for the attainment of equilibrium.It was found that the overall biosorption process was best described by pseudo second-order kinetic model.The crystals detected by scanning electron microscope and X-ray photoelectron spectroscopy suggested the precipitation was a possible mechanism of biosorption.The molecular genetics of silver resistance of B.cereus biomass was also detected and illustrated by a whole cell sensor tool.

  17. Challenges and advances in systems biology analysis of Bacillus spore physiology; molecular differences between an extreme heat resistant spore forming Bacillus subtilis food isolate and a laboratory strain

    NARCIS (Netherlands)

    Brul, S.; van Beilen, J.; Caspers, M.; O'Brien, A.; de Koster, C.; Oomes, S.; Smelt, J.; Kort, R.; ter Beek, A.

    2011-01-01

    Bacterial spore formers are prime organisms of concern in the food industry. Spores from the genus Bacillus are extremely stress resistant, most notably exemplified by high thermotolerance. This sometimes allows surviving spores to germinate and grow out to vegetative cells causing food spoilage and

  18. In situ detection of Bacillus anthracis spores using fully submersible, self-exciting, self-sensing PMN-PT/Sn piezoelectric microcantilevers.

    Science.gov (United States)

    McGovern, John-Paul; Shih, Wan Y; Shih, Wei-Heng

    2007-08-01

    In this study, we have demonstrated in situ, all-electrical detection of Bacillus anthracis (BA) spores using lead magnesium niobate-lead titanate/tin (PMN-PT/Sn) piezoelectric microcantilever sensors (PEMS) fabricated from PMN-PT freestanding films and electrically insulated with methyltrimethoxysilane (MTMS) coatings on the tin surface. Antibody specific to BA spore surface antigen was immobilized on the platinum electrode of the PMN-PT layer. In phosphate-buffered saline (PBS) solution, the PMN-PT/Sn PEMS exhibited quality (Q) values ranging from 50 to 75. The detection was carried out in a closed-loop flow cell with a liquid volume of 0.8 ml and a flow rate of 1 ml min(-1). It was shown that one sensor, "PEMS-A" (500 microm long, 800 microm wide, with a 22 microm thick PMN-PT layer, a 20 microm thick tin layer and a 1 +/- 0.5 x 10(-12) g Hz(-1) mass detection sensitivity) exhibited resonance frequency shifts of 2100 +/- 200, 1100 +/- 100 and 700 +/- 100 Hz at concentrations of 20,000, 2000, and 200 spores ml(-1) or 16,000, 1600, and 160 total spores, respectively. Additionally, "PEMS-B" (350 microm long, 800 microm wide, with an 8 microm thick PMN-PT layer, a 6 microm thick tin layer and a 2 +/- 1 x 10(-13) g Hz(-1) mass detection sensitivity) exhibited resonance frequency shifts of 2400 +/- 200, 1500 +/- 200, 500 +/- 150 and 200 +/- 100 Hz at concentrations of 20,000, 2000, 100, and 45 spores ml(-1) or 16,000, 1600, 80, and 36 total spores, respectively.

  19. Viable spore counts in biological controls pre-sterilization.

    Science.gov (United States)

    Brusca, María I; Bernat, María I; Turcot, Liliana; Nastri, Natalia; Nastri, Maria; Rosa, Alcira

    2005-01-01

    The aim of the present study was to evaluate the total count of viable spores in standardized inoculated carriers pre-sterilization. Samples of "Bacterial Spore Sterilization Strip" (R Biological Laboratories) (well before their expiry date) were divided into Group A (B. subtilis) and Group B (B. stearothermophylus). Twenty-four strips were tested per group. The strips were minced in groups of three, placed in chilled sterile water and vortexed for 5 minutes to obtain a homogenous suspension. Ten ml of the homogenous suspension were transferred to two sterile jars, i.e. one jar per group. The samples were then heated in a water bath at 95 degrees C (Group A) or 80 degrees C (Group B) for 15 minutes and cooled rapidly in an ice bath at 0- 4 degrees C during 15 minutes. Successive dilutions were performed until a final aliquot of 30 to 300 colony-forming units (CFU) was obtained. The inoculums were placed in Petri dishes with culture medium (soy extract, casein agar adapted for spores, melted and cooled to 45-50 degrees C) and incubated at 55 degrees C or 37 degrees C. Statistical analysis of the data was performed. A larger number of spores were found at 48 hours than at 24 hours. However, this finding did not hold true for all the groups. The present results show that monitoring viable spores pre-sterilization would guarantee the accuracy of the data. Total spore counts must be within 50 and 300% of the number of spores indicated in the biological control. The procedure is essential to guarantee the efficacy of the biological control.

  20. Characteristics and phylogeny of Bacillus cereus strains isolated from Maari, a traditional West African food condiment.

    Science.gov (United States)

    Thorsen, Line; Kando, Christine Kere; Sawadogo, Hagrétou; Larsen, Nadja; Diawara, Bréhima; Ouédraogo, Georges Anicet; Hendriksen, Niels Bohse; Jespersen, Lene

    2015-03-02

    Maari is a spontaneously fermented food condiment made from baobab tree seeds in West African countries. This type of product is considered to be safe, being consumed by millions of people on a daily basis. However, due to the spontaneous nature of the fermentation the human pathogen Bacillus cereus occasionally occurs in Maari. This study characterizes succession patterns and pathogenic potential of B. cereus isolated from the raw materials (ash, water from a drilled well (DW) and potash), seed mash throughout fermentation (0-96h), after steam cooking and sun drying (final product) from two production sites of Maari. Aerobic mesophilic bacterial (AMB) counts in raw materials were of 10(5)cfu/ml in DW, and ranged between 6.5×10(3) and 1.2×10(4)cfu/g in potash, 10(9)-10(10)cfu/g in seed mash during fermentation and 10(7) - 10(9) after sun drying. Fifty three out of total 290 AMB isolates were identified as B. cereus sensu lato by use of ITS-PCR and grouped into 3 groups using PCR fingerprinting based on Escherichia coli phage-M13 primer (M13-PCR). As determined by panC gene sequencing, the isolates of B. cereus belonged to PanC types III and IV with potential for high cytotoxicity. Phylogenetic analysis of concatenated sequences of glpF, gmk, ilvD, pta, pur, pycA and tpi revealed that the M13-PCR group 1 isolates were related to B. cereus biovar anthracis CI, while the M13-PCR group 2 isolates were identical to cereulide (emetic toxin) producing B. cereus strains. The M13-PCR group 1 isolates harboured poly-γ-D-glutamic acid capsule biosynthesis genes capA, capB and capC showing 99-100% identity with the environmental B. cereus isolate 03BB108. Presence of cesB of the cereulide synthetase gene cluster was confirmed by PCR in M13-PCR group 2 isolates. The B. cereus harbouring the cap genes were found in potash, DW, cooking water and at 8h fermentation. The "emetic" type B. cereus were present in DW, the seed mash at 48-72h of fermentation and in the final product