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Sample records for spore forming microorganisms

  1. Spore-Forming Bacteria that Resist Sterilization

    Science.gov (United States)

    LaDuc, Myron; Venkateswaran, Kasthuri

    2003-01-01

    A report presents a phenotypic and genotypic characterization of a bacterial species that has been found to be of the genus Bacillus and has been tentatively named B. odysseensis because it was isolated from surfaces of the Mars Odyssey spacecraft as part of continuing research on techniques for sterilizing spacecraft to prevent contamination of remote planets by terrestrial species. B. odysseensis is a Gram-positive, facultatively anaerobic, rod-shaped bacterium that forms round spores. The exosporium has been conjectured to play a role in the elevated resistance to sterilization. Research on the exosporium is proposed as a path toward improved means of sterilization, medical treatment, and prevention of biofouling.

  2. Formation and characterization of non-growth states in Clostridium thermocellum: spores and L-forms

    Directory of Open Access Journals (Sweden)

    Mearls Elizabeth B

    2012-08-01

    Full Text Available Abstract Background Clostridium thermocellum is an anaerobic thermophilic bacterium that exhibits high levels of cellulose solublization and produces ethanol as an end product of its metabolism. Using cellulosic biomass as a feedstock for fuel production is an attractive prospect, however, growth arrest can negatively impact ethanol production by fermentative microorganisms such as C. thermocellum. Understanding conditions that lead to non-growth states in C. thermocellum can positively influence process design and culturing conditions in order to optimize ethanol production in an industrial setting. Results We report here that Clostridium thermocellum ATCC 27405 enters non-growth states in response to specific growth conditions. Non-growth states include the formation of spores and a L-form-like state in which the cells cease to grow or produce the normal end products of metabolism. Unlike other sporulating organisms, we did not observe sporulation of C. thermocellum in low carbon or nitrogen environments. However, sporulation did occur in response to transfers between soluble and insoluble substrates, resulting in approximately 7% mature spores. Exposure to oxygen caused a similar sporulation response. Starvation conditions during continuous culture did not result in spore formation, but caused the majority of cells to transition to a L-form state. Both spores and L-forms were determined to be viable. Spores exhibited enhanced survival in response to high temperature and prolonged storage compared to L-forms and vegetative cells. However, L-forms exhibited faster recovery compared to both spores and stationary phase cells when cultured in rich media. Conclusions Both spores and L-forms cease to produce ethanol, but provide other advantages for C. thermocellum including enhanced survival for spores and faster recovery for L-forms. Understanding the conditions that give rise to these two different non-growth states, and the implications that

  3. LEVELS AND TYPES OF AEROBIC SPORE FORMING BACTERIA ...

    African Journals Online (AJOL)

    Limnothrissa miodon) had the product sourced from them analysed morphologically by a microscope and biochemically for levels of aerobic spore forming bacteria that could adversely affect safety of the product. The four companies whose packaged ...

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

    International Nuclear Information System (INIS)

    Koshikawa, Tomihiko

    1994-01-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1994-12-01

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

  6. Discrimination of Spore-Forming Bacilli Using spoIVA

    Science.gov (United States)

    Venkateswaran, Kasthuri; LaDuc, Myron; Stuecker, Tara

    2009-01-01

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

  7. DNA fingerprinting of spore-forming bacterial isolates, using Bacillus ...

    African Journals Online (AJOL)

    User

    Bc-repetitive extragenic palindromic polymerase chain reaction (Bc-Rep PCR) analysis was conducted on seven Bacillus thuringiensis isolates accessed from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) culture collection and on five local isolates of entomopathogenic spore- forming bacteria.

  8. DNA fingerprinting of spore-forming bacterial isolates, using Bacillus ...

    African Journals Online (AJOL)

    Bc-repetitive extragenic palindromic polymerase chain reaction (Bc-Rep PCR) analysis was conducted on seven Bacillus thuringiensis isolates accessed from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) culture collection and on five local isolates of entomopathogenic spore-forming bacteria.

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

    Energy Technology Data Exchange (ETDEWEB)

    Bagge, Elisabeth

    2009-07-01

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

  10. The structural bases of long-term anabiosis in non-spore-forming bacteria

    Science.gov (United States)

    Suzina, Natalia E.; Mulyukin, Andrey L.; Dmitriev, Vladimir V.; Nikolaev, Yury A.; Shorokhova, Anna P.; Bobkova, Yulia S.; Barinova, Ekaterina S.; Plakunov, Vladimir K.; El-Registan, Galina I.; Duda, Vitalii I.

    2006-01-01

    Peculiarities of the structural organization in non-spore-forming bacteria associated with long-term anabiosis were revealed both in laboratory cultures and in natural populations isolated from 1 3-Myr-old Eastern Siberian permafrost and tundra soil. Different advanced methods were used, including (a) high-resolution electron microscopy; (b) simulation of in situ conditions in the laboratory by varying the composition of growth medium and cultivation conditions; (c) low-temperature fractionation to isolate and concentrate microbial cells from natural soils; (d) comparative morphological analysis of microbial cells in model cultures and natural soils (in situ). Under laboratory conditions, the intense formation of resting cells by representatives of various taxa of eubacteria and halophilic archaea occurred in 2 9-month-old cultures grown in carbon-, nitrogen-, or phosphorus-limited media, in starved cell suspensions in the presence of sodium silicate, or on soil agar. Among resting cells, we revealed cystlike forms having a complicated structure and common features. These included a thick capsule; a thickened and multiprofile cell wall; the presence of large intramembrane particles on PF- and EF-fracture surfaces; fine-grained or lumpy cytoplasm; and a condensed nucleoid. The general morphological properties, ultrastructural organization, physiological features of cystlike cells, and their ability to germinate under the appropriate conditions suggest the existence of constitutive dormancy in non-spore-forming bacteria. It was found that the majority of microorganisms in permafrost and tundra soil are cystlike cells, very similar to those in laboratory cultures. Anabiotic (resting) cystlike cells are responsible for the survival of non-spore-formers in extreme Earth habitats and may be regarded as possible analogs of extraterrestrial forms of microbial life.

  11. The Prevalence and Control of Bacillus and Related Spore-Forming Bacteria in the Dairy Industry

    Science.gov (United States)

    Gopal, Nidhi; Hill, Colin; Ross, Paul R.; Beresford, Tom P.; Fenelon, Mark A.; Cotter, Paul D.

    2015-01-01

    Milk produced in udder cells is sterile but due to its high nutrient content, it can be a good growth substrate for contaminating bacteria. The quality of milk is monitored via somatic cell counts and total bacterial counts, with prescribed regulatory limits to ensure quality and safety. Bacterial contaminants can cause disease, or spoilage of milk and its secondary products. Aerobic spore-forming bacteria, such as those from the genera Sporosarcina, Paenisporosarcina, Brevibacillus, Paenibacillus, Geobacillus and Bacillus, are a particular concern in this regard as they are able to survive industrial pasteurization and form biofilms within pipes and stainless steel equipment. These single or multiple-species biofilms become a reservoir of spoilage microorganisms and a cycle of contamination can be initiated. Indeed, previous studies have highlighted that these microorganisms are highly prevalent in dead ends, corners, cracks, crevices, gaskets, valves and the joints of stainless steel equipment used in the dairy manufacturing plants. Hence, adequate monitoring and control measures are essential to prevent spoilage and ensure consumer safety. Common controlling approaches include specific cleaning-in-place processes, chemical and biological biocides and other novel methods. In this review, we highlight the problems caused by these microorganisms, and discuss issues relating to their prevalence, monitoring thereof and control with respect to the dairy industry. PMID:26733963

  12. The prevalence and control of Bacillus and related spore-forming bacteria in the dairy industry

    Directory of Open Access Journals (Sweden)

    Nidhi eGopal

    2015-12-01

    Full Text Available Milk produced in udder cells is sterile but due to its high nutrient content, it can be a good growth substrate for contaminating bacteria. The quality of milk is monitored via somatic cell counts and total bacterial counts, with prescribed regulatory limits to ensure quality and safety. Bacterial contaminants can cause disease, or spoilage of milk and its secondary products. Aerobic spore-forming bacteria, such as those from the genera Sporosarcina, Paenisporosarcina, Brevibacillus, Paenibacillus, Geobacillus and Bacillus, are a particular concern in this regard as they are able to survive industrial pasteurisation and form biofilms within pipes and stainless steel equipment. These single or multiple-species biofilms become a reservoir of spoilage microorganisms and a cycle of contamination can be initiated. Indeed, previous studies have highlighted that these microorganisms are highly prevalent in dead ends, corners, cracks, crevices, gaskets, valves and the joints of stainless steel equipment used in the dairy manufacturing plants. Hence, adequate monitoring and control measures are essential to prevent spoilage and ensure consumer safety. Common controlling approaches include specific cleaning-in-place processes, chemical and biological biocides and other novel methods. In this review, we highlight the problems caused by these microorganisms, and discuss issues relating to their prevalence, monitoring thereof and control with respect to the dairy industry.

  13. Isolation and genetic identification of spore-forming bacteria associated with concentrated-milk processing in Nebraska.

    Science.gov (United States)

    Martinez, Bismarck A; Stratton, Jayne; Bianchini, Andreia

    2017-02-01

    Spore-forming bacteria are heat-resistant microorganisms capable of surviving and germinating in milk after pasteurization. They have been reported to affect the quality of dairy products by the production of enzymes (lipolytic and proteolytic) under low-temperature conditions in fluid milk, and have become a limiting factor for milk powder in reaching some selective markets. The objective of this research was to isolate and identify the population of spore-forming bacteria (psychrotrophic and thermophilic strains) associated with concentrated milk processing in Nebraska. During 2 seasons, in-process milk samples from a commercial plant (raw, pasteurized, and concentrated) were collected and heat-treated (80°C/12 min) to recover only spore-formers. Samples were spread-plated using standard methods agar and incubated at 32°C to enumerate mesophilic spore counts. Heat-treated samples were also stored at 7°C and 55°C to recover spore-formers that had the ability to grow under those temperature conditions. Isolates obtained from incubation or storage conditions were identified using molecular techniques (16S or rpoB sequencing). Based on the identification of the isolates and their relatedness, strains found in raw, pasteurized, and concentrated milk were determined to be similar. Paenibacillus spp. were associated with both raw and concentrated milk. Due to their known ability to cause spoilage under refrigeration, this shows the potential risk associated with the transferring of these problematic organisms into other dairy products. Other Bacillus species found in concentrated milk included Bacillus clausii, Bacillus subtilis, Lysinibacillus sp., Bacillus safensis, Bacillus licheniformis, Bacillus sonorensis, and Brevibacillus sp., with the last 3 organisms being capable of growing at thermophilic temperatures. These strains can also be translocated to other dairy products, such as milk powder, representing a quality problem. The results of this research

  14. Optimization of Spore Forming Bacteria Flooding for Enhanced Oil Recovery in North Sea Chalk Reservoir

    DEFF Research Database (Denmark)

    Halim, Amalia Yunita; Nielsen, Sidsel Marie; Eliasson Lantz, Anna

    2015-01-01

    was used for this purpose. A spore forming bacterium, Bacillus licheniformis 421, was used as it was shown to be a good candidate in the previous study. Bacterial spore can penetrate deeper into the chalk rock, squeezing through the pore throats. Our results show that B. licheniformis 421 when injected...

  15. Microbial Enhanced Heavy Oil Recovery by the Aid of Inhabitant Spore-Forming Bacteria: An Insight Review

    Science.gov (United States)

    Shibulal, Biji; Al-Bahry, Saif N.; Al-Wahaibi, Yahya M.; Elshafie, Abdulkader E.; Al-Bemani, Ali S.; Joshi, Sanket J.

    2014-01-01

    Crude oil is the major source of energy worldwide being exploited as a source of economy, including Oman. As the price of crude oil increases and crude oil reserves collapse, exploitation of oil resources in mature reservoirs is essential for meeting future energy demands. As conventional recovery methods currently used have become less efficient for the needs, there is a continuous demand of developing a new technology which helps in the upgradation of heavy crude oil. Microbial enhanced oil recovery (MEOR) is an important tertiary oil recovery method which is cost-effective and eco-friendly technology to drive the residual oil trapped in the reservoirs. The potential of microorganisms to degrade heavy crude oil to reduce viscosity is considered to be very effective in MEOR. Earlier studies of MEOR (1950s) were based on three broad areas: injection, dispersion, and propagation of microorganisms in petroleum reservoirs; selective degradation of oil components to improve flow characteristics; and production of metabolites by microorganisms and their effects. Since thermophilic spore-forming bacteria can thrive in very extreme conditions in oil reservoirs, they are the most suitable organisms for the purpose. This paper contains the review of work done with thermophilic spore-forming bacteria by different researchers. PMID:24550702

  16. Microbial Enhanced Heavy Oil Recovery by the Aid of Inhabitant Spore-Forming Bacteria: An Insight Review

    Directory of Open Access Journals (Sweden)

    Biji Shibulal

    2014-01-01

    Full Text Available Crude oil is the major source of energy worldwide being exploited as a source of economy, including Oman. As the price of crude oil increases and crude oil reserves collapse, exploitation of oil resources in mature reservoirs is essential for meeting future energy demands. As conventional recovery methods currently used have become less efficient for the needs, there is a continuous demand of developing a new technology which helps in the upgradation of heavy crude oil. Microbial enhanced oil recovery (MEOR is an important tertiary oil recovery method which is cost-effective and eco-friendly technology to drive the residual oil trapped in the reservoirs. The potential of microorganisms to degrade heavy crude oil to reduce viscosity is considered to be very effective in MEOR. Earlier studies of MEOR (1950s were based on three broad areas: injection, dispersion, and propagation of microorganisms in petroleum reservoirs; selective degradation of oil components to improve flow characteristics; and production of metabolites by microorganisms and their effects. Since thermophilic spore-forming bacteria can thrive in very extreme conditions in oil reservoirs, they are the most suitable organisms for the purpose. This paper contains the review of work done with thermophilic spore-forming bacteria by different researchers.

  17. Characterization of Bacillus sporothermodurans IC4 spores; putative indicator microorganism for optimisation of thermal processes in food sterilisation

    NARCIS (Netherlands)

    van Zuijlen, A.; Periago, P.M.; Amézquita, A.; Palop, A.; Brul, S.; Fernández, P.S.

    2010-01-01

    Spore-forming bacteria with high heat resistance increasingly challenge industrial sterilisation processes in foods. To ensure stability of manufactured foods, generally worst case scenarios are applied often leading to unwanted over processing of foods. This means bigger requirements of energy and

  18. A Clostridium difficile-Specific, Gel-Forming Protein Required for Optimal Spore Germination

    Directory of Open Access Journals (Sweden)

    M. Lauren Donnelly

    2017-01-01

    Full Text Available Clostridium difficile is a Gram-positive spore-forming obligate anaerobe that is a leading cause of antibiotic-associated diarrhea worldwide. In order for C. difficile to initiate infection, its aerotolerant spore form must germinate in the gut of mammalian hosts. While almost all spore-forming organisms use transmembrane germinant receptors to trigger germination, C. difficile uses the pseudoprotease CspC to sense bile salt germinants. CspC activates the related subtilisin-like protease CspB, which then proteolytically activates the cortex hydrolase SleC. Activated SleC degrades the protective spore cortex layer, a step that is essential for germination to proceed. Since CspC incorporation into spores also depends on CspA, a related pseudoprotease domain, Csp family proteins play a critical role in germination. However, how Csps are incorporated into spores remains unknown. In this study, we demonstrate that incorporation of the CspC, CspB, and CspA germination regulators into spores depends on CD0311 (renamed GerG, a previously uncharacterized hypothetical protein. The reduced levels of Csps in gerG spores correlate with reduced responsiveness to bile salt germinants and increased germination heterogeneity in single-spore germination assays. Interestingly, asparagine-rich repeat sequences in GerG’s central region facilitate spontaneous gel formation in vitro even though they are dispensable for GerG-mediated control of germination. Since GerG is found exclusively in C. difficile, our results suggest that exploiting GerG function could represent a promising avenue for developing C. difficile-specific anti-infective therapies.

  19. Biocidal Energetic Materials for the Destruction of Spore Forming Bacteria

    Science.gov (United States)

    2015-07-01

    Ag2O samples show high amounts of Ag0.55Al0.35. Of the metal ions, silver exhibits the highest toxicity for microorganisms and the least toxicity to...Of the metal ions, silver typically exhibits the highest toxicity for microorganisms and the least toxicity to animal cells depending on the system...of metallic and metal oxide sponges , Nature Mater. 2: 386-390 (2003). 14. B. C. Tappan,* M. H. Huynh, M. A. Hiskey, D. E. Chavez, E. P. Luther, J. T

  20. 40 CFR 180.1011 - Viable spores of the microorganism Bacillus thuringiensis Berliner; exemption from the...

    Science.gov (United States)

    2010-07-01

    ... Bacillus thuringiensis Berliner; exemption from the requirement of a tolerance. 180.1011 Section 180.1011... microorganism Bacillus thuringiensis Berliner; exemption from the requirement of a tolerance. (a) For the... authentic strain of Bacillus thuringiensis Berliner conforming to the morphological and biochemical...

  1. Contamination pathways of spore-forming bacteria in a vegetable cannery.

    Science.gov (United States)

    Durand, Loïc; Planchon, Stella; Guinebretiere, Marie-Hélène; André, Stéphane; Carlin, Frédéric; Remize, Fabienne

    2015-06-02

    Spoilage of low-acid canned food during prolonged storage at high temperatures is caused by heat resistant thermophilic spores of strict or facultative bacteria. Here, we performed a bacterial survey over two consecutive years on the processing line of a French company manufacturing canned mixed green peas and carrots. In total, 341 samples were collected, including raw vegetables, green peas and carrots at different steps of processing, cover brine, and process environment samples. Thermophilic and highly-heat-resistant thermophilic spores growing anaerobically were counted. During vegetable preparation, anaerobic spore counts were significantly decreased, and tended to remain unchanged further downstream in the process. Large variation of spore levels in products immediately before the sterilization process could be explained by occasionally high spore levels on surfaces and in debris of vegetable combined with long residence times in conditions suitable for growth and sporulation. Vegetable processing was also associated with an increase in the prevalence of highly-heat-resistant species, probably due to cross-contamination of peas via blanching water. Geobacillus stearothermophilus M13-PCR genotypic profiling on 112 isolates determined 23 profile-types and confirmed process-driven cross-contamination. Taken together, these findings clarify the scheme of contamination pathway by thermophilic spore-forming bacteria in a vegetable cannery. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. PHYLOGENETIC ANALYSIS AND AUTECOLOGY OF SPORE-FORMING BACTERIA FROM HYPERSALINE ENVIRONMENTS.

    Science.gov (United States)

    Gladka, G V; Romanovskaya, V A; Tashyreva, H O; Tashyrev, O B

    2015-01-01

    Multi-resistant to extreme factors spore-forming bacteria of Bacillus genus are isolated from hypersaline environments of the Crimea (Ukraine) and the Dead Sea (Israel). Phylogenetic analysis showed distinction of dominating extremophilic culturable species in studied regions. In Crimean environments they are B. mojavensis and B. simplex, in the Dead Sea ecosystem--B. subtilis subsp. spizizenii, B. subtilis subsp. subtilis, B. licheniformis and B. simplex. Isolates are simultaneously halotolerant and resistant to UV radiation. Strains isolated from the Dead Sea and the Crimea environments were resistant to UV: LD90 and LD99.99 made 100-170 J/m2 and 750-1500 J/m2 respectively. Spores showed higher UV-resistance (LD99.99-2500 J/m2) than the vegetative cells. However the number of spores made 0.02-0.007% of the whole cell population, and should not significantly affect the UV LD99.99 value. Isolates of both environments were halotolerant in the range of 0.1-10% NaCl and thermotolerant in the range of 20-50 °C, and didn't grow at 15 °C. Survival strategy of spore-forming bacteria from hypersaline environments under high UV radiation level can be performed by spore formation which minimize cell damage as well as efficient DNA-repair systems that remove damages.

  3. Efficiency of boiling and four other methods for genomic DNA extraction of deteriorating spore-forming bacteria from milk

    Directory of Open Access Journals (Sweden)

    Jose Carlos Ribeiro Junior

    2016-10-01

    Full Text Available The spore-forming microbiota is mainly responsible for the deterioration of pasteurized milk with long shelf life in the United States. The identification of these microorganisms, using molecular tools, is of particular importance for the maintenance of the quality of milk. However, these molecular techniques are not only costly but also labor-intensive and time-consuming. The aim of this study was to compare the efficiency of boiling in conjunction with four other methods for the genomic DNA extraction of sporulated bacteria with proteolytic and lipolytic potential isolated from raw milk in the states of Paraná and Maranhão, Brazil. Protocols based on cellular lysis by enzymatic digestion, phenolic extraction, microwave-heating, as well as the use of guanidine isothiocyanate were used. This study proposes a method involving simple boiling for the extraction of genomic DNA from these microorganisms. Variations in the quality and yield of the extracted DNA among these methods were observed. However, both the cell lysis protocol by enzymatic digestion (commercial kit and the simple boiling method proposed in this study yielded sufficient DNA for successfully carrying out the Polymerase Chain Reaction (PCR of the rpoB and 16S rRNA genes for all 11 strains of microorganisms tested. Other protocols failed to yield sufficient quantity and quality of DNA from all microorganisms tested, since only a few strains have showed positive results by PCR, thereby hindering the search for new microorganisms. Thus, the simple boiling method for DNA extraction from sporulated bacteria in spoiled milk showed the same efficacy as that of the commercial kit. Moreover, the method is inexpensive, easy to perform, and much less time-consuming.

  4. Structure, diversity and evolution of protein toxins from spore-forming entomopathogenic bacteria

    NARCIS (Netherlands)

    Maagd, de R.A.; Bravo, A.; Berry, C.; Crickmore, N.; Schnepf, H.E.

    2003-01-01

    Gram-positive spore-forming entomopathogenic bacteria can utilize a large variety of protein toxins to help them invade, infect, and finally kill their hosts, through their action on the insect midgut. These toxins belong to a number of homology groups containing a diversity of protein structures

  5. Desulfotomaculum thermobenzoicum subsp. thermosyntrophicum subsp. nov., a thermophilic, syntrophic, propionate-oxidizing, spore-forming bacterium

    NARCIS (Netherlands)

    Plugge, C.M.; Balk, M.; Stams, A.J.M.

    2002-01-01

    From granular sludge from a laboratory-scale upflow anaerobic sludge bed reactor operated at 55 degrees C with a mixture of volatile fatty acids as feed, a novel anaerobic, moderately thermophilic, syntrophic, spore-forming bacterium, strain TPO, was enriched on propionate in co-culture with

  6. A simple identification method for spore-forming bacteria showing high resistance against γ-rays

    International Nuclear Information System (INIS)

    Koshikawa, Tomihiko; Sone, Koji; Kobayashi, Toshikazu

    1993-01-01

    A simple identification method was developed for spore-forming bacteria which are highly resistant against γ-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 γ-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 γ-rays. The method is specific for B. megaterium, B. thuringiensis and B. pumilus, and highly selective for B. aneurinolyticus and B. cereus. (author)

  7. Mosquitocidal toxins of spore forming bacteria: recent advancement ...

    African Journals Online (AJOL)

    Mosquito borne diseases form a major component of vector borne diseases from all over the world. Several control strategies have been adopted to control diseases transmitted by mosquitoes. The discovery of highly potential bacteriocides like Bacillus sphaericus (Bs) and Bacillus thuringiensis subsp. israelensis (Bti) have ...

  8. Microrganismos heterotróficos mesófilos e bactérias do grupo do Bacillus cereus em leite integral submetido a ultra alta temperatura Mesophilic heterotrophic microorganisms and spore forming bacteria from Bacillus cereus group in ultra high temperature milk

    Directory of Open Access Journals (Sweden)

    A.M.C. Vidal-Martins

    2005-06-01

    Full Text Available Cento e dez amostras de 11 diferentes marcas de leite ultra alta temperatura (UAT, comercializadas em São José do Rio Preto - SP, foram submetidas à contagem de microrganismos heterotróficos mesófilos viáveis e à pesquisa de bactérias do grupo do Bacillus cereus. A população de microrganismos mesófilos variou de ´10² UFC/ml a >1,0´10(6 UFC/ml. Bactérias do grupo do Bacillus cereus foram verificadas em 13 (11,8% amostras. Os resultados evidenciaram elevada população de microrganismos indicadores mesófilos.One hundred and ten ultra high temperature (UHT milk samples, from 11 different brands retailed in São José do Rio Preto - SP were analysed for mesophilic heterotrophic microorganisms counting, as well as for the presence of Bacillus cereus. Mesophilic heterotrophic microorganisms ranged from ´10² CFU/ml to >1.0´10(6 CFU/ml. Bacillus cereus was detected in 13 (11.8% samples. The results demonstrated high countings of mesophilic microorganisms.

  9. The Spoilage Yeast Zygosaccharomyces bailii Forms Mitotic Spores: a Screening Method for Haploidization

    OpenAIRE

    Rodrigues, Fernando; Ludovico, Paula; Sousa, Maria João; Steensma, H. Yde; Côrte-Real, Manuela; Leão, Cecília

    2003-01-01

    Zygosaccharomyces bailii ISA 1307 and the type strain of this spoilage yeast show a diploid DNA content. Together with a rather peculiar life cycle in which mitotic but no meiotic spores appear to be formed, the diploid DNA content explains the observed difficulties in obtaining auxotrophic mutants. Mitotic chromosome loss induced by benomyl and selection on canavanine media resulted in three haploid strains of Z. bailii. This new set of Z. bailii strains allows the easy isolation...

  10. 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

  11. The spoilage yeast Zygosaccharomyces bailii forms mitotic spores: a screening method for haploidization.

    Science.gov (United States)

    Rodrigues, Fernando; Ludovico, Paula; Sousa, Maria João; Steensma, H Yde; Côrte-Real, Manuela; Leão, Cecília

    2003-01-01

    Zygosaccharomyces bailii ISA 1307 and the type strain of this spoilage yeast show a diploid DNA content. Together with a rather peculiar life cycle in which mitotic but no meiotic spores appear to be formed, the diploid DNA content explains the observed difficulties in obtaining auxotrophic mutants. Mitotic chromosome loss induced by benomyl and selection on canavanine media resulted in three haploid strains of Z. bailii. This new set of Z. bailii strains allows the easy isolation of recessive mutants and is suitable for further molecular genetic studies.

  12. Infections with Spore-forming Bacteria in Persons Who Inject Drugs, 2000-2009.

    OpenAIRE

    Palmateer, NE; Hope, VD; Roy, K; Marongiu, A; White, JM; Grant, KA; Ramsay, CN; Goldberg, DJ; Ncube, F

    2013-01-01

    : Since 2000 in the United Kingdom, infections caused by spore-forming bacteria have been associated with increasing illness and death among persons who inject drugs (PWID). To assess temporal and geographic trends in these illnesses (botulism, tetanus, Clostridium novyi infection, and anthrax), we compared rates across England and Scotland for 2000-2009. Overall, 295 infections were reported: 1.45 per 1,000 PWID in England and 4.01 per 1,000 PWID in Scotland. The higher rate in Scotland was ...

  13. Comparative study on disinfection potency of spore forming bacteria by electron-beam irradiation and gamma-ray irradiation

    International Nuclear Information System (INIS)

    Takizawa, Hironobu; Suzuki, Satoru; Suzuki, Tetsuya; Takama, Kozo; Hayashi, Toru; Yasumoto, Kyoden.

    1990-01-01

    Along with gamma-ray irradiation, electron-beam irradiation (EB) is a method to disinfect microorganisms which cause food decomposition and food-poisoning. The present study was undertaken to compare sterilization efficacy of EB and gamma-ray irradiation on bacterial spores and vegetative cells under various conditions. Spores of Bacillus pumilus, a marker strain for irradiation study, and Bacillus stearothermophilus known as a thermophilic bacteria were irradiated by electron-beam and gamma-ray separately at irradiation dose of 0 to 10 kGy on combination of wet/dry and aerobic/anaerobic conditions. Sterilization effect of irradiation on spores was evaluated by colony counting on agar plates. Results showed that both EB and gamma-ray irradiation gave sufficient sterilization effect on spores, and the sterilization effect increased exponentially with irradiation dose. The sterilization effect of gamma-ray irradiation was higher than that of EB in all cases. Higher disinfection effect was observed under aerobic condition. The present study suggests that oxygen supply in EB is more important than gamma-ray irradiation. No results suggesting that chlorine ion at 0.1 ppm (as available chlorine concentration) enhanced the sterilization efficacy of either EB or gamma-ray irradiation was obtained under any conditions examined. (author)

  14. Microbial enhanced oil recovery—a modeling study of the potential of spore-forming bacteria

    DEFF Research Database (Denmark)

    Nielsen, Sidsel Marie; Nesterov, Igor; Shapiro, Alexander

    2016-01-01

    Microbial enhanced oil recovery (MEOR) utilizes microbes for enhancing the recovery by several mechanisms, among which the most studied are the following: (1) reduction of oil-water interfacial tension (IFT) by the produced biosurfactant and (2) selective plugging by microbes and metabolic products...... cause sporulation, reducing the risk of clogging. Substrate released during sporulation can be utilized by attached vegetative bacteria and they will continue growing and producing surfactant, which prolongs the effect of the injected substrate. The simulation scenarios show that application...... of the spore-forming bacteria gives a higher total production of surfactant and the reduced risk of clogging, leading to an increased period of production and a higher oil recovery....

  15. Experimental Test Of Whether Electrostatically Charged Micro-organisms And Their Spores Contribute To The Onset Of Arcs Across Vacuum Gaps

    Energy Technology Data Exchange (ETDEWEB)

    none,; Grisham, Larry R.

    2014-02-24

    Recently it was proposed [L.R. Grisham, A. vonHalle, A.F. Carpe, Guy Rossi, K.R. Gilton, E.D. McBride, E.P. Gilson, A. Stepanov, T.N. Stevenson, Physics of Plasma 19 023107 (2012)] that one of the initiators of vacuum voltage breakdown between condu cting electrodes might be micro-organisms and their spores, previously deposited during exposure to air, which tnen become electrostatically charged when an electric potential is applied across the vacuum gap. The note describes a simple experiment to compare the number of voltage-conditioning pulses required to reach the nominal maxium operating voltage across a gap between two metallic conductors in a vacuum, comparing cases in which biological cleaning was done just prior to pump-down with cases where this was not done, with each preceded by exposure to ambient air for three days. Based upon these results, it does not appear that air-deposited microbes and their spores constitute a major pathway for arc initiation, at least for exposure periods of a few days, and for vacuum gaps of a few millimeters, in the regime where voltage holding is usually observed to vary linearly with gap distance

  16. 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, Stanley; van Beilen, Johan; Caspers, Martien P M; O'Brien, Andrea; de Koster, Chris; Oomes, Suus; Smelt, Jan; Kort, Remco; Ter Beek, Alex

    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

  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. Spore-Forming Thermophilic Bacterium within Artificial Meteorite Survives Entry into the Earth's Atmosphere on FOTON-M4 Satellite Landing Module.

    Science.gov (United States)

    Slobodkin, Alexander; Gavrilov, Sergey; Ionov, Victor; Iliyin, Vyacheslav

    2015-01-01

    One of the key conditions of the lithopanspermia hypothesis is that microorganisms situated within meteorites could survive hypervelocity entry from space through the Earth's atmosphere. So far, all experimental proof of this possibility has been based on tests with sounding rockets which do not reach the transit velocities of natural meteorites. We explored the survival of the spore-forming thermophilic anaerobic bacterium, Thermoanaerobacter siderophilus, placed within 1.4-cm thick basalt discs fixed on the exterior of a space capsule (the METEORITE experiment on the FOTON-M4 satellite). After 45 days of orbital flight, the landing module of the space vehicle returned to Earth. The temperature during the atmospheric transit was high enough to melt the surface of basalt. T. siderophilus survived the entry; viable cells were recovered from 4 of 24 wells loaded with this microorganism. The identity of the strain was confirmed by 16S rRNA gene sequence and physiological tests. This is the first report on the survival of a lifeform within an artificial meteorite after entry from space orbit through Earth's atmosphere at a velocity that closely approached the velocities of natural meteorites. The characteristics of the artificial meteorite and the living object applied in this study can serve as positive controls in further experiments on testing of different organisms and conditions of interplanetary transport.

  19. Spore-Forming Thermophilic Bacterium within Artificial Meteorite Survives Entry into the Earth's Atmosphere on FOTON-M4 Satellite Landing Module.

    Directory of Open Access Journals (Sweden)

    Alexander Slobodkin

    Full Text Available One of the key conditions of the lithopanspermia hypothesis is that microorganisms situated within meteorites could survive hypervelocity entry from space through the Earth's atmosphere. So far, all experimental proof of this possibility has been based on tests with sounding rockets which do not reach the transit velocities of natural meteorites. We explored the survival of the spore-forming thermophilic anaerobic bacterium, Thermoanaerobacter siderophilus, placed within 1.4-cm thick basalt discs fixed on the exterior of a space capsule (the METEORITE experiment on the FOTON-M4 satellite. After 45 days of orbital flight, the landing module of the space vehicle returned to Earth. The temperature during the atmospheric transit was high enough to melt the surface of basalt. T. siderophilus survived the entry; viable cells were recovered from 4 of 24 wells loaded with this microorganism. The identity of the strain was confirmed by 16S rRNA gene sequence and physiological tests. This is the first report on the survival of a lifeform within an artificial meteorite after entry from space orbit through Earth's atmosphere at a velocity that closely approached the velocities of natural meteorites. The characteristics of the artificial meteorite and the living object applied in this study can serve as positive controls in further experiments on testing of different organisms and conditions of interplanetary transport.

  20. Investigation of spore forming bacterial flooding for enhanced oil recovery in a North Sea chalk Reservoir

    DEFF Research Database (Denmark)

    Halim, Amalia Yunita; Nielsen, Sidsel Marie; Eliasson Lantz, Anna

    2015-01-01

    Bacillus licheniformis 421 was used as it was shown to be a good candidate in a previous study. Bacterial spore can penetrate deeper into the chalk rock, squeezing through the pore throats. Our results showed that injection of B. licheniformis 421 as a tertiary oil recovery method, in the residual oil...

  1. The presence of biofilm forming microorganisms on hydrotherapy equipment and facilities.

    Science.gov (United States)

    Jarząb, Natalia; Walczak, Maciej

    2017-10-01

    Hydrotherapy equipment provides a perfect environment for the formation and growth of microbial biofilms. Biofilms may reduce the microbiological cleanliness of hydrotherapy equipment and harbour opportunistic pathogens and pathogenic bacteria. The aims of this study were to investigate the ability of microorganisms that colonize hydrotherapy equipment to form biofilms, and to assess the influence of temperature and nutrients on the rate of biofilm formation. Surface swab samples were collected from the whirlpool baths, inhalation equipment and submerged surfaces of a brine pool at the spa center in Ciechocinek, Poland. We isolated and identified microorganisms from the swab samples and measured their ability to form biofilms. Biofilm formation was observed at a range of temperatures, in both nutrient-deficient and nutrient-rich environments. We isolated and identified microorganisms which are known to form biofilms on medical devices (e.g. Stenotrophomonas maltophilia). All isolates were classified as opportunistic pathogens, which can cause infections in humans with weakened immunity systems. All isolates showed the ability to form biofilms in the laboratory conditions. The potential for biofilm formation was higher in the presence of added nutrients. In addition, the hydrolytic activity of the biofilm was connected with the presence of nutrients.

  2. Comparative analysis of the diversity of aerobic spore-forming bacteria in raw milk from organic and conventional dairy farms.

    Science.gov (United States)

    Coorevits, An; De Jonghe, Valerie; Vandroemme, Joachim; Reekmans, Rieka; Heyrman, Jeroen; Messens, Winy; De Vos, Paul; Heyndrickx, Marc

    2008-06-01

    Bacterial contamination of raw milk can originate from different sources: air, milking equipment, feed, soil, faeces and grass. It is hypothesized that differences in feeding and housing strategies of cows may influence the microbial quality of milk. This assumption was investigated through comparison of the aerobic spore-forming flora in milk from organic and conventional dairy farms. Laboratory pasteurized milk samples from five conventional and five organic dairy farms, sampled in late summer/autumn and in winter, were plated on a standard medium and two differential media, one screening for phospholipolytic and the other for proteolytic activity of bacteria. Almost 930 isolates were obtained of which 898 could be screened via fatty acid methyl ester analysis. Representative isolates were further analysed using 16S rRNA gene sequencing and (GTG)(5)-PCR. The majority of aerobic spore-formers in milk belonged to the genus Bacillus and showed at least 97% 16S rRNA gene sequence similarity with type strains of Bacillus licheniformis, Bacillus pumilus, Bacillus circulans, Bacillus subtilis and with type strains of species belonging to the Bacillus cereus group. About 7% of all isolates may belong to possibly new spore-forming taxa. Although the overall diversity of aerobic spore-forming bacteria in milk from organic vs. conventional dairy farms was highly similar, some differences between both were observed: (i) a relatively higher number of thermotolerant organisms in milk from conventional dairy farms compared to organic farms (41.2% vs. 25.9%), and (ii) a relatively higher number of B. cereus group organisms in milk from organic (81.3%) and Ureibacillus thermosphaericus in milk from conventional (85.7%) dairy farms. One of these differences, the higher occurrence of B. cereus group organisms in milk from organic dairy farms, may be linked to differences in housing strategy between the two types of dairy farming. However, no plausible clarification was found for

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

    OpenAIRE

    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 microorganisms such as Bacillus cereus, that may survive the use of milder heating regimes. Dormant spores produced by B. cereus can survive processing conditions and their subsequent outgrowth increases ...

  4. Bacteria, mould and yeast spore inactivation studies by scanning electron microscope observations.

    Science.gov (United States)

    Rozali, Siti N M; Milani, Elham A; Deed, Rebecca C; Silva, Filipa V M

    2017-12-18

    Spores are the most resistant form of microbial cells, thus difficult to inactivate. The pathogenic or food spoilage effects of certain spore-forming microorganisms have been the primary basis of sterilization and pasteurization processes. Thermal sterilization is the most common method to inactivate spores present on medical equipment and foods. High pressure processing (HPP) is an emerging and commercial non-thermal food pasteurization technique. Although previous studies demonstrated the effectiveness of thermal and non-thermal spore inactivation, the in-depth mechanisms of spore inactivation are as yet unclear. Live and dead forms of two food spoilage bacteria, a mould and a yeast were examined using scanning electron microscopy before and after the inactivation treatment. Alicyclobacillus acidoterrestris and Geobacillus stearothermophilus bacteria are indicators of acidic foods pasteurization and sterilization processes, respectively. Neosartorya fischeri is a phyto-pathogenic mould attacking fruits. Saccharomyces cerevisiae is a yeast with various applications for winemaking, brewing, baking and the production of biofuel from crops (e.g. sugar cane). Spores of the four microbial species were thermally inactivated. Spores of S. cerevisiae were observed in the ascus and free form after thermal and HPP treatments. Different forms of damage and cell destruction were observed for each microbial spore. Thermal treatment inactivated bacterial spores of A. acidoterrestris and G. stearothermophilus by attacking the inner core of the spore. The heat first altered the membrane permeability allowing the release of intracellular components. Subsequently, hydration of spores, physicochemical modifications of proteins, flattening and formation of indentations occurred, with subsequent spore death. Regarding N. fischeri, thermal inactivation caused cell destruction and leakage of intracellular components. Both thermal and HPP treatments of S. cerevisiae free spores attacked

  5. Killing effect of peppermint vapor against pink-slime forming microorganisms.

    Science.gov (United States)

    Ihara, Nozomi; Sakamoto, Jin; Yoshida, Munehiro; Tsuchido, Tetsuaki

    2015-01-01

    The killing effect of peppermint vapor (PMV) against pink-slime forming microorganisms, Methylobacterium mesophilicum as a bacterium and Rhodotorula mucilaginosa as a yeast, was investigated by the agar vapor assay. In this method, microbial cells were spread over the agar surface exposed to PMV in a petri dish, and then transferred into a recovery liquid. When 60μl of the peppermint liquid was added to a paper disc, a marked killing effect of PMV was observed after 48h against M. mesophilicum and after 168h against R. mucilaginosa. M. mesophilicum and R. mucilaginosa were found to be more resistant to PMV than Escherichia coli and Candida albicans, used as reference microorganisms, respectively. With the addition of 0.03% sodium pyruvate as a hydrogen peroxide scavenger in agar, the killing effect of PMV against E. coli and C. albicans was decreased, whereas it was little changed against M. mesophilicum and R. mucilaginosa. In fact, the properties of the killing effect of hydrogen peroxide solution at 0.2-1.0mM was in accord with those of PMV. M. mesophilicum and R. mucilaginosa were more resistant to the oxidant than E. coli and C. albicans, respectively. Results obtained suggested that reactive oxygen species (ROS) may be involved in the killing action of PMV and therefore pink-slime formers are more resistant to PMV than non-pink-slime formers because of the presence of carotenoids as an antioxidant in cells. We also suggest that the use of PMV appeared to be a potential tool for the control of pink-slime forming microorganisms occurring in wet areas of houses such as the bathroom and washing room.

  6. Desulfotomaculum arcticum sp. nov., a novel spore-forming, moderately thermophilic, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard

    DEFF Research Database (Denmark)

    Vandieken, Verona; Knoblauch, Christian; Jørgensen, Bo Barker

    2006-01-01

    Strain 15T is a novel spore-forming, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard. Sulfate could be replaced by sulfite or thiosulfate. Hydrogen, formate, lactate, propionate, butyrate, hexanoate, methanol, ethanol, propanol, butanol, pyruvate, malate...... growth occurred at 44 degrees C. Therefore, strain 15T apparently cannot grow at in situ temperatures of Arctic sediments from where it was isolated, and it was proposed that it was present in the sediment in the form of spores. The DNA G+C content was 48.9 mol%. Strain 15T was most closely related...

  7. Biofilm forming microorganisms on various substrata from greenhouse of Botanical Garden “Jevremovac”

    Directory of Open Access Journals (Sweden)

    Unković Nikola D.

    2017-01-01

    Full Text Available Diversity of subaerial biofilm forming cyanobacteria, algae and fungi was investigated on 10 different substrata from greenhouse of Botanical Garden “Jevremovac”. Out of 37 documented taxa, 16 cyanobacterial and 10 algal taxa were identified. Remaining 11 taxa belong to the Kingdom of Fungi. The highest diversity of biofilm forming microorganisms, a total of 24 taxa, was detected on the corroded metal surface, while significantly lower number of taxa was recorded on other examined substrata. Cyanobacterium Porphyrosiphon sp., diatom Achnanthes sp. and green algae Chlorella sp. and Chlorococcum minutum were the most frequently encountered photosynthetic components of biofilms. In all analyzed samples, Trichoderma sp., followed by Cladosporium sp. and Rhizopus stolonifer, were the most frequently identified fungi. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. OI176020, Grant no. OI176018, and Grant no. OI173032

  8. Metabolic potential of a Novel Gram-Negative, Spore-forming, and Putatively Sulfate-Reducing Bacterium in the Continental Subsurface

    Science.gov (United States)

    Lau, C. Y. M.; Becraft, E. D.; Cason, E. D.; Borgonie, G.; Kieft, T. L.; Li, L.; van Heerden, E.; Jarett, J.; Woyke, T.; Stepanauskas, R.; Onstott, T. C.

    2017-12-01

    Anaerobic sulfate reduction is among the most thermodynamically favorable biochemical reactions in the deep subsurface environments. Phylogenetically and functionally diverse sulfate-reducing bacteria (SRB) within Deltaproteobacteria and Firmicutes have been reported. However, only few of them have been isolated in pure cultures for detailed physiological characterization. Previous studies showed that fracture fluid samples from the 1 km-deep borehole DR5IPC (Driefontein gold mine, South Africa) harbored novel SRB, as indicated by the low percentages (84% and 90%) of identity of the 16S ribosomal RNA clone sequences to known SRB. To overcome the challenge of low cultivability, we employed next-generation sequencing to unveil the metabolic potential of these novel SRB. Metagenomic assembly and binning yielded seven >50% complete genomes including a methylotrophic SRB belonging to Deltaproteobacteria (DR5_3) and two draft genomes representing an uncultivated phylum, tentatively "Driefonteinae" (DR5_4 and DR5_5). They accounted for 3%, 2% and 18% of all metagenomic reads. Three single-cell assembled genomes (SAGs) sharing 99% of average nucleotide identity (ANI) with DR5_5 were obtained. Analysis of the protein-coding genes in DR5_5 and related SAGs indicated that "Driefonteinae" possesses dissimilatory sulfite reductase genes (dsrAB), suggesting that sulfate would be the terminal electron acceptor. Whereas it may use diverse electron acceptors such as carbon monoxide, acetate, lactate and formate. A near-complete collection of genes for Wood-Ljungdahl pathway and genes for partial pentose phosphate pathway, glycolysis and tricarboxylic acid cycle further showed that "Driefonteinae" may live a mixotrophic life style. It is evident that archaeal genes related to methanogens were acquired through horizontal gene transfer. Phenotypically, "Driefonteinae" has a Gram-negative cell wall and flagella. The ability of forming spores would enable this microorganism to endure

  9. Anaerosporomusa subterranea gen. nov., sp. nov., a spore-forming anaerobe belonging to the class Negativicutes isolated from saprolite.

    Science.gov (United States)

    Choi, Jessica K; Shah, Madhavi; Yee, Nathan

    2016-10-01

    A Gram-stain-negative, spore-forming, anaerobic bacterium designated strain RU4T was isolated from a saprolite core collected from Oak Ridge, Tennessee, USA. Cells were slightly curved rods and exhibited an outer membrane exterior to a thin cell wall. Strain RU4T formed heat-resistant endospores in late-log phase and stationary phase cultures. Under anaerobic conditions, strain RU4T grew by fermenting fumarate and maleate, but did not grow on glucose, glycerol, pyruvate, lactate, succinate, citrate, formate, acetate, propionate, butyrate or valerate. Strain RU4T did not reduce sulfate or ferric iron. The main cellular fatty acids were C17 : 0 cyclo, C16 : 0 and C15 : 0. The DNA G+C content was 52 mol%. Analysis of the 16S rRNA, rpoB, recA, infB, gyrB and atpD gene sequences indicated that the isolate is related to members of the family Sporomusaceae. Based on 92 % sequence similarity of the 16S rRNA gene to its closest relatives in the family Sporomusaceae and divergent physiological traits, the newly-cultivated isolate was assigned to a novel species of a new genus, Anaerosporomusa subterranea gen. nov., sp. nov. The type strain of Anaerosporomusa subterranea is RU4T (=DSM 29728T=ATCC BAA-2723T).

  10. Microorganisms form exocellular structures, trophosomes, to facilitate biodegradation of oil in aqueous media.

    Science.gov (United States)

    Dmitriev, Vladimir V; Crowley, David; Rogachevsky, Vadim V; Negri, Cristina Maria; Rusakova, Tatiana G; Kolesnikova, Svetlana A; Akhmetov, Lenar I

    2011-02-01

    Cytochemical staining and microscopy were used to study the trophic structures and cellular morphotypes that are produced during the colonization of oil-water interfaces by oil-degrading yeasts and bacteria. Among the microorganisms studied here, the yeasts (Schwanniomyces occidentalis, Torulopsis candida, Candida tropicalis, Candida lipolytica, Candida maltosa, Candida paralipolytica) and two representative bacteria (Rhodococcus sp. and Pseudomonas putida) produced exocellular structures composed of biopolymers during growth on petroleum hydrocarbons. Four of the yeasts including S. occidentalis, T. candida, C. tropicalis and C. maltosa excreted polymers through modified sites in their cell wall ('canals'), whereas C. lipolytica and C. paralipolytica and the two bacterial species secreted polymers over the entire cell surface. These polymers took the form of fibrils and films that clogged pores and cavities on the surfaces of the oil droplets. A three-dimensional reconstruction of the cavities using serial thin sections showed that the exopolymer films isolated the ambient aqueous medium together with microbial cells and oil to form both closed and open granules that contained pools of oxidative enzymes utilized for the degradation of the oil hydrocarbons. The formation of such granules, or 'trophosomes,' appears to be a fundamental process that facilitates the efficient degradation of oil in aqueous media. © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  11. Biomass, gas exchange, and nutrient contents in upland rice plants affected by application forms of microorganism growth promoters.

    Science.gov (United States)

    Nascente, Adriano Stephan; de Filippi, Marta Cristina Corsi; Lanna, Anna Cristina; de Souza, Alan Carlos Alves; da Silva Lobo, Valácia Lemes; da Silva, Gisele Barata

    2017-01-01

    Microorganisms are considered a genetic resource with great potential for achieving sustainable development of agricultural areas. The objective of this research was to determine the effect of microorganism application forms on the production of biomass, gas exchange, and nutrient content in upland rice. The experiment was conducted under greenhouse conditions in a completely randomized design in a factorial 7 × 3 + 1, with four replications. The treatments consisted of combining seven microorganisms with three application forms (microbiolized seed; microbiolized seed + soil drenched with a microorganism suspension at 7 and 15 days after sowing (DAS); and microbiolized seed + plant sprayed with a microorganism suspension at 7 and 15 DAS) and a control (water). Treatments with Serratia sp. (BRM32114), Bacillus sp. (BRM32110 and BRM32109), and Trichoderma asperellum pool provided, on average, the highest photosynthetic rate values and dry matter biomass of rice shoots. Plants treated with Burkolderia sp. (BRM32113), Serratia sp. (BRM32114), and Pseudomonas sp. (BRM32111 and BRM32112) led to the greatest nutrient uptake by rice shoots. Serratia sp. (BRM 32114) was the most effective for promoting an increase in the photosynthetic rate, and for the greatest accumulation of nutrients and dry matter at 84 DAS, in rice shoots, which differed from the control treatment. The use of microorganisms can bring numerous benefits of rice, such as improving physiological characteristics, nutrient uptake, biomass production, and grain yield.

  12. Microbiological method for radiation sterilization (III). Development of identification software of spore-forming bacteria by using BBL CRYSTAL GP identification kit

    International Nuclear Information System (INIS)

    Hironiwa, Takayuki; Yamamoto, Yoko; Koshikawa, Tomihiko

    2004-01-01

    The part III in this title series describes the development of software for identification of spore-forming bacteria using the commercially available BBL CRYSTAL GP Identification Kit (Becton, Dickinson and Co., Ltd.), which is essentially for identification of Gram positive bacteria and is not always suitable for the spore-former in the radiation sterilization of medical devices. Isolation and identification of a spore-forming bacterium have to be confirmed by phase-contrast microscopy. The bacteria cultured overnight are to be inoculated in the Kit and cultured for 18-24 hr at 35-37 deg C with the lid attached by substrates for identification. Here, 30 substrates and probability of positive reactions to the substrates have been tested for spore-formers to make the computer software for final identification. The system is possible to identify 13 spp. of Bacillus, 4 of Paenibacillus, 2 of Brevibaccilus and 1 of Virgibacillus, which are the usual bioburden. For possible misidentification, re-isolation of the bacterium, prolonged culture, concentrated inoculation and re-consideration for ranking of identification the software provides are necessary as well as other identification approaches. Thus, as described in this series, the radio-resistance of, and radiation dose for, the bioburden can be evaluated more easily than hitherto, with use of the kits in radiation sterilization. (N.I.)

  13. Hydrothermal vents in Lake Tanganyika harbor spore-forming thermophiles with extremely rapid growth

    DEFF Research Database (Denmark)

    Elsgaard, Lars; Prieur, Daniel

    2010-01-01

    endospores. Based on the 16 S rDNA sequence the novel strain was homologous to Thermobrachium celere and Caloramator indicus, which are closely related. The novel strain was strictly anaerobic, fermentative and had a doubling time as short as 10 min during growth on complex substrates, such as yeast extract......A thermophilic anaerobic bacterium was isolated from a sublacustrine hydrothermal vent site in Lake Tanganyika (East Africa) with recorded fluid temperatures of 66–103 °C and pH values of 7.7–8.9. The bacterium (strain TR10) was rod-shaped, about 1 by 5 μm in size, and readily formed distal...

  14. Efficacy of a marine bacterial nuclease against biofilm forming microorganisms isolated from chronic rhinosinusitis.

    Directory of Open Access Journals (Sweden)

    Robert C Shields

    Full Text Available BACKGROUND: The persistent colonization of paranasal sinus mucosa by microbial biofilms is a major factor in the pathogenesis of chronic rhinosinusitis (CRS. Control of microorganisms within biofilms is hampered by the presence of viscous extracellular polymers of host or microbial origin, including nucleic acids. The aim of this study was to investigate the role of extracellular DNA in biofilm formation by bacteria associated with CRS. METHODS/PRINCIPAL FINDINGS: Obstructive mucin was collected from patients during functional endoscopic sinus surgery. Examination of the mucous by transmission electron microscopy revealed an acellular matrix punctuated occasionally with host cells in varying states of degradation. Bacteria were observed in biofilms on mucosal biopsies, and between two and six different species were isolated from each of 20 different patient samples. In total, 16 different bacterial genera were isolated, of which the most commonly identified organisms were coagulase-negative staphylococci, Staphylococcus aureus and α-haemolytic streptococci. Twenty-four fresh clinical isolates were selected for investigation of biofilm formation in vitro using a microplate model system. Biofilms formed by 14 strains, including all 9 extracellular nuclease-producing bacteria, were significantly disrupted by treatment with a novel bacterial deoxyribonuclease, NucB, isolated from a marine strain of Bacillus licheniformis. Extracellular biofilm matrix was observed in untreated samples but not in those treated with NucB and extracellular DNA was purified from in vitro biofilms. CONCLUSION/SIGNIFICANCE: Our data demonstrate that bacteria associated with CRS form robust biofilms which can be reduced by treatment with matrix-degrading enzymes such as NucB. The dispersal of bacterial biofilms with NucB may offer an additional therapeutic target for CRS sufferers.

  15. Effect of Essential Oils on Germination and Growth of Some Pathogenic and Spoilage Spore-Forming Bacteria.

    Science.gov (United States)

    Voundi, Stève Olugu; Nyegue, Maximilienne; Lazar, Iuliana; Raducanu, Dumitra; Ndoye, Florentine Foe; Marius, Stamate; Etoa, François-Xavier

    2015-06-01

    The use of essential oils as a food preservative has increased due to their capacity to inhibit vegetative growth of some bacteria. However, only limited data are available on their effect on bacterial spores. The aim of the present study was to evaluate the effect of some essential oils on the growth and germination of three Bacillus species and Geobacillus stearothermophilus. Essential oils were chemically analyzed using gas chromatography and gas chromatography coupled to mass spectrometry. The minimal inhibitory and bactericidal concentrations of vegetative growth and spore germination were assessed using the macrodilution method. Germination inhibitory effect of treated spores with essential oils was evaluated on solid medium, while kinetic growth was followed using spectrophotometry in the presence of essential oils. Essential oil from Drypetes gossweileri mainly composed of benzyl isothiocyanate (86.7%) was the most potent, with minimal inhibitory concentrations ranging from 0.0048 to 0.0097 mg/mL on vegetative cells and 0.001 to 0.002 mg/mL on spore germination. Furthermore, essential oil from D. gossweileri reduced 50% of spore germination after treatment at 1.25 mg/mL, and its combination with other oils improved both bacteriostatic and bactericidal activities with additive or synergistic effects. Concerning the other essential oils, the minimal inhibitory concentration ranged from 5 to 0.63 mg/mL on vegetative growth and from 0.75 to 0.09 mg/mL on the germination of spores. Spectrophotometric evaluation showed an inhibitory effect of essential oils on both germination and outgrowth. From these results, it is concluded that some of the essential oils tested might be a valuable tool for bacteriological control in food industries. Therefore, further research regarding their use as food preservatives should be carried out.

  16. Tindallia californiensis sp. nov., a new anaerobic, haloalkaliphilic, spore-forming acetogen isolated from Mono Lake in California

    Science.gov (United States)

    Pikuta, E. V.; Hoover, R. B.; Bej, A. K.; Marsic, D.; Detkova, E. N.; Whitman, W. B.; Krader, P.

    2003-01-01

    A novel extremely haloalkaliphilic, strictly anaerobic, acetogenic bacterium strain APO was isolated from sediments of the athalassic, meromictic, alkaline Mono Lake in California. The Gram-positive, spore-forming, slightly curved rods with sizes 0.55- 0.7x1.7-3.0 microns were motile by a single laterally attached flagellum. Strain APO was mesophilic (range 10-48 C, optimum of 37 C); halophilic (NaCl range 1-20% (w/v) with optimum of 3-5% (w/v), and alkaliphilic (pH range 8.0-10.5, optimum 9.5). The novel isolate required sodium ions in the medium. Strain APO was an organotroph with a fermentative type of metabolism and used the substrates peptone, bacto-tryptone, casamino acid, yeast extract, L-serine, L-lysine, L-histidine, L-arginine, and pyruvate. The new isolate performed the Stickland reaction with the following amino acid pairs: proline + alanine, glycine + alanine, and tryptophan + valine. The main end product of growth was acetate. High activity of CO dehydrogenase and hydrogenase indicated the presence of a homoacetogenic, non-cycling acetyl-coA pathway. Strain APO was resistant to kanamycin but sensitive to chloramphenicol, tetracycline, and gentamycin. The G+C content of the genomic DNA was 44.4 mol% (by HPLC method). The sequence of the 16s rRNA gene of strain APO possessed 98.2% similarity with the sequence from Tindullia magadiensis Z-7934, but the DNA-DNA hybridization value between these organisms was only 55%. On the basis of these physiological and molecular properties, strain APO is proposed to be a novel species of the genus Tindallia with the name Tindallia californiensis sp. nov., (type strain APO = ATCC BAA-393 - DSM 14871).

  17. 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. © 2015 The Society for Applied Microbiology.

  18. Diversity of spore-forming bacteria and identification of Bacillus amyloliquefaciens as a species frequently associated with the ropy spoilage of bread.

    Science.gov (United States)

    Valerio, F; De Bellis, P; Di Biase, M; Lonigro, S L; Giussani, B; Visconti, A; Lavermicocca, P; Sisto, A

    2012-06-01

    This study examines the diversity of spore-forming bacteria isolated from raw materials/bread using molecular methods along with a rapid and innovative technology, the FT-NIR spectroscopy. Microbiological analysis showed that 23% of semolina and 42% of other raw materials (including grain, brewer yeast, improvers) contained more than 100 spores/g and more than 50% of each kind of sample was contaminated at a level ranging from 1 to 100 spores/g. A high bacterial diversity characterized raw materials. In total 176 isolates were collected and characterized: 13 bacterial species belonging to Bacillus (10) and Paenibacillus (3) genera were identified by sequencing of 16S rRNA, gyrA or gyrB genes. The two closely related species Bacillus amyloliquefaciens (strain N45.1) and Bacillus subtilis (strain S63) were also analyzed by the spectroscopic technique FT-NIR. This analysis gave clear discrimination between the strains in the score plot obtained by the PCA and allowed to identify the spectral region 5600-4000 cm(-1) as the information-rich region for discrimination. B. amyloliquefaciens, possibly misidentified as B. subtilis in previous studies, was recognized as the most frequent species, found also in ropy bread. Moreover, the screening test for rope production indicated that mainly B. amyloliquefaciens, together with B. subtilis and Bacillus pumilus, could cause spoilage in bread, even if the last two species were represented by a low number of isolates. The Bacillus cereus group and Bacillus megaterium showed a lower percentage (30-70%) of isolates potentially able to cause the rope, but considering the high number of B. cereus group isolates detected in this study, this bacterial group should also be considered important in rope spoilage. In conclusion, results demonstrate that raw materials used to produce bread represent a rich source of spore-forming bacteria, therefore their microbiological quality should be monitored before use. Moreover, this study

  19. 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

  20. 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

  1. Synergistic action of cinnamaldehyde with silver nanoparticles against spore-forming bacteria: a case for judicious use of silver nanoparticles for antibacterial applications

    Directory of Open Access Journals (Sweden)

    Ghosh IN

    2013-12-01

    Full Text Available Indro Neil Ghosh,1,* Supriya Deepak Patil,1,* Tarun Kumar Sharma,1,2 Santosh Kumar Srivastava,1 Ranjana Pathania,1 Naveen Kumar Navani11Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 2Center for Biodesign and Diagnostics, Translational Health Science and Technology Institute, Gurgaon Haryana, India*These authors contributed equally to this workAbstract: Silver has long been advocated as an effective antimicrobial. However, toxicity issues with silver have led to limited use of silver in nanoform, especially for food preservation. With the aim of exploring combinatorial options that could increase the antibacterial potency of silver nanoparticles and reduce the effective dosage of silver, we evaluated the extent of synergy that a combination of silver nanoparticles and an essential oil representative (cinnamaldehyde could offer. A battery of gram-positive and gram-negative bacterial strains was utilized for antibacterial assays, and extents of synergism were calculated from fractional inhibitory concentration indices. The activity of nanoparticles was greatly enhanced when utilized in the presence of cinnamaldehyde. We observed combinatorial effects that were strongly additive against all the bacterial strains tested, and genuine synergy was found against spore forming Bacillus cereus and Clostridium perfringens – bacterial strains associated with release of cytotoxins in contaminated food and known for their persistence. Bacterial kill curve analysis revealed a very fast bactericidal action when a combination of two agents was used. The electron and atomic force microscopy also revealed extensive damage to the bacterial cell envelop in the presence of both agents. We also performed hemolysis assays to investigate and approximate the extent of toxicity exhibited by the two agents, and observed no adverse effect at the concentrations required for synergy. This study shows that safe levels of silver in

  2. Lyophilized spore dispenser

    Science.gov (United States)

    Jessup, A. D. (Inventor)

    1973-01-01

    A lyophilized spore dispenser is provided which produces a finely divided, monoparticulate cloud of bacterial spores. The spores are contained within a tightly sealed chamber, and a turbulator orifice connected to an air supply source provides a jet of air which stirs up the spores and causes the spores to be suspended in eddy currents within the chamber. This air jet also produces a positive pressure within the chamber which forces the spores out of an injection orifice.

  3. Updates on Clostridium difficile spore biology.

    Science.gov (United States)

    Gil, Fernando; Lagos-Moraga, Sebastián; Calderón-Romero, Paulina; Pizarro-Guajardo, Marjorie; Paredes-Sabja, Daniel

    2017-06-01

    Clostridium difficile is a Gram-positive, anaerobic spore former, and an important nosocomial pathogenic bacterium. C. difficile spores are the morphotype of transmission and recurrence of the disease. The formation of C. difficile spores and their subsequent germination are essential processes during the infection. Recent in vitro and in vivo work has shed light on how spores are formed and the timing of in vivo sporulation in a mouse model. Advances have also been made in our understanding of the machineries involved in spore germination, and how antibiotic-induced dysbiosis affects the metabolism of bile salts and thus impacts C. difficile germination in vivo. Studies have also attempted to identify how C. difficile spores interact with the host's intestinal mucosa. Spore resistance has also been revisited by several groups highlighting the extreme resistance of this morphotype to traditional food processing regimes and disinfectants used in clinical settings. Therefore, the aim of this review is to summarize recent advances on spore formation/germination in vitro and in vivo, spore-host interactions, and spore resistance that contribute to our knowledge of the role of C. difficile spores in the infectious process. Copyright © 2017. Published by Elsevier Ltd.

  4. Survival of B. Horneckiae Spores Under Ground-simulated Space Conditions

    Science.gov (United States)

    Schanche, Bradley

    2012-01-01

    To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spore-forming microbes are highly resistant to various physical and chemical conditions, which include ionizing and UV radiation, desiccation and oxidative stress, and the harsh environment of outer space or planetary surfaces. Recently a radiation resistant, spore forming bacterial isolate, Bacillus horneckiae, was isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. The exceptionally high tolerance of extreme conditions demonstrated by sporeforming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. The proposed BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, will potentially be able to withstand harsh space conditions. As previously stated, the spore producing ability of these species gives them increased survivability to harsh conditions. Some of the spores will have the protective exosporium layer artificially removed before the test to determine if the existence of this layer significantly changes the survivability during the mission. In preparation for that experiment, we analyzed spores which were exposed during a ground simulation, the EXPOSE R2 Biofilm Organisms Surfing Space (BOSS). Previous to exposure, spores were deposited onto spacecraft grade aluminum coupons in a spore suspension calculated to contain between 10(exp 7) and 10(exp 8) spores. This precursor series will be used to establish a baseline survivability function for comparison with the future flight tests during EXPOSE-R. For each coupon, a 10% polyvinyl alcohol (PVA) film was applied and peeled

  5. 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.

  6. Bacillus subtilis Spore Inner Membrane Proteome

    NARCIS (Netherlands)

    Zheng, L.; Abhyankar, W.; Ouwerling, N.; Dekker, H.L.; van Veen, H.; van der Wel, N.N.; Roseboom, W.; de Koning, L.J.; Brul, S.; de Koster, C.G.

    2016-01-01

    The endospore is the dormant form of Bacillus subtilis and many other Firmicutes. By sporulation, these spore formers can survive very harsh physical and chemical conditions. Yet, they need to go through germination to return to their growing form. The spore inner membrane (IM) has been shown to

  7. 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.

  8. Spore Preparation Protocol for Enrichment of Clostridia from Murine Intestine.

    Science.gov (United States)

    Velazquez, Eric M; Rivera-Chávez, Fabian; Bäumler, Andreas J

    2017-05-20

    In recent years, many spore-forming commensal Clostridia found in the gut have been discovered to promote host physiology, immune development, and protection against infections. We provide a detailed protocol for rapid enrichment of spore-forming bacteria from murine intestine. Briefly, contents from the intestinal cecum are collected aerobically, diluted and finally treated with chloroform to enrich for Clostridia spores.

  9. 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

  10. Modeling to control spores in raw milk

    NARCIS (Netherlands)

    Vissers, M.

    2007-01-01

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

  11. Detection and differentiation of bacterial spores in a mineral matrix by Fourier transform infrared spectroscopy (FTIR and chemometrical data treatment

    Directory of Open Access Journals (Sweden)

    Brandes Ammann Andrea

    2011-07-01

    Full Text Available Abstract Background Fourier transform infrared spectroscopy (FTIR has been used as analytical tool in chemistry for many years. In addition, FTIR can also be applied as a rapid and non-invasive method to detect and identify microorganisms. The specific and fingerprint-like spectra allow - under optimal conditions - discrimination down to the species level. The aim of this study was to develop a fast and reproducible non-molecular method to differentiate pure samples of Bacillus spores originating from different species as well as to identify spores in a simple matrix, such as the clay mineral, bentonite. Results We investigated spores from pure cultures of seven different Bacillus species by FTIR in reflection or transmission mode followed by chemometrical data treatment. All species investigated (B. atrophaeus, B. brevis, B. circulans, B. lentus, B. megaterium, B. subtilis, B. thuringiensis are typical aerobic soil-borne spore formers. Additionally, a solid matrix (bentonite and mixtures of benonite with spores of B. megaterium at various wt/wt ratios were included in the study. Both hierarchical cluster analysis and principal component analysis of the spectra along with multidimensional scaling allowed the discrimination of different species and spore-matrix-mixtures. Conclusions Our results show that FTIR spectroscopy is a fast method for species-level discrimination of Bacillus spores. Spores were still detectable in the presence of the clay mineral bentonite. Even a tenfold excess of bentonite (corresponding to 2.1 × 1010 colony forming units per gram of mineral matrix still resulted in an unambiguous identification of B. megaterium spores.

  12. Aspects of tests and assessment of filtering materials used for respiratory protection against bioaerosols. Part II: sweat in the environment, microorganisms in the form of a bioaerosol.

    Science.gov (United States)

    Majchrzycka, Katarzyna; Gutarowska, Beata; Brochocka, Agnieszka

    2010-01-01

    The second part of the article presents the results of a study of antimicrobial activity of filter nonwovens with an addition of biocides, as a function of the presence of sweat in the environment and the method of microbe deposition on a nonwoven in the form of a liquid and a bioaerosol. At the same time, the filtration efficiency of nonwovens against microorganisms in the form of a bioaerosol was tested with the dynamic method. The results showed that the addition of sweat on the surface of a nonwoven resulted in an insignificant decrease of biological activity that still remained high. Moreover, an active nonwoven showed biostatic and biocidal activity only when microbes were deposited on the surface in the form of a solution. The nonwoven did not show any biological activity after deposition of microorganisms with the dynamical method in the form of a bioaerosol.

  13. What can spores do for us?

    NARCIS (Netherlands)

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

    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

  14. Microorganism immobilization

    Science.gov (United States)

    Compere, Alicia L.; Griffith, William L.

    1981-01-01

    Live metabolically active microorganisms are immobilized on a solid support by contacting particles of aggregate material with a water dispersible polyelectrolyte such as gelatin, crosslinking the polyelectrolyte by reacting it with a crosslinking agent such as glutaraldehyde to provide a crosslinked coating on the particles of aggregate material, contacting the coated particles with live microorganisms and incubating the microorganisms in contact with the crosslinked coating to provide a coating of metabolically active microorganisms. The immobilized microorganisms have continued growth and reproduction functions.

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

    African Journals Online (AJOL)

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

  16. Spoilage of Microfiltered and Pasteurized Extended Shelf Life Milk Is Mainly Induced by Psychrotolerant Spore-Forming Bacteria that often Originate from Recontamination.

    Science.gov (United States)

    Doll, Etienne V; Scherer, Siegfried; Wenning, Mareike

    2017-01-01

    Premature spoilage and varying product quality due to microbial contamination still constitute major problems in the production of microfiltered and pasteurized extended shelf life (ESL) milk. Spoilage-associated bacteria may enter the product either as part of the raw milk microbiota or as recontaminants in the dairy plant. To identify spoilage-inducing bacteria and their routes of entry, we analyzed end products for their predominant microbiota as well as the prevalence and biodiversity of psychrotolerant spores in bulk tank milk. Process analyses were performed to determine the removal of psychrotolerant spores at each production step. To detect transmission and recontamination events, strain typing was conducted with isolates obtained from all process stages. Microbial counts in 287 ESL milk packages at the end of shelf life were highly diverse ranging from shelf life is influenced only to a minor extent by raw-milk-associated factors. In contrast, recontamination with spores, particularly from the B. cereus complex, seems to occur. To enhance milk quality throughout the entire shelf life, improved plant sanitation and disinfection that target the elimination of spores are necessary.

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

    International Nuclear Information System (INIS)

    Fernandes, Joao C.; Eaton, Peter; Gomes, Ana M.; Pintado, Manuela E.; Xavier Malcata, F.

    2009-01-01

    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.

  18. [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.

  19. Microorganisms (Microbes), Role of

    DEFF Research Database (Denmark)

    Fenchel, Tom

    2013-01-01

    Microorganisms (microbes) are those life forms too small to be seen by the naked eye; that is, those that require a microscope or other form of magnification in order to be observed. The term microorganism is thus a functional description rather than a taxonomic one, and the grouping includes...... a wide variety of organisms. The article focuses on the functional role of microbes in the biosphere and in different types of habitats - especially in terms of flow of energy and matter....

  20. Adaptation of the spore discharge mechanism in the basidiomycota.

    Directory of Open Access Journals (Sweden)

    Jessica L Stolze-Rybczynski

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

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

    NARCIS (Netherlands)

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

    2013-01-01

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

  2. UV inactivation of pathogenic and indicator microorganisms

    Energy Technology Data Exchange (ETDEWEB)

    Chang, J.C.; Ossoff, S.F.; Lobe, D.C.; Dorfman, M.H.; Dumais, C.M.; Qualls, R.G.; Johnson, J.D.

    1985-06-01

    Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts.

  3. UV inactivation of pathogenic and indicator microorganisms

    International Nuclear Information System (INIS)

    Chang, J.C.; Ossoff, S.F.; Lobe, D.C.; Dorfman, M.H.; Dumais, C.M.; Qualls, R.G.; Johnson, J.D.

    1985-01-01

    Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts

  4. Phosphorescence In Bacillus Spores

    National Research Council Canada - National Science Library

    Reinisch, Lou; Swartz, Barry A; Bronk, Burt V

    2003-01-01

    .... Our present work attempts to build on this approach for environmental applications. We have measured a change in the fluorescence spectra of suspensions of Bacillus bacteria between the vegetative bacteria and their spores at room temperature...

  5. Aspects of tests and assessment of filtering materials used for respiratory protection against bioaerosols. Part I: type of active substance, contact time, microorganism species.

    Science.gov (United States)

    Majchrzycka, Katarzyna; Gutarowska, Beata; Brochocka, Agnieszka

    2010-01-01

    This paper presents the results of a study on antimicrobial activity of polymer filter nonwovens produced by needle-punching or melt-blowing with an addition of disinfecting agents. The first part of the paper discusses how the biocidal activity of nonwovens is a function of the active agent added to the nonwovens, the duration of the contact of microorganisms with nonwovens and the type of microorganisms. The types of fibres and disinfecting agents had a considerable effect on the biocidal activity of nonwovens. The biocidal effect of nonwovens increased with the duration of their contact with microorganisms. Fibre activity differed considerably depending on the species of the microorganism. The microorganisms most sensitive to biocidal activity of the active filter nonwoven were S. aureus, M. flavus and E. coli. There were no biocidal effects on spore-forming bacterium B. subtilis.

  6. Thermal inactivation kinetics of Bacillus coagulans spores in tomato juice.

    Science.gov (United States)

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

    2012-07-01

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

  7. Fossil Microorganisms in Archaean

    Science.gov (United States)

    Astafleva, Marina; Hoover, Richard; Rozanov, Alexei; Vrevskiy, A.

    2006-01-01

    Ancient Archean and Proterozoic rocks are the model objects for investigation of rocks comprising astromaterials. The first of Archean fossil microorganisms from Baltic shield have been reported at the last SPIE Conference in 2005. Since this confeence biomorphic structures have been revealed in Archean rocks of Karelia. It was determined that there are 3 types of such bion structures: 1. structures found in situ, in other words microorganisms even-aged with rock matrix, that is real Archean fossils biomorphic structures, that is to say forms inhabited early formed rocks, and 3. younger than Archean-Protherozoic minerali microorganisms, that is later contamination. We made attempt to differentiate these 3 types of findings and tried to understand of burial of microorganisms. The structures belongs (from our point of view) to the first type, or real Archean, forms were under examination. Practical investigation of ancient microorganisms from Green-Stone-Belt of Northern Karelia turns to be very perspective. It shows that even in such ancient time as Archean ancient diverse world existed. Moreover probably such relatively highly organized cyanobacteria and perhaps eukaryotic formes existed in Archean world.

  8. Spore analysis and tetrad dissection of Schizosaccharomyces pombe

    DEFF Research Database (Denmark)

    Ekwall, Karl; Thon, Genevieve

    2017-01-01

    Here we describe the processing of Schizosaccharomyces pombe spores in batches (random spore analysis) or through tetrad dissections. Spores are usually prepared from matings between haploid strains (producing zygotic asci) or from sporulating diploids (producing azygotic asci). In random spore...... analysis, a snail enzyme preparation is used to digest the walls of asci to release free spores that are diluted and plated to form colonies. In tetrad dissection, a needle attached to a micromanipulator is used to pick asci and separate spores. Tetrad dissection has traditionally been the method of choice...

  9. Bacterial Spores in Food : Survival, Emergence, and Outgrowth

    NARCIS (Netherlands)

    Wells-Bennik, Marjon H J; Eijlander, Robyn T; den Besten, Heidy M W; Berendsen, Erwin M; Warda, Alicja K; Krawczyk, Antonina O; Nierop Groot, Masja N; Xiao, Yinghua; Zwietering, Marcel H; Kuipers, Oscar P; Abee, Tjakko

    2016-01-01

    Spore-forming bacteria are ubiquitous in nature. The resistance properties of bacterial spores lie at the heart of their widespread occurrence in food ingredients and foods. The efficacy of inactivation by food-processing conditions is largely determined by the characteristics of the different types

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

    Science.gov (United States)

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

    2012-03-01

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

  11. Morpho-structural variations of bacterial spores after treatment in steam vacuum assisted autoclave.

    Science.gov (United States)

    Fonzi, M; Montomoli, E; Gasparini, R; Devanna, D; Fonzi, L

    1999-01-01

    This study intended to verify, through microbiological techniques and TEM investigations, the killing of bacterial spores after treatment in steam autoclave, and to propose strictly morphological considerations about the target of this sterilisation process. Autoclave is the most common device for sterilising instruments in order to prevent cross infections in dental offices. The autoclave efficiency has been improved in the last years and part of this improvement is related to both a better and more correct use of the autoclave system and to the technological innovations introduced in the last generation of devices. However, associations as ADA or CDC suggest to regularly verify the process of 'autoclaving' through biological indicators (BI). The most commonly used BI are made of spores strips or suspensions of Bacillus Subtilis (pb 168) and Bacillus Stearothermophilus (ATCC 10149). They visually prove, changing colours on enzymatic base, the death of micro-organism and if the physical parameters, necessary for sterilisation, have been achieved. These two strains of endospore-forming bacteria were processed and prepared following two different techniques: Karnovsky fixed and epon embedded--phosphotungstic acid fixed for direct observation. The kind and the extent of analysed modifications are extremely various: from deep lacerations, which changed the spore structure, to little clefts which let the cytoplasm go out.

  12. 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.

  13. Lysinibacillus louembei sp. nov., a spore-forming bacterium isolated from Ntoba Mbodi, alkaline fermented leaves of cassava from the Republic of the Congo

    DEFF Research Database (Denmark)

    Ouoba, Labia Irène I.; Mbozo, Alain B. Vouidibio; Thorsen, Line

    2015-01-01

    Investigation of the microbial diversity of Ntoba Mbodi, an African food made from the alkaline fermentation of cassava leaves, revealed the presence of a Gram-positive, catalase-positive, aerobic, motile and rod-shaped endospore-forming bacterium (NM73) with unusual phenotypic and genotypic...

  14. Bioplastics from microorganisms.

    Science.gov (United States)

    Luengo, José M; García, Belén; Sandoval, Angel; Naharro, Germán; Olivera, Elías R

    2003-06-01

    The term 'biomaterials' includes chemically unrelated products that are synthesised by microorganisms (or part of them) under different environmental conditions. One important family of biomaterials is bioplastics. These are polyesters that are widely distributed in nature and accumulate intracellularly in microorganisms in the form of storage granules, with physico-chemical properties resembling petrochemical plastics. These polymers are usually built from hydroxy-acyl-CoA derivatives via different metabolic pathways. Depending on their microbial origin, bioplastics differ in their monomer composition, macromolecular structure and physical properties. Most of them are biodegradable and biocompatible, which makes them extremely interesting from the biotechnological point of view.

  15. New pressure and temperature effects on bacterial spores

    International Nuclear Information System (INIS)

    Mathys, A; Knorr, D; Heinz, V

    2008-01-01

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

  16. 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

  17. 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

  18. Lignite microorganisms

    Energy Technology Data Exchange (ETDEWEB)

    Bulankina, M.A.; Lysak, L.V.; Zvyagintsev, D.G. [Moscow MV Lomonosov State University, Moscow (Russian Federation). Faculty of Soil Science

    2007-03-15

    The first demonstration that samples of lignite at a depth of 10 m are considerably enriched in bacteria is reported. According to direct microscopy, the abundance of bacteria was about 10{sup 7} cells/g. About 70% of cells had intact cell membranes and small size, which points to their anabiotic state. The fungal mycelium length was no more than 1 m. Lignite inoculation onto solid glucose-yeast-peptone medium allowed us to isolate bacteria of the genera Bacillus, Rhodococcus, Arthrobacter, Micrococcus, Spirillum, and Cytophaga. Representatives of the genera Penicillium and Trichoderma were identified on Czapek medium. Moistening of lignite powder increased the microbial respiration rate and microbial and fungal abundance but did not increase their generic diversity. This finding suggests that the studied microorganisms are autochthonous to lignite.

  19. Uses of irradiation for inactivation of microorganisms

    International Nuclear Information System (INIS)

    Brynjolfsson, A.

    1988-01-01

    The lethal effects of radiation on microorganisms was noted soon after the discovery of X rays in 1895. In 1904, it was shown that vegetative bacteria are more sensitive than spores; however, no industrial applications could be made as the radiation sources were too expensive. In the mid-1950s, it became economical and practical to sterilize medical products, and ever since sterilization has been a growing industry. Radiation sterilization technology has made possible users of new materials, such as plastics. Food irradiation is about to take off. Just as there was a resistance to pasteurization of milk when it was first introduced, there will be resistance to radpasteurization. Irradiated foods have been proven safe beyond reasonable doubt. Safety has been established through two independent methods: (1) through the most extensive multigeneration animal feeding studies ever carried out, and (2) by analyzing the radiolytic products formed and the chemical changes that take place when food is irradiated. The possible toxicity of these products has been evaluated by an independent group of toxicologists, who based their evaluation on the results of exposure of these products in large quantities either to humans or to animals

  20. Evaluating the Sporicidal Activity of Disinfectants againstClostridium difficileandBacillus amyloliquefaciensSpores by Using the Improved Methods Based on ASTM E2197-11.

    Science.gov (United States)

    Uwamahoro, Marie Christine; Massicotte, Richard; Hurtubise, Yves; Gagné-Bourque, François; Mafu, Akier Assanta; Yahia, L'Hocine

    2018-01-01

    Spore-forming pathogenic bacteria, such as Clostridium difficile , are associated with nosocomial infection, leading to the increased use of sporicidal disinfectants, which impacts socioeconomic costs. However, C. difficile can be prevented using microorganisms such as Bacillus amyloliquefaciens , a prophylactic agent that has been proven to be effective against it in recent tests or it can be controlled by sporicidal disinfectants. These disinfectants against spores should be evaluated according to a known and recommended standard. Unfortunately, some newly manufactured disinfectants like Bioxy products have not yet been tested. ASTM E2197-11 is a standard test that uses stainless steel disks (1 cm in diameter) as carriers, and the performance of the test formulation is calculated by comparing the number of viable test organisms to that on the control carriers. Surface tests are preferable for evaluating disinfectants with sporicidal effects on hard surfaces. This study applies improved methods, based on the ASTM E2197-11 standard, for evaluating and comparing the sporicidal efficacies of several disinfectants against spores of C. difficile and B. amyloliquefaciens , which are used as the test organisms. With the improved method, all spores were recovered through vortexing and membrane filtration. The results show that chlorine-based products are effective in 5 min and Bioxy products at 5% w/v are effective in 10 min. Although Bioxy products may take longer to prove their effectiveness, their non-harmful effects to hospital surfaces and people have been well established in the literature.

  1. Infrared signatures to discriminate viability of autoclaved Bacillus spores

    Science.gov (United States)

    Schneider, Matthew D. W.; Valentine, Nancy B.; Johnson, Timothy J.

    2011-11-01

    Optical methods can offer good sensitivity for detecting small amounts of chemicals and biologicals, and as these methods mature, are some of the few techniques that can offer true standoff detection. For detection of biological species, determining the viability is clearly important: Certain species of gram-positive bacteria are capable of forming endospores, specialized structures that arise when living conditions become unfavorable or little growth medium is available. Spores are also resistant to many chemicals as well as changes in heat or pH; such spores can remain dormant from months to years until more favorable conditions arise, resulting in germination back to the vegetative state. This persistence characteristic of bacterial spores allows for contamination of a surface (e.g. food or medical equipment) even after the surface has been nominally cleaned. Bacterial spores have also been used as biological weapons, as in the case of B. anthracis. Thus, having rapid analytical methods to determine a spore's viability after attempts to clean a given environment is crucial. The increasing availability of portable spectrometers may provide a key to such rapid onsite analysis. The present study was designed to determine whether infrared spectroscopy may be used to differentiate between viable vs. dead B. subtilis and B. atrophaeus spores. Preliminary results show that the reproducible differences in the IR signatures can be used to identify the viable vs. the autoclaved (dead) spores.

  2. Survival rates of some terrestrial microorganisms under simulated space conditions

    Science.gov (United States)

    Koike, J.; Oshima, T.; Koike, K. A.; Taguchi, H.; Tanaka, R.; Nishimura, K.; Miyaji, M.

    1992-10-01

    In connection with planetary quarantine, we have been studying the survival rates of nine species of terrestrial microorganisms (viruses, bacteria, yeasts, fungi, etc.) under simulated interstellar conditions. If common terrestrial microorganisms cannot survive in space even for short periods, we can greatly reduce expenditure for sterilizing space probes. The interstellar environment in the solar system has been simulated by low temperature, high vacuum (77 k, 4 × 10-6 torr), and protons irradiation from a Van de Graaff generator. After exposure to a barrage of protons corresponding to about 250 years of irradiation in solar space, Tobacco mosaic virus. Bacillus subtilis spores, Aspergillus niger spores, and Clostridium mangenoti spores showed survival rates of 82%, 45%, 28%, and 25%, respectively. Furthermore, pathogenic Candida albicans showed 7% survival after irradiation corresponding to about 60 years in space.

  3. Late Silurian trilete spores from northern Jiangsu, China.

    Science.gov (United States)

    Wang; Li

    2000-08-01

    The Late Silurian is generally considered to a particular significant key period in the study of early land vascular plants. A trilete spore assemblage of the Upper Silurian is described from northern Jiangsu, China. This assemblage comprises 11 genera and 20 species of trilete spores (including laevigate, apiculate, perinotrilite, patinate, rarely distally murornate and equatorially crassitate, and three indeterminate trilete miospores forms). It has similarities to those described from coeval assemblages from around the world (e.g., England and South Wales; Tripolitania, Libya; Cornwallis Island, Canadian Arctic; Northwest Spain). The rare cryptospore, only one specimen (Tetrahedraletes sp.) had been found to be associated with the Chinese trilete spore assemblage. The discovery of the trilete spores from Late Silurian rocks indicates the existence of early land plants, some possibly vascular, at that time in northern Jiangsu, China.

  4. 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.

  5. Identification and Characterization of Early Mission Phase Microorganisms Residing on the Mars Science Laboratory and Assessment of Their Potential to Survive Mars-like Conditions

    Science.gov (United States)

    Smith, Stephanie A.; Benardini, James N., III; Anderl, David; Ford, Matt; Wear, Emmaleen; Schrader, Michael; Schubert, Wayne; DeVeaux, Linda; Paszczynski, Andrzej; Childers, Susan E.

    2017-03-01

    Planetary protection is governed by the Outer Space Treaty and includes the practice of protecting planetary bodies from contamination by Earth life. Although studies are constantly expanding our knowledge about life in extreme environments, it is still unclear what the probability is for terrestrial organisms to survive and grow on Mars. Having this knowledge is paramount to addressing whether microorganisms transported from Earth could negatively impact future space exploration. The objectives of this study were to identify cultivable microorganisms collected from the surface of the Mars Science Laboratory, to distinguish which of the cultivable microorganisms can utilize energy sources potentially available on Mars, and to determine the survival of the cultivable microorganisms upon exposure to physiological stresses present on the martian surface. Approximately 66% (237) of the 358 microorganisms identified are related to members of the Bacillus genus, although surprisingly, 22% of all isolates belong to non-spore-forming genera. A small number could grow by reduction of potential growth substrates found on Mars, such as perchlorate and sulfate, and many were resistant to desiccation and ultraviolet radiation (UVC). While most isolates either grew in media containing ≥10% NaCl or at 4°C, many grew when multiple physiological stresses were applied. The study yields details about the microorganisms that inhabit the surfaces of spacecraft after microbial reduction measures, information that will help gauge whether microorganisms from Earth pose a forward contamination risk that could impact future planetary protection policy.

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

    International Nuclear Information System (INIS)

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

    2009-01-01

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

  7. Gene activity during germination of spores of the fern, Onoclea sensibilis. Cell-free translation analysis of mRNA of spores and the effect of alpha-amanitin on spore germination

    Science.gov (United States)

    Raghavan, V.

    1992-01-01

    Poly(A)-RNA fractions of dormant, dark-imbibed (non-germinating) and photoinduced (germinating) spores of Onoclea sensibilis were poor templates in the rabbit reticulocyte lysate protein synthesizing system, but the translational efficiency of poly(A)+RNA was considerably higher than that of unfractionated RNA. Poly(A)+RNA isolated from photoinduced spores had a consistently higher translational efficiency than poly(A)+RNA from dark-imbibed spores. Analysis of the translation products by one-dimensional polyacrylamide gel electrophoresis showed no qualitative differences in the mRNA populations of dormant, dark-imbibed, and photoinduced spores. However, poly(A)+RNA from dark-imbibed spores appeared to encode in vitro fewer detectable polypeptides at a reduced intensity than photoinduced spores. A DNA clone encoding the large subunit of maize ribulose bisphosphate carboxylase hybridized at strong to moderate intensity to RNA isolated from dark-imbibed spores, indicating the absence of mRNA degradation. Although alpha-amanitin did not inhibit the germination of spores, the drug prevented the elongation of the rhizoid and protonemal initial with a concomitant effect on the synthesis of poly(A)+RNA. These results are consistent with the view that some form of translational control involving stored mRNA operates during dark-imbibition and photoinduced germination of spores.

  8. 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.

  9. A mobile genetic element profoundly increases heat resistance of bacterial spores.

    Science.gov (United States)

    Berendsen, Erwin M; Boekhorst, Jos; Kuipers, Oscar P; Wells-Bennik, Marjon H J

    2016-11-01

    Bacterial endospores are among the most resilient forms of life on earth and are intrinsically resistant to extreme environments and antimicrobial treatments. Their resilience is explained by unique cellular structures formed by a complex developmental process often initiated in response to nutrient deprivation. Although the macromolecular structures of spores from different bacterial species are similar, their resistance to environmental insults differs widely. It is not known which of the factors attributed to spore resistance confer very high-level heat resistance. Here, we provide conclusive evidence that in Bacillus subtilis, this is due to the presence of a mobile genetic element (Tn1546-like) carrying five predicted operons, one of which contains genes that encode homologs of SpoVAC, SpoVAD and SpoVAEb and four other genes encoding proteins with unknown functions. This operon, named spoVA 2mob , confers high-level heat resistance to spores. Deletion of spoVA 2mob in a B. subtilis strain carrying Tn1546 renders heat-sensitive spores while transfer of spoVA 2mob into B. subtilis 168 yields highly heat-resistant spores. On the basis of the genetic conservation of different spoVA operons among spore-forming species of Bacillaceae, we propose an evolutionary scenario for the emergence of extremely heat-resistant spores in B. subtilis, B. licheniformis and B. amyloliquefaciens. This discovery opens up avenues for improved detection and control of spore-forming bacteria able to produce highly heat-resistant spores.

  10. The occurrence of Ganoderma spores in the air and its relationships with meteorological factors

    Directory of Open Access Journals (Sweden)

    Agnieszka Grinn-Gofroń

    2012-12-01

    Full Text Available According to a recent study, Ganoderma may be the third genus, after Alternaria and Cladosporium, whose spores cause symptoms of allergy and whose levels are directly related to meteorological factors. There are only few articles from different parts of the world about the relationships between Ganoderma spore count and meteorological factors. The aim of the study was to review all available publications about airborne Ganoderma spores and to compare the results in a short useful form.

  11. Infrared Signatures to Discriminate Viability of Autoclaved Bacillus Spores

    Energy Technology Data Exchange (ETDEWEB)

    Schneider, Matthew D.; Valentine, Nancy B.; Johnson, Timothy J.

    2011-10-06

    Optical methods can offer good sensitivity for detecting small amounts of chemicals and biologicals, and as these methods mature, are some of the few techniques that can offer true standoff detection. For detection of biological species, determining the viability is clearly important: Certain species of gram-positive bacteria are capable of forming endospores, specialized structures that arise when living conditions become unfavorable or little growth medium is available, being resistant to many chemicals as well as changes in heat or pH. Such spores can remain dormant from months to years until more favorable conditions arise, resulting in germination back to the vegetative state. This persistence characteristic of bacterial spores allows for contamination of a surface (e.g. food or medical equipment) even after the surface has been nominally cleaned. Bacterial spores have also been used as biological weapons, as in the case with B. anthracis. Thus, rapid analysis to determine a spore's viability in a given environment or after attempts to sterilize a given environment is crucial. The increasing availability of portable spectrometers may provide a key to such rapid onsite analysis. The present study was designed to determine whether infrared spectroscopy may be used to differentiate between viable vs. dead B. subtilis and B. atrophaeus spores. Preliminary results show that the reproducible differences in the IR signatures can be used to identify viable vs. autoclaved (dead) B. subtilis and B. atrophaeus bacterial spores.

  12. A Ratio of Spore to Viable Organisms: A Case Study of the JPL-SAF Cleanroom

    Science.gov (United States)

    Hendrickson, Ryan; Urbaniak, Camilla; Malli Mohan, Ganesh Babu; Aronson, Heidi; Venkateswaran, Kasthuri

    2017-01-01

    Spacecraft surfaces that are destined to land on potential life-harboring celestial bodies are required to be rigorously cleaned and continuously monitored for spore bioburden as a proxy for spacecraft cleanliness. The NASA standard assay (NSA), used for spacecraft bioburden estimates, specifically measures spores that are cultivable, aerobic, resistant to heat shock, and grow at 30 C in a nutrient-rich medium. Since the vast majority of microorganisms cannot be cultivated using the NSA, it is necessary to utilize state-of-the art molecular techniques to better understand the presence of all viable microorganisms, not just those measured with the NSA. In this study, the nutrient-deprived low biomass cleanrooms, where spacecraft are assembled, were used as a surrogate for spacecraft surfaces to measure the ratio of NSA spores in relation to the total viable microorganism population in order to make comparisons with the 2006 Space Studies Board (SSB) estimate of 1 spore per approximately 50,000 viable organisms. Ninety-eight surface wipe samples were collected from the Spacecraft Assembly Facility (SAF) cleanroom at the Jet Propulsion Laboratory (JPL) over a 6-month period. The samples were processed and analyzed using classical microbiology along with molecular methodology. Traditional microbiology plating methods were used to determine the cultivable bacterial, fungal, and spore populations. Molecular assays were used to determine the total organisms (TO, dead and live) and the viable organisms (VO, live). The TO was measured using adenosine triphosphate (ATP) and quantitative polymerase chain reaction (qPCR) assays. The VO was measured using internal ATP, propidium monoazide (PMA)-qPCR, and flow cytometry (after staining for viable microorganisms) assays. Based on the results, it was possible to establish a ratio between spore counts and VO for each viability assay. The ATP-based spore to VO ratio ranged from 149-746, and the bacterial PMA-qPCR assay-based ratio

  13. The role of heat resistance in thermorestoration of hydrated bacterial spores

    International Nuclear Information System (INIS)

    Friedman, Y.S.; Grecz, N.

    1973-01-01

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

  14. Sterilization of hydrogen peroxide resistant bacterial spores with stabilized chlorine dioxide.

    Science.gov (United States)

    Friedline, Anthony; Zachariah, Malcolm; Middaugh, Amy; Heiser, Matt; Khanna, Neeraj; Vaishampayan, Parag; Rice, Charles V

    2015-01-01

    Bacillus pumilus SAFR-032 spores isolated from a clean room environment are known to exhibit enhanced resistance to peroxide, desiccation, UV radiation and chemical disinfection than other spore-forming bacteria. The survival of B. pumilus SAFR-032 spores to standard clean room sterilization practices requires development of more stringent disinfection agents. Here, we report the effects of a stabilized chlorine dioxide-based biocidal agent against spores of B. pumilus SAFR-032 and Bacillus subtilis ATCC 6051. Viability was determined via CFU measurement after exposure. Chlorine dioxide demonstrated efficacy towards sterilization of spores of B. pumilus SAFR-032 equivalent or better than exposure to hydrogen peroxide. These results indicate efficacy of chlorine dioxide delivered through a stabilized chlorine dioxide product as a means of sterilization of peroxide- and UV-resistant spores.

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

    Directory of Open Access Journals (Sweden)

    Luz del Carmen Huesca-Espitia

    2017-10-01

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

  16. A highly redundant gene network controls assembly of the outer spore wall in S. cerevisiae.

    Directory of Open Access Journals (Sweden)

    Coney Pei-Chen Lin

    Full Text Available The spore wall of Saccharomyces cerevisiae is a multilaminar extracellular structure that is formed de novo in the course of sporulation. The outer layers of the spore wall provide spores with resistance to a wide variety of environmental stresses. The major components of the outer spore wall are the polysaccharide chitosan and a polymer formed from the di-amino acid dityrosine. Though the synthesis and export pathways for dityrosine have been described, genes directly involved in dityrosine polymerization and incorporation into the spore wall have not been identified. A synthetic gene array approach to identify new genes involved in outer spore wall synthesis revealed an interconnected network influencing dityrosine assembly. This network is highly redundant both for genes of different activities that compensate for the loss of each other and for related genes of overlapping activity. Several of the genes in this network have paralogs in the yeast genome and deletion of entire paralog sets is sufficient to severely reduce dityrosine fluorescence. Solid-state NMR analysis of partially purified outer spore walls identifies a novel component in spore walls from wild type that is absent in some of the paralog set mutants. Localization of gene products identified in the screen reveals an unexpected role for lipid droplets in outer spore wall formation.

  17. Work scheme to isolate the different microorganisms found in hydrogen-producing reactors: A study of effectiveness by pyrosequencing analysis.

    Science.gov (United States)

    Fuentes, Laura; Braga, Lucía; Castelló, Elena; Etchebehere, Claudia

    2018-03-24

    The aim of this research was to create a work scheme for the isolation of the different microorganisms commonly found in hydrogen-producing reactors and to test its effectiveness. Methods were selected to isolate anaerobic spore-forming fermenters, anaerobic fermenters that do not form spores, facultative aerobic fermenters, and lactic acid bacteria. The methods were tested in two samples taken from a hydrogen-producing reactor fed with cheese whey. 16S rRNA gene sequences from isolates were compared with pyrosequencing analysis from the same samples. The isolates represented more than 88% of the abundance detected by pyrosequencing. Organisms from the genera Clostridium, Rahnella, Megasphaera, Lactobacillus, Propionibacterium, Bifidobacterium, Chryseobacterium, and Acetobacter were isolated. Hydrogen-producing capacity was confirmed for the Clostridium, Rahnella, and Megasphaera isolates. Co-culture experiments indicate that Megasphaera prevented the total inhibition of Clostridium by Lactobacillus. The work scheme proposed was effective to isolate most of the microorganisms detected by pyrosequencing. Physiological studies suggested a key role of Megasphaera. We showed the high culturability of the microbial communities from hydrogen-producing bioreactors. The isolates can be used to perform physiological studies to understand the H 2 producing process. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  18. Radiosensitivity of spores of Paenibacillus larvae ssp. larvae in honey

    Energy Technology Data Exchange (ETDEWEB)

    Almeida, Wanderley Mendes de [Ministerio da Agricultura, Pecuaria e Abastecimento, Rio de Janeiro, RJ (Brazil). Servico de Inspecao de Produtos de Origem Animal]. E-mail: sipa-rj@agricultura.gov.br; Vital, Helio de Carvalho [Centro Tecnologico do Exercito CTEx, Rio de Janeiro, RJ (Brazil). Div. de Defesa Quimica, Biologica e Nuclear]. E-mail: vital@ctex.eb.br; Schuch, Dulce Maria Tocchetto [Ministerio da Agricultura, Pecuaria e Abastecimento, Porto Alegre, RS (Brazil)]. E-mail: micro-lara-rs@agricultura.gov.br

    2007-07-01

    Irradiation, usually used in combination with other conventional methods of conservation, has been proven to be an efficient tool to ensure the safety of many types of foods by destroying pathogenic microorganisms and extending their shelf-lives. This work has investigated the efficacy of gamma irradiation to inactivate spores of the bacterium Paenibacillus larvae that causes the 'American foulbrood', a highly contagious disease still exotic in Brazil that kills bees and contaminates honey, preventing its commercialization and causing great economical losses. In this study, 60 g samples of two types of honey inoculated with 3.5x10{sup 3} spores/mL of that bacterium were irradiated with doses of 0, 5, 7.5, 10, 12.5 and 15 kGy and counted. The analyses indicated a mean reduction of 97.5{+-}0.7% in the number of viable spores exposed to 5 kGy. The application of doses of 7.5 kGy or higher yielded no viable spores above the detection threshold (10/mL). In addition the value of D{sub 10} (3.1{+-}0.3 kGy) was estimated and the logarithm of the population of viable spores of Paenibacillus larvae subsp. larvae was determined as linear and quadratic polynomial functions of the radiation dose. The results indicated that the dose of 10 kGy could be insufficient to assure complete sterilization of honey in some cases while suggesting that 25 kGy would perform such task adequately. (author)

  19. A probabilistic modeling approach in thermal inactivation: estimation of postprocess Bacillus cereus spore prevalence and concentration

    NARCIS (Netherlands)

    Membre, J.M.; Amezquita, A.; Bassett, J.; Giavedoni, P.; Blackburn, W.; Gorris, L.G.M.

    2006-01-01

    The survival of spore-forming bacteria is linked to the safety and stability of refrigerated processed foods of extended durability (REPFEDs). A probabilistic modeling approach was used to assess the prevalence and concentration of Bacillus cereus spores surviving heat treatment for a semiliquid

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

    Science.gov (United States)

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

    2017-07-15

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

  1. The infrared spectral transmittance of Aspergillus niger spore aggregated particle swarm

    Science.gov (United States)

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

    2015-10-01

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

  2. Comparative characterization of silver nanoparticles synthesized by spore extract of Bacillus subtilis and Geobacillus stearothermophilus

    Directory of Open Access Journals (Sweden)

    Seyed Mahdi Ghasemi

    2018-01-01

    Full Text Available Objective(s: Silver nanostructures have gathered remarkable attention due to their applications in diversefields. Researchers have recently demonstrated that bacterial spores are capable of reducing silver ions toelemental silver leading to formation of nanoparticles.Materials and Methods: In this study, spores of Bacillus subtilis and Geobacillus stearothermophilus wereemployed to produce silver nanoparticles (SNPs from silver nitrate (AgNO3 through a green synthesismethod. The production of SNPs by spores, heat inactivated spores (microcapsule and spore extracts wasmonitored and compared at wavelengths between 300 to 700 nm. The biosynthesized SNPs by spore extractswere characterized and confirmed by XRD and TEM analyses.Results: UV-Visible spectroscopy showed that the spore extracts were able to synthesize more SNPs thanthe other forms. The XRD pattern also revealed that the silver nanometals have crystalline structure withvarious topologies. The TEM micrographs showed polydispersed nanocrystal with dimensions ranging from30 to 90 nm and 15 to 50 nm produced by spore extracts of B. subtilis and G. stearothermophilus, respectively.Moreover, these biologically synthesized nanoparticles exhibited antimicrobial activity against differentopportunistic pathogens.Conclusion: This study suggests the bacterial spore extract as a safe, efficient, cost effective and eco-friendlymaterial for biosynthesis of SNPs.

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

    Directory of Open Access Journals (Sweden)

    Rachna Pandey

    Full Text Available Spore-forming bacteria are a special problem for the food industry as some of them are able to survive preservation processes. Bacillus spp. spores can remain in a dormant, stress resistant state for a long period of time. Vegetative cells are formed by germination of spores followed by a more extended outgrowth phase. Spore germination and outgrowth progression are often very heterogeneous and therefore, predictions of microbial stability of food products are exceedingly difficult. Mechanistic details of the cause of this heterogeneity are necessary. In order to examine spore heterogeneity we made a novel closed air-containing chamber for live imaging. This chamber was used to analyze Bacillus subtilis spore germination, outgrowth, as well as subsequent vegetative growth. Typically, we examined around 90 starting spores/cells for ≥4 hours per experiment. Image analysis with the purposely built program "SporeTracker" allows for automated data processing from germination to outgrowth and vegetative doubling. In order to check the efficiency of the chamber, growth and division of B. subtilis vegetative cells were monitored. The observed generation times of vegetative cells were comparable to those obtained in well-aerated shake flask cultures. The influence of a heat stress of 85°C for 10 min on germination, outgrowth, and subsequent vegetative growth was investigated in detail. Compared to control samples fewer spores germinated (41.1% less and fewer grew out (48.4% less after the treatment. The heat treatment had a significant influence on the average time to the start of germination (increased and the distribution and average of the duration of germination itself (increased. However, the distribution and the mean outgrowth time and the generation time of vegetative cells, emerging from untreated and thermally injured spores, were similar.

  4. Potential of MALDI-TOF mass spectrometry as a rapid detection technique in plant pathology: identification of plant-associated microorganisms.

    Science.gov (United States)

    Ahmad, Faheem; Babalola, Olubukola O; Tak, Hamid I

    2012-09-01

    Plant diseases caused by plant pathogens substantially reduce crop production every year, resulting in massive economic losses throughout the world. Accurate detection and identification of plant pathogens is fundamental to plant pathogen diagnostics and, thus, plant disease management. Diagnostics and disease-management strategies require techniques to enable simultaneous detection and quantification of a wide range of pathogenic and non-pathogenic microorganisms. Over the past decade, rapid development of matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) techniques for characterization of microorganisms has enabled substantially improved detection and identification of microorganisms. In the biological sciences, MALDI-TOF MS is used to analyze specific peptides or proteins directly desorbed from intact bacteria, fungal spores, nematodes, and other microorganisms. The ability to record biomarker ions, in a broad m/z range, which are unique to and representative of individual microorganisms, forms the basis of taxonomic identification of microorganisms by MALDI-TOF MS. Recent advances in mass spectrometry have initiated new research, i.e. analysis of more complex microbial communities. Such studies are just beginning but have great potential for elucidation not only of the interactions between microorganisms and their host plants but also those among different microbial taxa living in association with plants. There has been a recent effort by the mass spectrometry community to make data from large scale mass spectrometry experiments publicly available in the form of a centralized repository. Such a resource could enable the use of MALDI-TOF MS as a universal technique for detection of plant pathogens and non-pathogens. The effects of experimental conditions are sufficiently understood, reproducible spectra can be obtained from computational database search, and microorganisms can be rapidly characterized by genus, species

  5. The effect of antagonistic micro-organisms on the brood of honeybees (Apis mellifera) and bumblebees (Bombus terrestris) 2003

    NARCIS (Netherlands)

    Steen, van der J.J.M.; Dik, A.J.

    2002-01-01

    Several plant pathogenic fungi enter the plant trough open flowers. Spores of antagonistic micro-organisms present on the flowers can successfully compete with the possible pathogens. Honeybees and bumblebees can be used for transporting these antagonistic micro-organisms from the hive into flowers

  6. The effect of metal ions commonly present in food on gene expression of sporulating Bacillus subtilis cells in relation to spore wet heat resistance.

    NARCIS (Netherlands)

    Oomes, S.J.C.M.; Brul, S.

    2004-01-01

    Bacillus subtilis is a food spoilage spore-forming bacterium. The spores can be very heat-resistant and may cause problems in the production of foods. Varying the metal concentration in the sporulation media is known to influence the heat resistance of the spores. The effect of changing the metal

  7. Biodiversity of the phosphate solubilizing microorganisms (PSMs ...

    African Journals Online (AJOL)

    The plant rhizosphere microorganisms having the phosphate solubilizing capacity can convert the insoluble soil organic and inorganic phosphates into a soluble form and make the phosphorus (P) available to the plant. With the objective of evaluating the phosphate solubilizing microorganism populations under the rice ...

  8. Development and fine structure of sclerotia and spores of the actinomycete Chainia olivacea.

    Science.gov (United States)

    Sharples, G P; Williams, S T

    1976-01-01

    Sclerotia and spores of Chainia olivacea were studied by transmission and scanning electron microscopy. Sclerotia formed by repeated branching of several hyphea. Branch tips were delimited by septa and increased in size, becoming filled with lipid-like inclusions. In mautre sclerotia, empty cells and intra-hyphal growth were observed. An electron-dense fibrillar material was deposited between hyphae and on the sclerotium surface. The similarities between these and the sclerotia of certain fungi are discussed. Spores were formed in a manner similar to that in Streptomyces species. Large inter-sporal pads were formed during ingrowth of the septa delimiting the spores.

  9. The proteome of spore surface layers in food spoiling bacteria

    NARCIS (Netherlands)

    Abhyankar, W.R.

    2014-01-01

    Endospores are dormant, multilayered, highly resistant cellular structures formed in response to stress by certain bacteria belonging to the genera Bacillus, Clostridium and other related organisms. In presence of nutrients and favorable conditions spores germinate and grow out as normal vegetative

  10. Spore Proteomics: The Past, Present and the Future

    NARCIS (Netherlands)

    Abhyankar, W.; de Koning, L.J.; Brul, S.; de Koster, C.G.

    2014-01-01

    Endospores are metabolically dormant, multi-layered cellular structures formed by Gram positive bacteria belonging to the genera Bacillus, Clostridium and related organisms. Their external layers are composed of proteins which in part play a role in resistance behaviour of spores to varied chemical

  11. Effect of irradiation of bacteria on the formation of spores

    International Nuclear Information System (INIS)

    Szulc, M.; Tropilo, J.; Olszewski, G.

    1980-01-01

    Studies were carried out on bacteria: Bac. subtilis, Bac. cereus, Cl. perfringens, Cl. botulinum which were irradiated in two media (PBS and broth containing 1% of protein) with 100, 1000, 5000 and 10 000 X-radiation doses. The results obtained show that: all bacteria species studied (vegetative forms) are characterized by a high sensitivity to X-radiation, though distinctly lower than the species of Enterobacteriaceae family; the bacteria species studied are characterized by various sporing rate. The highest sporing rate was shown by Bac. cereus, the following: Bac. subtilis, Cl. perfringens and Cl. botulinum; increased X-radiation doses weaken sporing of Bac. subtilis and Bac. cereus. This effect could not be observed in Cl. perfringens and Cl. botulinum. (author)

  12. Reduction of Clostridium sporogenes spore outgrowth in natural sausage casings using nisin.

    Science.gov (United States)

    Wijnker, J J; Weerts, E A W S; Breukink, E J; Houben, J H; Lipman, L J A

    2011-08-01

    Preservation of natural sausage casings using dry salt or saturated brine is regarded as sufficient to inactivate vegetative pathogenic non-spore-forming bacteria present on the casings. Although the outgrowth of bacterial spores is prevented by salt or saturated brine preservation, these spores will remain present and develop into vegetative cells when conditions are more favourable. To prevent subsequent outgrowth additional preservation measures should be implemented. In the experiments described the use of nisin was evaluated to reduce outgrowth of spores in desalinated casings. The bacteriocin nisin was chosen because of its known efficacy against spore-forming bacteria and their spores in various foodstuffs. Clostridium spore suspensions (Clostridium sporogenes, ATCC 3584) were used in two concentrations to inoculate three nisin concentrations (10, 50, 100 μg/mL) in water containing gamma-irradiated casings. Additionally, the binding of nisin to casings, using (14)C-labeled nisin Z and subsequent availability of nisin were evaluated. Results demonstrate that nisin is partly reversibly bound to casings and can reduce the outgrowth of Clostridium spores in the model used by approximately 1 log(10) (90%). However, the biological relevance of these results needs to be determined further by conducting industrial trials before any recommendation can be made on the practical implementation of nisin in the preservation of natural sausage casings. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Sorption of 241Am by Aspergillus niger spore and hyphae

    International Nuclear Information System (INIS)

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

    2004-01-01

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

  14. Anti-microorganism contamination measures for crude drugs utilizing radiation sterilization

    International Nuclear Information System (INIS)

    Kimura, Syojiro

    1998-01-01

    Crude drugs are manufactured by simple processing of natural mineral, animal or plant part materials, and are used in Chinese medicine. Because these components are originated by nature, they tend to have a much higher level of contaminating microorganisms than chemically synthesized compound. Many plant-derived crude drugs contain bacteria; 10 3 -10 5 cells/g and fungi; 10 2 -10 4 spores/g. Some animal-derived crude drugs contain bacteria levels of up to 10 8 cells/g, including dangerous varieties like E. coli or Salmonella. The survival rate and the required dose can be quantified as follows: N/No=e -kD and SD=D 10 x log (No/SAL), respectively. Where, k is the sterilization constant of microorganism, No is the initial count of bacteria, D is the absorbed dose, D 10 is the dose required to decrease the count to 1/10 and SAL is the count limit or sterilization assurance. The D 10 value for general microorganism is approximately 2 kGy for bacteria, 1 kGy for fungi, and 3-4 kGy for spore-forming bacteria. The results of our past studies have shown that the dose of 5-7 kGy is necessary to sterilize general microorganism that are attached to crude drugs. Besides, the D 10 value of specific bacteria ranges from 0.1 to 0.8 kGy, so the above dose should be sufficient for sterilizing the specific microorganism (SAL; 10 -6 ). The stability of crude drugs can be quantified be the following formula, C/Co=e -k'D . Where, C/Co is the residual rate of drug components, and k' is the loss coefficient. The value of k is 1-2, but that of k' is three to four figures smaller. The C/Co ratio of most drug components is more than 0.99 with a dose of 5-7 kGy. Furthermore, radiolysis materials that are decomposed by radiation are similar to compounds that are decomposed by ultraviolet rays or sunlight. 60 Co ray are suitable for sterilizing raw materials. On the other hand, electron beams are used to sterilize a thin layer of pulverized specimens, and can be incorporated into the

  15. Photometric immersion refractometry of bacterial spores.

    Science.gov (United States)

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

    1982-01-01

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

  16. A Clostridium difficile alanine racemase affects spore germination and accommodates serine as a substrate.

    Science.gov (United States)

    Shrestha, Ritu; Lockless, Steve W; Sorg, Joseph A

    2017-06-23

    Clostridium difficile has become one of the most common bacterial pathogens in hospital-acquired infections in the United States. Although C. difficile is strictly anaerobic, it survives in aerobic environments and transmits between hosts via spores. C. difficile spore germination is triggered in response to certain bile acids and glycine. Although glycine is the most effective co-germinant, other amino acids can substitute with varying efficiencies. Of these, l-alanine is an effective co-germinant and is also a germinant for most bacterial spores. Many endospore-forming bacteria embed alanine racemases into their spore coats, and these enzymes are thought to convert the l-alanine germinant into d-alanine, a spore germination inhibitor. Although the C. difficile Alr2 racemase is the sixth most highly expressed gene during C. difficile spore formation, a previous study reported that Alr2 has little to no role in germination of C. difficile spores in rich medium. Here, we hypothesized that Alr2 could affect C. difficile l-alanine-induced spore germination in a defined medium. We found that alr2 mutant spores more readily germinate in response to l-alanine as a co-germinant. Surprisingly, d-alanine also functioned as a co-germinant. Moreover, we found that Alr2 could interconvert l- and d-serine and that Alr2 bound to l- and d-serine with ∼2-fold weaker affinity to that of l- and d-alanine. Finally, we demonstrate that l- and d-serine are also co-germinants for C. difficile spores. These results suggest that C. difficile spores can respond to a diverse set of amino acid co-germinants and reveal that Alr2 can accommodate serine as a substrate. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Survival of microorganisms in space protected by meteorite material: results of the experiment 'EXOBIOLOGIE' of the PERSEUS mission.

    Science.gov (United States)

    Rettberg, P; Eschweiler, U; Strauch, K; Reitz, G; Horneck, G; Wanke, H; Brack, A; Barbier, B

    2002-01-01

    During the early evolution of life on Earth, before the formation of a protective ozone layer in the atmosphere, high intensities of solar UV radiation of short wavelengths could reach the surface of the Earth. Today the full spectrum of solar UV radiation is only experienced in space, where other important space parameters influence survival and genetic stability additionally, like vacuum, cosmic radiation, temperature extremes, microgravity. To reach a better understanding of the processes leading to the origin, evolution and distribution of life we have performed space experiments with microorganisms. The ability of resistant life forms like bacterial spores to survive high doses of extraterrestrial solar UV alone or in combination with other space parameters, e.g. vacuum, was investigated. Extraterrestrial solar UV was found to have a thousand times higher biological effectiveness than UV radiation filtered by stratospheric ozone concentrations found today on Earth. The protective effects of anorganic substances like artificial or real meteorites were determined on the MIR station. In the experiment EXOBIOLOGIE of the French PERSEUS mission (1999) it was found that very thin layers of anorganic material did not protect spores against the deleterious effects of energy-rich UV radiation in space to the expected amount, but that layers of UV radiation inactivated spores serve as a UV-shield by themselves, so that a hypothetical interplanetary transfer of life by the transport of microorganisms inside rocks through the solar system cannot be excluded, but requires the shielding of a substantial mass of anorganic substances. c2002 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

  18. Evaluation of the Performance of Iodine-Treated Biocide Filters Challenged with Bacterial Spores and Viruses

    National Research Council Canada - National Science Library

    Lee, Jin-Hwa; Wu, Chang-Yu

    2006-01-01

    Filter media coated with a cationic resin in triiodide form were challenged by Bacillus subtilis spores and MS2 bacteriophage aerosols delivered from a Collison nebulizer through air at 35% RH and 23 C...

  19. Ultraviolet-Resistant Bacterial Spores

    Science.gov (United States)

    Venkateswaran, Kasthuri; Newcombe, David; LaDuc, Myron T.; Osman, Shariff R.

    2007-01-01

    A document summarizes a study in which it was found that spores of the SAFR-032 strain of Bacillus pumilus can survive doses of ultraviolet (UV) radiation, radiation, and hydrogen peroxide in proportions much greater than those of other bacteria. The study was part of a continuing effort to understand the survivability of bacteria under harsh conditions and develop means of sterilizing spacecraft to prevent biocontamination of Mars that could interfere with the search for life there.

  20. Spore liberation in mosses revisited.

    Science.gov (United States)

    Gallenmüller, Friederike; Langer, Max; Poppinga, Simon; Kassemeyer, Hanns-Heinz; Speck, Thomas

    2018-02-01

    The ability to perform hygroscopic movements has evolved in many plant lineages and relates to a multitude of different functions such as seed burial, flower protection or regulation of diaspore release. In most mosses, spore release is controlled by hygroscopic movements of the peristome teeth and also of the spore capsule. Our study presents, for the first time, temporally and spatially well-resolved kinematic analyses of these complex shape changes in response to humidity conditions and provides insights into the sophisticated functional morphology and anatomy of the peristome teeth. In Brachythecium populeum the outer teeth of the peristome perform particularly complex hygroscopic movements during hydration and desiccation. Hydration induces fast inward dipping followed by partial re-straightening of the teeth. In their final shape, wet teeth close the capsule. During desiccation, the teeth perform an outward flicking followed by a re-straightening which opens the capsule. We present a kinematic analysis of these shape changes and of the underlying functional anatomy of the teeth. These teeth are shown to be composed of two layers which show longitudinal gradients in their material composition, structure and geometry. We hypothesize that these gradients result in (i) differences in swelling/shrinking capacity and velocity between the two layers composing the teeth, and in (ii) a gradient of velocity of swelling and shrinking from the tip to the base of the teeth. We propose these processes explain the observed movements regulating capsule opening or closing. This hypothesis is corroborated by experiments with isolated layers of peristome teeth. During hydration and desiccation, changes to the shape and mass of the whole spore capsule accompany the opening and closing. Results are discussed in relation to their significance for humidity-based regulation of spore release.

  1. In vitro mutagenesis of commercial fern, Asplenium nidus from spores

    International Nuclear Information System (INIS)

    Norazlina Noordin

    2004-01-01

    Asplenium is a largest, most diverse fern genera. One of the common species is Asplenium nidus, well known as Bird's-nest fern, a medium to large fern with erect, stout, unbranched rhizomes. In creating variability of ferns for the benefit of the ornamental plant industry, in vitro mutagenesis is used. In this study, spores of Asplenium nidus were collected from frond bearing mature sporangia. Spores were cultured in modified 1/2 MS basal medium supplemented with various combinations of 6-Benzylaminopurine (BAP) and Naphtalene Acetic Acid (NAA). Spore cultures were incubated in incubation room at 24 degree C with 16 hours photoperiod (3500 lux). It was found that, the most effective combinations were 1 mg/1 BAP + 0. 1 mg/1 NAA and 2mg/1 BAP + 0. 1 mg/1 NAA. Prothallus was formed after 10 days of cultures and gametophytes were formed 1 month later. These gametophytes were irradiated with Gamma ray at doses of 0, 20, 90, 120, 150 and 180 Gy. From the preliminary result obtained from this study, for generating variations and desired phenotypic expression for Asplenium nidus, recommended doses for in vitro mutagenesis using spores are between 90 Gy to 150 Gy. Gametophytes were subcultured at monthly interval to ensure further development and propagation. Frequent monitoring for any changes in the morphology of the irradiated Asplenium nidus plants were carried out. (Author)

  2. Effects of steam autoclave treatment on Geobacillus stearothermophilus spores.

    Science.gov (United States)

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

    2016-11-01

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

  3. Laboratory Investigations on the Survival of Bacillus subtilis Spores in Deliquescent Salt Mars Analog Environments

    Science.gov (United States)

    Nuding, Danielle L.; Gough, Raina V.; Venkateswaran, Kasthuri J.; Spry, James A.; Tolbert, Margaret A.

    2017-10-01

    Observed features such as recurring slope lineae suggest that liquid water may exist on the surface and near-subsurface of Mars today. The presence of this liquid water, likely in the form of a brine, has important implications for the present-day water cycle, habitability, and planetary protection policies. It is possible that this water is formed, at least partially, by deliquescence of salts, a process during which hygroscopic salts absorb water vapor from the atmosphere and form a saturated liquid brine. We performed laboratory experiments to examine the ability of Bacillus subtilis (B-168) spores, alone or mixed with calcium perchlorate salt (Ca(ClO4)2), to form liquid water via deliquescence under Mars-relevant conditions. Spore survival after exposure to these conditions was examined. An environmental chamber was used to expose the samples to temperature and relative humidity (RH) values similar to those found on Mars, and Raman microscopy was used to identify the phases of water and salt that were present and to confirm the presence of spores. We found that B-168 spores did not condense any detectable water vapor on their own during the diurnal cycle, even at 100% RH. However, when spores were mixed with perchlorate salt, the entire sample deliquesced at low RH values, immersing the spores in a brine solution during the majority of the simulated martian temperature and humidity cycle. After exposure to the simulated diurnal cycles and, in some cases, perchlorate brine, the impact of each environmental scenario on spore survival was estimated by standard plate assay. We found that, if there are deliquescent salts in contact with spores, there is a mechanism for the spores to acquire liquid water starting with only atmospheric water vapor as the H2O source. Also, neither crystalline nor liquid Ca(ClO4)2 is sporicidal despite the low water activity.

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

    Science.gov (United States)

    Gopal, Selvakumar; Shagol, Charlotte C; Kang, Yeongyeong; Chung, Bong Nam; Han, Seung Gab; Tong-Min, Sa

    2018-02-02

    The propagation of pure cultures of AMF is an essential requirement for their large scale agricultural application and commercialization as biofertilizers. The present study aimed to propagate AMF using the single spore inoculation technique and compare their propagation ability with the known reference spores. Arbuscular mycorrhizal fungal spores were collected from the salt-affected Saemangeum reclaimed soil in South Korea. The technique involved inoculation of Sorghum-Sudan grass (Sorghum bicolor L.) seedlings with single, healthy spores on filter paper followed by the transfer of successfully colonized seedlings to 1 kg capacity pots containing sterilized soil. After the first plant cycle, the contents were transferred to 2.5 kg capacity pots containing sterilized soil. Among the 150 inoculants, only 27 seedlings were colonized by AMF spores. After 240 days, five inoculants among the 27 seedlings resulted in the production of over 500 spores. The 18S rDNA sequencing of spores revealed that the spores produced through single spore inoculation method belonged to Gigaspora margarita, Claroideoglomus lamellosum, and Funneliformis mosseae. Furthermore, indigenous spore Funneliformis mosseae M-1 reported a higher spore count than the reference spores. The AMF spores produced using single spore inoculation technique may serve as potential bio-inoculants with an advantage of being more readily adopted by farmers due to the lack of requirement of a skilled technique in spore propagation. The results of the current study describes the feasible and cost effective method to mass produce AMF spores for large scale application. The AMF spores obtained from this method can effectively colonize plant roots and may be easily introduced to the new environment. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  5. Disinfection methods for spores of Bacillus atrophaeus, B. anthracis, Clostridium tetani, C. botulinum and C. difficile.

    Science.gov (United States)

    Oie, Shigeharu; Obayashi, Akiko; Yamasaki, Hirofumi; Furukawa, Hiroyuki; Kenri, Tsuyoshi; Takahashi, Motohide; Kawamoto, Keiko; Makino, Sou-ichi

    2011-01-01

    To evaluate disinfection methods for environments contaminated with bioterrorism-associated microorganism (Bacillus anthracis), we performed the following experiments. First, the sporicidal effects of sodium hypochlorite on spores of five bacterial species were evaluated. Bacillus atrophaeus was the most resistant to hypochlorite, followed in order by B. anthracis, Clostridium botulinum and Clostridium tetani, and Clostridium difficile. Subsequently, using B. atrophaeus spores that were the most resistant to hypochlorite, the sporicidal effects of hypochlorite at lower pH by adding vinegar were evaluated. Hypochlorite containing vinegar had far more marked sporicidal effects than hypochlorite alone. Cleaning with 0.5% (5000 ppm) hypochlorite containing vinegar inactivated B. atrophaeus spores attached to vinyl chloride and plywood plates within 15 s, while that not containing vinegar did not inactivate spores attached to cement or plywood plates even after 1 h. Therefore, the surfaces of cement or plywood plates were covered with gauze soaked in 0.5% hypochlorite containing vinegar, and the sporicidal effects were evaluated. B. atrophaeus spores attached to plywood plates were not inactivated even after 6 h, but those attached to cement plates were inactivated within 5 min. On the other hand, covering the surfaces of plywood plates with gauze soaked in 0.3% peracetic acid and gauze soaked in 2% glutaral inactivated B. atrophaeus spores within 5 min and 6 h, respectively. These results suggest that hypochlorite containing vinegar is effective for disinfecting vinyl chloride, tile, and cement plates contaminated with B. anthracis, and peracetic acid is effective for disinfecting plywood plates contaminated with such microorganism.

  6. Germination of Spores of Astrobiologically Relevant Bacillus Species in High-Salinity Environments

    Science.gov (United States)

    Nagler, Katja; Julius, Christina; Moeller, Ralf

    2016-07-01

    In times of increasing space exploration and search for extraterrestrial life, new questions and challenges for planetary protection, aiming to avoid forward contamination of different planets or moons with terrestrial life, are emerging. Spore-forming bacteria such as Bacillus species have a high contamination potential due to their spores' extreme resistance, enabling them to withstand space conditions. Spores require liquid water for their conversion into a growing cell (i.e., spore germination and subsequent growth). If present, water on extraterrestrial planets or moons is likely to be closely associated with salts (e.g., in salty oceans or brines), thus constituting high-salinity environments. Spores of Bacillus subtilis can germinate despite very high salt concentrations, although salt stress does exert negative effects on this process. In this study, germination and metabolic reactivation ("outgrowth") of spores of five astrobiologically relevant Bacillus species (B. megaterium, B. pumilus SAFR-032, B. nealsonii, B. mojavensis, and B. vallismortis) in high salinity (≤3.6 M NaCl) were investigated. Spores of different species exhibited different germination and outgrowth capabilities in high salinity, which strongly depended on germination conditions, especially the exact composition of the medium. In this context, a new "universal" germination trigger for Bacillus spores, named KAGE (KCl, L-alanine, D-glucose, ectoine), was identified, which will be very useful for future comparative germination and outgrowth studies on different Bacillus species. Overall, this study yielded interesting new insights on salt stress effects on spore germination and points out the difficulty of predicting the potential of spores to contaminate salty environments on extraterrestrial celestial bodies.

  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. Biosurfactants from marine microorganisms

    Directory of Open Access Journals (Sweden)

    Suppasil Maneerat

    2005-11-01

    Full Text Available Biosurfactants are the surface-active molecules synthesized by microorganisms. With the advantage of environmental compatibility, the demand for biosurfactants has been steadily increasing and may eventually replace their chemically synthesized counterparts. Marine biosurfactants produced by some marine microorganisms have been paid more attention, particularly for the bioremediation of the sea polluted by crude oil. This review describes screening of biosurfactant-producing microorganisms, the determination of biosurfactant activity as well as the recovery of marine surfactant. The uses of marine biosurfactants for bioremediation are also discussed.

  9. Evaluation of methods for enumerating microorganisms in filter samples from highly contaminated occupational environments.

    Science.gov (United States)

    Eduard, W; Lacey, J; Karlsson, K; Palmgren, U; Ström, G; Blomquist, G

    1990-08-01

    Scanning electron microscopy (SEM), light microscopy (LM), epifluorescence microscopy (FM), and culture were used to assess catches of microorganisms in parallel air samples on membrane filters from heavily contaminated working environments that differed in the relative abundance of bacteria, actinomycetes, and fungal spores. Except in pig houses, estimates by SEM and LM were similar, but those by FM and culture were smaller. However, in pig houses, the fluorescent stain enabled bacteria on skin scales, not seen by SEM or LM, to be counted. Although counts obtained by culturing were always smaller than those obtained by SEM or LM, they sometimes exceeded those obtained by FM. Counts suggested that 0.1-68% of bacteria + actinomycetes and 3-98% of fungal spores were viable. However, samples for culturing may have contained larger aggregates than parallel samples collected within a sampling apparatus. All spore types recognized by LM included aggregates--those of bacteria + actinomycetes sometimes exceeding 200 units, while Wallemia sebi spore aggregates were never larger than 3 spores. The size distributions of all types approximated to log-normal, although single spores and small aggregates of bacteria + actinomycetes were perhaps underrepresented. When spores were counted directly on the filter surface, as by SEM and LM, allowance was necessary for heavier deposition of particles near the center of filters by distributing counting fields systematically over the whole filter or a sector of it. Deposition was more uniform in graphite-filled polypropylene filter holders used open-faced. Losses within filter holders and during transportation from sampling site to laboratory were small. The precision of counting spore-containing particles by LM and SEM was better than that of counting individual spores. No such difference was found for FM because many large spore-containing particles were dispersed during preparation.

  10. Sporulation Temperature Reveals a Requirement for CotE in the Assembly of both the Coat and Exosporium Layers of Bacillus cereus Spores.

    Science.gov (United States)

    Bressuire-Isoard, Christelle; Bornard, Isabelle; Henriques, Adriano O; Carlin, Frédéric; Broussolle, Véronique

    2016-01-01

    The Bacillus cereus spore surface layers consist of a coat surrounded by an exosporium. We investigated the interplay between the sporulation temperature and the CotE morphogenetic protein in the assembly of the surface layers of B. cereus ATCC 14579 spores and on the resulting spore properties. The cotE deletion affects the coat and exosporium composition of the spores formed both at the suboptimal temperature of 20°C and at the optimal growth temperature of 37°C. Transmission electron microscopy revealed that ΔcotE spores had a fragmented and detached exosporium when formed at 37°C. However, when produced at 20°C, ΔcotE spores showed defects in both coat and exosporium attachment and were susceptible to lysozyme and mutanolysin. Thus, CotE has a role in the assembly of both the coat and exosporium, which is more important during sporulation at 20°C. CotE was more represented in extracts from spores formed at 20°C than at 37°C, suggesting that increased synthesis of the protein is required to maintain proper assembly of spore surface layers at the former temperature. ΔcotE spores formed at either sporulation temperature were impaired in inosine-triggered germination and resistance to UV-C and H2O2 and were less hydrophobic than wild-type (WT) spores but had a higher resistance to wet heat. While underscoring the role of CotE in the assembly of B. cereus spore surface layers, our study also suggests a contribution of the protein to functional properties of additional spore structures. Moreover, it also suggests a complex relationship between the function of a spore morphogenetic protein and environmental factors such as the temperature during spore formation. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  11. 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.

  12. Distribution of microorganisms in herb medicines and their decontamination by gamma-irradiation

    International Nuclear Information System (INIS)

    Ito, Hitoshi

    1999-01-01

    Herb medicines are traditional medicine in Japan and have been used for medical treatment. These herb medicines are contaminating frequently by microorganisms which has possibility to cause opportunistic diseases. Recently, hygienic standard of herb medicines become more strict than before, and it needs to decontaminate microorganisms by some treatments. However, chemical treatments such as by ethylene oxide fumigation leave toxic residues in the herbs while steam sterilization decease medicinal components. From study on the distribution of microorganisms in 31 samples of selected herb medicines, colony forming units of total aerobic bacteria were determined to be l.9 x 10 2 to l.4 x 10 8 per gram in 30 samples. Coliforms were also determined to be 6.9 x 10 2 to 4.3 x 10 6 per gram in 16 samples. The main aerobic bacteria were identified as Bacillus pumilus, B. circulans, B. megaterium, Erwinia, Enterobacter and Acinetobacter, whereas consisted mainly of Enterobacter in coliform counts. Molds were determined to be 6.3 x 10 1 to 1.9 x 10 5 per gram which consisted mainly Aspergillus glaucus group, A. restrictus group, A. flavus group, A. ostianus, A. phoenicis, Penicillium, Tricoderma, Rhizopus and Alternaria in 25 samples. A study on the inactivation of microorganisms at sample No. S18 showed that a gamma-irradiation dose of 20 kGy was required to reduce the total aerobic bacteria and the coliforms below a detectable level, while radiation-resistant bacteria were survived at high doses more than 10 kGy consisted with Acinetobacter and Enterobacter. Molds were inactivated below 8 kGy except Alternaria. However, a dose of 10 kGy should be effective for the sample No. S18 to reduce the spore-forming bacteria, the fecal coliforms and the molds below a detectable level per gram. On the study of inactivation of microorganisms in many samples except the No. 18, all kinds of microorganism were inactivated below a detectable level at 10 kGy irradiation. (author)

  13. Distribution of microorganisms in herb medicines and their decontamination by gamma-irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Ito, Hitoshi [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment; Kamakura, Hiroyuki; Sekita, Setuko [National Institute of Health Sciences, Kamiyoga, Tokyo (Japan)

    1999-09-01

    Herb medicines are traditional medicine in Japan and have been used for medical treatment. These herb medicines are contaminating frequently by microorganisms which has possibility to cause opportunistic diseases. Recently, hygienic standard of herb medicines become more strict than before, and it needs to decontaminate microorganisms by some treatments. However, chemical treatments such as by ethylene oxide fumigation leave toxic residues in the herbs while steam sterilization decease medicinal components. From study on the distribution of microorganisms in 31 samples of selected herb medicines, colony forming units of total aerobic bacteria were determined to be l.9 x 10{sup 2} to l.4 x 10{sup 8} per gram in 30 samples. Coliforms were also determined to be 6.9 x 10{sup 2} to 4.3 x 10{sup 6} per gram in 16 samples. The main aerobic bacteria were identified as Bacillus pumilus, B. circulans, B. megaterium, Erwinia, Enterobacter and Acinetobacter, whereas consisted mainly of Enterobacter in coliform counts. Molds were determined to be 6.3 x 10{sup 1} to 1.9 x 10{sup 5} per gram which consisted mainly Aspergillus glaucus group, A. restrictus group, A. flavus group, A. ostianus, A. phoenicis, Penicillium, Tricoderma, Rhizopus and Alternaria in 25 samples. A study on the inactivation of microorganisms at sample No. S18 showed that a gamma-irradiation dose of 20 kGy was required to reduce the total aerobic bacteria and the coliforms below a detectable level, while radiation-resistant bacteria were survived at high doses more than 10 kGy consisted with Acinetobacter and Enterobacter. Molds were inactivated below 8 kGy except Alternaria. However, a dose of 10 kGy should be effective for the sample No. S18 to reduce the spore-forming bacteria, the fecal coliforms and the molds below a detectable level per gram. On the study of inactivation of microorganisms in many samples except the No. 18, all kinds of microorganism were inactivated below a detectable level at 10 k

  14. Microorganisms involved in MIC

    Energy Technology Data Exchange (ETDEWEB)

    Sorensen, K. [Danish Technological Institute (Denmark)

    2011-07-01

    Microbiologically influenced corrosion (MIC) is a widespread problem that is difficult to detect and assess because of its complex mechanism. This paper presents the involvement of microorganisms in MIC. Some of the mechanisms that cause MIC include hydrogen consumption, production of acids, anode-cathode formation and electron shuttling. A classic bio-corrosive microorganism in the oil and gas industry is sulphate-reducing prokaryotes (SRP). Methanogens also increase corrosion rates in metals. Some of the phylogenetic orders detected while studying SRP and methanogens are archaeoglobales, clostridiales, methanosarcinales and methanothermococcus. There were some implications, such as growth of SRP not being correlated with growth of methanogens; methanogens were included in MIC risk assessment. A few examples are used to display how microorganisms are involved in topside corrosion and microbial community in producing wells. From the study, it can be concluded that, MIC risk assessment includes system data and empirical knowledge of the distribution and number of microorganisms in the system.

  15. Assessing the cleanliness of surfaces: Innovative molecular approaches vs. standard spore assays

    Energy Technology Data Exchange (ETDEWEB)

    Cooper, M.; Duc, M.T. La; Probst, A.; Vaishampayan, P.; Stam, C.; Benardini, J.N.; Piceno, Y.M.; Andersen, G.L.; Venkateswaran, K.

    2011-04-01

    A bacterial spore assay and a molecular DNA microarray method were compared for their ability to assess relative cleanliness in the context of bacterial abundance and diversity on spacecraft surfaces. Colony counts derived from the NASA standard spore assay were extremely low for spacecraft surfaces. However, the PhyloChip generation 3 (G3) DNA microarray resolved the genetic signatures of a highly diverse suite of microorganisms in the very same sample set. Samples completely devoid of cultivable spores were shown to harbor the DNA of more than 100 distinct microbial phylotypes. Furthermore, samples with higher numbers of cultivable spores did not necessarily give rise to a greater microbial diversity upon analysis with the DNA microarray. The findings of this study clearly demonstrated that there is not a statistically significant correlation between the cultivable spore counts obtained from a sample and the degree of bacterial diversity present. Based on these results, it can be stated that validated state-of-the-art molecular techniques, such as DNA microarrays, can be utilized in parallel with classical culture-based methods to further describe the cleanliness of spacecraft surfaces.

  16. Rates of molecular evolution in bacteria are relatively constant despite spore dormancy.

    Science.gov (United States)

    Maughan, Heather

    2007-02-01

    Rates of molecular evolution are known to vary considerably among lineages, partially due to differences in life-history traits such as generation time. The generation-time effect has been well documented in some eukaryotes, but its prevalence in prokaryotes is unknown. "Because many species of Firmicute bacteria spend long periods of time as metabolically dormant spores, which could result in fewer DNA substitutions per unit time, they present an excellent system for testing predictions of the molecular clock hypothesis." To test whether spore-forming bacteria evolve more slowly than their non-spore-forming relatives, I used phylogenetic methods to determine if there were differences in rates of amino acid substitution between spore-forming and non-spore-forming lineages of Firmicute bacteria. Although rates of evolution do vary among lineages, I find no evidence for an effect of spore-formation on evolutionary rate and, furthermore, evolutionary rates are similar to those calculated for enteric bacteria. These results support the notion that variation in generation time does not affect evolutionary rates in bacterial lineages.

  17. Caracterización morfológica y genética de las ectomicorrizas formadas entre Pinus montezumae y los hongos presentes en los bancos de esporas en la Faja Volcánica Transmexicana Morphologic and genetic characterization of ectomycorrhizae formed by Pinus montezumae and spore bank fungi in the Transmexican Volcanic Belt

    Directory of Open Access Journals (Sweden)

    Roberto Garibay-Orijel

    2013-03-01

    Full Text Available Los hongos ectomicorrízicos son indispensables para el establecimiento y funcionamiento de los bosques templados. Algunos de ellos tienen esporas u otros propágulos resistentes y longevos; éstos se acumulan en el suelo forestal formando bancos de propágulos que constituyen la fuente de inóculo más importante después de disturbios severos. En este trabajo caracterizamos morfológica y genéticamente las ectomicorrizas formadas entre Pinus montezumae y los hongos presentes en los bancos de esporas de la Faja Volcánica Transmexicana. Las micorrizas se obtuvieron por medio de un bioensayo del suelo de 8 de los volcanes más representativos, con plántulas de P. montezumae cultivadas durante 7 meses. La identidad taxonómica de los hongos se obtuvo por la similitud genética de la región de los ITS. Se presentan las descripciones de 27 ectomicorrizas; de éstas, 20 no habían sido publicadas previamente. Geopora sp., Hebeloma helodes, H. leucosarx, Peziza sp. 1, P. aff. ostracoderma, Pezizaceae sp. 1, sp. 2, sp. 4, Pulvinula constellatio, Sebacina sp. 1, sp. 2, Sordariales sp. 1, sp. 2 y Tuber separans, no se habían encontrado en bancos de propágulos. Estas especies podrían usarse para reforestar con plantas y hongos endémicos, lo que aumentaría la sobrevivencia, pues ambos simbiontes estarían adaptados a las condiciones ambientales locales.Ectomycorrhizal fungi are keystone in temperate forest establishment and functioning. Some of them have resistant and long living spores and propagules. These use to accumulate in soil, forming the so-called spore banks, which are the main inoculum resource after an intense disturbance. In this paper, we provide the morphological and genetic characterization of ectomycorrhizae formed by Pinus montezumae and the spore bank fungi from the Transmexican Volcanic Belt. We made a bioassay with P. montezumae and soil from 9 of the most representative volcanoes. Mycorrhizae were dissected after 7 months of

  18. Mechanisms of nickel toxicity in microorganisms

    OpenAIRE

    Macomber, Lee; Hausinger, Robert P.

    2011-01-01

    Nickel has long been known to be an important human toxicant, including having the ability to form carcinomas, but until recently nickel was believed to be an issue only to microorganisms living in nickel-rich serpentine soils or areas contaminated by industrial pollution. This assumption was overturned by the discovery of a nickel defense system (RcnR/RcnA) found in microorganisms that live in a wide range of environmental niches, suggesting that nickel homeostasis is a general biological co...

  19. Sporangium Exposure and Spore Release in the Peruvian Maidenhair Fern (Adiantum peruvianum, Pteridaceae.

    Directory of Open Access Journals (Sweden)

    Simon Poppinga

    Full Text Available We investigated the different processes involved in spore liberation in the polypod fern Adiantum peruvianum (Pteridaceae. Sporangia are being produced on the undersides of so-called false indusia, which are situated at the abaxial surface of the pinnule margins, and become exposed by a desiccation-induced movement of these pinnule flaps. The complex folding kinematics and functional morphology of false indusia are being described, and we discuss scenarios of movement initiation and passive hydraulic actuation of these structures. High-speed cinematography allowed for analyses of fast sporangium motion and for tracking ejected spores. Separation and liberation of spores from the sporangia are induced by relaxation of the annulus (the 'throwing arm' of the sporangium catapult and conservation of momentum generated during this process, which leads to sporangium bouncing. The ultra-lightweight spores travel through air with a maximum velocity of ~5 m s(-1, and a launch acceleration of ~6300 g is measured. In some cases, the whole sporangium, or parts of it, together with contained spores break away from the false indusium and are shed as a whole. Also, spores can stick together and form spore clumps. Both findings are discussed in the context of wind dispersal.

  20. Induction of Rhizopus oryzae germination under starvation using host metabolites increases spore susceptibility to heat stress.

    Science.gov (United States)

    Turgeman, Tidhar; Kakongi, Nathan; Schneider, Avishai; Vinokur, Yakov; Teper-Bamnolker, Paula; Carmeli, Shmuel; Levy, Maggie; Skory, Christopher D; Lichter, Amnon; Eshel, Dani

    2014-03-01

    Sweetpotato is a nutritional source worldwide. Soft rot caused by Rhizopus spp. is a major limiting factor in the storage of produce, rendering it potentially unsafe for human consumption. In this study, Rhizopus oryzae was used to develop a concept of postharvest disease control by weakening the pathogen through induction of spore germination under starvation conditions. We isolated the sweetpotato active fractions (SPAFs) that induce spore germination and used them at a low dose to enhance spore weakening caused by starvation. Germination in SPAF at 1 mg/ml weakened the pathogen spores by delaying their ability to form colonies on rich media and by increasing their sensitivity to heat stress. The weakening effect was also supported by reduced metabolic activity, as detected by Alarmar Blue fluorescent dye assays. Spores incubated with SPAF at 1 mg/ml showed DNA fragmentation in some of their nuclei, as observed by TUNEL assay. In addition, these spores exhibited changes in ultrastructural morphology (i.e., shrinkage of germ tubes, nucleus deformation, and vacuole formation) which are hallmarks of programmed cell death. We suggest that induction of spore germination under starvation conditions increases their susceptibility to stress and, therefore, might be considered a new strategy for pathogen control.

  1. Validated modified Lycopodium spore method development for ...

    African Journals Online (AJOL)

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

  2. Nano-Mechanical Properties of Heat Inactivated Bacillus anthracis and Bacillus thuringiensis Spores

    Science.gov (United States)

    2008-03-01

    spores were grown in plastic petri dishes on Criterion Dehydrated Culture Media , which contained per liter of formula 15 grams agar , 5 grams gelatin...became reality in 2001 when terrorists sent spores in a powdered form in letters to two senators and several news media offices, killing five people...is the causative agent of the disease anthrax. B. thuringiensis is often used in pesticides and bioengineering pest resistant crops because of its

  3. Plutonium uptake by a soil fungus and transport to its spores

    International Nuclear Information System (INIS)

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

    1976-01-01

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

  4. β-1,6-glucan synthesis-associated genes are required for proper spore wall formation in Saccharomyces cerevisiae.

    Science.gov (United States)

    Pan, Hua-Ping; Wang, Ning; Tachikawa, Hiroyuki; Nakanishi, Hideki; Gao, Xiao-Dong

    2017-11-01

    The yeast spore wall is an excellent model to study the assembly of an extracellular macromolecule structure. In the present study, mutants defective in β-1,6-glucan synthesis, including kre1∆, kre6∆, kre9∆ and big1∆, were sporulated to analyse the effect of β-1,6-glucan defects on the spore wall. Except for kre6∆, these mutant spores were sensitive to treatment with ether, suggesting that the mutations perturb the integrity of the spore wall. Morphologically, the mutant spores were indistinguishable from wild-type spores. They lacked significant sporulation defects partly because the chitosan layer, which covers the glucan layer, compensated for the damage. The proof for this model was obtained from the effect of the additional deletion of CHS3 that resulted in the absence of the chitosan layer. Among the double mutants, the most severe spore wall deficiency was observed in big1∆ spores. The majority of the big1∆chs3∆ mutants failed to form visible spores at a higher temperature. Given that the big1∆ mutation caused a failure to attach a GPI-anchored reporter, Cwp2-GFP, to the spore wall, β-1,6-glucan is involved in tethering of GPI-anchored proteins in the spore wall as well as in the vegetative cell wall. Thus, β-1,6-glucan is required for proper organization of the spore wall. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  5. Sphagnum moss disperses spores with vortex rings.

    Science.gov (United States)

    Whitaker, Dwight L; Edwards, Joan

    2010-07-23

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

  6. Ptaquiloside in bracken spores from Britain.

    Science.gov (United States)

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

    2013-03-01

    Secondary metabolites from bracken fern (Pteridium aquilinum (L.) Kuhn) are suspected of causing cancer in humans. The main carcinogen is the highly water-soluble norsesquiterpene glucoside ptaquiloside, which may be ingested by humans through food, e.g. via contaminated water, meat or milk. It has been postulated that carcinogens could also be ingested through breathing air containing bracken spores. Ptaquiloside has not previously been identified in bracken spores. The aim of the study was to determine whether ptaquiloside is present in bracken spores, and if so, to estimate its content in a collection of spores from Britain. Ptaquiloside was present in all samples, with a maximum of 29 μg g(-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 water. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Deposition of Bacteria and Bacterial Spores by Bathroom Hot Air Hand Dryers.

    Science.gov (United States)

    Del Carmen Huesca-Espitia, Luz; Aslanzadeh, Jaber; Feinn, Richard; Joseph, Gabrielle; Murray, Thomas S; Setlow, Peter

    2018-02-09

    Hot air hand dryers in multiple men's and women's bathrooms in 3 basic science research areas in an academic health center were screened for their deposition on plates of: i) total bacteria, some of which were identified; and ii) a kanamycin resistant Bacillus subtilis strain, PS533, spores of which are produced in large amounts in one basic science research laboratory. Plates exposed to hand dryer air for 30 seconds averaged 18-60 colonies/plate but interior hand dryer nozzle surfaces had minimal bacterial levels, plates exposed to bathroom air for 2 minutes with hand dryers off averaged ≤1 colony, and plates exposed to bathroom air moved by a small fan for 20 minutes had averages of 15 and 12 colonies/plate in two buildings tested. Retrofitting hand dryers with HEPA filters reduced bacterial deposition by hand dryers ∼4-fold, and potential human pathogens were recovered from plates exposed to hand dryer air whether or not a HEPA filter was present, and from bathroom air moved by a small fan. Spore-forming colonies, identified as B. subtilis PS533 averaged ∼2.5-5% of bacteria deposited by hand dryers throughout basic research areas examined regardless of distance from the spore forming laboratory, and these were almost certainly deposited as spores. Comparable results were obtained when bathroom air was sampled for spores. These results indicate that many kinds of bacteria, including potential pathogens and spores, can be deposited on hands exposed to bathroom hand dryers, and that spores could be dispersed throughout buildings and deposited on hands by hand dryers. Importance While there is evidence that bathroom hand dryers can disperse bacteria from hands or deposit bacteria on surfaces, including recently washed hands, there is less information on: i) the organisms dispersed by hand dryers; ii) if hand dryers provide a reservoir of bacteria or simply blow large amounts of bacterially contaminated air; and iii) if bacterial spores are deposited on

  8. Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms

    Directory of Open Access Journals (Sweden)

    Bazzle LJ

    2014-12-01

    97.5% of trials. The most common spore shapes observed were globose, spiked, elliptical, smooth and reticulate. Conclusion: Flotation can concentrate mushroom spores; however, false negative results can occur. Spore morphology could not be used to differentiate species of mushroom-forming fungi since the spore shape and surface characteristics seen in the present study were often observed with multiple species of mushroom-forming fungi. Keywords: gastrointestinal contents, mushroom spore identification, mushroom toxicity, Amanita spp.

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

    Directory of Open Access Journals (Sweden)

    Ozgur eCelebi

    2016-01-01

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

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

    International Nuclear Information System (INIS)

    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

  11. Systematic studies in the genus Mohria (Anemiaceae: Pteridophyta. III. Comparative sporangium and spore morphology

    Directory of Open Access Journals (Sweden)

    J. P. Roux

    1992-10-01

    Full Text Available The genera Mohria and Anemia (Anemiaceae: Pteridophyta can be separated on both their sporangia and spores. In Mohria the capsule is globose with an apical annulus but in Anemia it is ovate-globose to cylindrical with a subapical annulus. The spores of both genera are radially symmetrical, tetrahedral and trilete with near parallel muri. The exinal sculpture in Mohria is cicatricose and in Anemia it can be canaliculate or cicatricose. In both genera the mural sets anastomose to form common muri that extend from near the distal pole to the equatorial radial region. The muri in Mohria are hollow and differ from those in Anemia which are solid or microporate. Supramural sculpturing in Mohria and Anemia is perinous. Spores of the other schizaealean ferns show no or little taxonomic affinities with Mohria and Anemia. In Actinostachys and Schizaea the spores are monolete and in  Lygodium trilete but the exinal sculpture is smooth.

  12. Longitudinal assessment of dairy farm management practices associated with the presence of psychrotolerant Bacillales spores in bulk tank milk on 10 New York State dairy farms

    NARCIS (Netherlands)

    Masiello, S. N.; Kent, D.V.; Martin, N. H.; Schukken, Y. H.; Wiedmann, M.; Boor, K. J.

    2017-01-01

    The ability of certain spore-forming bacteria in the order Bacillales (e.g., Bacillus spp., Paenibacillus spp.) to survive pasteurization in spore form and grow at refrigeration temperatures results in product spoilage and limits the shelf life of high temperature, short time (HTST)-pasteurized

  13. Laboratory and Field Evidence for Long-Term Starvation Survival of Microorganisms in Subsurface Terrestrial Environments

    Energy Technology Data Exchange (ETDEWEB)

    Kieft, T.L. [Biology Dept., New Mexico Inst. of Mining and Technology, Socorro, NM (United States); Murphy, E.M. [Pacific Northwest National Lab., Richland, WA (United States); Amy, P.S.; Haldeman, D.L. [Department of Biological Sciences, University of Nevada, Las Vegas, NV (United States); Ringelberg, D. B. [Center for Environmental Biotechnology, University of Tennessee, Knoxville, TN (United States)]|[Oak Ridge National Lab., TN (United States)

    1997-12-31

    BIOGEOCHEMICAL MODELING OF GROUNDWATER FLOW AND NUTRIENT FLUX IN SUBSURFACE ENVIRONMENTS INDICATES THAT INHABITANT MICROORGANISMS EXPERIENCE SEVERE NUTRIENT LIMITATION. USING LABORATORY AND FIELD METHODS, WE HAVE BEEN TESTING STARVATION SURVIVAL IN SUBSURFACE MICROORGANISMS. IN MICROCOSM EXPERIMENTS, WE HAVE SHOWN THAT STRAINS OF TWO COMMONLY ISOLATED SUBSURFACE GENERA, ARTHROBACTER AND PSEUDOMONAS, ARE ABLE TO MAINTAIN VIABILITY IN LOW-NUTRIENT, NATURAL SUBSURFACE SEDIMENTS FOR OVER ONE YEAR. THESE NON-SPORE-FORMING BACTERIA UNDERGO RAPID INITIAL MINIATURIZATION FOLLOWED BY A STABILIZATION OF CELL SIZE. MEMBRANE LIPID PHOSPHOLIPID FATTY ACID (PLFA) PROFILES OF THE PSEUDOMONAS ARE CONSISTENT WITH ADAPTATION TO NUTRIENT STRESS; ARTHROBACTER APPARENTLY RESPONDS TO NUTRIENT DEPRIVATION WITHOUT ALTERING MEMBRANE PLFA. TO TEST SURVIVABILITY OF MICROORGANISMS OVER A GEOLOGIC TIME SCALE, WE CHARACTERIZED MICROBIAL COMMUNITIES IN A SEQUENCE OF UNSATURATED SEDIMENTS RANGING IN AGE FROM MODEM TO {gt}780,000 years. Sediments were relatively uniform silts in Eastern Washington State. Porewater ages at depth (measured by the chloride mass-balance approach) were as old as 3,600 years. Microbial abundance, biomass, and activities (measured by direct counts, culture counts, total PLFAs, and radiorespirometry) declined with sediment age. The pattern is consistent with laboratory microcosm studies of Microbial survival: rapid short-term change followed by long-term survival of a proportion of cells. Even the oldest sediments evinced a small but viable Microbial community. Microbial survival appeared to be a function of sediment age. Porewater age appeared to influence the markup of surviving communities, as indicated by PLFA profiles. Sites with different Porewater recharge rates and patterns of Pleistocene flooding had different communities.

  14. Low cost medium for spore production of Bacillus KKU02 and KKU03 and the effects of the produced spores on growth of giant freshwater prawn (Macrobrachium rosenbergii de Man).

    Science.gov (United States)

    Wangka-Orm, Charkrit; Deeseenthum, Sirirat; Leelavatcharamas, Vichai

    2014-08-01

    In order to extend the shelf life of 2 high potential Bacillus probiotic isolates which were Bacillus KKU02 and Bacillus KKU03, the spore forms of these 2 Bacillus isolates were studied for using as probiotic instead. The low cost medium for spore production of these 2 Bacillus isolates was examined in order to produce probiotic spores for feeding the shrimps. It was found that cassava at 100 g L(-1) and supplemented with 20.0 g L(-1) dextrose, 0.1 g L(-1) MgSO4 and 2.0 g L(-1) (NH4)2SO4 showed the highest spore concentration at about 1 x 10(8) CFU mL(-1). The effects of feeding these 2 Bacillus spores on the growth of giant freshwater prawns were further examined. The spores of Bacillus KKU02 and Bacillus KKU03 (-10(7) spore mL(-1)) in pure and mixed culture forms were mixed with commercial prawn feed (200 mL kg(-1)) to give six feed treatments. Body length and weight of the prawns in mixed spore culture tanks after rearing for 90 days (13.5 cm and 59.8 g, respectively) were significantly higher (p = 0.05) than other treatments. The treated prawns were further challenged with Aeromonas hydrophila for 7 days. The percentages of survival after the challenge in the prawns fed with the mixed spores (46.8%) were also found significantly higher (p = 0.05) than others groups, except the mixed live cell treatment (60%). These results indicated that the spores of Bacillus KKU02 and Bacillus KKU03 had a high potential for using as commercial probiotics.

  15. Sensitivity of thermally treated Bacillus subtilis spores to subsequent irradiation

    International Nuclear Information System (INIS)

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

    1986-01-01

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

  16. 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.

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

    International Nuclear Information System (INIS)

    Farkas, J.; Kiss, I.; Andrássy, E.

    1967-01-01

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

  18. Fungal spores overwhelm biogenic organic aerosols in a midlatitudinal forest

    Directory of Open Access Journals (Sweden)

    C. Zhu

    2016-06-01

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

  19. Mucosal vaccine delivery by non-recombinant spores of Bacillus subtilis.

    Science.gov (United States)

    Ricca, Ezio; Baccigalupi, Loredana; Cangiano, Giuseppina; De Felice, Maurilio; Isticato, Rachele

    2014-08-12

    Development of mucosal vaccines strongly relies on an efficient delivery system and, over the years, a variety of approaches based on phages, bacteria or synthetic nanoparticles have been proposed to display and deliver antigens. The spore of Bacillus subtilis displaying heterologous antigens has also been considered as a mucosal vaccine vehicle, and shown able to conjugate some advantages of live microrganisms with some of synthetic nanoparticles. Here we review the use of non-recombinant spores of B. subtilis as a delivery system for mucosal immunizations. The non-recombinant display is based on the adsorption of heterologous molecules on the spore surface without the need of genetic manipulations, thus avoiding all concerns about the use and environmental release of genetically modified microorganisms. In addition, adsorbed molecules are stabilized and protected by the interaction with the spore, suggesting that this system could reduce the rapid degradation of the antigen, often observed with other delivery systems and identified as a major drawback of mucosal vaccines.

  20. Population and function analysis of cultivable bacteria associated with spores of arbuscular mycorrhizal fungus Gigaspora margarita.

    Science.gov (United States)

    Long, Liangkun; Lin, Qunying; Yao, Qing; Zhu, Honghui

    2017-05-01

    This study was aimed to investigate the diversity and function of bacterial population associated with Gigaspora margarita spores. The fungus was propagated in sterilized sand/soil pots using alfalfa (Medicago sativa), grain sorghum (Sorghum bicolor), or maize (Zea mays) as host plants, or in sterilized vermiculite pots using alfalfa as host plants, respectively. Bacteria were isolated from the new-formed spores using diluted plate method, and typical bacterial isolates were identified according to 16S rRNA gene phylogenetic analysis. Total 43 bacterial isolates affiliated to three phyla and 23 genera were obtained. The spore-associated bacterial communities were obviously different among the four source spores, suggesting that plant species or substrates could influence the bacterial population. Bacillus and Streptomyces were most frequently associated with the fungal spores. Function analysis of these bacteria by plate tests, it was found that about 30.2% isolates stimulated the spore germination, five out of seven tested isolates improved the hyphal growth, total 57.5% of the tested isolates solubilized phosphorus at different levels, 15% isolates degraded chitin, and a few isolates suppressed the growth of Escherichia coli or Staphylococcus aureus. In pot experiment, three bacterial isolates (belonging to Curtobacterium, Ensifer, or Bacillus, respectively) displayed improvement effect on alfalfa growth and/or the colonization of roots by G. margarita.

  1. The nature of water within bacterial spores: protecting life in extreme environments

    Science.gov (United States)

    Rice, Charles V.; Friedline, Anthony; Johnson, Karen; Zachariah, Malcolm M.; Thomas, Kieth J., III

    2011-10-01

    The bacterial spore is a formidable container of life, protecting the vital contents from chemical attack, antimicrobial agents, heat damage, UV light degradation, and water dehydration. The exact role of the spore components remains in dispute. Nevertheless, water molecules are important in each of these processes. The physical state of water within the bacterial spore has been investigated since the early 1930's. The water is found two states, free or bound, in two different areas, core and non-core. It is established that free water is accessible to diffuse and exchange with deuterated water and that the diffusible water can access all areas of the spore. The presence of bound water has come under recent scrutiny and has been suggested the water within the core is mobile, rather than bound, based on the analysis of deuterium relaxation rates. Using an alternate method, deuterium quadrupole-echo spectroscopy, we are able to distinguish between mobile and immobile water molecules. In the absence of rapid motion, the deuterium spectrum of D2O is dominated by a broad line, whose line shape is used as a characteristic descriptor of molecular motion. The deuterium spectrum of bacterial spores reveals three distinct features: the broad peak of immobilized water, a narrow line of water in rapid motion, and a signal of intermediate width. This third signal is assigned this peak from partially deuterated proteins with the spore in which N-H groups have undergone exchange with water deuterons to form N-D species. As a result of these observations, the nature of water within the spore requires additional explanation to understand how the spore and its water preserve life.

  2. Informative communication of microorganisms

    Directory of Open Access Journals (Sweden)

    G. N. Kremenchutskу

    2010-06-01

    Full Text Available Macroorganism in combination with microbiota is considered as a “superorganism”. Microorganisms, belonging to the microbiota, are in dynamic equilibrium with a macroorganism. This balance is achieved through a molecular “language” of communication between prokaryotic and eukaryotic cells. Molecular communication between cells leads to positive and negative results. A large number of metabolites of microorganisms that carry the information load: autoinducers is revealed. Autoinducer affect on the immune systems, and variety of metabolic processes. This affects on practically all organs and systems of maсroorganism. Studied metabolites of aerococci affect on the immune system, regenerative cycles and other processes of macroorganism. The problem of informative communication between prokaryotes and eukaryotes provides new insights about vital functions of “superorganisms”.

  3. Growth and Survivability of Microorganisms at Martian Temperatures and Pressures

    Science.gov (United States)

    Mickol, Rebecca Lynne

    The discovery of methane in the martian atmosphere via numerous ground- and space-based sources has prompted the study of methanogens as models for life on Mars. Methanogens are microorganisms within the domain Archaea, many of which utilize carbon dioxide (CO2) and hydrogen to produce methane. The non-photosynthetic nature of methanogens indicates that they could exist in sub-surface environments, protected from harmful UV and ionizing radiation on the surface of Mars. These organisms also do not require organics, which are sparse on the planet. Additionally, the wide variety of environments we find life in on Earth, as well as evidence for liquid brines on the surface of Mars, suggest that habitable environments may still exist on the planet. However, there are a variety of conditions that any extant life on Mars would need to endure, including wide variations in temperature over one sol, a low-pressure atmosphere, and a limited availability of liquid water, among others. This dissertation encompasses various experiments that examined the ability of four species of methanogens (Methanosarcina barkeri, Methanobacterium formicicum, Methanococcus maripaludis, and Methanothermobacter wolfeii) to survive and/or grow under 1) low-pressure conditions and 2) freeze/thaw cycles. Low pressure studies include both survival and active growth experiments conducted between 7 mbar (the average surface pressure on Mars) and 143 mbar. Freeze/thaw experiments utilized short- and long-term cycles varying in temperature between the organisms' growth temperatures (22 °C, M. maripaludis; 37 °C, M. barkeri and M. formicicum; 55 °C, M. wolfeii ) and -80 °C, encompassing Mars-relevant temperature changes. As a comparison to methanogen growth and survivability, additional experiments were conducted using a non-spore-forming bacterium, Serratia liquefaciens , previously shown capable of growth at 7 mbar, 0 °C and within an anoxic CO2 atmosphere. The experiments described here assessed

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

    International Nuclear Information System (INIS)

    Hauser, P.M.; Karamata, D.

    1992-01-01

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

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

    NARCIS (Netherlands)

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

    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

  6. Sporulation dynamics and spore heat resistance in wet and dry biofilms of Bacillus cereus

    NARCIS (Netherlands)

    Hayrapetyan, Hasmik; Abee, Tjakko; Nierop Groot, Masja

    2016-01-01

    Environmental conditions and growth history can affect the sporulation process as well as subsequent properties of formed spores. The sporulation dynamics was studied in wet and air-dried biofilms formed on stainless steel (SS) and polystyrene (PS) for Bacillus cereus ATCC 10987 and the

  7. 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

  8. 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.

    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

  9. Bacillus cereus spore damage recovery and diversity in spore germination and carbohydrate utilisation

    NARCIS (Netherlands)

    Warda, Alicja K.

    2016-01-01

    Bacterial spores are extremely robust survival vehicles that are highly resistant towards environmental stress conditions including heat, UV radiation and other stresses commonly applied during food production and preservation. Spores, including those of the toxin-producing food-borne human pathogen

  10. Bacillus cereus spore damage recovery and diversity in spore germination and carbohydrate utilisation

    NARCIS (Netherlands)

    Warda, Alicja K.

    2016-01-01

    Bacterial spores are extremely robust survival vehicles that are highly resistant towards environmental stress conditions including heat, UV radiation and other stresses commonly applied during food production and preservation. Spores, including those of the toxin-producing food-borne human

  11. Final report for Assembling Microorganisms into Energy Converting Materials

    Energy Technology Data Exchange (ETDEWEB)

    Sahin, Ozgur

    2018-03-26

    The goal of this project was to integrate microorganisms capable of reversible energy transduction in response to changing relative humidity with non-biological materials to create hybrid energy conversion systems. While plants and many other biological organisms have developed structures that are extraordinarily effective in converting changes in relative humidity into mechanical energy, engineered energy transduction systems rarely take advantage of this powerful phenomenon. Rather than developing synthetic materials that can convert changes in relative humidity in to mechanical energy, we developed approaches to assemble bacterial spores into larger materials. These materials can convert energy from evaporation of water in dry atmospheric conditions, which we demonstrated by building energy harvesters from these materials. We have also developed experiments to investigate the interaction of water with the spore material, and to determine how this interaction imposes limits on energy conversion. In addition, we carried out theoretical calculations to investigate the limits imposed by the environmental conditions to the power available in the energy harvesting process. These calculations took into account heat and water vapor transfer in the atmosphere surrounding the spore based materials. Overall, our results suggest that biomolecular materials are promising candidates to convert energy from evaporation.

  12. Ecology of Hypersaline Microorganisms

    Digital Repository Service at National Institute of Oceanography (India)

    Kerkar, S.

    by microorganisms. A prime example is the Dead Sea, a lake in which the 37 concentration of divalent cations (about 1.9 M Mg+2 and 0.4 M Ca+2) exceeds that of monovalent cations (1.6 M Na+ and 0.14 M K+) and of which the pH is relatively low (around 6.0). Even... such a hostile environment periodically supports dense microbial blooms (Oren, 1988). The two largest and best-studied hypersaline lakes are the Great Salt Lake in the Western United States and the Dead Sea, in the Middle East. The Great Salt Lake...

  13. 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.

  14. Role of DNA repair in Bacillus subtilis spore resistance.

    OpenAIRE

    Setlow, B; Setlow, P

    1996-01-01

    Wet-heat or hydrogen peroxide treatment of wild-type Bacillus subtilis spores did not result in induction of lacZ fusions to three DNA repair-related genes (dinR, recA, and uvrC) during spore outgrowth. However, these genes were induced during outgrowth of wild-type spores treated with dry heat or UV. Wet-heat, desiccation, dry-heat, or UV treatment of spores lacking major DNA-binding proteins (termed alpha-beta- spores) also resulted in induction of the three DNA repair genes during spore ou...

  15. Isolation and Characterization of Cryptococcus neoformans Spores Reveal a Critical Role for Capsule Biosynthesis Genes in Spore Biogenesis▿

    Science.gov (United States)

    Botts, Michael R.; Giles, Steven S.; Gates, Marcellene A.; Kozel, Thomas R.; Hull, Christina M.

    2009-01-01

    Spores are essential particles for the survival of many organisms, both prokaryotic and eukaryotic. Among the eukaryotes, fungi have developed spores with superior resistance and dispersal properties. For the human fungal pathogens, however, relatively little is known about the role that spores play in dispersal and infection. Here we present the purification and characterization of spores from the environmental fungus Cryptococcus neoformans. For the first time, we purified spores to homogeneity and assessed their morphological, stress resistance, and surface properties. We found that spores are morphologically distinct from yeast cells and are covered with a thick spore coat. Spores are also more resistant to environmental stresses than yeast cells and display a spore-specific configuration of polysaccharides on their surfaces. Surprisingly, we found that the surface of the spore reacts with antibodies to the polysaccharide glucuronoxylomannan, the most abundant component of the polysaccharide capsule required for C. neoformans virulence. We explored the role of capsule polysaccharide in spore development by assessing spore formation in a series of acapsular strains and determined that capsule biosynthesis genes are required for proper sexual development and normal spore formation. Our findings suggest that C. neoformans spores may have an adapted cell surface that facilitates persistence in harsh environments and ultimately allows them to infect mammalian hosts. PMID:19181873

  16. Development of bioprocess for high density cultivation yield of the probiotic Bacillus coagulans and its spores

    Directory of Open Access Journals (Sweden)

    Kavita R. Pandey

    2016-09-01

    Full Text Available Bacillus coagulans is a spore forming lactic acid bacterium. Spore forming bacteria, have been extensively studied and commercialized as probiotics. Probiotics are produced by fermentation technology. There is a limitation to biomass produced by conventional modes of fermentation. With the great demand generated by range of probiotic products, biomass is becoming very valuable for several pharmaceutical, dairy and probiotic companies. Thus, there is a need to develop high cell density cultivation processes for enhanced biomass accumulation. The bioprocess development was carried out in 6.6 L bench top lab scale fermentor. Four different cultivation strategies were employed to develop a bioprocess for higher growth and sporulation efficiencies of probiotic B. coagulans. Batch fermentation of B. coagulans yielded 18 g L-1 biomass (as against 8.0 g L-1 productivity in shake flask with 60% spore efficiency. Fed-batch cultivation was carried out for glucose, which yielded 25 g L-1 of biomass. C/N ratio was very crucial in achieving higher spore titres. Maximum biomass yield recorded was 30 g L-1, corresponding to 3.8 × 1011 cells mL-1 with 81% of cells in sporulated stage. The yield represents increment of 85 times the productivity and 158 times the spore titres relative to the highest reported values for high density cultivation of B. coagulans.

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

    Science.gov (United States)

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

    2017-08-01

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

  18. Disinfection and regrowth potential of bacillus subtilis spores by ozone, ultraviolet rays and gamma irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hae Yeon; Lee, O Mi; Kim, Tae Hun; Lee, Myun Joo; Yu, Seung Ho [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

    2009-06-15

    Chlorination has been the most commonly adopted disinfection process for the treatment of drinking water. However, Cryptosporidium parvum oocysts and Giardia lamblia cysts were not treated effectively by the common chlorine-based disinfectants. Additionally the regrowth of pathogenic microorganisms is associated with hygienic and aesthetic problems for the consumers of drinking water. Study on alternative disinfection processes such as ozone, UV-C, VUV and gamma irradiation were conducted. Bacillus subtilis spores have been used as a surrogate microorganism for Cryptosporidium parvum oocysts and Giardia lamblia cyst. Inactivation efficiency by ozone was from 30% to 96% within the range of 5 min to 120 min exposures. Inactivation efficiencies by UV-C and VUV were 95.18%, 95.07% at 30 sec, respectively. Inactivation efficiency at gamma irradiation dose of 2 kGy was 99.4%. Microbial regrowths after ozone, UV-C, VUV and gamma irradiation disinfections were also evaluated for 4 days. Bacillus subtilis spores after ozone treatment for 120 min exposure at the rate of 1.68 mg {center_dot} min{sup -1} showed 96.02% disinfection efficiency and significant microbial regrowth. Bacillus subtilis spores after UV-C (99.25% disinfection efficiency) and VUV (99.67% disinfection efficiency) treatments for 5 min showed gradual regrowth. However, inactivation efficiency of gamma irradiation at dose of 1 kGy was 98.8% and the disinfected sample showed no microbial regrowth for 4 days. Therefore, gamma irradiation is the most effective process for the disinfection of pathogenic microorganisms such as oocysts of protozoan parasites among four disinfection process.

  19. Incidence of spore-forming bacteria in unsweetened evaporated ...

    African Journals Online (AJOL)

    brand C, to 0.54x105 for brand B and 1.67x105 for brand A. The organisms isolated from the three brands were identified as Bacillus cereus, Bacillus licheniformis, Bacillus coagulans, Bacillus stearothermophilus, Bacillus subtilis and Clostridium perfringens, with B. subtilis having the highest frequency of occurrence ...

  20. Determination of the Persistence of Non-Spore-Forming ...

    Science.gov (United States)

    Report This report presents the results of an investigation to evaluate the persistence (or natural attenuation) of Yersinia pestis (Y. pestis), Francisella tularensis (F. tularensis), and Burkholderia mallei (B. mallei) on glass and soil under multiple environmental conditions and time points.

  1. 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

  2. Assessing the Impact of Germination and Sporulation Conditions on the Adhesion of Bacillus Spores to Glass and Stainless Steel by Fluid Dynamic Gauging.

    Science.gov (United States)

    Xu Zhou, Ke; Li, Nan; Christie, Graham; Wilson, D Ian

    2017-11-01

    The adhesion of spores of 3 Bacillus species with distinctive morphologies to stainless steel and borosilicate glass was studied using the fluid dynamic gauging technique. Marked differences were observed between different species of spores, and also between spores of the same species prepared under different sporulation conditions. Spores of the food-borne pathogen B. cereus were demonstrated to be capable of withstanding shear stresses greater than 1500 Pa when adhered to stainless steel, in contrast to spores of Bacillus subtilis and Bacillus megaterium, which detached in response to lower shear stress. An extended DLVO model was shown to be capable of predicting the relative differences in spore adhesion between spores of different species and different culture conditions, but did not predict absolute values of force of adhesion well. Applying the model to germinating spores showed a significant reduction in adhesion force shortly after triggering germination, indicating a potential strategy to achieve enhanced removal of spores from surfaces in response to shear stress, such as during cleaning-in-place procedures. Spore-forming bacteria are a concern to the food industry because they have the potential to cause food-borne illness and product spoilage, while being strongly adhesive to processing surfaces and resistant to cleaning-in-place procedures. This work is of significance to the food processors and manufacturers because it offers insight to the properties of spore adhesion and identifies a potential strategy to facilitate the removal of spores during cleaning procedures. © 2017 The Authors. Journal of Food Science published by Wiley Periodicals, Inc. on behalf of Institute of Food Technologists.

  3. Bicarbonate and amino acids are co-germinants for spores of Clostridium perfringens type A isolates carrying plasmid-borne enterotoxin gene.

    Science.gov (United States)

    Alnoman, Maryam; Udompijitkul, Pathima; Banawas, Saeed; Sarker, Mahfuzur R

    2018-02-01

    Clostridium perfringens type A isolates carrying a chromosomal enterotoxin (cpe) gene (C-cpe) are generally linked to food poisoning, while isolates carrying cpe on a plasmid (P-cpe) are associated with non-food-borne gastrointestinal diseases. Both C-cpe and P-cpe isolates can form metabolically dormant spores, which through germination process return to actively growing cells to cause diseases. In our previous study, we showed that only 3 out of 20 amino acids (aa) in phosphate buffer (pH 7.0) triggered germination of spores of P-cpe isolates (P-cpe spores). We now found that 14 out of 20 individual aa tested induced germination of P-cpe spores in the presence of bicarbonate buffer (pH 7.0). However, no significant spore germination was observed with bicarbonate (pH 7.0) alone, indicating that aa and bicarbonate are co-germinants for P-cpe spores. P-cpe strain F4969 gerKC spores did not germinate, and gerAA spores germinated extremely poorly as compared to wild-type and gerKA spores with aa-bicarbonate (pH 7.0) co-germinants. The germination defects in gerKC and gerAA spores were partially restored by complementing gerKC or gerAA spores with wild-type gerKC or gerAA, respectively. Collectively, this study identified aa-bicarbonate as a novel nutrient germinant for P-cpe spores and provided evidence that GerKC and GerAA play major roles in aa-bicarbonate induced germination. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    National Research Council Canada - National Science Library

    Brittingham, Katherine C; Ruthel, Gordon; Panchal, Rekha G; Fuller, Claudette L; Ribot, Wilson J

    2005-01-01

    Phagocytosis of inhaled Bacillus anthracis spores and subsequent trafficking to lymph nodes are decisive events in the progression of inhaled anthrax because they initiate germination and dissemination of spores...

  5. Proteolysis in hyperthermophilic microorganisms

    Directory of Open Access Journals (Sweden)

    Donald E. Ward

    2002-01-01

    Full Text Available Proteases are found in every cell, where they recognize and break down unneeded or abnormal polypeptides or peptide-based nutrients within or outside the cell. Genome sequence data can be used to compare proteolytic enzyme inventories of different organisms as they relate to physiological needs for protein modification and hydrolysis. In this review, we exploit genome sequence data to compare hyperthermophilic microorganisms from the euryarchaeotal genus Pyrococcus, the crenarchaeote Sulfolobus solfataricus, and the bacterium Thermotoga maritima. An overview of the proteases in these organisms is given based on those proteases that have been characterized and on putative proteases that have been identified from genomic sequences, but have yet to be characterized. The analysis revealed both similarities and differences in the mechanisms utilized for proteolysis by each of these hyperthermophiles and indicated how these mechanisms relate to proteolysis in less thermophilic cells and organisms.

  6. Artifical Microorganism Infection in Aviation Kerosene

    Directory of Open Access Journals (Sweden)

    Dušan Vallo

    2004-12-01

    Full Text Available The fuel used in the aviation engineering has to be clean and dry, it may not contain mechanical impurities and water. Water inaviation kerosene may occur in soluble and insoluble form. The danger inheres in the insoluble form, which may drop out in the crystallineform and cause various failures, such as those caused by mechanical impurities. The water assists in the biological matter formation createdby various species of microorganisms (bacteria, mould fungi and yeast. The microorganisms, present in water phase occurring on thebottom of tanks or on the interface water phase – kerosene, grow and reproduce and subsequently may pollute (impair the fuel by thebiomass or by the products of their metabolism. There is a possibility to infect the fuel artificially by a selected reference microorganismstrain, which usually occur in contaminated fuel, or by microorganisms which cause a biological contamination of aviation kerosene.Out of the selected reference strains used in the experiments, the reference strains of Proteus vulgaris, Sacharamyces cerevisiae andClostridium perfringens were not cultivated in the sterile aviation kerosene and the propagating nutrient medium. The aviation kerosene actsas a biocide medium for the presented reference microorganism strains.

  7. Thermophilic microorganisms in biomining.

    Science.gov (United States)

    Donati, Edgardo Rubén; Castro, Camila; Urbieta, María Sofía

    2016-11-01

    Biomining is an applied biotechnology for mineral processing and metal extraction from ores and concentrates. This alternative technology for recovering metals involves the hydrometallurgical processes known as bioleaching and biooxidation where the metal is directly solubilized or released from the matrix for further solubilization, respectively. Several commercial applications of biomining can be found around the world to recover mainly copper and gold but also other metals; most of them are operating at temperatures below 40-50 °C using mesophilic and moderate thermophilic microorganisms. Although biomining offers an economically viable and cleaner option, its share of the world´s production of metals has not grown as much as it was expected, mainly considering that due to environmental restrictions in many countries smelting and roasting technologies are being eliminated. The slow rate of biomining processes is for sure the main reason of their poor implementation. In this scenario the use of thermophiles could be advantageous because higher operational temperature would increase the rate of the process and in addition it would eliminate the energy input for cooling the system (bioleaching reactions are exothermic causing a serious temperature increase in bioreactors and inside heaps that adversely affects most of the mesophilic microorganisms) and it would decrease the passivation of mineral surfaces. In the last few years many thermophilic bacteria and archaea have been isolated, characterized, and even used for extracting metals. This paper reviews the current status of biomining using thermophiles, describes the main characteristics of thermophilic biominers and discusses the future for this biotechnology.

  8. 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.

  9. 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

  10. The Cooperative and Interdependent Roles of GerA, GerK, and Ynd in Germination of Bacillus licheniformis Spores.

    Science.gov (United States)

    Borch-Pedersen, Kristina; Lindbäck, Toril; Madslien, Elisabeth H; Kidd, Shani W; O'Sullivan, Kristin; Granum, Per Einar; Aspholm, Marina

    2016-07-15

    When nutrients are scarce, Bacillus species form metabolically dormant and extremely resistant spores that enable survival over long periods of time under conditions not permitting growth. The presence of specific nutrients triggers spore germination through interaction with germinant receptors located in the spore's inner membrane. Bacillus licheniformis is a biotechnologically important species, but it is also associated with food spoilage and food-borne disease. The B. licheniformis ATCC 14580/DSM13 genome exhibits three gerA family operons (gerA, gerK, and ynd) encoding germinant receptors. We show that spores of B. licheniformis germinate efficiently in response to a range of different single l-amino acid germinants, in addition to a weak germination response seen with d-glucose. Mutational analyses revealed that the GerA and Ynd germination receptors function cooperatively in triggering an efficient germination response with single l-amino acid germinants, whereas the GerK germination receptor is essential for germination with d-glucose. Mutant spores expressing only GerA and GerK or only Ynd and GerK show reduced or severely impaired germination responses, respectively, with single l-amino acid germinants. Neither GerA nor Ynd could function alone in stimulating spore germination. Together, these results functionally characterize the germination receptor operons present in B. licheniformis We demonstrate the overlapping germinant recognition patterns of the GerA and Ynd germination receptors and the cooperative functionalities between GerA, Ynd, and GerK in inducing germination. To ensure safe food production and durable foods, there is an obvious need for more knowledge on spore-forming bacteria. It is the process of spore germination that ultimately leads to food spoilage and food poisoning. Bacillus licheniformis is a biotechnologically important species that is also associated with food spoilage and food-borne disease. Despite its importance, the

  11. 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

  12. Imaging bacterial spores by soft-x-ray microscopy

    International Nuclear Information System (INIS)

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

    1997-01-01

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

  13. 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.

  14. A Waking Review: Old and Novel Insights into the Spore Germination in Streptomyces

    Directory of Open Access Journals (Sweden)

    Jan Bobek

    2017-11-01

    Full Text Available The complex development undergone by Streptomyces encompasses transitions from vegetative mycelial forms to reproductive aerial hyphae that differentiate into chains of single-celled spores. Whereas their mycelial life – connected with spore formation and antibiotic production – is deeply investigated, spore germination as the counterpoint in their life cycle has received much less attention. Still, germination represents a system of transformation from metabolic zero point to a new living lap. There are several aspects of germination that may attract our attention: (1 Dormant spores are strikingly well-prepared for the future metabolic restart; they possess stable transcriptome, hydrolytic enzymes, chaperones, and other required macromolecules stabilized in a trehalose milieu; (2 Germination itself is a specific sequence of events leading to a complete morphological remodeling that include spore swelling, cell wall reconstruction, and eventually germ tube emergences; (3 Still not fully unveiled are the strategies that enable the process, including a single cell’s signal transduction and gene expression control, as well as intercellular communication and the probability of germination across the whole population. This review summarizes our current knowledge about the germination process in Streptomyces, while focusing on the aforementioned points.

  15. Pollen and spore monitoring in the world.

    Science.gov (United States)

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

    2018-01-01

    Ambient air quality monitoring is a governmental duty that is widely carried out in order to detect non-biological ("chemical") components in ambient air, such as particles of world and to create an interactive visualization of their distribution. The method employed to collect information was based on: (a) a review of the recent and historical bibliography related to pollen and fungal spore monitoring, and (b) personal surveys of the managers of national and regional monitoring networks. The interactive application was developed using the R programming language. We have created an inventory of the active pollen and spore monitoring stations in the world. There are at least 879 active pollen monitoring stations in the world, most of which are in Europe (> 500). The prevalent monitoring method is based on the Hirst principle (> 600 stations). The inventory is visualised as an interactive and on-line map. It can be searched, its appearance can be adjusted to the users' needs and it is updated regularly, as new stations or changes to those that already exist can be submitted online. The map shows the current situation of pollen and spore monitoring and facilitates collaboration among those individuals who are interested in pollen and spore counts. It might also help to improve the monitoring of biological particles up to the current level employed for non-biological components.

  16. Detecting bacterial spores in soup manufacturing

    NARCIS (Netherlands)

    van Zuijlen, A.C.M.; Oomes, S.J.C.M.; Vos, P.; Brul, S.

    2009-01-01

    Spores from mesophilic aerobic sporeforming bacteria (Bacillus) are sometimes able to survive the thermal process of commercial sterile products and sporadically cause spoilage or food poisoning. Because of an increasing demand for more fresh products, ideally the processing temperatures should be

  17. On some white-spored Geoglossaceae

    NARCIS (Netherlands)

    Maas Geesteranus, R.A.

    1964-01-01

    Some genera of Geoglossaceae, characterized by colourless spores and positive iodine reaction of the ascus pore, are compared with respect to the structure of the stipe. Ochroglossum is reduced to the synonymy of Microglossum. Mitrula is regarded as a monotypic genus. The generic name Heyderia is

  18. Paleozoic in situ spores and pollen. Lycopsida

    Czech Academy of Sciences Publication Activity Database

    Bek, Jiří

    2017-01-01

    Roč. 296, 1/6 (2017), s. 1-111 ISSN 0375-0299 R&D Projects: GA ČR GAP210/12/2053 Institutional support: RVO:67985831 Keywords : in situ spores * reproductive organs * Lycopsida * Paleozoic Subject RIV: DB - Geology ; Mineralogy OBOR OECD: Paleontology Impact factor: 1.333, year: 2016

  19. Results of studies on long-term exposition of dormant forms of various organisms in outer space environment

    Science.gov (United States)

    Novikova, Nataliya; Gusev, Oleg; Sugimoto, Manabu; Deshevaya, Elena; Levinskikh, Margarita; Sychev, Vladimir; Okuda, Takashi; Orlov, Oleg; Alekseev, Victor; Poddubko, Svetlana; Polikarpov, Nikolay

    The planetary quarantine is one of the key problems of deep space exploration. Risks of the possible transfer of biological objects across interplanetary space should be necessarily assessed during space exploration. The risks associated with a possible transfer of biological objects and primarily microorganisms in interplanetary space is a priority for space studies We can assume, that on the exterior side of both unmanned and manned space stations there can be millions of microbial cells, many of which are in spore forms, the stability of which towards the unfavorable factors is extremely high. However, direct evidence to support this assumption, obtained only in recent years. “Biorisk” is an apparatus designed for conduction of space experiments focused on long-term exposition of latent stages of different forms of organism on the outer side of Russian Segment of International Space Station was developed and used in SSC RF - Institute for Biomedical Problems RAS. The purpose of this experiment is to determine the principle capability of preservation of life capacity in test-cultures of microorganisms during long-term exposure (comparable with the term of interplanetary flight) in space. The first experiment was performed using spores of bacteria (Bacillus) and fungi (Penicillium, Aspergillus and Cladosporium) housed in 3 boxes that were exposed to outer space for 7, 12 or 18 months. It was for the first time demonstrated that bacterial and fungal spores could survive an exposure to outer space during the time period comparable with the duration of a return mission to Mars. Moreover, the microbial strains proved viable and highly active. The second experiment was expanded by flying, in addition to the above spores, dormant forms of higher plants, insects, lower crustaceans and vertebrates. The 31-month experiment showed that, in spite of harsher than in the first study temperatures, some specimens remained viable and capable of further multiplication. In

  20. Amplicon sequencing for the quantification of spoilage microbiota in complex foods including bacterial spores

    NARCIS (Netherlands)

    Boer, de P.; Caspers, M.; Sanders, J.W.; Kemperman, R.; Wijman, J.; Lommerse, G.; Roeselers, G.; Montijn, R.; Abee, T.; Kort, R.

    2015-01-01

    Background
    Spoilage of food products is frequently caused by bacterial spores and lactic acid bacteria. Identification of these organisms by classic cultivation methods is limited by their ability to form colonies on nutrient agar plates. In this study, we adapted and optimized 16S rRNA amplicon

  1. Removal of Bacillus anthracis sterne spore from commercial unpasteurized liquid egg white

    Science.gov (United States)

    Thermal pasteurization used by the egg industry for controlling vegetative cells of pathogens is ineffective for destroying endospores. There is a strong need in the agri-industries to develop effective intervention strategies to eliminate the possible bioterrorism threat from spore forming bacteria...

  2. Behaviour of individual spores of non proteolytic Clostridium botulinum as an element in quantitative risk assessment

    NARCIS (Netherlands)

    Smelt, J.P.; Stringer, S.C.; Brul, S.

    2013-01-01

    Botulism is a true food poisoning hazard. It is caused by a deadly neurotoxin produced in food by the spore forming organism Clostridium botulinum, which is found in soil all over the world. The frequency of botulism cases is rare, but each year a number of them occur. Ideally a quantitative

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

    NARCIS (Netherlands)

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

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

  4. Teaching Intelligent Design or Sparking Interest in Science? What Players Do with Will Wright's Spore

    Science.gov (United States)

    Owens, Trevor

    2012-01-01

    The 2008 commercial video game "Spore" allowed more than a million players to design their own life forms. Starting from single-celled organisms players played through a caricature of natural history. Press coverage of the game's release offer two frames for thinking about the implications of the game. Some scientists and educators saw the game as…

  5. Antifouling effects of the periostracum on algal spore settlement in the mussel Mytilus edulis

    Directory of Open Access Journals (Sweden)

    Ji-Young Kang

    2016-03-01

    Full Text Available Abstract In nature, marine mussels (Mytilus edulis suffer less fouling colonization on the newly formed sides of their shells. Using settlement assays with algal spores of Porphyra suborbiculata, we determined that spore attachment and germination on the periostracum decreased to 36.8 and 3.3 %, respectively. Additionally, the spore settlement was considerably diminished by periostracum dichloromethane extracts containing 19 % oleamide, a major antifouling compound. A scanning electron micrograph of the surface revealed a regular ripple structure with approximately 1.4 μm between ripples. Based on these results, mussel periostraca or their associated biomimetic materials may become environmentally friendly, antifouling agents for preventing the settlement of soft foulants.

  6. 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...display a currently valid OMB control number. 1. REPORT DATE 02 OCT 2015 2. REPORT TYPE N/A 3. DATES COVERED 4. TITLE AND SUBTITLE Inhibiting...inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores potential spore decontamination strategies 5a

  7. Antimicrobial efficacy of copper surfaces against spores and vegetative cells of Clostridium difficile: the germination theory.

    Science.gov (United States)

    Wheeldon, L J; Worthington, T; Lambert, P A; Hilton, A C; Lowden, C J; Elliott, T S J

    2008-09-01

    Persistent contamination of surfaces by spores of Clostridium difficile is a major factor influencing the spread of C. difficile-associated diarrhoea (CDAD) in the clinical setting. In recent years, the antimicrobial efficacy of metal surfaces has been investigated against microorganisms including methicillin-resistant Staphylococcus aureus. This study compared the survival of C. difficile on stainless steel, a metal contact surface widely used in hospitals, and copper surfaces. Antimicrobial efficacy was assessed using a carrier test method against dormant spores, germinating spores and vegetative cells of C. difficile (NCTC 11204 and ribotype 027) over a 3 h period in the presence and absence of organic matter. Copper metal eliminated all vegetative cells of C. difficile within 30 min, compared with stainless steel which demonstrated no antimicrobial activity (P or=2.5 log reduction (99.8% reduction) at 3 h. Organic material did not reduce the antimicrobial efficacy of the copper surface (P > 0.05). The use of copper surfaces within the clinical environment and application of a germination solution in infection control procedures may offer a novel way forward in eliminating C. difficile from contaminated surfaces and reducing CDAD.

  8. Polysaccharides from Extremophilic Microorganisms

    Science.gov (United States)

    Nicolaus, B.; Moriello, V. Schiano; Lama, L.; Poli, A.; Gambacorta, A.

    2004-02-01

    Several marine thermophilic strains were analyzed for exopolysaccharide production. The screening process revealed that a significant number of thermophilic microorganisms were able to produce biopolymers, and some of them also revealed interesting chemical compositions. We have identified four new polysaccharides from thermophilic marine bacteria, with complex primary structures and with different repetitive units: a galacto-mannane type from strain number 4004 and mannane type for the other strains. The thermophilic Bacillus thermantarcticus produces two exocellular polysaccharides (EPS 1, EPS 2) that give the colonies a typical mucous character. The exopolysaccharide fraction was produced with all substrates assayed, although a higher yield 400 mg liter-1 was obtained with mannose as carbon and energy source. NMR spectra confirmed that EPS 1 was a heteropolysaccharide of which the repeating unit was constituted by four different α-D-mannoses and three different β-D-glucoses. It seems to be close to some xantan polymers. EPS 2 was a mannan. Four different α-D-mannoses were found as the repeating unit. Production and chemical studies of biopolymers produced by halophilic archaea, Haloarcula species were also reported.

  9. Polysaccharides from extremophilic microorganisms.

    Science.gov (United States)

    Nicolaus, B; Schiano Moriello, V; Lama, L; Poli, A; Gambacorta, A

    2004-02-01

    Several marine thermophilic strains were analyzed for exopolysaccharide production. The screening process revealed that a significant number of thermophilic microorganisms were able to produce biopolymers, and some of them also revealed interesting chemical compositions. We have identified four new polysaccharides from thermophilic marine bacteria, with complex primary structures and with different repetitive units: a galacto-mannane type from strain number 4004 and mannane type for the other strains. The thermophilic Bacillus thermantarcticus produces two exocellular polysaccharides (EPS 1, EPS 2) that give the colonies a typical mucous character. The exopolysaccharide fraction was produced with all substrates assayed, although a higher yield 400 mg liter(-1) was obtained with mannose as carbon and energy source. NMR spectra confirmed that EPS 1 was a heteropolysaccharide of which the repeating unit was constituted by four different alpha-D-mannoses and three different beta-D-glucoses. It seems to be close to some xantan polymers. EPS 2 was a mannan. Four different alpha-D-mannoses were found as the repeating unit. Production and chemical studies of biopolymers produced by halophilic archaea, Haloarcula species were also reported.

  10. Proteins YlaJ and YhcN contribute to the efficiency of spore germination in Bacillus subtilis.

    Science.gov (United States)

    Johnson, Christian L; Moir, Anne

    2017-04-01

    The YlaJ and YhcN spore lipoproteins of Bacillus subtilis contain a common domain, and are of unknown function. Homologues of YlaJ or YhcN are widespread in Bacilli and are also encoded in those Clostridia that use cortex lytic enzymes SleB and CwlJ for cortex hydrolysis during germination. In B. subtilis, we report that single and double mutants lacking YlaJ and/or YhcN show a reduced rate of spore germination in L-alanine, with a delay in loss of heat resistance, release of dipicolinic acid and OD fall. If B. subtilis spores lack the cortex lytic enzyme CwlJ, spore cortex degradation and subsequent outgrowth to form colonies is strictly dependent on the other cortex lytic enzyme SleB, allowing a test of SleB function; in a cwlJ mutant background, the combined loss of both ylaJ and yhcN genes resulted in a spore population in which only 20% of spores germinated and outgrew to form colonies, suggesting that SleB activity is compromised. YlaJ and YhcN have a role in germination that is not yet well defined, but these proteins are likely to contribute, directly or indirectly, to early events in germination, including effective SleB function. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Mechanisms of nickel toxicity in microorganisms

    Science.gov (United States)

    Macomber, Lee

    2014-01-01

    Summary Nickel has long been known to be an important human toxicant, including having the ability to form carcinomas, but until recently nickel was believed to be an issue only to microorganisms living in nickel-rich serpentine soils or areas contaminated by industrial pollution. This assumption was overturned by the discovery of a nickel defense system (RcnR/RcnA) found in microorganisms that live in a wide range of environmental niches, suggesting that nickel homeostasis is a general biological concern. To date, the mechanisms of nickel toxicity in microorganisms and higher eukaryotes are poorly understood. In this review, we summarize nickel homeostasis processes used by microorganisms and highlight in vivo and in vitro effects of exposure to elevated concentrations of nickel. On the basis of this evidence we propose four mechanisms of nickel toxicity: 1) nickel replaces the essential metal of metalloproteins, 2) nickel binds to catalytic residues of non-metalloenzymes; 3) nickel binds outside the catalytic site of an enzyme to inhibit allosterically, and 4) nickel indirectly causes oxidative stress. PMID:21799955

  12. THE FACTORS FORMING QUALITY OF GRANULATED SUGAR

    Directory of Open Access Journals (Sweden)

    N. G. Kulneva

    2015-01-01

    Full Text Available Sugar, with good taste and high caloric, is one of the most popular human food. Consumers sugar must be sure that the sugar under normal conditions of use is of high quality and is not harmful to the health of the product. One reason for the decline in the quality of sugar is bacterial contamination. This is because the sugar industry products are good targets for the development of different groups of microorganisms, e.g., Bacillus subtilis, Clostridium perfringes, Leuconostoc dextranicum, Torula alba, Pseudomonas fluorescens, Sarcina lutea and others. These organisms are affected with beets, and then with sugar beet chips and diffusion juice fall into the processing line of sugar production. Their number in the diffusion juice varies and depends on many facto rs such as the quality of raw materials, the quality of cleaning beet root colonization of transporter-washing and the supply of water to the diffusion process, the temperature of the diffusion and others. In the diffusion unit has the most favorable conditions for the development of micro-organisms. Some of them, especially resistant bacteria and thermophilic bacteria or their spores, forming a capsule which protects against external influences occur in the final product sugar. When injected into the fresh crop of product (juice, syrup, they begin to multiply rapidly, causing difficulties in the process. The higher seeding beet microorganisms, the more they decompose and emit sucrose metabolism byproducts. To reduce the negative impact of microbiological and reduce losses from decomposition of sucrose conducted research on the possibility of using chlorine-containing substances in the sugar industry. It was established experimentally that the investigated chlorinated drug has bacteriostatic action and can be recommended for use in sugar beet production.

  13. Decontamination of carpet exposed to Microsporum canis hairs and spores.

    Science.gov (United States)

    Moriello, Karen A

    2017-04-01

    Objectives The objective of this study was to evaluate the efficacy of vacuuming and three carpet cleaning methods for the removal of Microsporum canis spores and hairs from experimentally contaminated carpets. Methods Sterile Berber carpeting was artificially contaminated with naturally infective M canis hairs and spores. Carpet swatches were vacuumed for 10 s, 30 s and 60 s, and then cultured. Three carpet cleaning methods were evaluated on area rugs experimentally contaminated with infective material: a beater brush carpet shampooing, beater brush carpet shampooing post-disinfectant application and hot water extraction. Home cleaning products labeled as having efficacy against Trichophyton species were used in addition to 1% potassium peroxymonosulfate. Carpets were cultured at 24 h, 48 h and 7 days after cleaning. Good efficacy was no detectable spores at post-cleaning culture. Results All pretreatment carpet samples were culture positive for M canis (>300 colony-forming units [cfu]/site). Vacuuming did not decontaminate carpets but did remove intact hairs. Spores were not detected by wipe samples after two washings with an upright beater brush carpet shampooer or pretreatment with a disinfectant prior to carpet shampooing. Carpets cleaned with one hot water extraction technique had a decrease from 300 cfu/site to a mean of 5.5 cfu/site at 24 and 48 h post-cleaning and 2 cfu/site at day 7. The use of disinfectants was associated with odor, even when dry, and permanent discoloration. Hot water extraction cleaning was associated with the fastest drying time and no discoloration. Conclusions and relevance Carpets exposed to M canis can be disinfected via carpet shampooing or hot water extraction cleaning. Vacuuming of carpets is recommended to remove infective hairs. For homes, exposed carpeting can be decontaminated by routine washing with a carpet shampooer (twice) or hot water extraction. Use of pretreatment with a disinfectant is recommended when a high level

  14. Doppler speedometer for micro-organisms

    International Nuclear Information System (INIS)

    Penkov, F.; Tuleushev, A.; Lisitsyn, V.; Kim, S.; Tuleushev, Yu.

    1996-01-01

    Objective of Investigations: Development and creation of the Doppler speedometer for micro-organisms which allows to evaluate, in a real temporal scale, variations in the state of water suspension of micro-organisms under the effect of chemical, physical and other external actions. Statement of the Problem The main problem is absence of reliable, accessible for users and simple, in view of application, Doppler speedometers for micro-organisms. Nevertheless, correlation Doppler spectrometry in the regime of heterodyning the supporting and cell-scattered laser radiation is welt known. The main idea is that the correlation function of photo-current pulses bears an information on the averages over the assembly of cell velocities. For solving the biological problems, construction of auto-correlation function in the real-time regime with the delay time values comprising, function in the real-time regime with the delay time values comprising, nearly, 100 me (10 khz) or higher is needed. Computers of high class manage this problem using but the program software. Due to this, one can simplify applications of the proposed techniques provided he creates the Doppler speedometer for micro-organism on a base of the P entium . Expected Result Manufactured operable mock-up of the Doppler speedometer for micro-organisms in a form of the auxiliary computer block which allows to receive an information, in the real time scale, on the results of external effects of various nature on the cell assembly in transparent medium with a small volume of the studied cell suspension

  15. Applicability of UV resistant Bacillus pumilus spore as a human adenovirus surrogate for evaluating the effectiveness of virus inactivation in low-pressure UV treatment systems

    Data.gov (United States)

    U.S. Environmental Protection Agency — Data set includes UV dose, and Bacillus pumilus spore plate counts in colony forming units. This dataset is associated with the following publication: Boczek , L.,...

  16. Bacillus cereus spore formation, structure and germination

    NARCIS (Netherlands)

    Vries, de Y.P.

    2006-01-01

    Bacterial spores arespecializeddifferentiated celltypes for

  17. Comparison of Bacillus atrophaeus spore viability following exposure to detonation of C4 and to deflagration of halogen-containing thermites

    International Nuclear Information System (INIS)

    Tringe, J. W.; Létant, S. E.; Dugan, L. C.; Levie, H. W.; Kuhl, A. L.; Murphy, G. A.; Alves, S. W.; Vandersall, K. S.; Pantoya, M. L.

    2013-01-01

    Energetic materials are being considered for the neutralization of spore-forming bacteria. In this study, the neutralization effects of a monomolecular explosive were compared to the effects of halogen-containing thermites. Bacillus atrophaeus spores were exposed to the post-detonation environment of a 100 g charge of the military explosive C-4 at a range of 50 cm. These tests were performed in the thermodynamically closed environment of a 506-l barometric calorimeter. Associated temperatures were calculated using a thermodynamic model informed by calculations with the Cheetah thermochemical code. Temperatures in the range of 2300–2800 K were calculated to persist for nearly the full 4 ms pressure observation time. After the detonation event, spores were characterized using optical microscopy and the number of viable spores was assessed. Results showed live spore survival rates in the range of 0.01%–1%. For the thermite tests, a similar, smaller-scale configuration was employed that examined the spore neutralization effects of two thermites: aluminum with iodine pentoxide and aluminum with potassium chlorate. Only the former mixture resulted in spore neutralization. These results indicate that the detonation environment produced by an explosive with no chemical biocides may provide effective spore neutralization similar to a deflagrating thermite containing iodine

  18. Comparison of Bacillus atrophaeus spore viability following exposure to detonation of C4 and to deflagration of halogen-containing thermites

    Energy Technology Data Exchange (ETDEWEB)

    Tringe, J. W.; Létant, S. E.; Dugan, L. C.; Levie, H. W.; Kuhl, A. L.; Murphy, G. A.; Alves, S. W.; Vandersall, K. S. [Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, California 94550 (United States); Pantoya, M. L. [Mechanical Engineering Department, Texas Tech University, Lubbock, Texas 79409 (United States)

    2013-12-21

    Energetic materials are being considered for the neutralization of spore-forming bacteria. In this study, the neutralization effects of a monomolecular explosive were compared to the effects of halogen-containing thermites. Bacillus atrophaeus spores were exposed to the post-detonation environment of a 100 g charge of the military explosive C-4 at a range of 50 cm. These tests were performed in the thermodynamically closed environment of a 506-l barometric calorimeter. Associated temperatures were calculated using a thermodynamic model informed by calculations with the Cheetah thermochemical code. Temperatures in the range of 2300–2800 K were calculated to persist for nearly the full 4 ms pressure observation time. After the detonation event, spores were characterized using optical microscopy and the number of viable spores was assessed. Results showed live spore survival rates in the range of 0.01%–1%. For the thermite tests, a similar, smaller-scale configuration was employed that examined the spore neutralization effects of two thermites: aluminum with iodine pentoxide and aluminum with potassium chlorate. Only the former mixture resulted in spore neutralization. These results indicate that the detonation environment produced by an explosive with no chemical biocides may provide effective spore neutralization similar to a deflagrating thermite containing iodine.

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

    Science.gov (United States)

    Flowers, R S; Adams, D M

    1976-02-01

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

  20. Method of obtaining enhanced oil recovery by the use of microorganisms; Fremgangsmaate for aa oppnaa forbedret oljeutvinning under anvendelse av mikroorganismer

    Energy Technology Data Exchange (ETDEWEB)

    Silver, R.S.; Moon, W.G.; Bunting, P.M.; Acheson, W.P.

    1995-03-27

    The invention deals with an method for improving the recovery from petroleum reservoirs by the use of microorganisms. According to the invention, there is to be injected a suspension of a biological clean breed of the Bacillus licheniformis or spores thereof in order to tighten selectively the relative strong penetrating zones of a reservoir. 2 figs.

  1. Microorganism mediated liquid fuels

    Energy Technology Data Exchange (ETDEWEB)

    Troiano, Richard

    2018-03-20

    Herein disclosed is a method for producing liquid hydrocarbon product, the method comprising disintegrating a hydrocarbon source; pretreating the disintegrated hydrocarbon source; solubilizing the disintegrated hydrocarbon source to form a slurry comprising a reactant molecule of the hydrocarbon source; admixing a biochemical liquor into the slurry, wherein the biochemical liquor comprises at least one conversion enzyme configured to facilitate bond selective photo-fragmentation of said reactant molecule of the hydrocarbon source, to form liquid hydrocarbons via enzyme assisted bond selective photo-fragmentation, wherein said conversion enzyme comprises reactive sites configured to restrict said reactant molecule such that photo-fragmentation favorably targets a preselected internal bond of said reactant molecule; separating the liquid hydrocarbons from the slurry, wherein contaminants remain in the slurry; and enriching the liquid hydrocarbons to form a liquid hydrocarbon product. Various aspects of such method/process are also discussed.

  2. Identification and characterization of a spore-like morphotype in chronically starved Mycobacterium avium subsp. paratuberculosis cultures.

    Directory of Open Access Journals (Sweden)

    Elise A Lamont

    Full Text Available Mycobacteria are able to enter into a state of non-replication or dormancy, which may result in their chronic persistence in soil, aquatic environments, and permissive hosts. Stresses such as nutrient deprivation and hypoxia provide environmental cues to enter a persistent state; however, a clear definition of the mechanism that mycobacteria employ to achieve this remains elusive. While the concept of sporulation in mycobacteria is not novel, it continues to spark controversy and challenges our perceptions of a non-replication. We investigated the potential role of sporulation in one-year old broth cultures of Mycobacterium subsp. paratuberculosis (MAP. We show that dormant cultures of MAP contain a mix of vegetative cells and a previously unknown morphotype resembling a spore. These spore-like structures can be enriched for using sporulating media. Furthermore, purified MAP spore forms survive exposure to heat, lysozyme and proteinase K. Heat-treated spores are positive for MAP 16SrRNA and IS900. MAP spores display enhanced infectivity as well as maintain acid-fast characteristics upon germination in a well-established bovine macrophage model. This is the first study to demonstrate a new MAP morphotype possessing spore-like qualities. Data suggest that sporulation may be a viable mechanism by which MAP accomplishes persistence in the host and/or environment. Thus, our current understanding of mycobacterial persistence, pathogenesis, epidemiology and rational drug and vaccine design may need to be reevaluated.

  3. Mucosal delivery of antigens using adsorption to bacterial spores.

    Science.gov (United States)

    Huang, Jen-Min; Hong, Huynh A; Van Tong, Hoang; Hoang, Tran H; Brisson, Alain; Cutting, Simon M

    2010-01-22

    The development of new-generation vaccines has followed a number of strategic avenues including the use of live recombinant bacteria. Of these, the use of genetically engineered bacterial spores has been shown to offer promise as both a mucosal as well as a heat-stable vaccine delivery system. Spores of the genus Bacillus are currently in widespread use as probiotics enabling a case to be made for their safety. In this work we have discovered that the negatively charged and hydrophobic surface layer of spores provides a suitable platform for adsorption of protein antigens. Binding can be promoted under conditions of low pH and requires a potent combination of electrostatic and hydrophobic interactions between spore and immunogen. Using appropriately adsorbed spores we have shown that mice immunised mucosally can be protected against challenge with tetanus toxin, Clostridium perfringens alpha toxin and could survive challenge with anthrax toxin. In some cases protection is actually greater than using a recombinant vaccine. Remarkably, killed or inactivated spores appear equally effective as live spores. The spore appears to present a bound antigen in its native conformation promoting a cellular (T(h)1-biased) response coupled with a strong antibody response. Spores then, should be considered as mucosal adjuvants, most similar to particulate adjuvants, by enhancing responses against soluble antigens. The broad spectrum of immune responses elicited coupled with the attendant benefits of safety suggest that spore adsorption could be appropriate for improving the immunogenicity of some vaccines as well as the delivery of biotherapeutic molecules.

  4. 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.

  5. Isolation of microorganisms from red pepper powder and their radiosensitivity

    International Nuclear Information System (INIS)

    Choi, E.H.; Kim, Y.B.; Lee, S.R.

    1977-01-01

    From samples of red pepper powder sold in Korea were isolated and identified 13 species of molds (Aspergillus amsteodami, Asp. chevalieri, Asp. clavatus, Asp. Flavus, Asp. janus var. effusus, Asp. oryzae, Asp. oryzae var. brevis, Asp. repens, Asp. sydowi, Asp. thomii, Asp. tubingensis, Penicillium thomii, Scopulariopsis brevicaulis) and 5 species of bacteria (Bacillus pumilus, Bac. subtilis, Micrococus luteus, M. varians, Staphylococcus aureus). Radiosensitivity of these microorganisms was examined to give D 10 values of 14-41 krad for molds, 11-24 krad for bacterial vegetative cells and 190-250 krad for bacterial spores. The red pepper powder was contaminated with 2-3x10 2 mold counts/g and 3-6x10 7 bacterial counts/g, which would be sufficiently destroyed by irradiating 200 krad r-rays. (Author)

  6. Identification of a Novel Lipoprotein Regulator of Clostridium difficile Spore Germination.

    Directory of Open Access Journals (Sweden)

    Kelly A Fimlaid

    2015-10-01

    Full Text Available Clostridium difficile is a Gram-positive spore-forming pathogen and a leading cause of nosocomial diarrhea. C. difficile infections are transmitted when ingested spores germinate in the gastrointestinal tract and transform into vegetative cells. Germination begins when the germinant receptor CspC detects bile salts in the gut. CspC is a subtilisin-like serine pseudoprotease that activates the related CspB serine protease through an unknown mechanism. Activated CspB cleaves the pro-SleC zymogen, which allows the activated SleC cortex hydrolase to degrade the protective cortex layer. While these regulators are essential for C. difficile spores to outgrow and form toxin-secreting vegetative cells, the mechanisms controlling their function have only been partially characterized. In this study, we identify the lipoprotein GerS as a novel regulator of C. difficile spore germination using targeted mutagenesis. A gerS mutant has a severe germination defect and fails to degrade cortex even though it processes SleC at wildtype levels. Using complementation analyses, we demonstrate that GerS secretion, but not lipidation, is necessary for GerS to activate SleC. Importantly, loss of GerS attenuates the virulence of C. difficile in a hamster model of infection. Since GerS appears to be conserved exclusively in related Peptostreptococcaeace family members, our results contribute to a growing body of work indicating that C. difficile has evolved distinct mechanisms for controlling the exit from dormancy relative to B. subtilis and other spore-forming organisms.

  7. Air pollution by allergenic spores of the genus Alternaria in the air of central and eastern Europe.

    Science.gov (United States)

    Kasprzyk, Idalia; Rodinkova, Victoria; Šaulienė, Ingrida; Ritenberga, Olga; Grinn-Gofron, Agnieszka; Nowak, Malgorzata; Sulborska, Aneta; Kaczmarek, Joanna; Weryszko-Chmielewska, Elzbieta; Bilous, Elena; Jedryczka, Malgorzata

    2015-06-01

    Spores of the genus Alternaria belong to one of the most prevailing constituents of the air in all regions of the world. They form infectious inoculum of numerous plant species as well as severe inhaled allergies. The aim of this study was to compare the biological pollution with Alternaria spores of the air of 12 cities located in central and eastern Europe. The experiment was done in 2010 and it covered the territory of Latvia (LV), Lithuania (LT), Poland (PL) and Ukraine (UA). The spores were counted using an identical method and standard equipment (7-day Lanzoni volumetric sampler) followed by extensive statistical calculations. The timing of the day of maximum concentration changed mainly along the N-S direction and had a positive correlation with latitude. The most important factor determining the increase in Alternaria spore concentration was the temperature, whereas other weather parameters were not related or of low significance. Regardless of geographical location, the first phase of the season (0-0.9 % of Alternaria spores in the air) was the longest (up to 60 days) and the last (97.5 to 99 %) was the shortest (22 days or less). The means of daily concentrations of Alternaria spores ranged from 11 spores m(-3) in Klaipeda (LT, Baltic Sea coast) to 187 in Poznan (west PL, agricultural plain). The threshold value of 80 spores m(-3) that triggers the first allergy symptoms was exceeded in 8 to 86 days (Vinnitsa, UA, temperate continental, forest-steppes region). There were considerable differences between the highest number of spores per cubic metre of air, varying from 139 in the north (Klaipeda, LT) to 2,295 in central west (Poznan, PL). The biological pollution by Alternaria spores in several places of central and eastern Europe was high; the number of days exceeding the threshold value of 300 spores m(-3) connected with serious health problems of atopic people ranged from 0 to 1 on the north (LV, LT) to 29 in central west (Poznan, PL).

  8. Amplicon sequencing for the quantification of spoilage microbiota in complex foods including bacterial spores.

    Science.gov (United States)

    de Boer, Paulo; Caspers, Martien; Sanders, Jan-Willem; Kemperman, Robèr; Wijman, Janneke; Lommerse, Gijs; Roeselers, Guus; Montijn, Roy; Abee, Tjakko; Kort, Remco

    2015-01-01

    Spoilage of food products is frequently caused by bacterial spores and lactic acid bacteria. Identification of these organisms by classic cultivation methods is limited by their ability to form colonies on nutrient agar plates. In this study, we adapted and optimized 16S rRNA amplicon sequencing for quantification of bacterial spores in a canned food matrix and for monitoring the outgrowth of spoilage microbiota in a ready-to-eat food matrix. The detection limit of bar-coded 16S rRNA amplicon sequencing was determined for the number of bacterial spores in a canned food matrix. Analysis of samples from a canned food matrix spiked with a mixture of equinumerous spores from the thermophiles, Geobacillus stearothermophilus and Geobacillus thermoglucosidans, and the mesophiles, Bacillus sporothermodurans, Bacillus cereus, and Bacillus subtilis, led to the detection of these spores with an average limit of 2 × 10(2) spores ml(-1). The data were normalized by setting the number of sequences resulting from DNA of an inactivated bacterial species, present in the matrix at the same concentration in all samples, to a fixed value for quantitative sample-to-sample comparisons. The 16S rRNA amplicon sequencing method was also employed to monitor population dynamics in a ready-to-eat rice meal, incubated over a period of 12 days at 7 °C. The most predominant outgrowth was observed by the genera Leuconostoc, Bacillus, and Paenibacillus. Analysis of meals pre-treated with weak acids showed inhibition of outgrowth of these three genera. The specificity of the amplicon synthesis was improved by the design of oligonucleotides that minimize the amplification of 16S rRNA genes from chloroplasts originating from plant-based material present in the food. This study shows that the composition of complex spoilage populations, including bacterial spores, can be monitored in complex food matrices by bar-coded amplicon sequencing in a quantitative manner. In order to allow sample

  9. Quantitative Proteomic Analysis of Germination of Nosema bombycis Spores under Extremely Alkaline Conditions.

    Science.gov (United States)

    Liu, Han; Chen, Bosheng; Hu, Sirui; Liang, Xili; Lu, Xingmeng; Shao, Yongqi

    2016-01-01

    The microsporidian Nosema bombycis is an obligate intracellular pathogen of the silkworm Bombyx mori , causing the epidemic disease Pebrine and extensive economic losses in sericulture. Although N. bombycis forms spores with rigid spore walls that protect against various environmental pressures, ingested spores germinate immediately under the extremely alkaline host gut condition (Lepidoptera gut pH > 10.5), which is a key developmental turning point from dormant state to infected state. However, to date this process remains poorly understood due to the complexity of the animal digestive tract and the lack of genetic tools for microsporidia. Here we show, using an in vitro spore germination model, how the proteome of N. bombycis changes during germination, analyse specific metabolic pathways employed in detail, and validate key functional proteins in vivo in silkworms. By a label-free quantitative proteomics approach that is directly based on high-resolution mass spectrometry (MS) data, a total of 1136 proteins were identified with high confidence, with 127 proteins being significantly changed in comparison to non-germinated spores. Among them, structural proteins including polar tube protein 1 and 3 and spore wall protein (SWP) 4 and 30 were found to be significantly down-regulated, but SWP9 significantly up-regulated. Some nucleases like polynucleotide kinase/phosphatase and flap endonucleases 1, together with a panel of hydrolases involved in protein degradation and RNA cleavage were overrepresented too upon germination, which implied that they might play important roles during spore germination. The differentially regulated trends of these genes were validated, respectively, by quantitative RT-PCR and 3 proteins of interest were confirmed by Western blotting analyses in vitro and in vivo . Furthermore, the pathway analysis showed that abundant up- and down-regulations appear involved in the glycolysis, pentose phosphate pathway, purine, and pyrimidine

  10. Quantitative proteomic analysis of germination of Nosema bombycis spores under extremely alkaline conditions

    Directory of Open Access Journals (Sweden)

    Yongqi Shao

    2016-09-01

    Full Text Available The microsporidian Nosema bombycis is an obligate intracellular pathogen of the silkworm Bombyx mori, causing the epidemic disease Pebrine and extensive economic losses in sericulture. Although N. bombycis forms spores with rigid spore walls that protect against various environmental pressures, ingested spores germinate immediately under the extremely alkaline host gut condition (Lepidoptera gut pH >10.5, which is a key developmental turning point from dormant state to infected state. However, to date this process remains poorly understood due to the complexity of the animal digestive tract and the lack of genetic tools for Microsporidia. Here we show, using an in vitro spore germination model, how the proteome of N. bombycis changes during germination, analyse specific metabolic pathways employed in detail, and validate key functional proteins in vivo in silkworms. By a label-free quantitative proteomics approach that is directly based on high-resolution mass spectrometry (MS data, a total of 1136 proteins were identified with high confidence, with 127 proteins being significantly changed in comparison to non-germinated spores. Among them, structural proteins including polar tube protein 1 and 3 and spore wall protein (SWP 4 and 30 were found to be significantly down-regulated, but SWP9 significantly up-regulated. Some nucleases like polynucleotide kinase/phosphatase and flap endonucleases 1, together with a panel of hydrolases involved in protein degradation and RNA cleavage were overrepresented too upon germination, which implied that they might play important roles during spore germination. The differentially regulated trends of these genes were validated respectively by quantitative RT-PCR and 3 proteins of interest were confirmed by Western blotting analyses in vitro and in vivo. Furthermore the pathway analysis showed that abundant up- and down-regulations appear involved in the glycolysis, pentose phosphate pathway, purine and

  11. The solar UV environment and bacterial spore UV resistance: considerations for Earth-to-Mars transport by natural processes and human spaceflight

    Science.gov (United States)

    Nicholson, Wayne L.; Schuerger, Andrew C.; Setlow, Peter

    2005-01-01

    The environment in space and on planets such as Mars can be lethal to microorganisms because of the high vacuum and high solar radiation flux, in particular UV radiation, in such environments. Spores of various Bacillus species are among the organisms most resistant to the lethal effects of high vacuum and UV radiation, and as a consequence are of major concern for planetary contamination via unmanned spacecraft or even natural processes. This review focuses on the spores of various Bacillus species: (i) their mechanisms of UV resistance; (ii) their survival in unmanned spacecraft, space flight and simulated space flight and Martian conditions; (iii) the UV flux in space and on Mars; (iv) factors affecting spore survival in such high UV flux environments.

  12. The solar UV environment and bacterial spore UV resistance: considerations for Earth-to-Mars transport by natural processes and human spaceflight

    International Nuclear Information System (INIS)

    Nicholson, Wayne L.; Schuerger, Andrew C.; Setlow, Peter

    2005-01-01

    The environment in space and on planets such as Mars can be lethal to microorganisms because of the high vacuum and high solar radiation flux, in particular UV radiation, in such environments. Spores of various Bacillus species are among the organisms most resistant to the lethal effects of high vacuum and UV radiation, and as a consequence are of major concern for planetary contamination via unmanned spacecraft or even natural processes. This review focuses on the spores of various Bacillus species: (i) their mechanisms of UV resistance; (ii) their survival in unmanned spacecraft, space flight and simulated space flight and Martian conditions; (iii) the UV flux in space and on Mars; (iv) factors affecting spore survival in such high UV flux environments

  13. Textiles for protection against microorganism

    Science.gov (United States)

    Sauperl, O.

    2016-04-01

    Concerning micro-organisms such as bacteria, viruses and fungi, there is a huge progress in the development of textile materials and procedures which should effectively protect against these various pathogens. In this sense there is especially problematic hospital environment, where it is necessary to take into account properly designed textile material which, when good selected and composed, act as a good barrier against transfer of micro-organisms through material mainly in its wet state. Respect to this it is necessary to be familiar with the rules regarding selection of the input material, the choice of proper yarn construction, the choice of the proper weaving mode, the rules regarding selection of antimicrobial-active compound suitable for (eco-friendly) treatment, and the choice of the most appropriate test method by which it is possible objectively to conclude on the reduction of selected microorganism. As is well known, fabrics are three-dimensional structures with void and non-void areas. Therefore, the physical-chemical properties of the textile material/fabric, the surface characteristics together with the shape of microorganism, and the carriers' characteristics contribute to control the transfer of microorganism through textile material. Therefore, careful planning of textile materials and treatment procedure with the compound which is able to reduce micro-organism satisfactory is particularly important, especially due to the fact that in hospital environment population with impaired immune system is mainly presented.

  14. No evidence for a Ganoderma spore dispersal mutualism in an obligate spore-feeding beetle Zearagytodes maculifer.

    Science.gov (United States)

    Kadowaki, Kohmei; Leschen, Richard A B; Beggs, Jacqueline R

    2011-08-01

    The role of spore dispersal mutualism remains equivocal in many fungus-insect assemblages. We tested experimentally whether an obligate spore-feeding beetle Zearagytodes maculifer has a mutualistic relationship with its host bracket fungus Ganoderma cf. applanatum via spore dispersal. We asked three specific questions: (1) whether or not Ganoderma spore germination rate is increased via beetle digestive activity and (2) is dependent on temperature and sporocarp identity. Spore germination rates were examined in 2×3×2 factorial experiments (spores consumed by beetles or not×temperature 20, 25, and 30°C×two independent pairs of sporocarp-beetle populations) replicated five times in an array of 60 experimental cultures. Analysis showed that consumption by beetles significantly reduced germination rate of Ganoderma spores. The effect of temperature was modulated by the effect of individual sporocarp, and was overridden by beetle feeding. Microscopic analysis revealed that spores from beetle faecal pellets exhibited extensive damage to their thin outer walls (pellicles) and thick inner walls, as well as significant loss of cytoplasm, while control spores were intact. The overall evidence argued against our spore dispersal mutualism hypothesis, suggesting that Z. maculifer can potentially exert a negative, if vanishingly small, fitness effect on its host fungus G. cf. applanatum. Copyright © 2011 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  15. Identification of Differentially Expressed Genes during Bacillus subtilis Spore Outgrowth in High-Salinity Environments Using RNA Sequencing

    NARCIS (Netherlands)

    Nagler, Katja; Krawczyk, Antonina O; De Jong, Anne; Madela, Kazimierz; Hoffmann, Tamara; Laue, Michael; Kuipers, Oscar P; Bremer, Erhard; Moeller, Ralf

    2016-01-01

    In its natural habitat, the soil bacterium Bacillus subtilis often has to cope with fluctuating osmolality and nutrient availability. Upon nutrient depletion it can form dormant spores, which can revive to form vegetative cells when nutrients become available again. While the effects of salt stress

  16. 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.

  17. Modeling Radiation Effectiveness for Inactivation of Bacillus Spores

    Science.gov (United States)

    2015-09-17

    MODELING RADIATION EFFECTIVENESS FOR INACTIVATION OF BACILLUS SPORES DISSERTATION Emily A. Knight, Major, USAF AFIT-ENC-DS-15-S-001 DEPARTMENT OF THE...not subject to copyright protection in the United States. AFIT-ENC-DS-15-S-001 MODELING RADIATION EFFECTIVENESS FOR INACTIVATION OF BACILLUS SPORES...EFFECTIVENESS FOR INACTIVATION OF BACILLUS SPORES Emily A. Knight, B.A., M.S. Major, USAF Committee Membership: Dr. William P. Baker Chair Dr. Larry W

  18. Chitinolytic activity in viable spores of encephalitozoon species

    OpenAIRE

    Schottelius,J; Hünger,F; Schüler,Th; Gonçalves da Costa,SC

    2000-01-01

    By employing 4-methylumbelliferyl-beta-D-NN',N"-triacetylchitotriose substrate in a semi quantitative assay, chitinolytic activity in viable spores of Encephalitozoon cuniculi and E. intestinalis was detected and dependence on reaction time, spore concentration, concentration of substrate and temperature were demonstrated. It was possible to block the chitinolytic activity by chitin hydrolysate. By incubation at 80°C for 10 min or at 55°C for 20 min the spores were loosing the chitinolytic ac...

  19. Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Jeong, Youngsu; Lee, Jonghee; Kim, Seongsoo [Agency for Defense Development, Daejeon (Korea, Republic of)

    2013-09-15

    The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field.

  20. Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

    International Nuclear Information System (INIS)

    Jeong, Youngsu; Lee, Jonghee; Kim, Seongsoo

    2013-01-01

    The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field

  1. Chitinolytic activity in viable spores of encephalitozoon species

    Directory of Open Access Journals (Sweden)

    J Schottelius

    2000-10-01

    Full Text Available By employing 4-methylumbelliferyl-beta-D-NN',N"-triacetylchitotriose substrate in a semi quantitative assay, chitinolytic activity in viable spores of Encephalitozoon cuniculi and E. intestinalis was detected and dependence on reaction time, spore concentration, concentration of substrate and temperature were demonstrated. It was possible to block the chitinolytic activity by chitin hydrolysate. By incubation at 80°C for 10 min or at 55°C for 20 min the spores were loosing the chitinolytic activity. Incubation of the spores in trypsin reduced the chitinolytic activity. Cellulase activity could not be detected.

  2. Chitinolytic activity in viable spores of Encephalitozoon species.

    Science.gov (United States)

    Schottelius, J; Hünger, F; Schüler, T; Gonçalves da Costa, S C

    2000-01-01

    By employing 4-methylumbelliferyl-beta-D-NN',N"-triacetylchitotriose substrate in a semi quantitative assay, chitinolytic activity in viable spores of Encephalitozoon cuniculi and E. intestinalis was detected and dependence on reaction time, spore concentration, concentration of substrate and temperature were demonstrated. It was possible to block the chitinolytic activity by chitin hydrolysate. By incubation at 80 degrees C for 10 min or at 55 degrees C for 20 min the spores were loosing the chitinolytic activity. Incubation of the spores in trypsin reduced the chitinolytic activity. Cellulase activity could not be detected.

  3. Effects of Chlorine Dioxide on Spore Structural and Functional Properties

    National Research Council Canada - National Science Library

    Leighton, Terrance

    2003-01-01

    .... The experimental results described in this report were designed to test this hypothesis. Dormant bacterial endospores are highly birefringent due to the anhydrous nature of the spore cytoplasm...

  4. Bioemulsan Production by Iranian Oil Reservoirs Microorganisms

    Directory of Open Access Journals (Sweden)

    A Amiriyan, M Mazaheri Assadi, VA Saggadian, A Noohi

    2004-10-01

    Full Text Available The biosurfactants are believed to be surface active components that are shed into the surrounding medium during the growth of the microorganisms. The oil degrading microorganism Acinetobacter calcoaceticus RAG-1 produces a poly-anionic biosurfactant, hetero-polysaccharide bioemulsifier termed as emulsan which forms and stabilizes oil-water emulsions with a variety of hydrophobic substrates. In the present paper results of the possibility of biosurfactant (Emulsan production by microorganisms isolated from Iranian oil reservoirs is presented. Fourthy three gram negative and gram positive, non fermentative, rod bacilli and coccobacilli shaped baceria were isolated from the oil wells of Bibi Hakimeh, Siri, Maroon, Ilam , East Paydar and West Paydar. Out of the isolated strains, 39 bacterial strains showed beta haemolytic activity, further screening revealed the emulsifying activity and surface tension. 11 out of 43 tested emulsifiers were identified as possible biosurfactant producers and two isolates produced large surface tension reduction, indicating the high probability of biosurfactant production. Further investigation revealed that, two gram negative, oxidase negative, aerobic and coccoid rods isolates were the best producers and hence designated as IL-1, PAY-4. Whole culture broth of isolates reduced surface tension from 68 mN /m to 30 and 29.1mN/m, respectively, and were stable during exposure to high salinity (10%NaCl and elevated temperatures(120C for 15 min .

  5. Development of an eco-friendly approach for biogenesis of silver nanoparticles using spores of Bacillus athrophaeus.

    Science.gov (United States)

    Hosseini-Abari, Afrouzossadat; Emtiazi, Giti; Ghasemi, Seyed Mahdi

    2013-12-01

    The biological synthesis methods have been emerging as a promising new approach for production of nanoparticles due to their simplicity and non-toxicity. In the present study, spores of Bacillus athrophaeus were used to achieve the objective of developing a green synthesis method of silver nanoparticles. Enzyme assay revealed that the spores and their heat inactivated forms (microcapsules) were highly active and their enzymatic contents differed from the vegetative cells. Laccase, glucose oxidase, and alkaline phosphatase activities were detected in the dormant forms, but not in the vegetative cells. Although no nanoparticle was produced by active cells of B. athrophaeus, both spores and microcapsules were efficiently capable of reducing the silver ions (Ag⁺) to elemental silver (Ag⁰) leading to the formation of nanoparticles from silver nitrate (AgNO₃). The presence of biologically synthesized silver nanoparticles was determined by obtaining broad spectra with maximum absorbance at 400 nm in UV-visible spectroscopy. The X-ray diffraction analysis pattern revealed that the nanoscale particles have crystalline nature with various topologies, as confirmed by transmission electron microscopy (TEM). The TEM micrograph showed the nanocrystal structures with dimensions ranging from 5 to 30 nm. Accordingly, the spore mixture could be employed as a factory for detoxification of heavy metals and subsequent production of nanoparticles. This research introduces an environmental friendly and cost effective biotechnological process for the extracellular synthesis of silver nanoparticles using the bacterial spores.

  6. Dynamics of Bacillus thuringiensis var. israelensis and Lysinibacillus sphaericus spores in urban catch basins after simultaneous application against mosquito larvae.

    Science.gov (United States)

    Guidi, Valeria; Lehner, Angelika; Lüthy, Peter; Tonolla, Mauro

    2013-01-01

    Bacillus thuringiensis var. israelensis (Bti) and Lysinibacillus sphaericus (Lsph) are extensively used in mosquito control programs. These biocides are the active ingredients of a commercial larvicide. Quantitative data on the fate of both Bti and Lsph applied together for the control of mosquitoes in urban drainage structures such as catch basins are lacking. We evaluated the dynamics and persistence of Bti and Lsph spores released through their concomitant application in urban catch basins in southern Switzerland. Detection and quantification of spores over time in water and sludge samples from catch basins were carried out using quantitative real-time PCR targeting both cry4A and cry4B toxin genes for Bti and the binA gene for Lsph. After treatment, Bti and Lsph spores attained concentrations of 3.76 (± 0.08) and 4.13 (± 0.09) log ml(-1) in water, then decreased progressively over time, reaching baseline values. For both Bti and Lsph, spore levels in the order of 10(5) g(-1) were observed in the bottom sludge two days after the treatment and remained constant for the whole test period (275 days). Indigenous Lsph strains were isolated from previously untreated catch basins. A selection of those was genotyped using pulsed field gel electrophoresis of SmaI-digested chromosomal DNA, revealing that a subset of isolates were members of the clonal population of strain 2362. No safety issues related to the use of this biopesticide in the environment have been observed during this study, because no significant increase in the number of spores was seen during the long observation period. The isolation of native Lysinibacillus sphaericus strains belonging to the same clonal population as strain 2362 from catch basins never treated with Lsph-based products indicates that the use of a combination of Bti and Lsph for the control of mosquitoes does not introduce non-indigenous microorganisms in this area.

  7. Effect of liner and core materials of plasterboard on microbial growth, spore-induced inflammatory responses, and cytotoxicity in macrophages.

    Science.gov (United States)

    Murtoniemi, Timo; Nevalainen, Aino; Suutari, Merja; Hirvonen, Maija-Riitta

    2002-11-01

    Microorganisms, when grown on wetted plasterboards, can produce bioactive compounds capable of inducing inflammatory and toxic reactions in mammalian cells. The paper liner of plasterboard is commonly regarded as the major substrate for microbial growth. In this study, we cultured Stachybotrys chartarum, Aspergillus versicolor, Penicillium spinulosum, and Streptomyces californicus on liners and cores of plasterboards in order to examine the role of these main plasterboard components on microbial growth and the resulting bioactivity, which was assessed as the ability of microbial spores to induce inflammatory responses and to evoke cytotoxicity in mouse macrophages. The microbes, isolated from mold problem buildings, were grown under saturated humidity conditions on wetted liners and cores of six different plasterboards. The spores were collected, applied to RAW264.7 macrophages at different doses, and evaluated 24 h after exposure for their ability to evoke cytotoxicity and to stimulate production of nitric oxide (NO), tumor necrosis factor alpha (TNFalpha), and interleukin-6 (IL-6). In general, microbial growth was better on the cores than on the liners. All of the studied microbes collected from cores induced a dose-dependent production of TNFalpha in macrophages. The TNFalpha production stimulated by spores of Stachybotrys, Aspergillus, and Streptomyces paralleled their cytotoxicity. Spores of Streptomyces and Aspergillus collected from liners were among the most potent inducers of NO and IL-6. Good growth of Stachybotrys on cores was associated with high cytotoxicity. Penicillium grew only on cores, but it did not induce major inflammatory mediator productions, nor was it significantly cytotoxic. These results indicate that previously reported microbial growth on plasterboards and spore-induced production of important inflammatory mediators and cell death in macrophages is not only due to the paper liner of plasterboard, but the core material also has a crucial

  8. Dynamics of Bacillus thuringiensis var. israelensis and Lysinibacillus sphaericus spores in urban catch basins after simultaneous application against mosquito larvae.

    Directory of Open Access Journals (Sweden)

    Valeria Guidi

    Full Text Available Bacillus thuringiensis var. israelensis (Bti and Lysinibacillus sphaericus (Lsph are extensively used in mosquito control programs. These biocides are the active ingredients of a commercial larvicide. Quantitative data on the fate of both Bti and Lsph applied together for the control of mosquitoes in urban drainage structures such as catch basins are lacking. We evaluated the dynamics and persistence of Bti and Lsph spores released through their concomitant application in urban catch basins in southern Switzerland. Detection and quantification of spores over time in water and sludge samples from catch basins were carried out using quantitative real-time PCR targeting both cry4A and cry4B toxin genes for Bti and the binA gene for Lsph. After treatment, Bti and Lsph spores attained concentrations of 3.76 (± 0.08 and 4.13 (± 0.09 log ml(-1 in water, then decreased progressively over time, reaching baseline values. For both Bti and Lsph, spore levels in the order of 10(5 g(-1 were observed in the bottom sludge two days after the treatment and remained constant for the whole test period (275 days. Indigenous Lsph strains were isolated from previously untreated catch basins. A selection of those was genotyped using pulsed field gel electrophoresis of SmaI-digested chromosomal DNA, revealing that a subset of isolates were members of the clonal population of strain 2362. No safety issues related to the use of this biopesticide in the environment have been observed during this study, because no significant increase in the number of spores was seen during the long observation period. The isolation of native Lysinibacillus sphaericus strains belonging to the same clonal population as strain 2362 from catch basins never treated with Lsph-based products indicates that the use of a combination of Bti and Lsph for the control of mosquitoes does not introduce non-indigenous microorganisms in this area.

  9. UV air cleaners and upper-room air ultraviolet germicidal irradiation for controlling airborne bacteria and fungal spores.

    Science.gov (United States)

    Kujundzic, Elmira; Matalkah, Fatimah; Howard, Cody J; Hernandez, Mark; Miller, Shelly L

    2006-10-01

    In-room air cleaners (ACs) and upper-room air ultraviolet germicidal irradiation (UVGI) are engineering control technologies that can help reduce the concentrations of airborne bacteria and fungal spores in the indoor environment. This study investigated six different types of ACs and quantified their ability to remove and/or inactivate airborne bacteria and fungal spores. Four of the air cleaners incorporated UV lamp(s) into their flow path. In addition, the efficacy of combining ACs with upper-room air UVGI was investigated. With the ventilation system providing zero or six air changes per hour, the air cleaners were tested separately or with the upper-room air UVGI system in operation in an 87-m3 test room. Active bacteria cells and fungal spores were aerosolized into the room such that their numbers and physiologic state were comparable both with and without air cleaning and upper-room air UVGI. In addition, the disinfection performance of a UV-C lamp internal to one of the ACs was evaluated by estimating the percentage of airborne bacteria cells and fungal spores captured on the air filter medium surface that were inactivated with UV exposure. Average airborne microbial clean air delivery rates (CADRm) varied between 26-981 m3 hr-1 depending on the AC, and between 1480-2370 m3 hr-1, when using air cleaners in combination with upper-room air UVGI. Culturing, direct microscopy, and optical particle counting revealed similar CADRm. The ACs performed similarly when challenged with three different microorganisms. Testing two of the ACs showed that no additional air cleaning was provided with the operation of an internal UV-C lamp; the internal UV-C lamps, however, inactivated 75% of fungal spores and 97% of bacteria cells captured in the air filter medium within 60 min.

  10. Airborne fungal spores of Alternaria, meteorological parameters and predicting variables

    Science.gov (United States)

    Filali Ben Sidel, Farah; Bouziane, Hassan; del Mar Trigo, Maria; El Haskouri, Fatima; Bardei, Fadoua; Redouane, Abdelbari; Kadiri, Mohamed; Riadi, Hassane; Kazzaz, Mohamed

    2015-03-01

    Alternaria is frequently found as airborne fungal spores and is recognized as an important cause of respiratory allergies. The aerobiological monitoring of fungal spores was performed using a Burkard volumetric spore traps. To establish predicting variables for daily and weakly spore counts, a stepwise multiple regression between spore concentrations and independent variables (meteorological parameters and lagged values from the series of spore concentrations: previous day or week concentration (Alt t - 1) and mean concentration of the same day or week in other years ( C mean)) was made with data obtained during 2009-2011. Alternaria conidia are present throughout the year in the atmosphere of Tetouan, although they show important seasonal fluctuations. The highest levels of Alternaria spores were recorded during the spring and summer or autumn. Alternaria showed maximum daily values in April, May or October depending on year. When the spore variables of Alternaria, namely C mean and Alt t - 1, and meteorological parameters were included in the equation, the resulting R 2 satisfactorily predict future concentrations for 55.5 to 81.6 % during the main spore season and the pre-peak 2. In the predictive model using weekly values, the adjusted R 2 varied from 0.655 to 0.676. The Wilcoxon test was used to compare the results from the expected values and the pre-peak spore data or weekly values for 2012, indicating that there were no significant differences between series compared. This test showed the C mean, Alt t - 1, frequency of the wind third quadrant, maximum wind speed and minimum relative humidity as the most efficient independent variables to forecast the overall trend of this spore in the air.

  11. Airborne fungal spores of Alternaria, meteorological parameters and predicting variables.

    Science.gov (United States)

    Filali Ben Sidel, Farah; Bouziane, Hassan; Del Mar Trigo, Maria; El Haskouri, Fatima; Bardei, Fadoua; Redouane, Abdelbari; Kadiri, Mohamed; Riadi, Hassane; Kazzaz, Mohamed

    2015-03-01

    Alternaria is frequently found as airborne fungal spores and is recognized as an important cause of respiratory allergies. The aerobiological monitoring of fungal spores was performed using a Burkard volumetric spore traps. To establish predicting variables for daily and weakly spore counts, a stepwise multiple regression between spore concentrations and independent variables (meteorological parameters and lagged values from the series of spore concentrations: previous day or week concentration (Alt t - 1) and mean concentration of the same day or week in other years (C mean)) was made with data obtained during 2009-2011. Alternaria conidia are present throughout the year in the atmosphere of Tetouan, although they show important seasonal fluctuations. The highest levels of Alternaria spores were recorded during the spring and summer or autumn. Alternaria showed maximum daily values in April, May or October depending on year. When the spore variables of Alternaria, namely C mean and Alt t - 1, and meteorological parameters were included in the equation, the resulting R (2) satisfactorily predict future concentrations for 55.5 to 81.6 % during the main spore season and the pre-peak 2. In the predictive model using weekly values, the adjusted R (2) varied from 0.655 to 0.676. The Wilcoxon test was used to compare the results from the expected values and the pre-peak spore data or weekly values for 2012, indicating that there were no significant differences between series compared. This test showed the C mean, Alt t - 1, frequency of the wind third quadrant, maximum wind speed and minimum relative humidity as the most efficient independent variables to forecast the overall trend of this spore in the air.

  12. Resistance of soil microorganisms to starvation.

    Science.gov (United States)

    Chen, M.; Alexander, M.

    1972-01-01

    Most groups of soil microorganisms died when exposed to prolonged starvation in a carbon-free solution, but the relative abundance of Bacillus and actinomycetes increased with time. Certain nonspore-forming bacteria also persisted. The ability of individual soil isolates to endure starvation in solution was not correlated with their glycogen content or rate of endogenous respiration. However, cells of the resistant populations were rich in poly-beta-hydroxybutyrate, whereas the starvation-susceptible bacteria generally contained little of this substance. Poly-beta-hydroxybutyrate was used rapidly in cells deprived of exogenous sources of carbon.

  13. CHITINASE AND CHITINOLYTIC MICROORGANISM : ISOLATION, CHARACTERIZATION AND POTENTIAL

    Directory of Open Access Journals (Sweden)

    Nuniek Herdyastuti

    2010-06-01

    Full Text Available Chitinase is enzyme that hydrolyzes chitin, a polimer of b-1,4-N-Acetilglucosamine which is the most abundant natural resource after cellulose. Chitinolytic microorganism can be found in  environment like soil and water that contain chitin, or in extreme environment which is known as thermofilic microorganism. Chitinolytic microorganism is identified by recognizing the morphological and physiological properties based on Bergey's Manual of Systematic Bacteriology. The sequence data of the 16S rRNA genes is determinated in the GeneBank nucleotide sequence database. Chitinase activity can be qualitatively determined from the clearance zone around the colony formed in agar medium containing colloidal chitin. Chitinase can be utilized as biocontrol agent and derivate chitin as the result of chitinase degradation which can be used in the fieltd of health, food, industry and waste management   Keywords: Biocontrol, chitin, chitinase,  chitinolytic microorganism

  14. Irradiation Effects on the Decontamination of Microorganisms in Honey

    International Nuclear Information System (INIS)

    Jo, Cheorun; Kim, Jae Kyung; Kang, Ho Jin; Lee Eun Young; Byun, Myung Woo

    2005-09-01

    Honey is usually contaminated with numerous microorganisms. Especially, the presence of the spore of Clostridium in honey is dangerous to infants and small children. To determine the effect of a gamma irradiation on the microorganisms in honey, our kinds of honeys (acacia flower, poly floral flower, native A, and native B) were irradiated (0, 5, and 10 kGy) and the total aerobic bacterial count, yeasts and molds, coliform, and Clostridium spp. were detected. In all the honeys, the coliform bacterial, yeast and molds counts were below. The limit of detection (< 101 cfu/g). Total aerobic bacterial and Clostridium spp counts ranged from 85 to 450 cfu/g and 0 to 450 cfu/g in the samples respectively. Acacia and poly floral flower honey were completely sterilized by a 5 kGy irradiation, and both the native honey A and B showed decreased populations of the total aerobic bacteria and Clostridium spp. with a 5 kGy irradiation. Therefore it is concluded that an irradiation treatment could be an effective tool to disinfect honey

  15. Contribution of fungal spores to particulate matter in a tropical rainforest

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Ting; Chan Chuenyu; Zhang Yinan; Zhang Zhisheng; Lin Mang; Sang Xuefang [School of Environmental Science and Engineering, Sun Yat-Sen University, Guangzhou (China); Engling, Guenter [Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan (China); Li, Y D [Institute of Subtropical Forestry, Bureau of Forestry, Guangzhou (China); Li, Yok-Sheung, E-mail: guenter@gate.sinica.edu.tw, E-mail: chzy@mail.sysu.edu.cn [Department of Civil and Structural Engineering, Hong Kong Polytechnic University (Hong Kong)

    2010-04-15

    The polyols arabitol and mannitol, recently proposed as source tracers for fungal spores, were used in this study to estimate fungal contributions to atmospheric aerosol. Airborne particulate matter (PM{sub 2.5} and PM{sub 10}) was collected at Jianfengling Mountain, a tropical rainforest on Hainan Island situated off the south China coast, during spring and analyzed for arabitol and mannitol by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The average concentrations of arabitol and mannitol exhibited high values with averages of 7.0 and 16.0 ng m{sup -3} respectively in PM{sub 2.5} and 44.0 and 71.0 ng m{sup -3} in PM{sub 10}. The two tracers correlated well with each other, especially in the coarse mode aerosol (PM{sub 2.5-10}), indicating they were mainly associated with coarse aerosol particles and had common sources. Arabitol and mannitol in PM{sub 10} showed significant positive correlations with relative humidity, as well as positive correlations with average temperature, suggesting a wet emissions mechanism of biogenic aerosol in the form of fungal spores. We made estimations of the contribution of fungal spores to ambient PM mass and to organic carbon, based on the observed ambient concentrations of these two tracers. The relative contributions of fungal spores to the PM{sub 10} mass were estimated to range from 1.6 to 18.2%, with a rather high mean value of 7.9%, and the contribution of fungal spores to organic carbon in PM{sub 10} ranged from 4.64 to 26.1%, with a mean value of 12.1%, implying that biological processes are important sources of atmospheric aerosol.

  16. Infection of Tribolium castaneum with Bacillus thuringiensis: Quantification of Bacterial Replication within Cadavers, Transmission via Cannibalism, and Inhibition of Spore Germination

    Science.gov (United States)

    Milutinović, Barbara; Höfling, Christina; Futo, Momir; Scharsack, Jörn P.

    2015-01-01

    Reproduction within a host and transmission to the next host are crucial for the virulence and fitness of pathogens. Nevertheless, basic knowledge about such parameters is often missing from the literature, even for well-studied bacteria, such as Bacillus thuringiensis, an endospore-forming insect pathogen, which infects its hosts via the oral route. To characterize bacterial replication success, we made use of an experimental oral infection system for the red flour beetle Tribolium castaneum and developed a flow cytometric assay for the quantification of both spore ingestion by the individual beetle larvae and the resulting spore load after bacterial replication and resporulation within cadavers. On average, spore numbers increased 460-fold, showing that Bacillus thuringiensis grows and replicates successfully in insect cadavers. By inoculating cadaver-derived spores and spores from bacterial stock cultures into nutrient medium, we next investigated outgrowth characteristics of vegetative cells and found that cadaver-derived bacteria showed reduced growth compared to bacteria from the stock cultures. Interestingly, this reduced growth was a consequence of inhibited spore germination, probably originating from the host and resulting in reduced host mortality in subsequent infections by cadaver-derived spores. Nevertheless, we further showed that Bacillus thuringiensis transmission was possible via larval cannibalism when no other food was offered. These results contribute to our understanding of the ecology of Bacillus thuringiensis as an insect pathogen. PMID:26386058

  17. Infection of Tribolium castaneum with Bacillus thuringiensis: quantification of bacterial replication within cadavers, transmission via cannibalism, and inhibition of spore germination.

    Science.gov (United States)

    Milutinović, Barbara; Höfling, Christina; Futo, Momir; Scharsack, Jörn P; Kurtz, Joachim

    2015-12-01

    Reproduction within a host and transmission to the next host are crucial for the virulence and fitness of pathogens. Nevertheless, basic knowledge about such parameters is often missing from the literature, even for well-studied bacteria, such as Bacillus thuringiensis, an endospore-forming insect pathogen, which infects its hosts via the oral route. To characterize bacterial replication success, we made use of an experimental oral infection system for the red flour beetle Tribolium castaneum and developed a flow cytometric assay for the quantification of both spore ingestion by the individual beetle larvae and the resulting spore load after bacterial replication and resporulation within cadavers. On average, spore numbers increased 460-fold, showing that Bacillus thuringiensis grows and replicates successfully in insect cadavers. By inoculating cadaver-derived spores and spores from bacterial stock cultures into nutrient medium, we next investigated outgrowth characteristics of vegetative cells and found that cadaver-derived bacteria showed reduced growth compared to bacteria from the stock cultures. Interestingly, this reduced growth was a consequence of inhibited spore germination, probably originating from the host and resulting in reduced host mortality in subsequent infections by cadaver-derived spores. Nevertheless, we further showed that Bacillus thuringiensis transmission was possible via larval cannibalism when no other food was offered. These results contribute to our understanding of the ecology of Bacillus thuringiensis as an insect pathogen. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. Determination of fungal spore release from wet building materials

    DEFF Research Database (Denmark)

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

    2003-01-01

    The release and transport of fungal spores from water-damaged building materials is a key factor for understanding the exposure to particles of fungal origin as a possible cause of adverse health effects associated to growth of fungi indoors. In this study, the release of spores from nine species...

  19. 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.

  20. Survival of Clostridium difficile spores at low water activity.

    Science.gov (United States)

    Deng, Kai; Talukdar, Prabhat K; Sarker, Mahfuzur R; Paredes-Sabja, Daniel; Torres, J Antonio

    2017-08-01

    Clostridium difficile is frequently found in meat and meat products. Germination efficiency, defined as colony formation, was previously investigated at temperatures found in meat handling and processing for spores of strain M120 (animal isolate), R20291 (human isolate), and DK1 (beef isolate). In this study, germination efficiency of these spore strains was assessed in phosphate buffered saline (PBS, a w ∼1.00), commercial beef jerky (a w ∼0.82/0.72), and a w -adjusted PBS (a w ∼0.82/0.72). Surface hydrophobicity was followed for spores stored in PBS. After three months and for all PBS a w levels tested, M120 and DK1 spores showed a ∼1 decimal reduction in colony formation but this was not the case when kept in beef jerky suggesting a protective food matrix effect. During storage, and with no significant a w effect, an increase in colony formation was observed for R20291 spores kept in PBS (∼2 decimal log increase) and beef jerky (∼1 decimal log increase) suggesting a loss of spore superdormancy. For all strains, no significant changes in spore surface hydrophobicity were observed after storage. Collectively, these results indicate that depending on the germination properties of C. difficile spores and the media properties, their germination efficiency may increase or decrease during long term food storage. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Germination of Bacillus cereus spores adhered to stainless steel

    NARCIS (Netherlands)

    Hornstra, L.M.; Leeuw, de P.P.L.A.; Moezelaar, R.; Wolbert, E.J.H.; Vries, de Y.P.; Vos, de W.M.; Abee, T.

    2007-01-01

    Adhered spores of Bacillus cereus represent a significant part of the surface-derived contamination in processing equipment used in the dairy industry. As germinated spores lose their resistance capacities instantaneously, efficient germination prior to a cleaning in place treatment could aid to the

  2. 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.

  3. Macroalgal spore dysfunction: ocean acidification delays and weakens adhesion.

    Science.gov (United States)

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

    2018-04-01

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

  4. 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

  5. Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes

    Science.gov (United States)

    2014-10-30

    effect of SWNTs in combination with antimicrobial chemicals on inactivation of B. anthracis spores; 4) the effect of CNTs coated surfaces on the...2010 31-May-2014 Approved for Public Release; Distribution Unlimited Final Report: (Life Science Division/ Biochemistry ) Inactivation of Bacillus... Biochemistry ) Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes Report Title The Specific Aims of the project were to investigate: 1) the

  6. Breaking the spores of Ganoderma lucidum by fermentation with ...

    African Journals Online (AJOL)

    In this paper, fermentation of G. lucidum with Lactobacillus plantarum was applied to break down the sporoderm. Scanning electron microscope (SEM) was used to characterize the spores. The broken spores were found on the 3rd day and complete breaking on the 5th day of fermentation. Lactic acid, acetic acid and ...

  7. Sensitivity of Dormant and Germinating B, Anthracis Spores to Polycationic Compound

    Science.gov (United States)

    2005-06-01

    practical problems such as food spoilage , degradation of industrial materials, and "sick building syndrome" resulting from colonization of ventilation systems...species of Clostridium and Bacillus are of concern to the food industry due to their potential for spoilage of or toxin production in improperly...1.2. While most spore-forming bacterial species are classified as non-pathogenic or opportunistically pathogenic, Bacillus anthracis is well-known a

  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. Biofuel production by recombinant microorganisms

    Energy Technology Data Exchange (ETDEWEB)

    Liao, James C.; Atsumi, Shota; Cann, Anthony F.

    2017-07-04

    Provided herein are metabolically-modified microorganisms useful for producing biofuels. More specifically, provided herein are methods of producing high alcohols including isobutanol, 1-butanol, 1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol and 2-phenylethanol from a suitable substrate.

  10. Airborne microorganisms from waste containers.

    Science.gov (United States)

    Jedlicka, Sabrina S; Stravitz, David M; Lyman, Charles E

    2012-01-01

    In physician's offices and biomedical labs, biological waste is handled every day. This waste is disposed of in waste containers designed for holding red autoclave bags. The containers used in these environments are closed hands-free containers, often with a step pedal. While these containers protect the user from surface-borne microorganisms, the containers may allow airborne microorganisms to escape via the open/close mechanism because of the air current produced upon open/close cycles. In this study, the air current was shown to be sufficient to allow airborne escape of microorganisms held in the container, including Aspergillus niger. However, bacterial cultures, such as Escherichia coli and Lactococcus lactis did not escape. This may be due to the choice of bacterial cultures and the absence of solid waste, such as dust or other particulate matter in the waste containers, that such strains of bacteria could travel on during aerosolization. We compared these results to those obtained using a re-designed receptacle, which mimimizes air currents, and detected no escaping microorganisms. This study highlights one potential source of airborne contamination in labs, hospitals, and other environments that dispose of biological waste.

  11. Seasonal Trends in Airborne Fungal Spores in Coastal California Ecosystems

    Science.gov (United States)

    Morfin, J.; Crandall, S. G.; Gilbert, G. S.

    2014-12-01

    Airborne fungal spores cause disease in plants and animals and may trigger respiratory illnesses in humans. In terrestrial systems, fungal sporulation, germination, and persistence are strongly regulated by local meteorological conditions. However, few studies investigate how microclimate affects the spatio-temporal dynamics of airborne spores. We measured fungal aerospora abundance and microclimate at varying spatial and time scales in coastal California in three habitat-types: coast redwood forest, mixed-evergreen forest, and maritime chaparral. We asked: 1) is there a difference in total airborne spore concentration between habitats, 2) when do we see peak spore counts, and 3) do spore densities correlate with microclimate conditions? Fungal spores were caught from the air with a volumetric vacuum air spore trap during the wet season (January - March) in 2013 and 2014, as well as monthly in 2014. Initial results suggest that mixed-evergreen forests exhibit the highest amounts of spore abundance in both years compared to the other habitats. This may be due to either a higher diversity of host plants in mixed-evergreen forests or a rich leaf litter layer that may harbor a greater abundance of saprotrophic fungi. Based on pilot data, we predict that temperature and to a lesser degree, relative humidity, will be important microclimate predictors for high spore densities. These data are important for understanding when and under what weather conditions we can expect to see high levels of fungal spores in the air; this can be useful information for managers who are interested in treating diseased plants with fungicides.

  12. 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.

  13. Role played by exosporium glycoproteins in the surface properties of Bacillus cereus spores and in their adhesion to stainless steel.

    Science.gov (United States)

    Lequette, Yannick; Garénaux, Estelle; Tauveron, Grégoire; Dumez, Sylvain; Perchat, Stéphane; Slomianny, Christian; Lereclus, Didier; Guérardel, Yann; Faille, Christine

    2011-07-01

    Bacillus cereus spores are surrounded by a loose-fitting layer called the exosporium, whose distal part is mainly formed from glycoproteins. The role played by the exosporium glycoproteins of B. cereus ATCC 14579 (BclA and ExsH) was investigated by considering hydrophobicity and charge, as well as the properties of spore adhesion to stainless steel. The absence of BclA increased both the isoelectric point (IEP) and hydrophobicity of whole spores while simultaneously reducing the interaction between spores and stainless steel. However, neither the hydrophobicity nor the charge associated with BclA could explain the differences in the adhesion properties. Conversely, ExsH, another exosporium glycoprotein, did not play a significant role in spore surface properties. The monosaccharide analysis of B. cereus ATCC 14579 showed different glycosylation patterns on ExsH and BclA. Moreover, two specific glycosyl residues, namely, 2-O-methyl-rhamnose (2-Me-Rha) and 2,4-O-methyl-rhamnose (2,4-Me-Rha), were attached to BclA, in addition to the glycosyl residues already reported in B. anthracis.

  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. Volatilization of Po by microorganisms at laboratory culture experiments

    International Nuclear Information System (INIS)

    Momoshima, N.; Ishida, A.; Yoshinaga, C.; Fukuda, A.

    2005-01-01

    The previous experiments proved the volatility of polonium form culture medium in which microorganisms were propagated from seed of seawater, river water or pond water, therefore we did not know what kind of species are responsible to Po volatility. To search microorganisms, which concerned with Po emission we carried out culture experiments using known microorganisms. Three microorganisms were examined; Escherichia coli K-12, Bacillus subtilis and Chromobacterium violaceum. The microorganisms were pre-cultured in LB medium at 30 degree C and a small portion of the pre-cultured was transferred to a culture bottle in which LB medium and 208 Po tracer were contained. The culture was done at 30 degree C with shaking the culture bottle and air passed through a filter was introduced. The Po volatilized was transferred into the trap vials in which scintillator for liquid scintillation counting (LSC) was contained. The Po activity was measured by LSC. All of the microorganisms examined volatilized Po but their ability was quite different each other. Highest ability was observed on Chromobacterium violaceum and then Escherichia coli K-12 followed by Bacillus subtilis, the relative magnitude of the ability was 10 2 , 10, 1, respectively. Chromobacterium violaceum and Escherichia coli K-12 showed high volatility for the first 24 h but Escherichia coli K-12 showed a decrease thereafter. However high volatility was continued on Chromobacterium violaceum during the culture. The low culture temperature suppressed Po volatility, supporting biologically mediated Po emission from the culture.

  16. Functional characterisation of germinant receptors in Clostridium botulinum and Clostridium sporogenes presents novel insights into spore germination systems.

    Science.gov (United States)

    Brunt, Jason; Plowman, June; Gaskin, Duncan J H; Itchner, Manoa; Carter, Andrew T; Peck, Michael W

    2014-09-01

    Clostridium botulinum is a dangerous pathogen that forms the highly potent botulinum toxin, which when ingested causes a deadly neuroparalytic disease. The closely related Clostridium sporogenes is occasionally pathogenic, frequently associated with food spoilage and regarded as the non-toxigenic equivalent of Group I C. botulinum. Both species form highly resistant spores that are ubiquitous in the environment and which, under favourable growth conditions germinate to produce vegetative cells. To improve the control of botulinum neurotoxin-forming clostridia, it is imperative to comprehend the mechanisms by which spores germinate. Germination is initiated following the recognition of small molecules (germinants) by a specific germinant receptor (GR) located in the spore inner membrane. The present study precisely defines clostridial GRs, germinants and co-germinants. Group I C. botulinum ATCC3502 contains two tricistronic and one pentacistronic GR operons, while C. sporogenes ATCC15579 has three tricistronic and one tetracistronic GR operons. Insertional knockout mutants, allied with characterisation of recombinant GRs shows for the first time that amino acid stimulated germination in C. botulinum requires two tri-cistronic encoded GRs which act in synergy and cannot function individually. Spore germination in C. sporogenes requires one tri-cistronic GR. Two other GRs form part of a complex involved in controlling the rate of amino-acid stimulated germination. The suitability of using C. sporogenes as a substitute for C. botulinum in germination studies and food challenge tests is discussed.

  17. Microbiological analysis of coliforms and mesophilic aerobic spore formers in gamma irradiated cocoa beans (Theobroma cacao L.)

    International Nuclear Information System (INIS)

    Barata, Anderson Demetrio; Mansur Netto, Elias

    1995-01-01

    The presence of coliforms in processed foods is an useful indicator of post-sanitization and post processing contamination, and members of the mesophilic aerobic spore formers have great importance in food spoilage. Spore - forming aerobic bacilli have been observed in fermenting cocoa in Jamaica and West Africa. The results of this work has shown a considerable reduction of the mesophilic aerobic spore formers in irradiated Brazilian Comun Cocoa beans as long as the irradiation dose was increased from 1.05 to 3.99 kGy. The presence of coliforms irradiated has not been found even in the coroa beans with the dose of 1.05 kGy. (author). 4 refs., 1 tab

  18. Germination Requirements of Bacillus macerans Spores

    Science.gov (United States)

    Sacks, L. E.; Thompson, P. A.

    1971-01-01

    2-Phenylacetamide is an effective germinant for spores of five strains of Bacillus macerans, particularly in the presence of fructose. Benzyl penicillin, the phenyl acetamide derivative of penicillin, and phenylacetic acid are also good germinants. l-Asparagine is an excellent germinant for four strains. α-Amino-butyric acid is moderately effective. Pyridoxine, pyridoxal, adenine, and 2,6-diaminopurine are potent germinants for NCA strain 7X1 only. d-Glucose is a powerful germinant for strain B-70 only. d-Fructose and d-ribose strongly potentiate germination induced by other germinants (except l-asparagine) but have only weak activity by themselves. Niacinamide and nicotinamide-adenine dinucleotide, inactive by themselves, are active in the presence of fructose or ribose. Effects of pH, ion concentration, and temperature are described. PMID:4251279

  19. Using Spores for Fusarium spp. Classification by MALDI-Based Intact Cell/Spore Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Wolfgang Winkler

    2012-01-01

    Full Text Available Fusarium is a widespread genus of filamentous fungi and a member of the soil microbial community. Certain subspecies are health threatening because of their mycotoxin production that affects the human and animal food chain. Thus, for early and effective pest control, species identification is of particular interest; however, differentiation on the subspecies level is challenging and time-consuming for this fungus. In the present study, we show the possibilities of intact cell mass spectrometry for spore analysis of 22 different Fusarium strains belonging to six Fusarium subspecies. We found that species differentiation is possible if mass spectrometric analyses are performed under well-defined conditions with fixed parameters. A critical point for analysis is a proper sample preparation of spores, which increases the quality of mass spectra with respect to signal intensity and m/z value variations. It was concluded that data acquistion has to be performed automatically; otherwise, user-specific variations are introduced generating data which cannot fit the existing datasets. Data that show clearly that matrix-assisted laser desorption ionization-based intact cell/intact spore mass spectrometry (IC/ISMS can be applied to differentiate closely related Fusarium spp. are presented. Results show a potential to build a database on Fusarium species for accurate species identification, for fast response in the case of infections in the cornfield. We furthermore demonstrate the high precision of our approach in classification of intact Fusarium species according to the location of their collection.

  20. Microorganisms .

    African Journals Online (AJOL)

    ) and heat/pH-shift treatments. This technique resulted in 47% enzyme yield with a purification fac- tor of 12. Technique II which involved two extraction steps by' aqueous two - phase system. (APS) coupled with UF resulted in 62 % enzyme ...

  1. Germination and inactivation of Bacillus coagulans and Alicyclobacillus acidoterrestris spores by high hydrostatic pressure treatment in buffer and tomato sauce.

    Science.gov (United States)

    Vercammen, Anne; Vivijs, Bram; Lurquin, Ine; Michiels, Chris W

    2012-01-16

    Acidothermophilic bacteria like Alicyclobacillus acidoterrestris and Bacillus coagulans can cause spoilage of heat-processed acidic foods because they form spores with very high heat resistance and can grow at low pH. The objective of this work was to study the germination and inactivation of A. acidoterrestris and B. coagulans spores by high hydrostatic pressure (HP) treatment at temperatures up to 60°C and both at low and neutral pH. In a first experiment, spores suspended in buffers at pH 4.0, 5.0 and 7.0 were processed for 10min at different pressures (100-800MPa) at 40°C. None of these treatments caused any significant inactivation, except perhaps at 800MPa in pH 4.0 buffer where close to 1 log inactivation of B. coagulans was observed. Spore germination up to about 2 log was observed for both bacteria but occurred mainly in a low pressure window (100-300MPa) for A. acidoterrestris and only in a high pressure window (600-800MPa) for B. coagulans. In addition, low pH suppressed germination in A. acidoterrestris, but stimulated it in B. coagulans. In a second series of experiments, spores were treated in tomato sauce of pH 4.2 and 5.0 at 100 - 800MPa at 25, 40 and 60°C for 10min. At 40°C, results for B. coagulans were similar as in buffer. For A. acidoterrestris, germination levels in tomato sauce were generally higher than in buffer, and showed little difference at low and high pressure. Remarkably, the pH dependence of A. acidoterrestris spore germination was reversed in tomato sauce, with more germination at the lowest pH. Furthermore, HP treatments in the pH 4.2 sauce caused between 1 and 1.5 log inactivation of A. acidoterrestris. Germination of spores in the high pressure window was strongly temperature dependent, whereas germination of A. acidoterrestris in the low pressure window showed little temperature dependence. When HP treatment was conducted at 60°C, most of the germinated spores were also inactivated. For the pH 4.2 tomato sauce, this

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

    Science.gov (United States)

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

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

  3. Water Behavior in Bacterial Spores by Deuterium NMR Spectroscopy

    Science.gov (United States)

    2015-01-01

    Dormant bacterial spores are able to survive long periods of time without nutrients, withstand harsh environmental conditions, and germinate into metabolically active bacteria when conditions are favorable. Numerous factors influence this hardiness, including the spore structure and the presence of compounds to protect DNA from damage. It is known that the water content of the spore core plays a role in resistance to degradation, but the exact state of water inside the core is a subject of discussion. Two main theories present themselves: either the water in the spore core is mostly immobile and the core and its components are in a glassy state, or the core is a gel with mobile water around components which themselves have limited mobility. Using deuterium solid-state NMR experiments, we examine the nature of the water in the spore core. Our data show the presence of unbound water, bound water, and deuterated biomolecules that also contain labile deuterons. Deuterium–hydrogen exchange experiments show that most of these deuterons are inaccessible by external water. We believe that these unreachable deuterons are in a chemical bonding state that prevents exchange. Variable-temperature NMR results suggest that the spore core is more rigid than would be expected for a gel-like state. However, our rigid core interpretation may only apply to dried spores whereas a gel core may exist in aqueous suspension. Nonetheless, the gel core, if present, is inaccessible to external water. PMID:24950158

  4. Water behavior in bacterial spores by deuterium NMR spectroscopy.

    Science.gov (United States)

    Friedline, Anthony W; Zachariah, Malcolm M; Johnson, Karen; Thomas, Kieth J; Middaugh, Amy N; Garimella, Ravindranath; Powell, Douglas R; Vaishampayan, Parag A; Rice, Charles V

    2014-07-31

    Dormant bacterial spores are able to survive long periods of time without nutrients, withstand harsh environmental conditions, and germinate into metabolically active bacteria when conditions are favorable. Numerous factors influence this hardiness, including the spore structure and the presence of compounds to protect DNA from damage. It is known that the water content of the spore core plays a role in resistance to degradation, but the exact state of water inside the core is a subject of discussion. Two main theories present themselves: either the water in the spore core is mostly immobile and the core and its components are in a glassy state, or the core is a gel with mobile water around components which themselves have limited mobility. Using deuterium solid-state NMR experiments, we examine the nature of the water in the spore core. Our data show the presence of unbound water, bound water, and deuterated biomolecules that also contain labile deuterons. Deuterium-hydrogen exchange experiments show that most of these deuterons are inaccessible by external water. We believe that these unreachable deuterons are in a chemical bonding state that prevents exchange. Variable-temperature NMR results suggest that the spore core is more rigid than would be expected for a gel-like state. However, our rigid core interpretation may only apply to dried spores whereas a gel core may exist in aqueous suspension. Nonetheless, the gel core, if present, is inaccessible to external water.

  5. Practical aspects of temperature intervention in germination and post-germination development of bacterial spores

    International Nuclear Information System (INIS)

    Stastna, J.; Vinter, V.; Babicka, J.

    1974-01-01

    Temperature dependence of germination and post-germination growth was studied in the spores of B a c i l l u s c e r e u s NCIB 8122. It was found that a temperature of 5 0 C slowed down germination, with the cells showing the capacity of synthetizing only a limited amount of proteins. The synthesis of the cellular wall, however, went on for another few hours. Thick-walled, less permeable and less metabolically active cells formed having an altered ultrastructure. A prolonged cultivation at 30 0 C resulted in the reduction of living cells while the low cultivation temperature (5 0 C) was found to have a protective effect. Pre-irradiation with 30g krad of gamma radiation increased the sensitivity of surviving cells to the cultivation conditions. Spores in the post-germination period were found to be much more resistent and alternating use of low and higher temperatures had little effect on growth

  6. Evaluation of the Efficacy of Methyl Bromide in the Decontamination of Building and Interior Materials Contaminated with Bacillus anthracis Spores

    Science.gov (United States)

    Wendling, Morgan; Richter, William; Lastivka, Andrew; Mickelsen, Leroy

    2016-01-01

    The primary goal of this study was to determine the conditions required for the effective inactivation of Bacillus anthracis spores on materials by using methyl bromide (MeBr) gas. Another objective was to obtain comparative decontamination efficacy data with three avirulent microorganisms to assess their potential for use as surrogates for B. anthracis Ames. Decontamination tests were conducted with spores of B. anthracis Ames and Geobacillus stearothermophilus, B. anthracis NNR1Δ1, and B. anthracis Sterne inoculated onto six different materials. Experimental variables included temperature, relative humidity (RH), MeBr concentration, and contact time. MeBr was found to be an effective decontaminant under a number of conditions. This study highlights the important role that RH has when fumigation is performed with MeBr. There were no tests in which a ≥6-log10 reduction (LR) of B. anthracis Ames was achieved on all materials when fumigation was done at 45% RH. At 75% RH, an increase in the temperature, the MeBr concentration, or contact time generally improved the efficacy of fumigation with MeBr. This study provides new information for the effective use of MeBr at temperatures and RH levels lower than those that have been recommended previously. The study also provides data to assist with the selection of an avirulent surrogate for B. anthracis Ames spores when additional tests with MeBr are conducted. PMID:26801580

  7. Phosphate Biomineralization of Cambrian Microorganisms

    Science.gov (United States)

    McKay, David S.; Rozanov, Alexei Yu.; Hoover, Richard B.; Westall, Frances

    1998-01-01

    As part of a long term study of biological markers (biomarkers), we are documenting a variety of features which reflect the previous presence of living organisms. As we study meteorites and samples returned from Mars, our main clue to recognizing possible microbial material may be the presence of biomarkers rather than the organisms themselves. One class of biomarkers consists of biominerals which have either been precipitated directly by microorganisms, or whose precipitation has been influenced by the organisms. Such microbe-mediated mineral formation may include important clues to the size, shape, and environment of the microorganisms. The process of fossilization or mineralization can cause major changes in morphologies and textures of the original organisms. The study of fossilized terrestrial organisms can help provide insight into the interpretation of mineral biomarkers. This paper describes the results of investigations of microfossils in Cambrian phosphate-rich rocks (phosphorites) that were found in Khubsugul, Northern Mongolia.

  8. Antitumor effects and mechanisms of Ganoderma extracts and spores oil

    Science.gov (United States)

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

    2016-01-01

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

  9. 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

  10. Sterilization Resistance of Bacterial Spores Explained with Water Chemistry.

    Science.gov (United States)

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

    2015-11-05

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

  11. 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

  12. 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

  13. Antitumor effects and mechanisms of Ganoderma extracts and spores oil.

    Science.gov (United States)

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

    2016-11-01

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

  14. Microorganism Utilization for Synthetic Milk

    Science.gov (United States)

    Morford, Megan A.; Khodadad, Christina L.; Caro, Janicce I.; Spencer, LaShelle E.; Richards, Jeffery T.; Strayer, Richard F.; Birmele, Michele N.; Wheeler, Raymond M.

    2014-01-01

    A desired architecture for long duration spaceflight, like aboard the International Space Station or for future missions to Mars, is to provide a supply of fresh food crops for the astronauts. However, some crops can create a high proportion of inedible plant waste. The main goal of the Synthetic Biology project, Cow in a Column, was to produce the components of milk (sugar, lipid, protein) from inedible plant waste by utilizing microorganisms (fungi, yeast, bacteria). Of particular interest was utilizing the valuable polysaccharide, cellulose, found in plant waste, to naturally fuel-through microorganism cellular metabolism- the creation of sugar (glucose), lipid (milk fat), and protein (casein) in order to produce a synthetic edible food product. Environmental conditions such as pH, temperature, carbon source, aeration, and choice microorganisms were optimized in the laboratory and the desired end-products, sugars and lipids, were analyzed. Trichoderma reesei, a known cellulolytic fungus, was utilized to drive the production of glucose, with the intent that the produced glucose would serve as the carbon source for milk fat production and be a substitute for the milk sugar lactose. Lipid production would be carried out by Rhodosporidium toruloides, yeast known to accumulate those lipids that are typically found in milk fat. Results showed that glucose and total lipid content were below what was expected during this phase of experimentation. In addition, individual analysis of six fatty acids revealed that the percentage of each fatty acid was lower than naturally produced bovine milk. Overall, this research indicates that microorganisms could be utilized to breakdown inedible solid waste to produce useable products. For future work, the production of the casein protein for milk would require the development of a genetically modified organism, which was beyond the scope of the original project. Additional trials would be needed to further refine the required

  15. Microorganisms as sources of oils

    Directory of Open Access Journals (Sweden)

    Thevenieau France

    2013-11-01

    Full Text Available A number of microorganism belonging to the genera of yeast, fungi, bacteria and microalgae have ability to accumulate substantial amounts of oil, sometimes up to an even in excess of 70% of their biomass weight under specific cultivation conditions. For nearly 100 years, the commercial opportunities of using microorganisms as sources of oils have been continuously examined. Although it was evident that microbial oils could never compete commercially with the major commodity plant oils, there were commercially opportunities for the production of some of the higher valued oils. Today, with the great progress of metabolic and genetic engineering, the developments are focus on the high value oils containing important polyunsaturated or specific fatty acids. Such oils have the potential to be used in different applications area as food, feed and oleochemistry. This review is covering the related researches about different oleaginous microorganisms for lipids production and microbial oils biosynthesis process. In add, the lipid metabolism, metabolic engineering strategies to increase lipid production and the economics of microbial oils production are introduced.

  16. Secondary metabolites from marine microorganisms

    Directory of Open Access Journals (Sweden)

    KELECOM ALPHONSE

    2002-01-01

    Full Text Available After 40 years of intensive research, chemistry of marine natural products has become a mature field. Since 1995, there are signals of decreased interest in the search of new metabolites from traditional sources such as macroalgae and octocorals, and the number of annual reports on marine sponges stabilized. On the contrary, metabolites from microorganisms is a rapidly growing field, due, at least in part, to the suspicion that a number of metabolites obtained from algae and invertebrates may be produced by associated microorganisms. Studies are concerned with bacteria and fungi, isolated from seawater, sediments, algae, fish and mainly from marine invertebrates such as sponges, mollusks, tunicates, coelenterates and crustaceans. Although it is still to early to define tendencies, it may be stated that the metabolites from microorganisms are in most cases quite different from those produced by the invertebrate hosts. Nitrogenated metabolites predominate over acetate derivatives, and terpenes are uncommon. Among the latter, sesquiterpenes, diterpenes and carotenes have been isolated; among nitrogenated metabolites, amides, cyclic peptides and indole alkaloids predominate.

  17. PROBIOTICS BASED ON TRANSGENIC MICROORGANISMS

    Directory of Open Access Journals (Sweden)

    S. А. Starovoitova

    2012-02-01

    Full Text Available Modern tendencies of recombinant microorganisms creation for obtaining on their basis a new effective biopreparations (probiotics with wider spectrum of biological and therapeutic properties were considered. A lot of attention was focused on the main genera of perspective bacteria for creation of recombinant probiotics particularly: Lactococcus, Bifidobac terium,Bacillus, Escherichia. The main created Ukrainian and foreign gene-modified strains, that are widely used today in creation of effective recombinant biopreparations were characterized. Some fundamental directions and methods of gene-modified strains obtaining, which are used in getting effective biopreparations that used for therapy and prophylactic illness were reported, under which this group of pharmaceutical drugs were not used earlier. The safety matters of probiotics using on basis of genemodified strains were examined. Medical and veterinary biopreparations on basis of recombinant microorganisms could be used directly and effectively for therapy and prophylaxis of different illness, beginning from disbacteriosis up to cardiovascular diseases. It is related with some probiotic microorganisms ability for lowering of serum cholesterol at the host organism.

  18. Studying marine microorganisms from space.

    Science.gov (United States)

    Pedrós-Alió, C; Simó, R

    2002-12-01

    Microorganisms are but a few micrometers in diameter and are not visible to the naked eye. Yet, the large numbers of microorganisms present in the oceans and the global impact of their activities make it possible to observe them from space. Here a few examples of how microorganisms can be studied from satellites are presented. The first case is the best known: the main pigment used in photosynthesis (chlorophyll a) can be determined from satellites. These kinds of studies have contributed a tremendous amount of understanding about the distribution and dynamics of primary production in the oceans. Two other examples will concern analysis of heterotrophic prokaryotic production and estimates of dimethyl sulfide (DMS) concentration and flux to the atmosphere. These three processes are of fundamental importance for the functioning of the biosphere. Marine microbes carry out about half of the total primary production in the planet. A substantial fraction of the respiration in the oceans is due to the activity of heterotrophic prokaryotes. Finally, the flux of DMS to the atmosphere is believed to constitute one of the mechanisms by which the biota can regulate climate. The global implications of microbial processes in the oceans can only be addressed with the help of satellites.

  19. Inactivation of microorganisms within collagen gel biomatrices using pulsed electric field treatment.

    Science.gov (United States)

    Griffiths, Sarah; Maclean, Michelle; Anderson, John G; MacGregor, Scott J; Grant, M Helen

    2012-02-01

    Pulsed electric field (PEF) treatment was examined as a potential decontamination method for tissue engineering biomatrices by determining the susceptibility of a range of microorganisms whilst within a collagen gel. High intensity pulsed electric fields were applied to collagen gel biomatrices containing either Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, Candida albicans, Saccharomyces cerevisiae or the spores of Aspergillus niger. The results established varying degrees of microbial PEF susceptibility. When high initial cell densities (10(6)-10(7) CFU ml(-1)) were PEF treated with 100 pulses at 45 kV cm(-1), the greatest log reduction was achieved with S. cerevisiae (~6.5 log(10) CFU ml(-1)) and the lowest reduction achieved with S. epidermidis (~0.5 log(10) CFU ml(-1)). The results demonstrate that inactivation is influenced by the intrinsic properties of the microorganism treated. Further investigations are required to optimise the microbial inactivation kinetics associated with PEF treatment of collagen gel biomatrices.

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

    Directory of Open Access Journals (Sweden)

    Shinkafi Sa’adatu Aliyu

    2014-07-01

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

  1. DRY COMPOSITION OF STARTER CULTURES FORM LACTIC ACID MICROORGANISMS

    Directory of Open Access Journals (Sweden)

    At. Kraevska

    2014-03-01

    Full Text Available The purpose of this work is to investigate the possibility of producing of lyophilized lactic acid starter Lb. plantarum strain 226/1, designed for a starter culture in the production of pickles. The results of our studies demonstrate achievement of this goal by the specified process parameters.

  2. Fossil Microorganisms in Archaean deposits of Northern Karelia

    Science.gov (United States)

    Astafieva, M. M.; Hoover, R. B.; Rozanov, A. Y.; Vrevskiy, A. B.

    2005-01-01

    Newly found biomorphic microstructures from the Upper Archaean (lopian) rocks from Northern Karelia are described. The presence of various microorganisms of bacterial nature and even cyanobacteria (and possibly eukaryotic forms) is suggested. The necessity of employing methods of electron microscopy, as well as traditional methods, while studying the very early manifestations of life in Archaean and Early Proterozoic is noted.

  3. Mitigating cyanobacterial blooms: how effective are 'effective microorganisms'?

    NARCIS (Netherlands)

    Lürling, M.F.L.L.W.; Tolman, Y.; Euwe, M.

    2009-01-01

    This study examined the effects of 'Effective Microorganisms (EM)' on the growth of cyanobacteria, and their ability to terminate cyanobacterial blooms. The EM was tested in the form of 'mudballs' or 'Bokashi-balls', and as a suspension (EM-A) in laboratory experiments. No growth inhibition was

  4. Microsphere fluoroimmunoassay for microorganisms: An update

    Energy Technology Data Exchange (ETDEWEB)

    Whitten, W.B.; Ramsey, J.M. [Oak Ridge National Lab., TN (United States); Bronk, B.V. [Edgewood Research, Development and Engineering Center, Aberdeen Proving Ground, MD (United States)

    1994-12-31

    Microspheres are used as labels in a fluorescence immunoassay for individual microorganisms. The diameter of a sphere that has reacted with microorganisms is determined from measurements of the optical resonance frequencies. The spheres have been coated with antibodies so that each microsphere diameter corresponds to a different species of microorganism. Further experiments on specificity and on optical resonance measurements are presented.

  5. 40 CFR 725.85 - Microorganism identity.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Microorganism identity. 725.85 Section... to Information § 725.85 Microorganism identity. (a) Claims applicable to the period prior to... specific microorganism identity at the time of submission of the information. This claim will apply only to...

  6. Development of a selective medium for the determination of the spore concentrations of Botrytis cinerea in the air.

    Science.gov (United States)

    Gielen, S; Aerts, R; Seels, B

    2003-01-01

    Gray mold, caused by Botrytis cinerea, is an important disease that causes world-wide extensive damage to a wide range of economically important crops. When it is necessary to determine the spore concentration of Botrytis cinerea in a certain area, it is important to develop a method that can capture the spores of Botrytis cinerea and that can identify them. For the identification and enumeration of the plant pathogen Botrytis cinerea in the environment the easiest method available for the moment is the use of a selective medium. Several selective media for the isolation of Botrytis spp. have been developed by other research groups. All these media contain fungicides that are usually non-toxic towards Botrytis species and tannic acid, which is oxidized to produce a brown pigment that visualises the growth of Botrytis cinerea on the selective media. It seemed that different isolates of Botrytis cinerea that are found in nature have different sensitivities towards the different fungicide concentrations that are used in the selective media. Making the "optimal" selective media for Botrytis cinerea, we have to take in consideration that so many as possible Botrytis cinerea isolates must be able to germinate and grow on this selective medium and that the contamination of other micro-organisms on the selective medium must be minimized. Before the final composition of our selective medium for Botrytis cinerea, different combinations of fungicide concentrations were tried out of the following three fungicides: Rubigan, maneb and PCNB (pentachloronitrobenzene). All these selective media with different fungicides concentrations were tested out for spore germination and mycelium growth of Botrytis cinerea. Because it was obvious that the percentage Botrytis cinerea that germinated on PDA (Potato Dextrose Agar) was higher than on the selective medium a few experiments were executed in which the percentage of spore germination on PDA was compared with the percentage of spore

  7. Inactivation Strategies for Clostridium perfringens Spores and Vegetative Cells.

    Science.gov (United States)

    Talukdar, Prabhat K; Udompijitkul, Pathima; Hossain, Ashfaque; Sarker, Mahfuzur R

    2017-01-01

    Clostridium perfringens is an important pathogen to human and animals and causes a wide array of diseases, including histotoxic and gastrointestinal illnesses. C. perfringens spores are crucial in terms of the pathogenicity of this bacterium because they can survive in a dormant state in the environment and return to being live bacteria when they come in contact with nutrients in food or the human body. Although the strategies to inactivate C. perfringens vegetative cells are effective, the inactivation of C. perfringens spores is still a great challenge. A number of studies have been conducted in the past decade or so toward developing efficient inactivation strategies for C. perfringens spores and vegetative cells, which include physical approaches and the use of chemical preservatives and naturally derived antimicrobial agents. In this review, different inactivation strategies applied to control C. perfringens cells and spores are summarized, and the potential limitations and challenges of these strategies are discussed. Copyright © 2016 American Society for Microbiology.

  8. Analysis of Bacillus Globigii Spores Using the BioDetector

    National Research Council Canada - National Science Library

    Lee, William

    1999-01-01

    .... An automated immunoassay instrument capable of providing rapid identification of biological agents was used to analyses laboratory and field trial samples containing the field trial simulants Bacillus globigii (BG) spores...

  9. Small Probes for Orbital Return of Experiments (SPORE), Phase I

    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...

  10. 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...

  11. Detection of Bacillus anthracis spores by super-paramagnetic lateral-flow immunoassays based on "Road Closure".

    Science.gov (United States)

    Wang, Dian-Bing; Tian, Bo; Zhang, Zhi-Ping; Wang, Xu-Ying; Fleming, Joy; Bi, Li-Jun; Yang, Rui-Fu; Zhang, Xian-En

    2015-05-15

    Detection of Bacillus anthracis in the field, whether as a natural infection or as a biothreat remains challenging. Here we have developed a new lateral-flow immunochromatographic assay (LFIA) for B. anthracis spore detection based on the fact that conjugates of B. anthracis spores and super-paramagnetic particles labeled with antibodies will block the pores of chromatographic strips and form retention lines on the strips, instead of the conventionally reported test lines and control lines in classic LFIA. As a result, this new LFIA can simultaneously realize optical, magnetic and naked-eye detection by analyzing signals from the retention lines. As few as 500-700 pure B. anthracis spores can be recognized with CV values less than 8.31% within 5 min of chromatography and a total time of 20 min. For powdery sample tests, this LFIA can endure interference from 25% (w/v) milk, 10% (w/v) baking soda and 10% (w/v) starch without any sample pre-treatment, and has a corresponding detection limit of 6×10(4) spores/g milk powder, 2×10(5) spores/g starch and 5×10(5) spores/g baking soda. Compared with existing methods, this new approach is very competitive in terms of sensitivity, specificity, cost and ease of operation. This proof-of-concept study can also be extended for detection of many other large-sized analytes. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  12. Architecture and Assembly of the Bacillus subtilis Spore Coat

    Science.gov (United States)

    Plomp, Marco; Carroll, Alicia Monroe; Setlow, Peter; Malkin, Alexander J.

    2014-01-01

    Bacillus spores are encased in a multilayer, proteinaceous self-assembled coat structure that assists in protecting the bacterial genome from stresses and consists of at least 70 proteins. The elucidation of Bacillus spore coat assembly, architecture, and function is critical to determining mechanisms of spore pathogenesis, environmental resistance, immune response, and physicochemical properties. Recently, genetic, biochemical and microscopy methods have provided new insight into spore coat architecture, assembly, structure and function. However, detailed spore coat architecture and assembly, comprehensive understanding of the proteomic composition of coat layers, and specific roles of coat proteins in coat assembly and their precise localization within the coat remain in question. In this study, atomic force microscopy was used to probe the coat structure of Bacillus subtilis wild type and cotA, cotB, safA, cotH, cotO, cotE, gerE, and cotE gerE spores. This approach provided high-resolution visualization of the various spore coat structures, new insight into the function of specific coat proteins, and enabled the development of a detailed model of spore coat architecture. This model is consistent with a recently reported four-layer coat assembly and further adds several coat layers not reported previously. The coat is organized starting from the outside into an outermost amorphous (crust) layer, a rodlet layer, a honeycomb layer, a fibrous layer, a layer of “nanodot” particles, a multilayer assembly, and finally the undercoat/basement layer. We propose that the assembly of the previously unreported fibrous layer, which we link to the darkly stained outer coat seen by electron microscopy, and the nanodot layer are cotH- and cotE- dependent and cotE-specific respectively. We further propose that the inner coat multilayer structure is crystalline with its apparent two-dimensional (2D) nuclei being the first example of a non-mineral 2D nucleation crystallization

  13. Architecture and assembly of the Bacillus subtilis spore coat.

    Science.gov (United States)

    Plomp, Marco; Carroll, Alicia Monroe; Setlow, Peter; Malkin, Alexander J

    2014-01-01

    Bacillus spores are encased in a multilayer, proteinaceous self-assembled coat structure that assists in protecting the bacterial genome from stresses and consists of at least 70 proteins. The elucidation of Bacillus spore coat assembly, architecture, and function is critical to determining mechanisms of spore pathogenesis, environmental resistance, immune response, and physicochemical properties. Recently, genetic, biochemical and microscopy methods have provided new insight into spore coat architecture, assembly, structure and function. However, detailed spore coat architecture and assembly, comprehensive understanding of the proteomic composition of coat layers, and specific roles of coat proteins in coat assembly and their precise localization within the coat remain in question. In this study, atomic force microscopy was used to probe the coat structure of Bacillus subtilis wild type and cotA, cotB, safA, cotH, cotO, cotE, gerE, and cotE gerE spores. This approach provided high-resolution visualization of the various spore coat structures, new insight into the function of specific coat proteins, and enabled the development of a detailed model of spore coat architecture. This model is consistent with a recently reported four-layer coat assembly and further adds several coat layers not reported previously. The coat is organized starting from the outside into an outermost amorphous (crust) layer, a rodlet layer, a honeycomb layer, a fibrous layer, a layer of "nanodot" particles, a multilayer assembly, and finally the undercoat/basement layer. We propose that the assembly of the previously unreported fibrous layer, which we link to the darkly stained outer coat seen by electron microscopy, and the nanodot layer are cotH- and cotE- dependent and cotE-specific respectively. We further propose that the inner coat multilayer structure is crystalline with its apparent two-dimensional (2D) nuclei being the first example of a non-mineral 2D nucleation crystallization

  14. Purification and Properties of Clostridium perfringens Spore Lytic Enzymes.

    Science.gov (United States)

    1983-01-01

    reaction mixture at 550C gave the opti- mum response although the activity of initiation protein remained high at 65"C and 75"C. Denaturation of the...CASSIER M. and Sebald M. 1969. Germination Iysozyme-ddpendente des spores de aCnerilim perfringeno ATCC 3624 sprks tralitment thermique . Ann. Inst. Pasteur...activity upon prolonged extraction -of spores in GME was not surprising, since this compound is an active protein denaturant . Urea acts in the same

  15. Compost supplementation with nutrients and microorganisms in composting process.

    Science.gov (United States)

    Sánchez, Óscar J; Ospina, Diego A; Montoya, Sandra

    2017-11-01

    The composting is an aerobic, microorganism-mediated, solid-state fermentation process by which different organic materials are transformed into more stable compounds. The product obtained is the compost, which contributes to the improvement of physical, chemical and microbiological properties of the soil. However, the compost usage in agriculture is constrained because of its long-time action and reduced supply of nutrients to the crops. To enhance the content of nutrients assimilable by the plants in the compost, its supplementation with nutrients and inoculation with microorganisms have been proposed. The objective of this work was to review the state of the art on compost supplementation with nutrients and the role played by the microorganisms involved (or added) in their transformation during the composting process. The phases of composting are briefly compiled and different strategies for supplementation are analyzed. The utilization of nitrogenous materials and addition of microorganisms fixing nitrogen from the atmosphere or oxidizing ammonia into more assimilable for plants nitrogenous forms are analyzed. Several strategies for nitrogen conservation during composting are presented as well. The supplementation with phosphorus and utilization of microorganisms solubilizing phosphorus and potassium are also discussed. Main groups of microorganisms relevant during the composting process are described as well as most important strategies to identify them. In general, the development of this type of nutrient-enriched bio-inputs requires research and development not only in the supplementation of compost itself, but also in the isolation and identification of microorganisms and genes allowing the degradation and conversion of nitrogenous substances and materials containing potassium and phosphorus present in the feedstocks undergoing the composting process. In this sense, most important research trends and strategies to increase nutrient content in the compost

  16. Comparison of the disinfection effects of vacuum-UV (VUV) and UV light on Bacillus subtilis spores in aqueous suspensions at 172, 222 and 254 nm.

    Science.gov (United States)

    Wang, Ding; Oppenländer, Thomas; El-Din, Mohamed Gamal; Bolton, James R

    2010-01-01

    The efficacy of UV and vacuum-UV (VUV) disinfection of Bacillus subtilis spores in aqueous suspensions at wavelengths of 172, 222 and 254 nm was evaluated. A Xe(2)* excilamp, a KrCl* excilamp and a low-pressure mercury lamp were used as almost monochromatic light sources at these three wavelengths. The first-order inactivation rate constants at 172, 222 and 254 nm were 0.0023, 0.122 and 0.069 cm(2) mJ(-1), respectively. Therefore, a 2 log reduction of B. subtilis spores was reached with fluences (UV doses) of 870, 21.6 and 40.4 mJ cm(-2) at these individual wavelengths. Consequently, for the inactivation of B. subtilis spores, VUV exposure at 172 nm is much less efficient than exposure at the other two wavelengths, while exposure at 222 nm is more efficient than that at 254 nm, which is probably because triplet energy transfer from DPA to thymine bases at 222 nm is higher than that at 254 nm. This research indicated quantitatively that VUV light is not practicable for microorganism disinfection in water and wastewater treatment. However, in comparison with other advanced oxidation processes (e.g. UV/TiO(2), UV/H(2)O(2) or O(3)/H(2)O(2)) the VUV-initiated photolysis of water is likely more efficient in generating hydroxyl radicals and more effective for the inactivation of microorganisms.

  17. 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.

  18. Tip-enhanced Raman scattering of bacillus subtilis spores

    Science.gov (United States)

    Rusciano, G.; Zito, G.; Pesce, G.; Sasso, A.; Isticato, R.; Ricca, E.

    2015-07-01

    Understanding of the complex interactions of molecules at biological interfaces is a fundamental issue in biochemistry, biotechnology as well as biomedicine. A plethora of biological processes are ruled by the molecular texture of cellular membrane: cellular communications, drug transportations and cellular recognition are just a few examples of such chemically-mediated processes. Tip-Enhanced Raman Scattering (TERS) is a novel, Raman-based technique which is ideally suited for this purpose. TERS relies on the combination of scanning probe microscopy and Raman spectroscopy. The basic idea is the use of a metalled tip as a sort of optical nano-antenna, which gives place to SERS effect close to the tip end. Herein, we present the application of TERS to analyze the surface of Bacillus subtilis spores. The choice of this biological systems is related to the fact that a number of reasons support the use of spores as a mucosal delivery system. The remarkable and well-documented resistance of spores to various environmental and toxic effects make them clear potentials as a novel, surface-display system. Our experimental outcomes demonstrate that TERS is able to provide a nano-scale chemical imaging of spore surface. Moreover, we demonstrate that TERS allows differentiation between wilde-type spore and genetically modified strains. These results hold promise for the characterization and optimization of spore surface for drug-delivery applications.

  19. Reversible Hydrolysis Reaction with the Spore Photoproduct under Alkaline Conditions.

    Science.gov (United States)

    Adhikari, Surya; Lin, Gengjie; Li, Lei

    2016-09-16

    DNA lesions may reduce the electron density at the nucleobases, making them prone to further modifications upon the alkaline treatment. The dominant DNA photolesion found in UV-irradiated bacterial endospores is a thymine dimer, 5-thyminyl-5,6-dihydrothymine, i.e., the spore photoproduct (SP). Here we report a stepwise addition/elimination reaction in the SP hydrolysis product under strong basic conditions where a ureido group is added to the carboxyl moiety to form a cyclic amide, regenerating SP after eliminating a hydroxide ion. Direct amidation of carboxylic acids by reaction with amines in the presence of a catalyst is well documented; however, it is very rare for an amidation reaction to occur without activation. This uncatalyzed SP reverse reaction in aqueous solution is even more surprising because the carboxyl moiety is not a good electrophile due to the negative charge it carries. Examination of the base-catalyzed hydrolyses of two other saturated pyrimidine lesions, 5,6-dihydro-2'-deoxyuridine and pyrimidine (6-4) pyrimidone photoproduct, reveals that neither reaction is reversible even though all three hydrolysis reactions may share the same gem-diol intermediate. Therefore, the SP structure where the two thymine residues maintain a stacked conformation likely provides the needed framework enabling this highly unusual carboxyl addition/elimination reaction.

  20. 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 study, we tested the hypothesis that maternal effects on offspring production and quality are greater than paternal effects in both offspring number (fertility) and offspring viability (mortality). We used the model filamentous fungus Neurospora crassa. This fungus is anisogamous......, 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...

  1. Is arsenic biotransformation a detoxification mechanism for microorganisms?

    Energy Technology Data Exchange (ETDEWEB)

    Rahman, M. Azizur, E-mail: Mohammad.Rahman@uts.edu.au [Centre for Environmental Sustainability, School of the Environment, Faculty of Science, University of Technology, P.O. Box 123, Broadway, Sydney, NSW 2007 (Australia); Hassler, Christel [Marine and Lake Biogeochemistry, Institute F. A. Forel, University of Geneva, 10 rte de Suisse, Versoix, 1290 Switzerland (Switzerland)

    2014-01-15

    Arsenic (As) is extremely toxic to living organisms at high concentration. In aquatic systems, As exists in different chemical forms. The two major inorganic As (iAs) species are As{sup V}, which is thermodynamically stable in oxic waters, and As{sup III}, which is predominant in anoxic conditions. Photosynthetic microorganisms (e.g., phytoplankton and cyanobacteria) take up As{sup V}, biotransform it to As{sup III}, then biomethylate it to methylarsenic (MetAs) forms. Although As{sup III} is more toxic than As{sup V}, As{sup III} is much more easily excreted from the cells than As{sup V}. Therefore, majority of researchers consider the reduction of As{sup V} to As{sup III} as a detoxification process. The biomethylation process results in the conversion of toxic iAs to the less toxic pentavalent MetAs forms (monomethylarsonate; MMA{sup V}, dimethylarsonate; DMA{sup V}, and trimethylarsenic oxide; TMAO{sup V}) and trimethylarsine (TMAO{sup III}). However, biomethylation by microorganisms also produces monomethylarsenite (MMA{sup III}) and dimethylarsenite (DMA{sup III}), which are more toxic than iAs, as a result of biomethylation by the microorganisms, demonstrates the need to reconsider to what extent As biomethylation contributes to a detoxification process. In this review, we focused on the discussion of whether the biotransformation of As species in microorganisms is really a detoxification process with recent data.

  2. Is arsenic biotransformation a detoxification mechanism for microorganisms?

    International Nuclear Information System (INIS)

    Rahman, M. Azizur; Hassler, Christel

    2014-01-01

    Arsenic (As) is extremely toxic to living organisms at high concentration. In aquatic systems, As exists in different chemical forms. The two major inorganic As (iAs) species are As V , which is thermodynamically stable in oxic waters, and As III , which is predominant in anoxic conditions. Photosynthetic microorganisms (e.g., phytoplankton and cyanobacteria) take up As V , biotransform it to As III , then biomethylate it to methylarsenic (MetAs) forms. Although As III is more toxic than As V , As III is much more easily excreted from the cells than As V . Therefore, majority of researchers consider the reduction of As V to As III as a detoxification process. The biomethylation process results in the conversion of toxic iAs to the less toxic pentavalent MetAs forms (monomethylarsonate; MMA V , dimethylarsonate; DMA V , and trimethylarsenic oxide; TMAO V ) and trimethylarsine (TMAO III ). However, biomethylation by microorganisms also produces monomethylarsenite (MMA III ) and dimethylarsenite (DMA III ), which are more toxic than iAs, as a result of biomethylation by the microorganisms, demonstrates the need to reconsider to what extent As biomethylation contributes to a detoxification process. In this review, we focused on the discussion of whether the biotransformation of As species in microorganisms is really a detoxification process with recent data

  3. Mini-review: Inhibition of biofouling by marine microorganisms.

    Science.gov (United States)

    Dobretsov, Sergey; Abed, Raeid M M; Teplitski, Max

    2013-01-01

    Any natural or artificial substratum exposed to seawater is quickly fouled by marine microorganisms and later by macrofouling species. Microfouling organisms on the surface of a substratum form heterogenic biofilms, which are composed of multiple species of heterotrophic bacteria, cyanobacteria, diatoms, protozoa and fungi. Biofilms on artificial structures create serious problems for industries worldwide, with effects including an increase in drag force and metal corrosion as well as a reduction in heat transfer efficiency. Additionally, microorganisms produce chemical compounds that may induce or inhibit settlement and growth of other fouling organisms. Since the last review by the first author on inhibition of biofouling by marine microbes in 2006, significant progress has been made in the field. Several antimicrobial, antialgal and antilarval compounds have been isolated from heterotrophic marine bacteria, cyanobacteria and fungi. Some of these compounds have multiple bioactivities. Microorganisms are able to disrupt biofilms by inhibition of bacterial signalling and production of enzymes that degrade bacterial signals and polymers. Epibiotic microorganisms associated with marine algae and invertebrates have a high antifouling (AF) potential, which can be used to solve biofouling problems in industry. However, more information about the production of AF compounds by marine microorganisms in situ and their mechanisms of action needs to be obtained. This review focuses on the AF activity of marine heterotrophic bacteria, cyanobacteria and fungi and covers publications from 2006 up to the end of 2012.

  4. Inactivation of Spores of Bacillus Species by Wet Heat: Studies on Single Spores Using Laser Tweezers Taman Spectroscopy

    Science.gov (United States)

    2013-02-01

    13/2011 22.00 Keren K. Griffiths, Jingqiao Zhang, Ann E. Cowan, Ji Yu, Peter Setlow. Germination proteins in the inner membrane of dormant Bacillus...that this technique can be used to rapidly identify single airborne particles or bacteria collected on a slide and to monitor germination dynamics of...the environment of dipicolinic acid in the core of superdormant spores is different from that in dormant spores [J. Bacteriol., 191, 5584 (2009

  5. Improvement of Biological Indicators by Uniformly Distributing Bacillus subtilis Spores in Monolayers To Evaluate Enhanced Spore Decontamination Technologies.

    Science.gov (United States)

    Raguse, Marina; Fiebrandt, Marcel; Stapelmann, Katharina; Madela, Kazimierz; Laue, Michael; Lackmann, Jan-Wilm; Thwaite, Joanne E; Setlow, Peter; Awakowicz, Peter; Moeller, Ralf

    2016-01-22

    Novel decontamination technologies, including cold low-pressure plasma and blue light (400 nm), are promising alternatives to conventional surface decontamination methods. However, the standardization of the assessment of such sterilization processes remains to be accomplished. Bacterial endospores of the genera Bacillus and Geobacillus are frequently used as biological indicators (BIs) of sterility. Ensuring standardized and reproducible BIs for reliable testing procedures is a significant problem in industrial settings. In this study, an electrically driven spray deposition device was developed, allowing fast, reproducible, and homogeneous preparation of Bacillus subtilis 168 spore monolayers on glass surfaces. A detailed description of the structural design as well as the operating principle of the spraying device is given. The reproducible formation of spore monolayers of up to 5 × 10(7) spores per sample was verified by scanning electron microscopy. Surface inactivation studies revealed that monolayered spores were inactivated by UV-C (254 nm), low-pressure argon plasma (500 W, 10 Pa, 100 standard cubic cm per min), and blue light (400 nm) significantly faster than multilayered spores were. We have thus succeeded in the uniform preparation of reproducible, highly concentrated spore monolayers with the potential to generate BIs for a variety of nonpenetrating surface decontamination techniques. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  6. Inactivation of Bacterial Spores and Vegetative Bacterial Cells by Interaction with ZnO-Fe2O3 Nanoparticles and UV Radiation

    Directory of Open Access Journals (Sweden)

    José Luis Sánchez-Salas

    2017-09-01

    Full Text Available ZnO-Fe2O3 nanoparticles (ZnO-Fe NPs were synthesized and characterized by scanning electron microscopy (SEM, energy dispersive X-ray spectroscopy (EDS and dynamic light scattering (DLS. The generation of chemical reactive hydroxyl radicals (•OH was measured spectrophotometrically (UV-Vis by monitoring of p-nitrosodimethylaniline (pNDA bleaching. Inactivation of E. coli and B. subtilis spores in the presence of different concentrations of ZnO-Fe NPs, under UV365nm or visible radiation, was evaluated. We observed the best results under visible light, of which inactivation of E. coli of about 90% was accomplished in 30 minutes, while B. subtilis inactivation close to 90% was achieved in 120 minutes. These results indicate that the prepared photocatalytic systems are promising for improving water quality by reducing the viability of water-borne microorganisms, including bacterial spores.

  7. Airway inflammation among compost workers exposed to actinomycetes spores.

    Science.gov (United States)

    Heldal, Kari Kulvik; Madsø, Lene; Eduard, Wijnand

    2015-01-01

    To study the associations between exposure to bioaerosols and work-related symptoms, lung function and biomarkers of airway inflammation in compost workers. Personal full-shift exposure measurements were performed on 47 workers employed at five windrow plants (n=20) and five reactor plants (n=27). Samples were analyzed for endotoxins, bacteria, fungal and actinomycetes spores. Health examinations were performed on workers and 37 controls before and after work on the day exposure was measured. The examinations included symptoms recorded by questionnaire, lung function by spirometry and nasal dimensions by acoustic rhinometry (AR). The pneumoproteins CC16, SP-D and SP-A were measured in a blood sample drawn at the end of the day. The levels of endotoxins (median 3 EU/m(3), range 0-730 EU/m(3)) and actinomycetes spores (median 0.2 × 10(6) spores/m(3), range 0-590 × 10(6) spores/m(3)) were significantly higher in reactor plants compared to windrow plants. However, windrow composting workers reported more symptoms than reactor composting workers, probably due to use of respiratory protection. Exposure-response relationships between actinomycetes spores exposure and respiratory effects, found as cough and nose irritation during a shift, was significantly increased (OR 4.3, 95% CI 1.1-16, OR 6.1, 95% CI 1.5-25, respectively, pactinomycetes spores/m3, and FEV1/FVC% decreased cross shift (b=-3.2, SE=1.5%, pactinomycetes spores which was associated with work related cough symptoms and work-shift lung function decrease.

  8. The sensitivity of the most common microorganisms of pig liquid manure to the effect of 60Co irradiation

    International Nuclear Information System (INIS)

    Szemeredy, Gy.; Simon, J.

    1979-01-01

    In model experiments the most common representatives of pathogens and facultative pathogens of pig liquid manure contaminating bacterial flora were examined for their sensitivity to different doses of 60 Co, using liquid medium supplemented with 1% glucose, as well as Korthof-medium and different kinds of selective media. Erysipelothrix rhusiopathiea, Brucella suis, Mycoplasma hyorhinis and Pasteurella haemolytica proved to be the most senstive ones to irradiation, as they did not survive 100 krad dose. The highest resistances were observed in the cases of the following microorganisms: the spore containing Bacillus anthracis, killed by 700 krad irradiation dose, the saprophytic and pathogenic Mycobacteria and the spore containing Clostridium perfringens, killed by 1000-1500 krad irradiation. (author)

  9. 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.

  10. A comparative study of the disinfection efficacy of H2O2/ferrate and UV/H2O2/ferrate processes on inactivation of Bacillus subtilis spores by response surface methodology for modeling and optimization.

    Science.gov (United States)

    Matin, Atiyeh Rajabi; Yousefzadeh, Samira; Ahmadi, Ehsan; Mahvi, Amirhossein; Alimohammadi, Mahmood; Aslani, Hassan; Nabizadeh, Ramin

    2018-04-03

    Although chlorination can inactivate most of the microorganisms in water but protozoan parasites like C. parvum oocysts and Giardia cysts can resist against it. Therefore, many researches have been conducted to find a novel method for water disinfection. Present study evaluated the synergistic effect of H2O2 and ferrate followed by UV radiation to inactivate Bacillus subtilis spores as surrogate microorganisms. Response surface methodology(RSM) was employed for the optimization for UV/H2O2/ferrate and H2O2/ferrate processes. By using central composite design(CCD), the effect of three main parameters including time, hydrogen peroxide, and ferrate concentrations was examined on process performance. The results showed that the combination of UV, H2O2 and ferrate was the most effective disinfection process in compare with when H2O2 and ferrate were used. This study indicated that by UV/H2O2/ferrate, about 5.2 log reductions of B. subtilis spores was inactivated at 9299 mg/l of H2O2 and 0.4 mg/l of ferrate concentrations after 57 min of contact time which was the optimum condition, but H2O2/ferrate can inactivate B. subtilis spores about 4.7 logs compare to the other process. Therefore, the results of this research demonstrated that UV/H2O2 /ferrate process is a promising process for spore inactivation and water disinfection. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. Desulfotomaculum arcticum sp nov., a novel spore-formin, moderately thermophilic, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard

    DEFF Research Database (Denmark)

    Vandieken, V.; Knoblauch, C.; Jørgensen, BB

    2006-01-01

    Strain 15 T is a novel spore-forming, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard. Sulfate could be replaced by sulfite or thiosulfate. Hydrogen, formate, lactate, propionate, butyrate, hexanoate, methanol, ethanol, propanol, butanol, pyruvate, malate...... and optimal growth occurred at 44 degrees C. Therefore, strain 15 T apparently cannot grow at in situ temperatures of Arctic sediments from where it was isolated, and it was proposed that it was present in the sediment in the form of spores. The DNA G+C content was 48(.)9 mol%. Strain 15 T was most closely...

  12. Functional Basis of Microorganism Classification.

    Science.gov (United States)

    Zhu, Chengsheng; Delmont, Tom O; Vogel, Timothy M; Bromberg, Yana

    2015-08-01

    Correctly identifying nearest "neighbors" of a given microorganism is important in industrial and clinical applications where close relationships imply similar treatment. Microbial classification based on similarity of physiological and genetic organism traits (polyphasic similarity) is experimentally difficult and, arguably, subjective. Evolutionary relatedness, inferred from phylogenetic markers, facilitates classification but does not guarantee functional identity between members of the same taxon or lack of similarity between different taxa. Using over thirteen hundred sequenced bacterial genomes, we built a novel function-based microorganism classification scheme, functional-repertoire similarity-based organism network (FuSiON; flattened to fusion). Our scheme is phenetic, based on a network of quantitatively defined organism relationships across the known prokaryotic space. It correlates significantly with the current taxonomy, but the observed discrepancies reveal both (1) the inconsistency of functional diversity levels among different taxa and (2) an (unsurprising) bias towards prioritizing, for classification purposes, relatively minor traits of particular interest to humans. Our dynamic network-based organism classification is independent of the arbitrary pairwise organism similarity cut-offs traditionally applied to establish taxonomic identity. Instead, it reveals natural, functionally defined organism groupings and is thus robust in handling organism diversity. Additionally, fusion can use organism meta-data to highlight the specific environmental factors that drive microbial diversification. Our approach provides a complementary view to cladistic assignments and holds important clues for further exploration of microbial lifestyles. Fusion is a more practical fit for biomedical, industrial, and ecological applications, as many of these rely on understanding the functional capabilities of the microbes in their environment and are less concerned with

  13. Comparison of four commercial DNA extraction kits for the recovery of Bacillus spp. spore DNA from spiked powder samples.

    Science.gov (United States)

    Mölsä, Markos; Kalin-Mänttäri, Laura; Tonteri, Elina; Hemmilä, Heidi; Nikkari, Simo

    2016-09-01

    Bacillus spp. include human pathogens such as Bacillus anthracis, the causative agent of anthrax and a biothreat agent. Bacillus spp. form spores that are physically highly resistant and may remain active over sample handling. We tested four commercial DNA extraction kits (QIAamp DNA Mini Kit, RTP Pathogen Kit, ZR Fungal/Bacterial DNA MiniPrep, and genesig Easy DNA/RNA Extraction kit) for sample inactivation and DNA recovery from two powders (icing sugar and potato flour) spiked with Bacillus thuringiensis spores. The DNA was analysed using a B. thuringiensis-specific real-time PCR assay. The detection limit was 3×10(1)CFU of spiked B. thuringiensis spores with the QIAamp DNA Mini, RTP Pathogen, and genesig Easy DNA/RNA Extraction kits, and 3×10(3)CFU with the ZR Fungal/Bacterial DNA MiniPrep kit. The results showed that manual extraction kits are effective and safe for fast and easy DNA extraction from powder samples even in field conditions. Adding a DNA filtration step to the extraction protocol ensures the removal of Bacillus spp. spores from DNA samples without affecting sensitivity. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Survival of yeast spores in hypervelocity impact events up to velocities of 7.4 km s-1

    Science.gov (United States)

    Price, M. C.; Solscheid, C.; Burchell, M. J.; Josse, L.; Adamek, N.; Cole, M. J.

    2013-01-01

    We report on the survivability in hypervelocity impacts of yeast in spore form, and as mature cultures, at impact velocities from 1 to 7.4 km s-1, corresponding to an estimated peak shock pressure of ˜43 GPa. Spores from a yeast strain (BY4743), deficient in an enzyme required for uracil production, were fired into water (to simulate oceanic impact from space) using a light gas gun. The water was then retrieved and filtered and the resulting retentate and filtrate cultured to determine viability and survival rates of remnant spores. Yeast growth (confirmed as coming from the original sample as it had the same enzyme deficiency) was found in recovered samples at all impact speeds, albeit in smaller quantities at the higher speeds. The survival probabilities were measured as ˜50% at 1 km s-1, falling to ˜10-3% at 7.4 km s-1. This follows the pattern observed in previous work on survival of microbial life and spores exposed to extreme shock loading, where there is reasonable survival at low peak shock pressures with more severe lethality above a critical shock pressure at the GPa scale (here between 2 and 10 GPa). These results are explained in the context of a general model for survival against extreme shock and are relevant to the hypotheses of panspermia and litho-panspermia, showing that extreme shocks during transfer across space are not necessarily sterilising.

  15. Fighting Ebola with novel spore decontamination technologies for the military

    Science.gov (United States)

    Doona, Christopher J.; Feeherry, Florence E.; Kustin, Kenneth; Olinger, Gene G.; Setlow, Peter; Malkin, Alexander J.; Leighton, Terrance

    2015-01-01

    Recently, global public health organizations such as Doctors without Borders (MSF), the World Health Organization (WHO), Public Health Canada, National Institutes of Health (NIH), and the U.S. government developed and deployed Field Decontamination Kits (FDKs), a novel, lightweight, compact, reusable decontamination technology to sterilize Ebola-contaminated medical devices at remote clinical sites lacking infra-structure in crisis-stricken regions of West Africa (medical waste materials are placed in bags and burned). The basis for effectuating sterilization with FDKs is chlorine dioxide (ClO2) produced from a patented invention developed by researchers at the US Army Natick Soldier RD&E Center (NSRDEC) and commercialized as a dry mixed-chemical for bacterial spore decontamination. In fact, the NSRDEC research scientists developed an ensemble of ClO2 technologies designed for different applications in decontaminating fresh produce; food contact and handling surfaces; personal protective equipment; textiles used in clothing, uniforms, tents, and shelters; graywater recycling; airplanes; surgical instruments; and hard surfaces in latrines, laundries, and deployable medical facilities. These examples demonstrate the far-reaching impact, adaptability, and versatility of these innovative technologies. We present herein the unique attributes of NSRDEC’s novel decontamination technologies and a Case Study of the development of FDKs that were deployed in West Africa by international public health organizations to sterilize Ebola-contaminated medical equipment. FDKs use bacterial spores as indicators of sterility. We review the properties and structures of spores and the mechanisms of bacterial spore inactivation by ClO2. We also review mechanisms of bacterial spore inactivation by novel, emerging, and established non-thermal technologies for food preservation, such as high pressure processing, irradiation, cold plasma, and chemical sanitizers, using an array of Bacillus

  16. Scanning Surface Potential Microscopy of Spore Adhesion on Surfaces

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-01-01

    The adhesion of spores of Bacillus anthracis - the cause of anthrax and a likely biological threat - to solid surfaces is an important consideration in cleanup after an accidental or deliberate release. However, because of safety concerns, directly studying B. anthracis spores with advanced instrumentation is problematic. As a first step, we are examining the electrostatic potential of Bacillus thuringiensis (Bt), which is a closely related species that is often used as a simulant to study B. anthracis. Scanning surface potential microscopy (SSPM), also known as Kelvin probe force microscopy (KPFM), was used to investigate the influence of relative humidity (RH) on the surface electrostatic potential of Bt that had adhered to silica, mica, or gold substrates. AFM/SSPM side-by-side images were obtained separately in air, at various values of RH, after an aqueous droplet with spores was applied on each surface and allowed to dry before measurements. In the SSPM images, a negative potential on the surface of the spores was observed compared with that of the substrates. The surface potential decreased as the humidity increased. Spores were unable to adhere to a surface with an extremely negative potential, such as mica.

  17. Scanning surface potential microscopy of spore adhesion on surfaces.

    Science.gov (United States)

    Lee, I; Chung, E; Kweon, H; Yiacoumi, S; Tsouris, C

    2012-04-01

    The adhesion of spores of Bacillus anthracis - the cause of anthrax and a likely biological threat - to solid surfaces is an important consideration in cleanup after an accidental or deliberate release. However, because of safety concerns, directly studying B. anthracis spores with advanced instrumentation is problematic. As a first step, we are examining the electrostatic potential of Bacillus thuringiensis (Bt), which is a closely related species that is often used as a simulant to study B. anthracis. Scanning surface potential microscopy (SSPM), also known as Kelvin probe force microscopy (KPFM), was used to investigate the influence of relative humidity (RH) on the surface electrostatic potential of Bt that had adhered to silica, mica, or gold substrates. AFM/SSPM side-by-side images were obtained separately in air, at various values of RH, after an aqueous droplet with spores was applied on each surface and allowed to dry before measurements. In the SSPM images, a negative potential on the surface of the spores was observed compared with that of the substrates. The surface potential decreased as the humidity increased. Spores were unable to adhere to a surface with an extremely negative potential, such as mica. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Detection of Bacillus spores using PCR and FTA filters.

    Science.gov (United States)

    Lampel, Keith A; Dyer, Deanne; Kornegay, Leroy; Orlandi, Palmer A

    2004-05-01

    Emphasis has been placed on developing and implementing rapid detection systems for microbial pathogens. We have explored the utility of expanding FTA filter technology for the preparation of template DNA for PCR from bacterial spores. Isolated spores from several Bacillus spp., B. subtilis, B. cereus, and B. megaterium, were applied to FTA filters, and specific DNA products were amplified by PCR. Spore preparations were examined microscopically to ensure that the presence of vegetative cells, if any, did not yield misleading results. PCR primers SRM86 and SRM87 targeted a conserved region of bacterial rRNA genes, whereas primers Bsub5F and Bsub3R amplified a product from a conserved sequence of the B. subtilis rRNA gene. With the use of the latter set of primers for nested PCR, the sensitivity of the PCR-based assay was increased. Overall, 53 spores could be detected after the first round of PCR, and the sensitivity was increased to five spores by nested PCR. FTA filters are an excellent platform to remove PCR inhibitors and have universal applications for environmental, clinical, and food samples.

  19. Germination of Bacillus cereus spores adhered to stainless steel.

    Science.gov (United States)

    Hornstra, L M; de Leeuw, P L A; Moezelaar, R; Wolbert, E J; de Vries, Y P; de Vos, W M; Abee, T

    2007-05-30

    Adhered spores of Bacillus cereus represent a significant part of the surface-derived contamination in processing equipment used in the dairy industry. As germinated spores lose their resistance capacities instantaneously, efficient germination prior to a cleaning in place treatment could aid to the disinfecting effect of such a treatment. Therefore, spores of B. cereus ATCC 14579 and that of the environmental isolate B. cereus CMCC 3328 were assessed for their germination behaviour when adhered to a stainless steel surface. A mixture of l-alanine and inosine initiated germination of adhered spores efficiently, resulting in 3.2 decimal logarithms of germination. Notably, implementation of a germination-inducing step prior to a representative cleaning in place procedure reduced the number of survivors with over 3 decimal log units, while an alkali treatment alone, as part of the cleaning in place procedure, did not show any effect on B. cereus spore viability. These results show that implementation of a germination step enhances the disinfection effect of currently used cleaning in place procedures.

  20. Daily variations of Alternaria spores in the city of Murcia (semi-arid southeastern Spain)

    Science.gov (United States)

    Munuera Giner, M.; Carrión García, J. S.

    1995-12-01

    Annual variations in the abundance of Alternaria spores were related to the length of the spore period for data from Murcia (southeastern Spain). To understand the relationship between the number of spores and climatic factors, Alternaria spore counts for March 1993 to February 1994 were examined by means of correlation and regression analyses with fourteen different weather parameters. The results indicated that there was a tendency for Alternaria spore concentrations to increase with increases in temperature, wind speed and hours of sunshine. Negative correlations were observed with air pressure, wind direction and humidity. Theoretical curves for Alternaria spore counts are given in relation to temperatures during the period studied.

  1. Survival of microorganisms in space: a review.

    Science.gov (United States)

    Horneck, G

    1981-01-01

    Spores of Bacillus subtilis were exposed to selected factors of space (vacuum, solar UV radiation, heavy ions of cosmic radiation), and their response was studied after recovery. These investigations were supplemented by ground-based studies under simulated space conditions. The vacuum of space did not inactivate the spores. However, vacuum-induced structural changes in the DNA, and probably in the proteins, caused a supersensitivity to solar UV radiation. This phenomenon is caused by the production of specific photoproducts in DNA and protein, which cannot be removed by normal cellular repair processes. In vegetative bacterial cells, exposed to vacuum, cell dehydration led to damage of the cell membrane, which could be partly repaired during subsequent incubation. The high local effectiveness of the cosmic heavy ions further decreases the chance that spores can survive for any length of time in space. Nonetheless, a spore travelling through space and protected from ultraviolet radiation could possibly survive an interplanetary journey. Such a situation favors panspermia as a possible explanation for the origin of life.

  2. SURVIVAL OF MICROORGANISMS FROM MODERN PROBIOTICS IN MODEL CONDITIONS OF THE INTESTINE

    Directory of Open Access Journals (Sweden)

    Kabluchko TV

    2017-03-01

    .3, later after centrifugation and washing, they were incubated for 3 hours in intestinal model environment (bile salts 3% pancreatin 0.1%, pH 7.0. Inoculation was performed before incubation, after incubation in the gastric medium and after incubation in intestinal medium. We used the medium corresponding to the studied genus of bacteria - MRS-environment for lactobacilli, bifidum for Bifidobacterium, sabouraud medium for the isolation of yeasts and fungi and endo agar for the isolation of Enterobacteriaceae. We assessed the quantity of CFU before and after impact. Results and discussion. After incubation in a simulated gastric environment, bacteria of the type Lactobacillus and Bifidobacterium did not survive and were not defined. Only Bacillus coagulans and Saccharomyces boulardii were resistant. These microorganisms grew after incubation in the same amount as before incubation - 105-6 and 107-8 CFU respectively. Bacillus clausii also survived in these conditions, but to a lesser extent: initially - 107 CFU, after incubation - 105 CFU. After staying in model environment of the duodenum Bacillus coagulans and Saccharomyces boulardii were still fully viable, and the number of germinating Bacillus clausii bacteria decreased by an order - up to 104 CFU. Conclusion. The probiotics containing Bacillus coagulans and Saccharomyces boulardii showed complete resistance to the impact of the model environment of the stomach and duodenum, Bacillus clausii was partially resistant. It leads to conclusion that probiotic drugs containing lactobacilli and bifidobacteria, cannot withstand the aggressive environmental influence of the stomach and duodenum and become inactivated under their influence. Probiotic drugs Enterol containing yeast Saccharomyces boulardii, and Laktovit Forte containing the spore-forming bacterium Bacillus coagulans are completely resistant to the action of the model environment of the stomach and duodenum.

  3. Food Targeting: A Real-Time PCR Assay Targeting 16S rDNA for Direct Quantification of Alicyclobacillus spp. Spores after Aptamer-Based Enrichment.

    Science.gov (United States)

    Hünniger, Tim; Felbinger, Christine; Wessels, Hauke; Mast, Sophia; Hoffmann, Antonia; Schefer, Anna; Märtlbauer, Erwin; Paschke-Kratzin, Angelika; Fischer, Markus

    2015-05-06

    Spore-forming Alicyclobacillus spp. are able to form metabolites that induce even in small amounts an antiseptical or medicinal off-flavor in fruit juices. Microbial contaminations could occur by endospores, which overcame the pasteurization process. The current detection method for Alicyclobacillus spp. can take up to 1 week because of microbiological enrichment. In a previous study, DNA aptamers were selected and characterized for an aptamer-driven rapid enrichment of Alicyclobacillus spp. spores from orange juice by magnetic separation. In the present work, a direct quantification assay for Alicyclobacillus spp. spores was developed to complete the two-step approach of enrichment and detection. After mechanical treatment of the spores, the isolated DNA was quantified in a real-time PCR-assay targeting 16S rDNA. The assay was evaluated by the performance requirements of the European Network of Genetically Modified Organisms Laboratories (ENGL). Hence, the presented method is applicable for direct spore detection from orange juice in connection with an enrichment step.

  4. Cyanide utilization and degradation by microorganisms.

    Science.gov (United States)

    Knowles, C J

    1988-01-01

    Various microorganisms can produce (cyanogenesis) or degrade cyanide. They degrade cyanide either to detoxify it, or to use it as a source of nitrogen for growth. Significant amounts of cyanide are formed as a secondary metabolite by a wide range of fungi and a few bacteria by decarboxylation of glycine. When cyanide has been formed by the snow mould fungus it is degraded by conversion to carbon dioxide and ammonia via an unknown pathway. In contrast, cyanogenic bacteria either do not further catabolize cyanide or they convert it into beta-cyanoalanine by addition to cysteine or O-acetylserine. Several non-cyanogenic fungi that are pathogens of cyanogenic plants are known to degrade cyanide by hydration to formamide by the enzyme cyanide hydratase. Such fungi can be immobilized and used in packed-cell columns to continuously detoxify cyanide. ICI Biological Products Business market a preparation of spray-dried fungal mycelia, 'CYCLEAR', to detoxify industrial wastes. Novo Industri have also introduced a cyanidase preparation to convert cyanide directly into formate and ammonia. Bacteria have been isolated that use cyanide as a source of nitrogen for growth. Because cyanide, as KCN or NaCN, is toxic for growth, the bacteria (Pseudomonas fluorescens) have to be grown in fed-batch culture with cyanide as the limiting nutrient. Cyanide is converted to carbon dioxide and ammonia (which is then assimilated) by an NADH-linked cyanide oxygenase system.

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

    Directory of Open Access Journals (Sweden)

    Michelle Marie Nerandzic

    2017-10-01

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

  6. Pollen and spores as a passive monitor of ultraviolet radiation

    Directory of Open Access Journals (Sweden)

    Wesley Toby Fraser

    2014-04-01

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

  7. Lipoxygenase Activity Accelerates Programmed Spore Germination in Aspergillus fumigatus

    Directory of Open Access Journals (Sweden)

    Gregory J. Fischer

    2017-05-01

    Full Text Available The opportunistic human pathogen Aspergillus fumigatus initiates invasive growth through a programmed germination process that progresses from dormant spore to swollen spore (SS to germling (GL and ultimately invasive hyphal growth. We find a lipoxygenase with considerable homology to human Alox5 and Alox15, LoxB, that impacts the transitions of programmed spore germination. Overexpression of loxB (OE::loxB increases germination with rapid advance to the GL stage. However, deletion of loxB (ΔloxB or its signal peptide only delays progression to the SS stage in the presence of arachidonic acid (AA; no delay is observed in minimal media. This delay is remediated by the addition of the oxygenated AA oxylipin 5-hydroxyeicosatetraenoic acid (5-HETE that is a product of human Alox5. We propose that A. fumigatus acquisition of LoxB (found in few fungi enhances germination rates in polyunsaturated fatty acid-rich environments.

  8. Mutagenic effect of tritated water on spores of Bacillus subtilis

    International Nuclear Information System (INIS)

    Tanooka, H.; Munakata, N.

    1978-01-01

    The mutagenic effect of tritiated water was observed with spores of Bacillus subtilis polA strain suspended in 50 mCi/ml of tritiated water for various intervals. Dose rate given by tritium beta particles to spore core was estimated to be 400 rad/hr from some assumptions and E. coli data computed by Bockrath et al. and Sands et al. The initial mutation rate was 4.2 x 10 -9 mutants/rad, as compared with 2.4 x 10 -9 mutants/rad for 60 Co γ rays and 3.3 x 10 -9 mutants/rad for 30-kVp x rays. The mutagenic effect of tritiated water on spores is most likely due to beta particle ionizing radiation damage

  9. Evolutionary stasis of sporopollenin biochemistry revealed by unaltered Pennsylvanian spores.

    Science.gov (United States)

    Fraser, W T; Scott, A C; Forbes, A E S; Glasspool, I J; Plotnick, R E; Kenig, F; Lomax, B H

    2012-10-01

    The biopolymer sporopollenin present in the spore/pollen walls of all land plants is regarded as one of the most recalcitrant biomacromolecules (biopolymers), providing protection against a range of abiotic stresses. This long-term stability is demonstrated by the near-ubiquitous presence of pollen and spores in the fossil record with spores providing the first evidence for the colonization of the land. Here, we report for the first time chemical analyses of geologically unaltered sporopollenin from Pennsylvanian (c. 310 million yr before present (MyBP)) cave deposits. Our data show that Pennsylvanian Lycophyta megaspore sporopollenin has a strong chemical resemblance to extant relatives and indicates that a co-polymer model of sporopollenin formation is the most likely configuration. Broader comparison indicates that extant sporopollenin structure is similar across widely spaced phylogenetic groups and suggests land plant sporopollenin structure has remained stable since embryophytes invaded land. © 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.

  10. Large-spored Alternaria pathogens in section Porri disentangled

    Science.gov (United States)

    Woudenberg, J.H.C.; Truter, M.; Groenewald, J.Z.; Crous, P.W.

    2014-01-01

    The omnipresent fungal genus Alternaria was recently divided into 24 sections based on molecular and morphological data. Alternaria sect. Porri is the largest section, containing almost all Alternaria species with medium to large conidia and long beaks, some of which are important plant pathogens (e.g. Alternaria porri, A. solani and A. tomatophila). We constructed a multi-gene phylogeny on parts of the ITS, GAPDH, RPB2, TEF1 and Alt a 1 gene regions, which, supplemented with morphological and cultural studies, forms the basis for species recognition in sect. Porri. Our data reveal 63 species, of which 10 are newly described in sect. Porri, and 27 species names are synonymised. The three known Alternaria pathogens causing early blight on tomato all cluster in one clade, and are synonymised under the older name, A. linariae. Alternaria protenta, a species formerly only known as pathogen on Helianthus annuus, is also reported to cause early blight of potato, together with A. solani and A. grandis. Two clades with isolates causing purple blotch of onion are confirmed as A. allii and A. porri, but the two species cannot adequately be distinguished based on the number of beaks and branches as suggested previously. This is also found among the pathogens of Passifloraceae, which are reduced from four to three species. In addition to the known pathogen of sweet potato, A. bataticola, three more species are delineated of which two are newly described. A new Alternaria section is also described, comprising two large-spored Alternaria species with concatenate conidia. PMID:25492985

  11. Developmentally-regulated excision of the SPβ prophage reconstitutes a gene required for spore envelope maturation in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Kimihiro Abe

    2014-10-01

    Full Text Available Temperate phages infect bacteria by injecting their DNA into bacterial cells, where it becomes incorporated into the host genome as a prophage. In the genome of Bacillus subtilis 168, an active prophage, SPβ, is inserted into a polysaccharide synthesis gene, spsM. Here, we show that a rearrangement occurs during sporulation to reconstitute a functional composite spsM gene by precise excision of SPβ from the chromosome. SPβ excision requires a putative site-specific recombinase, SprA, and an accessory protein, SprB. A minimized SPβ, where all the SPβ genes were deleted, except sprA and sprB, retained the SPβ excision activity during sporulation, demonstrating that sprA and sprB are necessary and sufficient for the excision. While expression of sprA was observed during vegetative growth, sprB was induced during sporulation and upon mitomycin C treatment, which triggers the phage lytic cycle. We also demonstrated that overexpression of sprB (but not of sprA resulted in SPβ prophage excision without triggering the lytic cycle. These results suggest that sprB is the factor that controls the timing of phage excision. Furthermore, we provide evidence that spsM is essential for the addition of polysaccharides to the spore envelope. The presence of polysaccharides on the spore surface renders the spore hydrophilic in water. This property may be beneficial in allowing spores to disperse in natural environments via water flow. A similar rearrangement occurs in Bacillus amyloliquefaciens FZB42, where a SPβ-like element is excised during sporulation to reconstitute a polysaccharide synthesis gene, suggesting that this type of gene rearrangement is common in spore-forming bacteria because it can be spread by phage infection.

  12. Developmentally-Regulated Excision of the SPβ Prophage Reconstitutes a Gene Required for Spore Envelope Maturation in Bacillus subtilis

    Science.gov (United States)

    Abe, Kimihiro; Kawano, Yuta; Iwamoto, Keito; Arai, Kenji; Maruyama, Yuki; Eichenberger, Patrick; Sato, Tsutomu

    2014-01-01

    Temperate phages infect bacteria by injecting their DNA into bacterial cells, where it becomes incorporated into the host genome as a prophage. In the genome of Bacillus subtilis 168, an active prophage, SPβ, is inserted into a polysaccharide synthesis gene, spsM. Here, we show that a rearrangement occurs during sporulation to reconstitute a functional composite spsM gene by precise excision of SPβ from the chromosome. SPβ excision requires a putative site-specific recombinase, SprA, and an accessory protein, SprB. A minimized SPβ, where all the SPβ genes were deleted, except sprA and sprB, retained the SPβ excision activity during sporulation, demonstrating that sprA and sprB are necessary and sufficient for the excision. While expression of sprA was observed during vegetative growth, sprB was induced during sporulation and upon mitomycin C treatment, which triggers the phage lytic cycle. We also demonstrated that overexpression of sprB (but not of sprA) resulted in SPβ prophage excision without triggering the lytic cycle. These results suggest that sprB is the factor that controls the timing of phage excision. Furthermore, we provide evidence that spsM is essential for the addition of polysaccharides to the spore envelope. The presence of polysaccharides on the spore surface renders the spore hydrophilic in water. This property may be beneficial in allowing spores to disperse in natural environments via water flow. A similar rearrangement occurs in Bacillus amyloliquefaciens FZB42, where a SPβ-like element is excised during sporulation to reconstitute a polysaccharide synthesis gene, suggesting that this type of gene rearrangement is common in spore-forming bacteria because it can be spread by phage infection. PMID:25299644

  13. Genetic barcoding of dark-spored myxomycetes (Amoebozoa)-Identification, evaluation and application of a sequence similarity threshold for species differentiation in NGS studies

    DEFF Research Database (Denmark)

    Borg Dahl, Mathilde; Brejnrod, Asker D; Unterseher, Martin

    2017-01-01

    Unicellular, eukaryotic organisms (protists) play a key role in soil food webs as major predators of microorganisms. However, due to the polyphyletic nature of protists, no single universal barcode can be established for this group, and the structure of many protistean communities remains...... unresolved. Plasmodial slime moulds (Myxogastria or Myxomycetes) stand out among protists by their formation of fruit bodies, which allow for a morphological species concept. By Sanger sequencing of a large collection of morphospecies, this study presents the largest database to date of dark...... match, thus thought to represent undiscovered diversity of dark-spored myxomycetes....

  14. Effect of synthetic detergents on germination of fern spores

    Energy Technology Data Exchange (ETDEWEB)

    Devi, Y.; Devi, S.

    1986-12-01

    Synthetic detergents constitute one of the most important water pollutants by contaminating the lakes and rivers through domestic and industrial use. Considerable information is now available for the adverse effects of detergents an aquatic fauna including fish, algae, and higher aquatic plants. Marked inhibition of germination in orchids and brinjals and of seedlings growth in raddish suggest that rapidly growing systems could be sensitive to detergent polluted water. The present study of the effect of linear alkyl benzene sulphonate on germination of the spores of a fern, Diplazium esculentum aims at the understanding of the effects of water pollution on pteridophytes and the development of spore germination assay for phytoxicity evaluation.

  15. Architecture and Assembly of the Bacillus subtilis Spore Coat

    Science.gov (United States)

    2014-09-26

    483 489. 15. Abhyankar W, Ter Beek A, Dekker H, Kort R, Brul S, et al. (2011) Gel-free proteomic identification of the Bacillus subtilis insoluble coat... identification of additional sporulation genes in Bacillus subtilis. J Mol Biol 327: 945 972. AFM of Spore Coat Architecture PLOS ONE | www.plosone.org 16 September 2014 | Volume 9 | Issue 9 | e108560 ...1ITLE AND SUBTITLE 5a CONTRACTNUMBER Architecture and assembly of the Bacillus subtilis spore coat W911NF-09-l-0286 5b. GRANT NUMBER 5c. PROGRAM

  16. Physical determinants of radiation sensitivity in bacterial spores

    International Nuclear Information System (INIS)

    Powers, E.L.

    1982-01-01

    Several factors modifying radiation sensitivity in dry bacterial spores are described and discussed. Vacuum inducing the loss of critical structural water, very low dose rates of radiation from which the cell may recover, radiations of high linear energy transfer, and the action of temperature over long periods of time on previously irradiated cells are recognized from extensive laboratory work as important in determining survival of spores exposed to low radiation doses at low temperatures for long periods of time. Some extensions of laboratory work are proposed

  17. Systems Biology of Industrial Microorganisms

    Science.gov (United States)

    Papini, Marta; Salazar, Margarita; Nielsen, Jens

    The field of industrial biotechnology is expanding rapidly as the chemical industry is looking towards more sustainable production of chemicals that can be used as fuels or building blocks for production of solvents and materials. In connection with the development of sustainable bioprocesses, it is a major challenge to design and develop efficient cell factories that can ensure cost efficient conversion of the raw material into the chemical of interest. This is achieved through metabolic engineering, where the metabolism of the cell factory is engineered such that there is an efficient conversion of sugars, the typical raw materials in the fermentation industry, into the desired product. However, engineering of cellular metabolism is often challenging due to the complex regulation that has evolved in connection with adaptation of the different microorganisms to their ecological niches. In order to map these regulatory structures and further de-regulate them, as well as identify ingenious metabolic engineering strategies that full-fill mass balance constraints, tools from systems biology can be applied. This involves both high-throughput analysis tools like transcriptome, proteome and metabolome analysis, as well as the use of mathematical modeling to simulate the phenotypes resulting from the different metabolic engineering strategies. It is in fact expected that systems biology may substantially improve the process of cell factory development, and we therefore propose the term Industrial Systems Biology for how systems biology will enhance the development of industrial biotechnology for sustainable chemical production.

  18. [Genome editing of industrial microorganism].

    Science.gov (United States)

    Zhu, Linjiang; Li, Qi

    2015-03-01

    Genome editing is defined as highly-effective and precise modification of cellular genome in a large scale. In recent years, such genome-editing methods have been rapidly developed in the field of industrial strain improvement. The quickly-updating methods thoroughly change the old mode of inefficient genetic modification, which is "one modification, one selection marker, and one target site". Highly-effective modification mode in genome editing have been developed including simultaneous modification of multiplex genes, highly-effective insertion, replacement, and deletion of target genes in the genome scale, cut-paste of a large DNA fragment. These new tools for microbial genome editing will certainly be applied widely, and increase the efficiency of industrial strain improvement, and promote the revolution of traditional fermentation industry and rapid development of novel industrial biotechnology like production of biofuel and biomaterial. The technological principle of these genome-editing methods and their applications were summarized in this review, which can benefit engineering and construction of industrial microorganism.

  19. Impact of temperature, nutrients, pH and cold storage on the germination, growth and resistance of Bacillus cereus spores in egg white.

    Science.gov (United States)

    Soni, Aswathi; Oey, Indrawati; Silcock, Patrick; Bremer, Phil J

    2018-04-01

    B. cereus spores are a concern to the food industry, especially to the producers of heat sensitive food products like egg white or precooked and stored food such as fried rice. This study investigated the impact of nutrients, temperature (4, 8, 15 and 25 °C), pH (4, 5, 7 and 9), and cold storage on the germination, growth and resistance of B. cereus spores. In egg white held at 4 °C for 12 days spore germination was not apparent, however the addition of egg yolk (5%) resulted in a 2 Log colony forming units (CFU)/mL increase in vegetative cells (p egg white did not induce germination unless the spores were simultaneously heat activated at 70 °C for 30 min. On incubation at 15 or 25 °C in egg white, spore germination increased by 3.0 Log and 3.7 Log CFU/mL on day 12. The presence of 5% yolk further enhanced germination and subsequent sporulation during storage at 15 and 25 °C. Acidification (pH 4) of 10% egg white solution prevented germination at 4, 8, 15 and 25 °C. Spores held at 4 °C for 6 days in phosphate buffer (50 mM, pH 4) had visible deformations on their surface (scanning electron microscopy) and a significant reduction in D 88, and D 92 values of 13.9 and 8.2 min respectively. A better understanding of how spores sense and respond to changing environmental conditions will help in the development of processing strategies, involving multiple hurdles to ensure the prevention of germination and subsequent toxin production. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. Physical Pre-Treatment Improves Efficient DNA Extraction and qPCR Sensitivity from Clostridium Difficile Spores in Faecal Swine Specimens.

    Science.gov (United States)

    Grześkowiak, Łukasz; Zentek, Jürgen; Vahjen, Wilfried

    2016-11-01

    A considerable fraction of the faecal microbiota is spore-forming. Molecular quantification of bacteria may be underestimated if preceded with nucleic acid extraction without special treatment to extract recalcitrant bacterial spores. The objective of this study was to improve the DNA extraction regarding the presence of Clostridium difficile spores in faecal swine specimens. Sow faeces were inoculated with spores of C. difficile (10(6) CFU), frozen at - 30 °C overnight and subjected to DNA extraction. As a preceding step to a standard DNA extraction method (QIAamp DNA stool Mini kit), different physical treatments such as microwave oven heating and repeated bead-beating techniques and a combination of both were applied and compared with each other by means of qPCR. Using a standard DNA extraction method only, C. difficile spores were quantified at 4.96 log copy number/200 mg of faeces. A repeated bead-beating at 6 m/s for 10 min followed by a standard DNA extraction resulted in 5.77 log copy number of spores in inoculated faeces. Heating in a microwave oven at 800 W for 1, 3, 5 and 10 min followed by a standard DNA extraction resulted in a gene quantification of up to 4.89 log copy number. A combination of both methods resulted in the bacterial gene quantity of 5.37 log copy number. Pre-treatment with repeated bead-beating led to the highest quantification of bacteria, and therefore it can be applied for more efficient DNA extraction from spores of C. difficile in faecal specimens.

  1. Toxicity of Tolyltriazole to Bacillus Microorganisms.

    Science.gov (United States)

    2000-03-01

    Bacillus coagulans Microbacterium lacticum Jupiter Bacillus thuringiensis Bacillus thuringiensis Bacillus cereus Bacillus Bacillus thuringiensis...TOXICITY OF TOLYLTRIAZOLE TO BACILLUS MICROORGANISMS THESIS Christopher J. Leonard, First Lieutenant, USAF AFIT/GEE/ENV/OOM-12 Approved for...AND DATES COVERED Master’s Thesis 4. TITLE AND SUBTITLE TOXICITY OF TOLYLTRIAZOLE TO BACILLUS MICROORGANISMS 6. AUTHOR(S) Christopher J

  2. Fossil Microorganisms and Formation of Early Precambrian Weathering Profiles

    Science.gov (United States)

    Rozanov, A. Yu; Astafieva, M. M.; Vrevsky, A. B.; Alfimova, N. A.; Matrenichev, V. A.; Hoover, R. B.

    2009-01-01

    Weathering crusts are the only reliable evidences of the existence of continental conditions. Often they are the only source of information about exogenous processes and subsequently about conditions under which the development of the biosphere occurred. A complex of diverse fossil microorganisms was discovered as a result of Scanning Electron Microscope investigations. The chemical composition of the discovered fossils is identical to that of the host rocks and is represented by Si, Al, Fe, Ca and Mg. Probably, the microorganisms fixed in rocks played the role of catalyst. The decomposition of minerals comprising the rocks and their transformation into clayey (argillaceous) minerals, most likely occurred under the influence of microorganisms. And may be unique weathering crusts of Early Precambrian were formed due to interaction between specific composition of microorganism assemblage and conditions of hypergene transformations. So it is possible to speak about colonization of land by microbes already at that time and about existence of single raw from weathering crusts (Primitive soils) to real soils.

  3. Spoilage microorganisms in milk and dairy products

    Directory of Open Access Journals (Sweden)

    Andrea Skelin

    2007-12-01

    Full Text Available Spoilage microorganisms cause changes of primary characteristics and properties of milk and dairy products. The product defects depends on the specific species and number of microorganisms involved in pre- and post- technological processing. Most often, these changes are related to single undesirable sensory characteristic, smell, flavour or conistency. However, in the case of heavier microbial contamination all these undesirable characteristics can occur simultaneously. Besides, even small changes caused by presence of spoilage microorganisms lead to decreased quality of milk and various dairy products. Despite of the importance for the overall quality, the control of spoilage microorganisms for dairy industry is not obligated and therefore, only a few producers control them. Therefore, the present study describes the undesirable effect of spoilage microorganisms on quality of raw, pasteurized and sterilized milk, fermented milk, butter, sour cream and cheeses with the intention to emphasize the importance and significance of their control in the dairy industry.

  4. A Novel Spectroscopic Methodology for the Investigation of Individual Bacillus Spores

    National Research Council Canada - National Science Library

    Alexander, Troy A; Pellegrino, Paul; Gillespie, James B

    2005-01-01

    A methodology has been developed for the investigation of bacterial spores. Specifically, this method has been used to probe the spore coat composition of two different Bacillus stearothermophilus variants...

  5. Influence of heat and radiation on the germinability and viability of B. cereus BIS-59 spores

    International Nuclear Information System (INIS)

    Kamat, A.S.; Lewis, N.F.

    1983-01-01

    Spores of Bicillus cereus BIS-59, isolated in this laboratory from shrimps, exhibited an exponential gamma radiation survival curve with a d 10 value of 400 krad as compared with a D 10 value of 30 krad for the vegetative cells. The D 10 value of DPA-depleted spores was also 400 krad indicating that DPA does not influence the radiation response of these spores. Maximum germination monitored with irradiated spores was 60 percent as compared with 80 percent in case of unirradiated spores. Radiation-induced inhibition of the germination processes was not dose dependent. Heat treatment (15 min at 80 C) to spores resulted in activation of the germination process; however, increase in heating time (30 min and 60 min) increased the germination lag period. DPA-depleted spores were less heat resistant than normal spores and exhibited biphasic exponential inactivation. (author)

  6. FORMALDEHYDE GAS INACTIVATION OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACE MATERIALS.

    Science.gov (United States)

    Research evaluated the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface material using formaldehyde gas. Spores were dried on seven types of indoor surfaces and exposed to 1100 ppm formaldehyde gas for 10 hr. Fo...

  7. Levels of glycine betaine in growing cells and spores of Bacillus species and lack of effect of glycine betaine on dormant spore resistance.

    Science.gov (United States)

    Loshon, Charles A; Wahome, Paul G; Maciejewski, Mark W; Setlow, Peter

    2006-04-01

    Bacteria of various Bacillus species are able to grow in media with very high osmotic strength in part due to the accumulation of low-molecular-weight osmolytes such as glycine betaine (GB). Cells of Bacillus species grown in rich and minimal media contained low levels of GB, but GB levels were 4- to 60-fold higher in cells grown in media with high salt. GB levels in Bacillus subtilis cells grown in minimal medium were increased approximately 7-fold by GB in the medium and 60-fold by GB plus high salt. GB was present in spores of Bacillus species prepared in media with or without high salt but at lower levels than in comparable growing cells. With spores prepared in media with high salt, GB levels were highest in B. subtilis spores and > or =20-fold lower in B. cereus and B. megaterium spores. Although GB levels in B. subtilis spores were elevated 15- to 30-fold by GB plus high salt in sporulation media, GB levels did not affect spore resistance. GB levels were similar in wild-type B. subtilis spores and spores that lacked major small, acid-soluble spore proteins but were much lower in spores that lacked dipicolinic acid.

  8. 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

    processes also have significant applied interest, since (i) spores are major agents of food spoilage and food poisoning and (ii) spores’ extreme...by U N IV O F C O N N E C T IC U T http://aem .asm .org/ D ow nloaded from needed in conjunction with HP to inactivate bacterial spores and...achieve the commercial sterility of low-acid foods (22). While HP is probably most often used as a single treatment, a number of studies have demonstrated

  9. Observations on the migration of bacillus spores outside a contaminated facility during a decontamination efficacy study

    Science.gov (United States)

    Silvestri, Erin E.; Perkins, Sarah; Lordo, Robert; Kovacik, William; Nichols, Tonya L.; Bowling, Charlena Yoder; Griffin, Dale W.; Schaefer, Frank W.

    2015-01-01

    The potential for an intentional wide-area or indoor release of Bacillus anthracis spores remains a concern, but the fate and transport of B. anthracis spores in indoor and outdoor environments are not well understood. Some studies have examined the possibility of spore transport within ventilation systems and in buildings and transport into a building following an outdoor release. Little research exists regarding the potential for spores to migrate to the outside of a building following an indoor release.

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

    Digital Repository Service at National Institute of Oceanography (India)

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

    of fungal spores in the deep sea may face several obstacles like the mycostatic effect of seawater (Kirk, 1980), low temperature, elevated hydrostatic pressure and low nutrient conditions. A defining characteristic of spores is their ability to develop... hyphal colony. The first step in this is the spore germina- tion, which can be defined as the sequence of events that converts the resting/dormant spore into a rapidly growing germ tube from which the myce- lium is produced by elongation, septum formation...

  11. Size matters for violent discharge height and settling speed of Sphagnum spores: important attributes for dispersal potential.

    Science.gov (United States)

    Sundberg, Sebastian

    2010-02-01

    Initial release height and settling speed of diaspores are biologically controlled components which are key to modelling wind dispersal. Most Sphagnum (peat moss) species have explosive spore liberation. In this study, how capsule and spore sizes affect the height to which spores are propelled were measured, and how spore size and spore number of discharged particles relate to settling speed in the aspherical Sphagnum spores. Spore discharge and spore cloud development were filmed in a closed chamber (nine species). Measurements were taken from snapshots at three stages of cloud development. Settling speed of spores (14 species) and clusters were timed in a glass tube. The maximum discharge speed measured was 3.6 m s(-1). Spores reached a maximum height of 20 cm (average: 15 cm) above the capsule. The cloud dimensions at all stages were related positively to capsule size (R(2) = 0.58-0.65). Thus species with large shoots (because they have large capsules) have a dispersal advantage. Half of the spores were released as singles and the rest as clusters (usually two to four spores). Single spores settled at 0.84-1.86 cm s(-1), about 52 % slower than expected for spherical spores with the same diameters. Settling speed displayed a positive curvilinear relationship with spore size, close to predictions by Stokes' law for spherical spores with 68 % of the actual diameters. Light-coloured spores settled slower than dark spores. Settling speed of spore clusters agrees with earlier studies. Effective spore discharge and small, slowly settling spores appear particularly important for species in forested habitats. The spore discharge heights in Sphagnum are among the greatest for small, wind-dispersed propagules. The discharge heights and the slow settling of spores affect dispersal distances positively and may help to explain the wide distribution of most boreal Sphagnum species.

  12. Induction of prophages in spores of Bacillus subtilis by ultraviolet irradiation from synchrotron orbital radiation

    Energy Technology Data Exchange (ETDEWEB)

    Sadaie, Y.; Kada, T.; Ohta, Y. (National Inst. of Genetics, Mishima, Shizuoka (Japan)); Kobayashi, K.; Hieda, K.; Ito, T.

    1984-06-01

    Prophages were induced from Bacillus subtilis spores lysogenic with SP02 by ultraviolet (160 nm to 240 nm) irradiation from synchrotron orbital radiation (SR UV). SR UV at around 220 nm was most effective in the inactivation of spores and prophage induction from lysogenic spores, suggesting that the lesions are produced on the DNA molecule which eventually induces signals to inactivate the phage repressor.

  13. Anthrax surrogate spores are destroyed by PDT mediated by phenothiazinium dyes

    Science.gov (United States)

    Demidova, Tatiana N.; Hamblin, Michael R.

    2005-04-01

    Some Gram-positive bacteria (including the causative agent of anthrax - Bacillus anthracis) survive conditions of stress and starvation by producing dormant stage spores. The spore"s multilayered capsule consists of inner and outer membranes, cortex, proteinaceous spore coat, and in some species an exosporium. These outer layers enclose dehydrated and condensed DNA, saturated with small, acid-soluble proteins. These protective structures make spores highly resistant to damage by heat, radiation, and commonly employed anti-bacterial agents. Previously Bacillus spores have been shown to be resistant to photodynamic inactivation (PDI) using dyes and light that easily destroy the corresponding vegetative bacteria, but recently we have discovered that they are susceptible to PDI. Photoinactivation, however, is only possible if phenothiazinium dyes are used. Dimethylmethylene blue, methylene blue, new methylene blue and toluidine blue O are all effective photosensitizers. Alternative photosensitizers such as Rose Bengal, polylysine chlorin(e6) conjugate, a tricationic porphyrin and benzoporphyrin derivative are ineffective against spores even though they can easily kill vegetative cells. Spores of B. cereus and B. thuringiensis are most susceptible, B. subtilis and B. atrophaeus are also killed, while B. megaterium is resistant. Photoinactivation is most effective when excess dye is washed from the spores showing that the dye binds to the spores and that excess dye in solution can quench light delivery. The relatively mild conditions needed for spore killing could have applications for treating wounds contaminated by anthrax spores and for which conventional sporicides would have unacceptable tissue toxicity.

  14. Spore-killing meiotic drive factors in a natural population of the fungus Podospora anserina

    NARCIS (Netherlands)

    Gaag, van der M.; Debets, A.J.M.; Oosterhof, J.; Slakhorst, S.M.; Thijssen, J.A.G.M.; Hoekstra, R.F.

    2000-01-01

    In fungi, meiotic drive is observed as spore killing. In the secondarily homothallic ascomycete Podospora anserina it is characterized by the abortion of two of the four spores in the ascus. We have identified seven different types of meiotic drive elements (Spore killers). Among 99 isolates from

  15. SporeWeb : an interactive journey through the complete sporulation cycle of Bacillus subtilis

    NARCIS (Netherlands)

    Eijlander, Robyn T.; Jong, Anne de; Krawczyk, Antonina O.; Holsappel, Siger; Kuipers, Oscar P.

    2014-01-01

    Bacterial spores are a continuous problem for both food-based and health-related industries. Decades of scientific research dedicated towards understanding molecular and gene regulatory aspects of sporulation, spore germination and spore properties have resulted in a wealth of data and information.

  16. Role Played by Exosporium Glycoproteins in the Surface Properties of Bacillus cereus Spores and in Their Adhesion to Stainless Steel ▿

    Science.gov (United States)

    Lequette, Yannick; Garénaux, Estelle; Tauveron, Grégoire; Dumez, Sylvain; Perchat, Stéphane; Slomianny, Christian; Lereclus, Didier; Guérardel, Yann; Faille, Christine

    2011-01-01

    Bacillus cereus spores are surrounded by a loose-fitting layer called the exosporium, whose distal part is mainly formed from glycoproteins. The role played by the exosporium glycoproteins of B. cereus ATCC 14579 (BclA and ExsH) was investigated by considering hydrophobicity and charge, as well as the properties of spore adhesion to stainless steel. The absence of BclA increased both the isoelectric point (IEP) and hydrophobicity of whole spores while simultaneously reducing the interaction between spores and stainless steel. However, neither the hydrophobicity nor the charge associated with BclA could explain the differences in the adhesion properties. Conversely, ExsH, another exosporium glycoprotein, did not play a significant role in spore surface properties. The monosaccharide analysis of B. cereus ATCC 14579 showed different glycosylation patterns on ExsH and BclA. Moreover, two specific glycosyl residues, namely, 2-O-methyl-rhamnose (2-Me-Rha) and 2,4-O-methyl-rhamnose (2,4-Me-Rha), were attached to BclA, in addition to the glycosyl residues already reported in B. anthracis. PMID:21622795

  17. Microbial modeling of thermal resistance of Alicyclobacillus acidoterrestris CRA7152 spores in concentrated orange juice with nisin addition

    Science.gov (United States)

    Peña, Wilmer Edgard Luera; de Massaguer, Pilar Rodriguez; Teixeira, Luciano Quintão

    2009-01-01

    The nisin effect on thermal death of Alicyclobacillus acidoterrestris CRA 7152 spores in concentrated orange juice (64°Brix) was studied. Concentrations of 0, 50, 75 and 100 IU of nisin/ml juice, at temperatures of 92, 95, 98 and 102°C were evaluated. The quadratic polynomial model was used to analyze the effects of the factors and their interaction. Verification of surviving spores was carried out through plating in K medium (pH 3.7). The results showed that the D values without nisin addition were 25.5, 12.9, 6.1 and 2.3 min for 92, 95, 98 and 102°C respectively. With addition of nisin into the juice there was a drop of heat resistance as the concentration was increased at a same temperature. With 30, 50, 75, 100 and 150 IU/ml at 95°C, the D values were 12.34, 11.38, 10.49, 9.49 and 9.42 min respectively, showing that a decrease in the D value up to 27% can be obtained. The second order polynomial model established with r2 = 0.995 showed that the microorganism resistance was affected by the action of temperature followed by the nisin concentration. Nisin therefore is an alternative for reducing the rigor of the A. acidoterrestris CRA 7152 thermal treatment. PMID:24031405

  18. DNA capturing machinery through spore-displayed proteins.

    Science.gov (United States)

    Park, T J; Lee, S J; Pan, J-G; Jung, H-C; Park, J Y; Park, J P; Lee, S Y

    2011-10-01

    The purpose of this study was to develop a general method for the facile development of a new DNA biosensor which utilizes streptavidin-displayed spores as a molecular machinery. Fluorescence spectroscopy was used as a monitoring tool for the streptavidin displayed on the surface of Bacillus thuringiensis spores and as a diagnosis method for DNA detection. As a proof-of-concept, four pathogenic bacteria including Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli and Klebsiella pneumonia were used for the detection of pathogenic species. In addition, a set of mutant variants of Wilson's disease were also used for the detection of single nucleotide polymorphism (SNP) in this system. This strategy, utilizing streptavidin-displayed spores, is capable of capturing DNA targets for the detection of pathogenic bacteria and for mutation analysis in Wilson's disease. This approach could be useful as a simple platform for developing sensitive spore-based biosensors for any desired DNA targets in diagnostic applications. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  19. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores.

    Directory of Open Access Journals (Sweden)

    Jason Edmonds

    Full Text Available The release of biological agents, including those which could be used in biowarfare or bioterrorism in large urban areas, has been a concern for governments for nearly three decades. Previous incidents from Sverdlosk and the postal anthrax attack of 2001 have raised questions on the mechanism of spread of Bacillus anthracis spores as an aerosol or contaminant. Prior studies have demonstrated that Bacillus atrophaeus is easily transferred through simulated mail handing, but no reports have demonstrated this ability with Bacillus anthracis spores, which have morphological differences that may affect adhesion properties between spore and formite. In this study, equipment developed to simulate interactions across three generations of envelopes subjected to tumbling and mixing was used to evaluate the potential for cross-contamination of B. anthracis spores in simulated mail handling. In these experiments, we found that the potential for cross-contamination through letter tumbling from one generation to the next varied between generations while the presence of a fluidizer had no statistical impact on the transfer of material. Likewise, the presence or absence of a fluidizer had no statistically significant impact on cross-contamination levels or reaerosolization from letter opening.

  20. Stem rust spores elicit rapid RPG1 phosphorylation

    Science.gov (United States)

    Stem rust threatens cereal production worldwide. Understanding the mechanism by which durable resistance genes, such as Rpg1, function is critical. We show that the RPG1 protein is phosphorylated within 5 min by exposure to spores from avirulent but not virulent races of stem rust. Transgenic mutant...

  1. Increased resistance of environmental anaerobic spores to inactivation by UV

    NARCIS (Netherlands)

    Hijnen, W.A.M.; Veer, A.J. van der; Beerendonk, E.F.; Medema, Gerriet Jan

    2004-01-01

    Water Company Europoort started a pilot plant (MP)UV study to determine the UV-fluence to meet the Dutch drinking water standards. The results of large volume sampling of this pilot plant demonstrated that environmental spores of sulphite-reducing clostridia (SSRC) were highly resistant against UV.

  2. Changes in spore chemistry and appearance with increasing maturity

    NARCIS (Netherlands)

    Fraser, W.T.; Watson, J.S.; Sephton, M.A.; Lomax, B.H.; Harrington, G.; Gosling, W.D.; Self, S.

    2014-01-01

    Sporopollenin is the primary biopolymer found in the walls of pollen and spores; during maturation sporopollenin undergoes a number of discrete chemical changes, despite maintaining identifiable morphological features which can be exploited for palynological study. Here we report the results of

  3. Decontamination of Bacillus spores adhered to iron and ...

    Science.gov (United States)

    Journal Article This study examines the effectiveness of decontaminating Bacillus globigii spores attached to corroded iron and cement-mortar coupons with free chlorine at two pH levels, monochloramine, chlorine dioxide, ozone, peracetic acid (PAA) and acidified nitrite, followed by flushing.

  4. Biomarkers of Aspergillus spores: Strain typing and protein identification

    Czech Academy of Sciences Publication Activity Database

    Šulc, Miroslav; Pešlová, Kateřina; Žabka, Martin; Hajdúch, M.; Havlíček, Vladimír

    2009-01-01

    Roč. 280, 1-3 (2009), s. 162-168 ISSN 1387-3806 R&D Projects: GA MŠk LC07017; GA ČR GP203/05/P575 Institutional research plan: CEZ:AV0Z50200510 Keywords : aspergillus * spore * protein Subject RIV: EE - Microbiology, Virology Impact factor: 2.117, year: 2009

  5. Effects of Ingesting Bacillus Thuringiensis (Berliner) Spores on ...

    African Journals Online (AJOL)

    Bacillus thuringiensis Berliner was isolated from dead Sesamia calamistis Hampson (Lepidoptera: Noctuidae) larvae collected from maize farms in Cape Coast, Ghana. Spores produced from the vegetative cells were incorporated into an artificial diet and fed to 2nd instar S. calamistis larvae. The duration of larval and pupal ...

  6. Effects of Ingesting Bacillus Thuringiensis (Berliner) Spores on ...

    African Journals Online (AJOL)

    Effects of Ingesting Bacillus Thuringiensis (Berliner) Spores on Developmental Stages and Fecundity of Surviving Sesamia Calamistis (Hampson) (Lepidoptera: ... The PDF file you selected should load here if your Web browser has a PDF reader plug-in installed (for example, a recent version of Adobe Acrobat Reader).

  7. Spore survival during batch dry rendering of abattoir waste.

    Science.gov (United States)

    Lowry, P D; Fernando, T; Gill, C O

    1979-01-01

    Normal batch dry rendering practice does not ensure sterile products, because bacterial spores are protected against thermal denaturation by the high fat-low water content environment which results from drying the materials at temperatures below those required for sterilization. PMID:117753

  8. In vitro spore germination and gametophytic growth development of ...

    African Journals Online (AJOL)

    The effects of sucrose, pH and plant growth hormones on spore germination percentage and gametophyte growths of Pteris tripartita were studied. Various morphological structures of gametophytes were observed namely, filamentous, spatulate and heart stages in the MS culture medium with hormones. After 15 days, the ...

  9. Application of flow cytometry to wine microorganisms.

    Science.gov (United States)

    Longin, Cédric; Petitgonnet, Clément; Guilloux-Benatier, Michèle; Rousseaux, Sandrine; Alexandre, Hervé

    2017-04-01

    Flow cytometry (FCM) is a powerful technique allowing detection and enumeration of microbial populations in food and during food process. Thanks to the fluorescent dyes used and specific probes, FCM provides information about cell physiological state and allows enumeration of a microorganism in a mixed culture. Thus, this technique is increasingly used to quantify pathogen, spoilage microorganisms and microorganisms of interest. Since one decade, FCM applications to the wine field increase greatly to determine population and physiological state of microorganisms performing alcoholic and malolactic fermentations. Wine spoilage microorganisms were also studied. In this review we briefly describe FCM principles. Next, a deep revision concerning enumeration of wine microorganisms by FCM is presented including the fluorescent dyes used and techniques allowing a yeast and bacteria species specific enumeration. Then, the last chapter is dedicated to fluorescent dyes which are used to date in fluorescent microscopy but applicable in FCM. This chapter also describes other interesting "future" techniques which could be applied to study the wine microorganisms. Thus, this review seeks to highlight the main advantages of the flow cytometry applied to wine microbiology. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Airway inflammation among compost workers exposed to actinomycetes spores

    Directory of Open Access Journals (Sweden)

    Kari Kulvik Heldal

    2015-05-01

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

  11. 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

  12. DEVELOPMENT OF MICROORGANISMS WITH IMPROVED TRANSPORT AND BIOSURFACTANT ACTIVITY FOR ENHANCED OIL RECOVERY

    Energy Technology Data Exchange (ETDEWEB)

    M.J. McInerney; N. Youssef; T. Fincher; S.K. Maudgalya; M.J. Folmsbee; R. Knapp; D. Nagle

    2004-05-31

    Diverse microorganisms were screened for biosurfactant production and anaerobic growth at elevated salt concentrations to obtain candidates most suitable for microbial oil recovery. Seventy percent of the 205 strains tested, mostly strains of Bacillus mojavensis, Bacillus subtilis, Bacillus licheniformis, and Bacillus sonorensis, produced biosurfactants aerobically and 41% of the strains had biosurfactant activity greater than Bacillus mojavensis JF-2, the current candidate for oil recovery. Biosurfactant activity varied with the percentage of the 3-hydroxy-tetradecanoate isomers in the fatty acid portion of the biosurfactant. Changing the medium composition by incorporation of different precursors of 3-hydroxy tetradecanoate increased the activity of biosurfactant. The surface tension and critical micelle concentration of 15 different, biosurfactant-producing Bacillus strains was determined individually and in combination with other biosurfactants. Some biosurfactant mixtures were found to have synergistic effect on surface tension (e.g. surface tension was lowered from 41 to 31 mN/m in some cases) while others had a synergistic effect on CMD-1 values. We compared the transport abilities of spores from three Bacillus strains using a model porous system to study spore recovery and transport. Sand-packed columns were used to select for spores or cells with the best transport abilities through brine-saturated sand. Spores of Bacillus mojavensis strains JF-2 and ROB-2 and a natural recombinant, strain C-9, transported through sand at very high efficiencies. The earliest cells/spores that emerged from the column were re-grown, allowed to sporulate, and applied to a second column. This procedure greatly enhanced the transport of strain C-9. Spores with enhanced transport abilities can be easily obtained and that the preparation of inocula for use in MEOR is feasible. Tertiary oil recovery experiments showed that 10 to 40 mg/l of JF-2 biosurfactant in the presence of 0

  13. Achieving consistent multiple daily low-dose Bacillus anthracis spore inhalation exposures in the rabbit model

    Directory of Open Access Journals (Sweden)

    Roy E Barnewall

    2012-06-01

    Full Text Available Repeated low-level exposures to Bacillus anthracis could occur before or after the remediation of an environmental release. This is especially true for persistent agents such as Bacillus anthracis spores, the causative agent of anthrax. Studies were conducted to examine aerosol methods needed for consistent daily low aerosol concentrations to deliver a low-dose (less than 106 colony forming units (CFU of B. anthracis spores and included a pilot feasibility characterization study, acute exposure study, and a multiple fifteen day exposure study. This manuscript focuses on the state-of-the-science aerosol methodologies used to generate and aerosolize consistent daily low aerosol concentrations and resultant low inhalation doses. The pilot feasibility characterization study determined that the aerosol system was consistent and capable of producing very low aerosol concentrations. In the acute, single day exposure experiment, targeted inhaled doses of 1 x 102, 1 x 103, 1 x 104, and 1 x 105 CFU were used. In the multiple daily exposure experiment, rabbits were exposed multiple days to targeted inhaled doses of 1 x 102, 1 x 103, and 1 x 104 CFU. In all studies, targeted inhaled doses remained fairly consistent from rabbit to rabbit and day to day. The aerosol system produced aerosolized spores within the optimal mass median aerodynamic diameter particle size range to reach deep lung alveoli. Consistency of the inhaled dose was aided by monitoring and recording respiratory parameters during the exposure with real-time plethysmography. Overall, the presented results show that the animal aerosol system was stable and highly reproducible between different studies and multiple exposure days.

  14. Fighting Ebola through Novel Spore Decontamination Technologies for the Military

    Directory of Open Access Journals (Sweden)

    Christopher J. Doona

    2015-08-01

    Full Text Available AbstractRecently, global public health organizations such as Doctors without Borders (MSF, the World Health Organization (WHO, Public Health Canada, National Institutes of Health (NIH, and the U.S. government developed and deployed Field Decontamination Kits (FDKs, a novel, lightweight, compact, reusable decontamination technology to sterilize Ebola-contaminated medical devices at remote clinical sites lacking infra-structure in crisis-stricken regions of West Africa (medical waste materials are placed in bags and burned. The basis for effectuating sterilization with FDKs is chlorine dioxide (ClO2 produced from a patented invention developed by researchers at the US Army – Natick Soldier RD&E Center (NSRDEC and commercialized as a dry mixed-chemical for bacterial spore decontamination. In fact, the NSRDEC research scientists developed an ensemble of ClO2 technologies designed for different applications in decontaminating fresh produce; food contact and handling surfaces; personal protective equipment; textiles used in clothing, uniforms, tents, and shelters; graywater recycling; airplanes; surgical instruments; and hard surfaces in latrines, laundries, and deployable medical facilities. These examples demonstrate the far-reaching impact, adaptability, and versatility of these innovative technologies. We present herein the unique attributes of NSRDEC’s novel decontamination technologies and a Case Study of the development of FDKs that were deployed in West Africa by international public health organizations to sterilize Ebola-contaminated medical equipment. FDKs use bacterial spores as indicators of sterility. We review the properties and structures of spores and the mechanisms of bacterial spore inactivation by ClO2. We also review mechanisms of bacterial spore inactivation by novel, emerging, and established nonthermal technologies for food preservation, such as high pressure processing, irradiation, cold plasma, and chemical sanitizers

  15. 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. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Studies on the radiosensitivity and DNA damage by ionizing radiation in microorganisms

    International Nuclear Information System (INIS)

    Choi, E.H.

    1980-01-01

    Sensitivity of food-borne microorganisms to 60 Co gamma radiation and their radiosensitization by heat and chemical preservatives in the presence of nitrogen or oxygen gas were investigated with eight species of bacteria and twelve species of molds. Radiosensitivity of all tested bacteria was examined to give D 10 values of 11-30 krad for vegetative cells and 185-470 krad for spores. Conidia of molds showed dose-survival curves of sigmoidal type which had D 10 values of 15-37 krad and induction doses of 12-75 krad. Sal. typhimurium (10 min at 45 0 C) and spores of B. subtilis 168 (30 min at 95 0 C) showed a synergistic effect of inactivation with postirradiation heating, whereas E. coli B/r (15 min at 50 0 C) and Pen. expansum (10 min at 50 0 C) were synergistically inactivated by both pre- and post-irradiation heating. On the other hand vegetative cells (10 min at 45 0 C) and spores (10 min at 80 0 C) of B. megaterium NRRL B-1368 did not show any synergistic effects with the pre- or postirradiation heating. Heating before or after irradiation induced more breaks of DNA single strands of E. coil B/r than the irradiation alone. However the repair process of the broken DNA was not observed. Irradiation in the presence of oxygen gas caused more increase in break and repair of DNA single strands whereas the treatment in the presence of nitrogen gas resulted in a decrease. (author)

  17. Response surface methodology as a tool for modeling and optimization of Bacillus subtilis spores inactivation by UV/ nano-Fe0process for safe water production.

    Science.gov (United States)

    Yousefzadeh, Samira; Matin, Atiyeh Rajabi; Ahmadi, Ehsan; Sabeti, Zahra; Alimohammadi, Mahmood; Aslani, Hassan; Nabizadeh, Ramin

    2018-04-01

    One of the most important aspects of environmental issues is the demand for clean and safe water. Meanwhile, disinfection process is one of the most important steps in safe water production. The present study aims at estimating the performance of UV, nano Zero-Valent Iron particles (nZVI, nano-Fe 0 ), and UV treatment with the addition of nZVI (combined process) for Bacillus subtilis spores inactivation. Effects of different factors on inactivation including contact time, initial nZVI concentration, UV irradiance and various aerations conditions were investigated. Response surface methodology, based on a five-level, two variable central composite design, was used to optimize target microorganism reduction and the experimental parameters. The results indicated that the disinfection time had the greatest positive impact on disinfection ability among the different selected independent variables. According to the results, it can be concluded that microbial reduction by UV alone was more effective than nZVI while the combined UV/nZVI process demonstrated the maximum log reduction. The optimum reduction of about 4 logs was observed at 491 mg/L of nZVI and 60 min of contact time when spores were exposed to UV radiation under deaerated condition. Therefore, UV/nZVI process can be suggested as a reliable method for Bacillus subtilis spores inactivation. Copyright © 2018. Published by Elsevier Ltd.

  18. Tolerance of spores to ionizing radiation: mechanisms of inactivation, injury and repair

    International Nuclear Information System (INIS)

    Farkas, J.

    1994-01-01

    Radiation resistance of bacterial spores is of great practical importance both in radiation preservation of food and in radiation sterilization of medicine products. This paper attempts to review selected aspects of the effects of ionizing radiation on bacterial spores. It focuses on irradiation in the high-moisture environments that are the usual characteristic of food irradiation, with less emphasis on dry systems in radiation sterilization of medical products. Topics covered include the tolerance of bacterial spores to ionizing radiation, the mechanism of radiation resistance of spores, the effect of environmental factors on radiation resistance, and radiation injury of spores and its consequences. (UK)

  19. Stingless bees (Hymenoptera, Meliponini feeding on stinkhorn spores (Fungi, Phallales: robbery or dispersal?

    Directory of Open Access Journals (Sweden)

    Marcio L. Oliveira

    2000-09-01

    Full Text Available Records about stingless bee-fungi interaction are very rare. In Brazilian Amazonia, workers of Trigona crassipes (Fabricius, 1793 and Trigona fulviventris Guérin, 1835 visiting two stinkhorn species, Dictyophora sp. and Phallus sp., respectively, were observed. The workers licked the fungi gleba, a mucilaginous mass of spores covering the pileum. Neither gleba residue nor spores were found on the body surface of these bee workers. These observations indicate that these bee species include spores as a complement in their diet. On the other hand, they also suggest that these stingless bees can, at times, facilitale spore dispersal, in case intact spores are eliminated with the feces.

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

    Science.gov (United States)

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

    1978-01-01

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

  1. Germination, outgrowth and vegetative growth kinetics of dry heat-treated individual spores ofBacillusspecies.

    Science.gov (United States)

    He, Lin; Chen, Zhan; Wang, Shiwei; Wu, Muying; Setlow, Peter; Li, Yong-Qing

    2018-01-12

    DNA damage kills dry-heated spores of Bacillus subtilis , but dry heat-treatment effects on spore germination and outgrowth have not been studied. This is important, since if dry heat-killed spores germinate and undergo outgrowth, toxic proteins could be synthesized. Here, Raman spectroscopy and differential interference contrast microscopy were used to study germination and outgrowth of individual dry heat-treated B. subtilis and Bacillus megaterium spores. Major findings in this work were as follows. 1) Spores dry heat-treated at 140°C for 20 min nearly all lost viability but retained their Ca 2+ -dipicolinic acid (CaDPA) depot. 2) In most cases, dry heat treatment increased the average times of and variability in all major events in B. subtilis spore germination with nutrient germinants or CaDPA, and one nutrient germination event with B. megaterium spores. 3) B. subtilis spore germination with dodecylamine, which activates spores' CaDPA release channel, was unaffected by dry heat treatment. 4) These results indicate that dry heat treatment likely damages spore proteins important in nutrient germinant recognition and cortex peptidoglycan hydrolysis, but not CaDPA release itself. 5) Analysis of single spores incubated on nutrient-rich agar showed that while dry heat-treated spores that are dead can complete germination, they cannot proceed into outgrowth thus not to vegetative growth. The results of this study provide new information on effects of dry heat on bacterial spores, and indicate that dry heat sterilization regimens should give spores that cannot outgrow and thus cannot synthesize potentially dangerous proteins. IMPORTANCE Much research has shown that high temperature dry heat is a promising means for the inactivation of spores on medical devices and spacecraft decontamination. Dry heat is known to kill Bacillus subtilis spores by DNA damage. However, knowledge about effects of dry heat treatment on spore germination and outgrowth is limited

  2. Training Program Organic Fertilizer Production by Microorganisms Efficient

    Directory of Open Access Journals (Sweden)

    Diana Carolina Cuevas Ordoñez

    2017-02-01

    Full Text Available This research aims to develop a training program for Organic Fertilizer Production by Efficient Microorganisms aimed at teachers of the Bolivarian National School "Jose Vicente Unda" Barinitas Parish, Municipality Bolivar, Barinas State, 2015-2016 school year. The nature of research is framed in qualitative methodology, in the form of participatory action research, with a field design under the following phases: diagnosis, planning, implementation and evaluation, supported by a depth interview, using a log audiovisual face to face and through generating scripts questions, you will have three key informants. As for the technical analysis of the information box categories in order to make comparisons and draw contrasts possible. Among the final considerations it is estimated that the creation of practical activities for the production of organic fertilizer through efficient microorganisms represents an important contribution to strengthening the socio spaces in the institution, maintaining biodiversity and reducing the negative impact to the environment.

  3. Detection of extracellular enzymatic activity in microorganisms ...

    African Journals Online (AJOL)

    Detection of extracellular enzymatic activity in microorganisms isolated from waste vegetable oil contaminated soil using plate methodologies. Eugenia G. Ortiz Lechuga, Isela Quintero Zapata, Katiushka Arévalo Niño ...

  4. Growth and extracellular enzyme production by microorganisms ...

    African Journals Online (AJOL)

    Ugba', an indigenous Nigerian fermented food condiment. The isolated microorganisms were screened for amylase, protease and lipase production, the activity and specific activity of the enzymes were also determined. The effect of pH and ...

  5. Evaluation of microorganisms transmissible through handshake ...

    African Journals Online (AJOL)

    Microorganisms transmissible through handshake were experimentally isolated from samples collected from primary and secondary school students as well as undergraduates and staff of the Federal University of Technology, Akure. Bacteria isolated include Staphylococcus aureus, S. epididimis, Bacillus subtilis, ...

  6. Defensive properties of pyrrolizidine alkaloids against microorganisms

    NARCIS (Netherlands)

    Joosten, L.; Van Veen, J.A.

    2011-01-01

    The understanding of the selection factors that drive chemical diversification of secondary metabolites of constitutive defence systems in plants, such as pyrrolizidine alkaloids (PAs), is still incomplete. Historically, plants always have been confronted with microorganisms. Long before herbivores

  7. Novel Industrial Enzymes from Uncultured Arctic Microorganisms

    DEFF Research Database (Denmark)

    Vester, Jan Kjølhede

    % of the microorganisms in an environmental sample can be cultured in the laboratory with standard techniques, which is also the case for the ikaite columns. Thus, there is an enormous potential in the uncultured microorganisms, which cannot be accessed through cultivation based methods. This PhD thesis presents studies......, and reduce the risk of contaminations. Cold- and alkaline-active enzymes can be found in microorganisms adapted to living in natural environments with these conditions, which are extremely rare but found in the unique ikaite columns from SW Greenland (4-6 °C, pH >10). It is estimated that less than 1...... on the diversity of microorganisms from the ikaite columns as well as bioprospecting for enzyme activities using both culture dependent and independent methods. Two cold-active β-galactosidases and one extremely cold-active α-amylase, all related to Clostridia, were characterized in more details....

  8. growth and extracellular enzyme production by microorganisms

    African Journals Online (AJOL)

    Okorie

    2013-06-26

    Ugba', an indigenous. Nigerian fermented food condiment. The isolated microorganisms were screened for amylase, protease and lipase production, the activity and specific activity of the enzymes were also determined.

  9. Predicting Bacillus coagulans spores inactivation in tomato pulp under nonisothermal heat treatments.

    Science.gov (United States)

    Zimmermann, Morgana; Longhi, Daniel A; Schaffner, Donald W; Aragão, Gláucia M F

    2014-05-01

    The knowledge and understanding of Bacillus coagulans inactivation during a thermal treatment in tomato pulp, as well as the influence of temperature variation during thermal processes are essential for design, calculation, and optimization of the process. The aims of this work were to predict B. coagulans spores inactivation in tomato pulp under varying time-temperature profiles with Gompertz-inspired inactivation model and to validate the model's predictions by comparing the predicted values with experimental data. B. coagulans spores in pH 4.3 tomato pulp at 4 °Brix were sealed in capillary glass tubes and heated in thermostatically controlled circulating oil baths. Seven different nonisothermal profiles in the range from 95 to 105 °C were studied. Predicted inactivation kinetics showed similar behavior to experimentally observed inactivation curves when the samples were exposed to temperatures in the upper range of this study (99 to 105 °C). Profiles that resulted in less accurate predictions were those where the range of temperatures analyzed were comparatively lower (inactivation profiles starting at 95 °C). The link between fail prediction and both lower starting temperature and magnitude of the temperature shift suggests some chemical or biological mechanism at work. Statistical analysis showed that overall model predictions were acceptable, with bias factors from 0.781 to 1.012, and accuracy factors from 1.049 to 1.351, and confirm that the models used were adequate to estimate B. coagulans spores inactivation under fluctuating temperature conditions in the range from 95 to 105 °C. How can we estimate Bacillus coagulans inactivation during sudden temperature shifts in heat processing? This article provides a validated model that can be used to predict B. coagulans under changing temperature conditions. B. coagulans is a spore-forming bacillus that spoils acidified food products. The mathematical model developed here can be used to predict the spoilage

  10. Pathogenic and opportunistic microorganisms in caves

    Directory of Open Access Journals (Sweden)

    Sanchez-Moral Sergio

    2010-01-01

    Full Text Available With today’s leisure tourism, the frequency of visits to many caves makes it necessary to know about possible potentially pathogenic microorganisms in caves, determine their reservoirs, and inform the public about the consequences of such visits. Our data reveal that caves could be a potential danger to visitors because of the presence of opportunistic microorganisms, whose existence and possible development in humans is currently unknown.

  11. A two-step transport pathway allows the mother cell to nurture the developing spore in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Fernando H Ramírez-Guadiana

    2017-09-01

    Full Text Available One of the hallmarks of bacterial endospore formation is the accumulation of high concentrations of pyridine-2,6-dicarboxylic acid (dipicolinic acid or DPA in the developing spore. This small molecule comprises 5-15% of the dry weight of dormant spores and plays a central role in resistance to both wet heat and desiccation. DPA is synthesized in the mother cell at a late stage in sporulation and must be translocated across two membranes (the inner and outer forespore membranes that separate the mother cell and forespore. The enzymes that synthesize DPA and the proteins required to translocate it across the inner forespore membrane were identified over two decades ago but the factors that transport DPA across the outer forespore membrane have remained mysterious. Here, we report that SpoVV (formerly YlbJ is the missing DPA transporter. SpoVV is produced in the mother cell during the morphological process of engulfment and specifically localizes in the outer forespore membrane. Sporulating cells lacking SpoVV produce spores with low levels of DPA and cells engineered to express SpoVV and the DPA synthase during vegetative growth accumulate high levels of DPA in the culture medium. SpoVV resembles concentrative nucleoside transporters and mutagenesis of residues predicted to form the substrate-binding pocket supports the idea that SpoVV has a similar structure and could therefore function similarly. These findings provide a simple two-step transport mechanism by which the mother cell nurtures the developing spore. DPA produced in the mother cell is first translocated into the intermembrane space by SpoVV and is then imported into the forespore by the SpoVA complex. This pathway is likely to be broadly conserved as DPA synthase, SpoVV, and SpoVA proteins can be found in virtually all endospore forming bacteria.

  12. A two-step transport pathway allows the mother cell to nurture the developing spore in Bacillus subtilis.

    Science.gov (United States)

    Ramírez-Guadiana, Fernando H; Meeske, Alexander J; Rodrigues, Christopher D A; Barajas-Ornelas, Rocío Del Carmen; Kruse, Andrew C; Rudner, David Z

    2017-09-01

    One of the hallmarks of bacterial endospore formation is the accumulation of high concentrations of pyridine-2,6-dicarboxylic acid (dipicolinic acid or DPA) in the developing spore. This small molecule comprises 5-15% of the dry weight of dormant spores and plays a central role in resistance to both wet heat and desiccation. DPA is synthesized in the mother cell at a late stage in sporulation and must be translocated across two membranes (the inner and outer forespore membranes) that separate the mother cell and forespore. The enzymes that synthesize DPA and the proteins required to translocate it across the inner forespore membrane were identified over two decades ago but the factors that transport DPA across the outer forespore membrane have remained mysterious. Here, we report that SpoVV (formerly YlbJ) is the missing DPA transporter. SpoVV is produced in the mother cell during the morphological process of engulfment and specifically localizes in the outer forespore membrane. Sporulating cells lacking SpoVV produce spores with low levels of DPA and cells engineered to express SpoVV and the DPA synthase during vegetative growth accumulate high levels of DPA in the culture medium. SpoVV resembles concentrative nucleoside transporters and mutagenesis of residues predicted to form the substrate-binding pocket supports the idea that SpoVV has a similar structure and could therefore function similarly. These findings provide a simple two-step transport mechanism by which the mother cell nurtures the developing spore. DPA produced in the mother cell is first translocated into the intermembrane space by SpoVV and is then imported into the forespore by the SpoVA complex. This pathway is likely to be broadly conserved as DPA synthase, SpoVV, and SpoVA proteins can be found in virtually all endospore forming bacteria.

  13. Nanoscale Structural and Mechanical Analysis of Bacillus anthracis Spores Inactivated with Rapid Dry Heating

    Science.gov (United States)

    Felker, Daniel L.; Burggraf, Larry W.

    2014-01-01

    Effective killing of Bacillus anthracis spores is of paramount importance to antibioterrorism, food safety, environmental protection, and the medical device industry. Thus, a deeper understanding of the mechanisms of spore resistance and inactivation is highly desired for developing new strategies or improving the known methods for spore destruction. Previous studies have shown that spore inactivation mechanisms differ considerably depending upon the killing agents, such as heat (wet heat, dry heat), UV, ionizing radiation, and chemicals. It is believed that wet heat kills spores by inactivating critical enzymes, while dry heat kills spores by damaging their DNA. Many studies have focused on the biochemical aspects of spore inactivation by dry heat; few have investigated structural damages and changes in spore mechanical properties. In this study, we have inactivated Bacillus anthracis spores with rapid dry heating and performed nanoscale topographical and mechanical analysis of inactivated spores using atomic force microscopy (AFM). Our results revealed significant changes in spore morphology and nanomechanical properties after heat inactivation. In addition, we also found that these changes were different under different heating conditions that produced similar inactivation probabilities (high temperature for short exposure time versus low temperature for long exposure time). We attributed the differences to the differential thermal and mechanical stresses in the spore. The buildup of internal thermal and mechanical stresses may become prominent only in ultrafast, high-temperature heat inactivation when the experimental timescale is too short for heat-generated vapor to efficiently escape from the spore. Our results thus provide direct, visual evidences of the importance of thermal stresses and heat and mass transfer to spore inactivation by very rapid dry heating. PMID:24375142

  14. Role of the gerA operon in L-alanine germination of Bacillus licheniformis spores

    Directory of Open Access Journals (Sweden)

    Løvdal Irene S

    2012-03-01

    Full Text Available Abstract Background The genome of Bacillus licheniformis DSM 13 harbours three neighbouring open reading frames showing protein sequence similarities to the proteins encoded from the Bacillus subtilis subsp. subtilis 168 gerA operon, GerAA, GerAB and GerAC. In B. subtilis, these proteins are assumed to form a germinant receptor involved in spore germination induced by the amino acid L-alanine. Results In this study we show that disruption of the gerAA gene in B. licheniformis MW3 hamper L-alanine and casein hydrolysate-triggered spore germination, measured by absorbance at 600 nm and confirmed by phase contrast microscopy. This ability was restored by complementation with a plasmid-borne copy of the gerA locus. Addition of D-alanine in the casein hydrolysate germination assay abolished germination of both B. licheniformis MW3 and the complementation mutant. Germination of both B. licheniformis MW3 and the gerA disruption mutant was induced by the non-nutrient germinant Ca2+-Dipicolinic acid. Conclusions These results demonstrate that the B. licheniformis MW3 gerA locus is involved in germination induced by L-alanine and potentially other components present in casein hydrolysate.

  15. Role of the gerA operon in L-alanine germination of Bacillus licheniformis spores

    Science.gov (United States)

    2012-01-01

    Background The genome of Bacillus licheniformis DSM 13 harbours three neighbouring open reading frames showing protein sequence similarities to the proteins encoded from the Bacillus subtilis subsp. subtilis 168 gerA operon, GerAA, GerAB and GerAC. In B. subtilis, these proteins are assumed to form a germinant receptor involved in spore germination induced by the amino acid L-alanine. Results In this study we show that disruption of the gerAA gene in B. licheniformis MW3 hamper L-alanine and casein hydrolysate-triggered spore germination, measured by absorbance at 600 nm and confirmed by phase contrast microscopy. This ability was restored by complementation with a plasmid-borne copy of the gerA locus. Addition of D-alanine in the casein hydrolysate germination assay abolished germination of both B. licheniformis MW3 and the complementation mutant. Germination of both B. licheniformis MW3 and the gerA disruption mutant was induced by the non-nutrient germinant Ca2+-Dipicolinic acid. Conclusions These results demonstrate that the B. licheniformis MW3 gerA locus is involved in germination induced by L-alanine and potentially other components present in casein hydrolysate. PMID:22420404

  16. Resistance of Spores of Clostridium botulinum 33A to Combinations of Ultraviolet and Gamma Rays1

    Science.gov (United States)

    Durban, E.; Grecz, N.

    1969-01-01

    Spores of Clostridium botulinum 33A exhibit a sigmoidal survival curve if subjected to gamma radiation. The present investigation was concerned with two questions: (i) what is the form of an ultraviolet (UV)-survival curve and (ii) what is the combined effect of UV- and gamma radiation? The UV-survival curve was found to be of sigmoidal type with a “shoulder” width of 675 ergs/mm2 and a D10 (exp) of 2,950 ergs/mm2. To test the combination effect, spores were subjected to UV doses of 225, 450, 675, and 900 ergs/mm2 followed by a series of increasing doses of gamma rays from 200 to 2,000 krad in 200-krad steps. The gamma ray-survival curves showed that increasing UV pretreatment caused a gradual loss of the “Prodiginine” yielding straight line exponential survival curves after preirradiation with UV doses of 675 ergs/mm2 and above. Simultaneously the D10 value for gamma-ray irradiation was reduced, e.g. UV preirradiation with 900 ergs/mm2 reduced the D10 by 40%. This observation emphasizes the potential practical advantage of combining UV and gamma rays for sterilization of heat-sensitive commodities. PMID:4896102

  17. Teaching Form as Form

    DEFF Research Database (Denmark)

    Keiding, Tina Bering

    2012-01-01

    understanding of form per se, or, to use an expression from this text, of form as form. This challenge can be reduced to one question: how can design teaching support students in achieving not only the ability to recognize and describe different form-related concepts in existing design (i.e. analytical...... and concepts from real designs by studying form in abstract contexts. The challenge for the first approach is how to support students in decoupling form from the work as a whole. The challenge for the second approach is how to translate general form into real design. Hence, choosing between the two approaches...

  18. 21 CFR 600.10 - Personnel.

    Science.gov (United States)

    2010-04-01

    ... necessary to protect the product from contamination. (3) Pathogenic viruses and spore-forming organisms. Persons working with viruses pathogenic for man or with spore-forming microorganisms, and persons engaged...

  19. 14C Analysis of protein extracts from Bacillus spores.

    Science.gov (United States)

    Cappuccio, Jenny A; Falso, Miranda J Sarachine; Kashgarian, Michaele; Buchholz, Bruce A

    2014-07-01

    Investigators of bioagent incidents or interdicted materials need validated, independent analytical methods that will allow them to distinguish between recently made bioagent samples versus material drawn from the archives of a historical program. Heterotrophic bacteria convert the carbon in their food sources, growth substrate or culture media, into the biomolecules they need. The F(14)C (fraction modern radiocarbon) of a variety of media, Bacillus spores, and separated proteins from Bacillus spores was measured by accelerator mass spectrometry (AMS). AMS precisely measures F(14)C values of biological materials and has been used to date the synthesis of biomaterials over the bomb pulse era (1955 to present). The F(14)C of Bacillus spores reflects the radiocarbon content of the media in which they were grown. In a survey of commercial media we found that the F(14)C value indicated that carbon sources for the media were alive within about a year of the date of manufacture and generally of terrestrial origin. Hence, bacteria and their products can be dated using their (14)C signature. Bacillus spore samples were generated onsite with defined media and carbon free purification and also obtained from archived material. Using mechanical lysis and a variety of washes with carbon free acids and bases, contaminant carbon was removed from soluble proteins to enable accurate (14)C bomb-pulse dating. Since media is contemporary, (14)C bomb-pulse dating of isolated soluble proteins can be used to distinguish between historical archives of bioagents and those produced from recent media. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  20. Characterization of spore laccase from Bacillus subtilis WD23 and ...

    African Journals Online (AJOL)

    The strain was identified as Bacillus subtilis based on its morphological and physiological properties, and 16S rDNA sequence analysis. The optimum pH and temperature for the spore-bound laccase were 6.8 and 60°C, respectively. The temperature half-life of the laccase was 2.5 h at 80°C and 68 h at 60°C. It also showed ...

  1. Antitumor effects and mechanisms of Ganoderma extracts and spores oil

    OpenAIRE

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

    2016-01-01

    Ganoderma lucidum is a popular herbal medicine used in China to promote health. Modern studies have disclosed that the active ingredients of Ganoderma can exhibit several effects, including antitumor effects and immunomodulation. The present study evaluated the antitumor effects of self-prepared Ganoderma extracts and spores oil, and investigated the possible underlying mechanisms by observing the effects of the extracts and oil on topoisomerases and the cell cycle. The results showed that Ga...

  2. Activation of inoculum microorganism from dairy cattle feces

    Science.gov (United States)

    Ayuningtyas, Widya D.; Ridwan, Roni; Joni, I. M.; Marlina, E. T.; Harlia, Ellin

    2018-02-01

    Coal produces Coal Bed Methane (CBM) which is formed both biogenically and thermogenically. Lignite is not utilized optimally because it has low heat content and productivity time limit that decreases CBM production. In order to utilize lignite waste, adding inoculum consortium microorganism from dairy cattle waste as starter for biogas process can be a solution. This study aimed to produce inoculum consortium microorganism as biogas starter from dairy cattle feces through in vitro activation process by Theoudorou modification method. The research used complete randomized design with 3 replications. The treatments were blank (R0), 100% concentrate (R1), 70% concentrate+30% grass (R2), 70% grass+30% concentrate (R3) and 100% grass (R4). All treatments were added by buffer solution and feces with ratio of 2:1 into 100 ml serum injection bottle with anaerobic conditions. The parameters observed were gas production, pH and gas kinetics (orskov's equation) for 2, 4, 6, 8, 10, 12, 24 and 48 hours. The results showed that the treatment had significant effect (P biogas starter selected based on the observed parameters and potential availability of proteolytic and fibrocytic microorganisms is R2 (70% concentrate +30% grass).

  3. Muricholic acids inhibit Clostridium difficile spore germination and growth.

    Directory of Open Access Journals (Sweden)

    Michael B Francis

    Full Text Available Infections caused by Clostridium difficile have increased steadily over the past several years. While studies on C. difficile virulence and physiology have been hindered, in the past, by lack of genetic approaches and suitable animal models, newly developed technologies and animal models allow these processes to be studied in detail. One such advance is the generation of a mouse-model of C. difficile infection. The development of this system is a major step forward in analyzing the genetic requirements for colonization and infection. While important, it is equally as important in understanding what differences exist between mice and humans. One of these differences is the natural bile acid composition. Bile acid-mediated spore germination is an important step in C. difficile colonization. Mice produce several different bile acids that are not found in humans. These muricholic acids have the potential to impact C. difficile spore germination. Here we find that the three muricholic acids (α-muricholic acid, β-muricholic acid and ω-muricholic acid inhibit C. difficile spore germination and can impact the growth of vegetative cells. These results highlight an important difference between humans and mice and may have an impact on C. difficile virulence in the mouse-model of C. difficile infection.

  4. Feasibility of a combined carrier test for disinfectants: studies with a mixture of five types of microorganisms.

    Science.gov (United States)

    Best, M; Springthorpe, V S; Sattar, S A

    1994-06-01

    There is mounting concern regarding the efficacy of many germicides on the market because officially recognized germicidal tests for various classes of microorganisms vary widely and often lack reproducibility and proper quantitation. We report here a carrier method for simultaneously and quantitatively assessing the efficacy of liquid chemical germicides against a mixture of microorganisms of varying degrees of resistance. In the test, each small glass cup (10 mm wide x 14 mm long) was contaminated with 10 microliters of a standardized mixture of Staphylococcus aureus, Mycobacterium bovis bacille Calmette-Guérin, Trichophyton mentagrophytes spores, Sabin poliovirus type 1, and Bacillus stearothermophilus spores in 5% fetal bovine serum. The inoculum was dried for 60 minutes under ambient conditions and covered with 60 microliters of the disinfectant under test or a balanced salt solution control for the desired contact time. The carrier was then placed in 2940 microliters of an eluent and the eluates assayed separately for the five microorganisms. Tap water was used to dilute the test product as needed. Of the 11 products tested, 2% alkaline glutaraldehyde, 0.6% sodium hypochlorite (about 5000 ppm free chlorine), and a 0.4% quarternary ammonium compound containing 23% hydrochloric acid were effective against all five challenge organisms. A hard-surface spray containing 0.1% o-phenylphenol with 79% ethanol was effective against all but bacterial spores; 70% (volume/volume) ethanol alone and povidone-iodine (1% available iodine) were effective against S. aureus, the mycobacterium, and the fungus; a 3% solution of peroxygen compounds was effective only against S. aureus and the poliovirus; 1.5% chlorhexidine gluconate, 0.06% quaternary ammonia compound, and 0.03% o-phenylphenol + 0.03% p-tertiary amylphenol could inactivate nothing but S. aureus; and 3% hydrogen peroxide was ineffective in all tests. This method shows promise for use with various classes of

  5. Morphology and peculiar features of spores of fern species occurring in Poland

    Directory of Open Access Journals (Sweden)

    Elżbieta Zenkteler

    2012-12-01

    Full Text Available The morphology of fern spores collected from natural sites in Poland was examined under light microscopy. Spore samples represented 44 species in 18 genera and in 13 families. Only spores of Ophioglossaceae were obtained from the herbarium of the Adam Mickiewicz University in Poznań while the remaining samples were obtained from living plants. Spore size ranges between 20 to 75 μm and the spores of Osmunda regalis and Polypodium interjectum were found to have remarkably large dimensions. The spores are ellipsoidal, tetrahedral and spherical/globoid in shape. Their apertures are monolete or trilete types. The exine surface patterns are baculate, cristate, granulate, reticulate, tuberculate and verrucate. Pictures of the analyzed spores are collected in a contrasting (size, colour table to make it easier to distinguish between species. The peculiar characters of fern spores are described after a review of major articles concerning the allergenic features of fern spores with special attention to Pteridium aquilinum whose spores and vegetative tissues revealed mutagenic and carcinogenic activity.

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

    Science.gov (United States)

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

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

  7. Bacillus amyloliquefaciens Spore Production Under Solid-State Fermentation of Lignocellulosic Residues.

    Science.gov (United States)

    Berikashvili, Violet; Sokhadze, Kakha; Kachlishvili, Eva; Elisashvili, Vladimir; Chikindas, Michael L

    2017-12-16

    This study was conducted to elucidate cultivation conditions determining Bacillus amyloliquefaciens B-1895 growth and enhanced spore formation during the solid-state fermentation (SSF) of agro-industrial lignocellulosic biomasses. Among the tested growth substrates, corncobs provided the highest yield of spores (47 × 10 10 spores g -1 biomass) while the mushroom spent substrate and sunflower oil mill appeared to be poor growth substrates for spore formation. Maximum spore yield (82 × 10 10 spores g -1 biomass) was achieved when 15 g corncobs were moistened with 60 ml of the optimized nutrient medium containing 10 g peptone, 2 g KH 2 PO 4 , 1 g MgSO 4 ·7H 2 O, and 1 g NaCl per 1 l of distilled water. The cheese whey usage for wetting of lignocellulosic substrate instead water promoted spore formation and increased the spore number to 105 × 10 10 spores g -1 . Addition to the cheese whey of optimized medium components favored sporulation process. The feasibility of developed medium and strategy was shown in scaled up SSF of corncobs in polypropylene bags since yield of 10 × 10 11 spores per gram of dry biomass was achieved. In the SSF of lignocellulose, B. amyloliquefaciens B-1895 secreted comparatively high cellulase and xylanase activities to ensure good growth of the bacterial culture.

  8. Genetic Diversity and Association Characters of Bacteria Isolated from Arbuscular Mycorrhizal Fungal Spore Walls.

    Directory of Open Access Journals (Sweden)

    Gopal Selvakumar

    Full Text Available Association between arbuscular mycorrhizal fungi (AMF and bacteria has long been studied. However, the factors influencing their association in the natural environment is still unknown. This study aimed to isolate bacteria associated with spore walls of AMF and identify their potential characters for association. Spores collected from coastal reclamation land were differentiated based on their morphology and identified by 18S rDNA sequencing as Funneliformis caledonium, Racocetra alborosea and Funneliformis mosseae. Bacteria associated with AMF spore walls were isolated after treating them with disinfection solution at different time intervals. After 0, 10 and 20 min of spore disinfection, 86, 24 and 10 spore associated bacteria (SAB were isolated, respectively. BOX-PCR fingerprinting analysis showed that diverse bacterial communities were associated to AMF spores. Bacteria belonging to the same genera could associate with different AMF spores. Gram positive bacteria were more closely associated with AMF spores. Isolated SAB were characterized and tested for spore association characters such as chitinase, protease, cellulase enzymes and exopolysaccharide production (EPS. Among the 120 SAB, 113 SAB were able to show one or more characters for association and seven SAB did not show any association characters. The 16S rDNA sequence of SAB revealed that bacteria belonging to the phyla Firmicutes, Proteobacteria, Actinobacteria and Bactereiodes were associated with AMF spore walls.

  9. Application of thermotolerant microorganisms for biofertilizer preparation.

    Science.gov (United States)

    Chen, Kuo-Shu; Lin, Yann-Shying; Yang, Shang-Shyng

    2007-12-01

    Intensive agriculture is practised in Taiwan, and compost application is very popular as a means of improving the soil physical properties and supplying plant nutrition. We tested the potential of inoculation with thermotolerant microorganisms to shorten the maturity and improve the quality of biofertilizer prepared by composting. Thermotolerant microorganisms were isolated from compost and reinoculated for the preparation of biofertilizer. The physical, chemical and biological properties of the biofertilizer were determined during composting. The effects of biofertilizer application on the growth and yield of rape were also studied. Among 3823 colonies of thermotolerant microorganisms, Streptomyces thermonitrificans NTU-88, Streptococcus sp. NTU-130 and Aspergillus fumigatus NTU-132 exhibited high growth rates and cellulolytic and proteolytic activities. When a mixture of rice straw and swine manure were inoculated with these isolates and composted for 61 days, substrate temperature increased initially and then decreased gradually during composting. Substrate pH increased from 7.3 to 8.5. Microbial inoculation enhanced the rate of maturity, and increased the content of ash and total and immobilized nitrogen, improved the germination rate of alfalfa seed, and decreased the content of total organic carbon and the carbon/nitrogen ratio. Biofertilizer application increased the growth and yield of rape. Inoculation of thermotolerant and thermophilic microorganisms to agricultural waste for biofertilizer preparation enhances the rate of maturity and improves the quality of the resulting biofertilizer. Inoculation of appropriate microorganisms in biofertilizer preparation might be usefully applied to agricultural situations.

  10. Isolation of Electrogenic Microorganisms with Potential to Reduce Hexavalent Chromium

    Directory of Open Access Journals (Sweden)

    Alexander Mora Collazos

    2017-01-01

    Full Text Available Isolation of cultivable microorganisms was made from the biofilm formed on the anode of a microbial fuel cell put into operation for 30 days; isolated microorganisms were evaluated for their ability to produce energy and reduce the hexavalent chromium Cr (VI. Five microorganisms were isolated, which were characterized by analysis of 16S rRNA gene, placing them in four bacterial genera: Exiguobacterium (CrMFC1, Acinetobacter (CrMFC2, Aeromonas (CrMFC3 and CrMFC5 and Serratia (CrMFC4. All isolates showed electrogenic activity and ability to reduce hexavalent chromium; the Acinetobacter CrMFC1 strain showed the best electrochemical performance registering a maximum power density of 18.61 mW/m2; the other strains showed values of maximum power density between 4.6 mW/m2and 7.1 mW/m2. Strains Aeromonas CrMFC5 and Exiguobacterium CrMFC1 showed the best rates of chromium reduction being able to reduce 100 % of the Cr (VI in less than 24 hours, the Aeromonas CrMFC5 strain was the most efficient, reducing 100 % of Cr (VI in 10 hours; the other strains reduced 100% of the contaminant after 28 to 30 hours. The microorganisms isolated in this study are hardly known for their electrogenic capacity and for reducing Cr (VI; however, show promise for their use in combined systems involving energy production system coupled to bioremediation of chromium contaminated water.

  11. Novel Industrial Enzymes from Uncultured Arctic Microorganisms

    DEFF Research Database (Denmark)

    Vester, Jan Kjølhede

    Many industrial and biotechnological processes make use of cold-active enzymes or could benefit from the use, as the reduced temperature can be beneficial in multiple ways. Such processes may save energy and production costs, improve hygiene, maintain taste and other organoleptic properties......, and reduce the risk of contaminations. Cold- and alkaline-active enzymes can be found in microorganisms adapted to living in natural environments with these conditions, which are extremely rare but found in the unique ikaite columns from SW Greenland (4-6 °C, pH >10). It is estimated that less than 1......% of the microorganisms in an environmental sample can be cultured in the laboratory with standard techniques, which is also the case for the ikaite columns. Thus, there is an enormous potential in the uncultured microorganisms, which cannot be accessed through cultivation based methods. This PhD thesis presents studies...

  12. Physiologically anaerobic microorganisms of the deep subsurface

    Energy Technology Data Exchange (ETDEWEB)

    Stevens, S.E. Jr.; Chung, K.T.

    1991-06-01

    This study seeks to determine numbers, diversity, and morphology of anaerobic microorganisms in 15 samples of subsurface material from the Idaho National Engineering Laboratory, in 18 samples from the Hanford Reservation and in 1 rock sample from the Nevada Test Site; set up long term experiments on the chemical activities of anaerobic microorganisms based on these same samples; work to improve methods for the micro-scale determination of in situ anaerobic microbial activity;and to begin to isolate anaerobes from these samples into axenic culture with identification of the axenic isolates.

  13. Functional Properties of Microorganisms in Fermented Foods

    Science.gov (United States)

    Tamang, Jyoti P.; Shin, Dong-Hwa; Jung, Su-Jin; Chae, Soo-Wan

    2016-01-01

    Fermented foods have unique functional properties imparting some health benefits to consumers due to presence of functional microorganisms, which possess probiotics properties, antimicrobial, antioxidant, peptide production, etc. Health benefits of some global fermented foods are synthesis of nutrients, prevention of cardiovascular disease, prevention of cancer, gastrointestinal disorders, allergic reactions, diabetes, among others. The present paper is aimed to review the information on some functional properties of the microorganisms associated with fermented foods and beverages, and their health-promoting benefits to consumers. PMID:27199913

  14. Involvement of alanine racemase in germination of Bacillus cereus spores lacking an intact exosporium.

    Science.gov (United States)

    Venir, Elena; Del Torre, Manuela; Cunsolo, Vincenzo; Saletti, Rosaria; Musetti, Rita; Stecchini, Mara Lucia

    2014-02-01

    The L-alanine mediated germination of food isolated Bacillus cereus DSA 1 spores, which lacked an intact exosporium, increased in the presence of D-cycloserine (DCS), which is an alanine racemase (Alr) inhibitor, reflecting the activity of the Alr enzyme, capable of converting L-alanine to the germination inhibitor D-alanine. Proteomic analysis of the alkaline extracts of the spore proteins, which include exosporium and coat proteins, confirmed that Alr was present in the B. cereus DSA 1 spores and matched to that encoded by B. cereus ATCC 14579, whose spore germination was strongly affected by the block of conversion of L- to D-alanine. Unlike ATCC 14579 spores, L-alanine germination of B. cereus DSA 1 spores was not affected by the preincubation with DCS, suggesting a lack of restriction in the reactant accessibility.

  15. Diurnal variations of airborne fungal spores concentration in the town and rural area

    Directory of Open Access Journals (Sweden)

    Idalia Kasprzyk

    2012-12-01

    Full Text Available Airborne fungal spores were monitored in 2001-2002 in Rzeszów (town and its neighborhood. The aim of investigations was to ascertain if there were differences in diurnal variations of airborne fungal spores concentration between town and rural area. The sampling was carried out using volumetric method. Traps were located at the same heights - app. 12 m. Airborne spores were sampled continuously. Microscopical slides were prepared for each day. Analysis was carried out on one longitudinal band of 48 mm long divided into 24 segments corresponding following hours of day. The results were expressed as mean number of fungal spores per cubic meter per 24 hours. For this survey, five geni of allergenic fungi were selected: Alternaria, Botrytis, Cladosporium, Epicoccum, Ganoderma. The concentrations of their airborne spores were high or very high. It was calculated theoretical day, where the hourly count was the percentage mean of number of spores at that time every chosen day without rainfall from 2001 and 2001 years. The diurnal periodicity of Alternaria, Cladosporium, Epicoccum and Ganoderma showed one peak, while Botrytis two. Anamorphic spores peaked in the afternoon, while their minima occurred in the morning. The highest concentrations of Ganoderma basidiospores were at down or at night, but minima during the day. There were no clear differences in the peak values between two studied sites. The results indicate that maximum concentrations of all spores generally occurred a few hour earlier in the rural area than in the town. Probably, in the rural area airborne spores came from many local sources and their diurnal periodicity reflected rhythm of spore liberation. Towns are characterized by specific microclimate with higher temperature and wind blowing to the centre. In Rzeszów fungal spores could be transported outside and carried out by wind from distant sources. This study showed, among others, that habitat conditions are an important factors

  16. Effect of Ultrasonic Waves on the Heat Resistance of Bacillus cereus and Bacillus licheniformis Spores

    Science.gov (United States)

    Burgos, J.; Ordóñez, J. A.; Sala, F.

    1972-01-01

    Heat resistance of Bacillus cereus and Bacillus licheniformis spores in quarter-strength Ringer solution decreases markedly after ultrasonic treatments which are unable to kill a significant proportion of the spore population. This effect does not seem to be caused by a loss of Ca2+ or dipicolinic acid. The use of ultrasonics to eliminate vegetative cells or to break aggregates in Bacillus spore suspensions to be used subsequently in heat resistance experiments appears to be unadvisable. PMID:4627969

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

    Science.gov (United States)

    Farenhorst, Marit; Knols, Bart G J

    2010-01-20

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

  18. 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

  19. Computer-assisted image processing to detect spores from the fungus Pandora neoaphidis

    OpenAIRE

    Korsnes, Reinert; Westrum, Karin; Fløistad, Erling; Klingen, Ingeborg

    2016-01-01

    This contribution demonstrates an example of experimental automatic image analysis to detect spores prepared on microscope slides derived from trapping. The application is to monitor aerial spore counts of the entomopathogenic fungus Pandora neoaphidis which may serve as a biological control agent for aphids. Automatic detection of such spores can therefore play a role in plant protection. The present approach for such detection is a modification of traditional manual microscopy of prepared s...

  20. Fecundity, spore recruitment and size in Gelidium sesquipedale (Gelidiales,Rhodophyta)

    OpenAIRE

    Santos, R.; Duarte, Pedro

    1996-01-01

    Gelidium sesquipedale fecundity was quantified by counting tetrasporangial sori and cystocarps per meter squared and by estimating the number of spores contained inside them . These were obtained by regression on a size metric of reproductive structures . Tetrasporangial sori length and cystocarp thickness were the best estimators of spore number. To assess spore recruitment, 12 pottery tiles were fixed to the bottom, and the appearance of small fronds was monitored. No clear s...

  1. Airborne pollen and fungal spores in Garki, Abuja (North-Central Nigeria)

    OpenAIRE

    Ezike, Dimphna Nneka; Nnamani, Catherine V.; Ogundipe, Oluwatoyin T.; Adekanmbi, Olushola H.

    2016-01-01

    The ambient atmosphere is dominated with pollen and spores, which trigger allergic reactions and diseases and impact negatively on human health. A survey of pollen and fungal spores constituents of the atmosphere of Garki, Abuja (North-Central Nigeria) was carried out for 1?year (June 1, 2011?May 31, 2012). The aim of the study was to determine the prevalence and abundance of pollen and fungal spores in the atmosphere and their relationship with meteorological parameters. Airborne samples wer...

  2. Neutrophil chemotactic responses induced by fresh and swollen Rhizopus oryzae spores and Aspergillus fumigatus conidia.

    OpenAIRE

    Waldorf, A R; Diamond, R D

    1985-01-01

    With the induction of germination, Rhizopus oryzae spores and Aspergillus fumigatus conidia activate the complement system and induce neutrophil chemotaxis. In contrast, freshly isolated R. oryzae spores did not induce neutrophil migration into lung tissue of mice after intranasal inoculation. Moreover, in microchemotaxis assays neither fresh R. oryzae spores nor A. fumigatus conidia activated sera to stimulate human neutrophil chemotaxis above control migration until at least 10(7) or 10(8) ...

  3. Environmental microbiology as related to planetary quarantine. [water activity and temperature effects on bacterial spore survival

    Science.gov (United States)

    Pflug, I. J.

    1972-01-01

    The survival of Bacillus subtilis var. niger spores suspended in solutions of sucrose and glycerol at calculated water activities and varying temperatures was studied. The overall results indicated that as the water activity of the liquid decreased from .99 to .85, the heat resistance of the spores increased. The nature of the substance controlling the water activity, and the history of the spores prior to treatment also had an affect on their heat resistance.

  4. Particularities of pathogenic microorganism development at anthropogenic influence and estimate of their adaptation potential by means of radiobiological method

    International Nuclear Information System (INIS)

    Shilina, Yu.V.; Gushcha, N.I.; Dyachenko, A.I.; Dmitriev, A.P.; Molozhava, O.S.; Romashko, V.M.

    2008-01-01

    Influence of anthropogenic factors on ecosystems causes their structure disturbance and reduction of species variety. Some resistance nonspecific forms of pathogenic microorganisms, which have high adaptation potential, become dominant. Thus their aggressiveness can increase. (authors)

  5. Experimental modeling of aerosols produced by microorganisms in working area air as risk factor exerting hazardous impacts on health of workers employed at biotechnological production

    Directory of Open Access Journals (Sweden)

    N.V. Dudchik

    2017-09-01

    Full Text Available Scientific foundation and practices in the sphere of hygienic and ecological standardization concerning biological factors of the environment have a number of peculiarities and are methodically less developed than chemical factors standardization. Efficient industrial control over maximum permissible concentrations of standardized microorganisms-producers in working area air is based on validated instrumental techniques of quantitative assessment. Our goal was to create experimental models for microorganisms-producers' aerosols of a multi-component microbe specimen in working area air as a risk factor causing impacts on health of workers employed at biotechnological production; another task was to work out a procedure for measuring Pseudomonas aurantiaca B-162/255.17concentration and cells and spores of Bacillus sp. BB58-3 strain in working area air. We gave grounds for a technology aimed at quantitative determination of microorganisms-producers in working area air in a modeling experiment; it was based on conventional stages and tech-niques accepted in microbiological practices, namely air samples taking via aspiration technique allowing for a volume taken; cultivation under conditions which are optimal for examined microorganisms-producers in a nutrient medium with reduced composition; calculation of evolved colonies with specific morphological features; morphologic identification of microorganisms and colonies; calculation of microorganisms' quantity on dishes with recalculation per 1 m3 of air. Bas-ing on the detected regular concentration dependences of microbe contamination dynamics in air we worked out a proce-dure for quantitative determination of microorganisms-producers; we also performed metrological estimate of opera-tional properties for assessing microorganisms-producers of a multi-component microbe specimen as a risk factor caus-ing hazardous impacts on health of workers employed at biotechnological production. We validated our

  6. Ecology and metagenomics of soil microorganisms

    Czech Academy of Sciences Publication Activity Database

    Baldrian, Petr; Head, I. M.; Prosser, J. I.; Schloter, M.; Smalla, K.; Tebbe, C. C.

    2011-01-01

    Roč. 78, č. 1 (2011), s. 1-2 ISSN 0168-6496 R&D Projects: GA MŠk LC06066; GA MŠk(CZ) LA10001 Institutional research plan: CEZ:AV0Z50200510 Keywords : microorganism * bioremediation * biogenesis of soil Subject RIV: EE - Microbiology, Virology Impact factor: 3.408, year: 2011

  7. Microorganisms' mediated reduction of β-ketoesters

    African Journals Online (AJOL)

    STORAGESEVER

    2008-10-20

    Oct 20, 2008 ... Several microorganism strains of genera Saccharomyces, Candida, Hansenula, Aspergillus and. Lactobacillus were screened for their ability to perform the reduction of γ-chloro-β-ketobutyric acid ethyl ester to γ-chloro-β-hydroxybutyric acid ethyl ester. The optimal conditions for both stages of the.

  8. Airborne microorganisms and dust from livestock houses

    NARCIS (Netherlands)

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

    2011-01-01

    The objective of this study was to evaluate the efficiencies and suitability of samplers for airborne microorganisms and dust, which could be used in practical livestock houses. Two studies were performed: 1) Testing impaction and cyclone pre-separators for dust sampling in livestock houses; 2)

  9. Modelling the morphology of filamentous microorganisms

    DEFF Research Database (Denmark)

    Nielsen, Jens Bredal

    1996-01-01

    The rapid development in image analysis techniques has made it possible to study the growth kinetics of filamentous microorganisms in more detail than previously, However, owing to the many different processes that influence the morphology it is important to apply mathematical models to extract...

  10. Novel genome alteration system for microorganisms

    NARCIS (Netherlands)

    Daran, J.G.; Geertman, J.M.; Bolat, I.

    2015-01-01

    The invention relates to a set of targeting constructs, comprising a first construct comprising a recognition site for an endonuclease, a first region of homology with a target gene of a microorganism, and a first part of a selection marker, and a second construct comprising a second part of the

  11. Atmospheric Sampling of Microorganisms with UAS

    Science.gov (United States)

    Schmale, D. G., III

    2017-12-01

    Many microorganisms relevant to crops, domestic animals, and humans are transported over long distances through the atmosphere. Some of these atmospheric microbes catalyze the freezing of water at higher temperatures and facilitate the onset of precipitation. A few have crossed continents. New technologies are needed to study the movement of microorganisms in the atmosphere. We have used unmanned aircraft systems (UAS) to study the transport of microorganisms tens to hundreds of meters above the ground. These UAS are equipped with unique devices for collecting microbes in the atmosphere during flight. Autonomous systems enable teams of UAS to perform complex atmospheric sampling tasks, and coordinate flight missions with one another. Data collected with UAS can be used to validate and improve disease forecasting models along highways in the sky, connecting transport scales across farms, states, and continents. Though terrestrial environments are often considered a major contributor to atmospheric microbial aerosols, little is known about aquatic sources of microbial aerosols. Droplets containing microorganisms can aerosolize from the water surface, liberating them into the atmosphere. We are using teams of unmanned surface vehicles (USVs) and UAS to study the aerosolization of microbes from aquatic environments. Controlled flume studies using highspeed video have allowed us to observe unique aerosolization phenomena that can launch microbes out of the water and into the air. Unmanned systems may be used to excite the next generation of biologists and engineers, and raise important ethical considerations about the future of human-robot interactions.

  12. Host Defense Against Opportunist Microorganisms Following Trauma

    Science.gov (United States)

    1989-06-30

    Include Security Clasification ) (U) Host Defense Against Opportunist Microorganisms Following Trauma 12. PERSONAL AUTHOR(S) Bjornson, A. B., Bjornson...the course of bacterial and viral infec- tions. Infect. Immun. 30:824-831. 25. Zimmerli, W., B. Seligmann, and J. I. Gallin. 1986. Exudation primes

  13. Risk Assessment of Genetically Modified Microorganisms

    DEFF Research Database (Denmark)

    Jacobsen, B. L.; Wilcks, Andrea

    2001-01-01

    the industry, national administration and research institutions were gathered to discuss which elements should be considered in a risk assessment of genetically modified microorganisms used as food or food ingredients. The existing EU and national regulations were presented, together with the experiences...

  14. Genetic fingerprint of microorganisms associated with the ...

    Indian Academy of Sciences (India)

    adapting molecular techniques to identify microorganisms. Molecular ... The under- standing achieved from molecular identification of the mi- .... cillium brevicompactum associated with the different species of Bacillus. It is been reported that the death and lyses of such bacteria would promote the growth of fungi (Garg et al.

  15. False identification of other microorganisms as Staphylococcus ...

    African Journals Online (AJOL)

    Methods: 507 microorganisms which have been previously identified as S. aureus in 8 States in Southern Nigeria through characteristic morphology on blood agar, Gram staining, growth and fermentation on Mannitol Salt Agar and coagulase formation were collected. All the isolates were identified in this study through ...

  16. Pesticides in Soil: Effects on Microorganisms

    Directory of Open Access Journals (Sweden)

    Ljiljana Radivojević

    2007-01-01

    Full Text Available Since their discovery to the present day, pesticides have been an inevitable segment of agricultural production and efforts have been made to synthesize compounds that would share a required efficacy along with selectivity, sufficient persistence on the object of protection and favourable toxicological and ecotoxicological characteristics so as to minimize their effect on the environment.When a pesticide gets into soil after application, it takes part in a number of physical, chemical and biological processes that depend not only on the compound itself, but a number of other factors as well, such as: physical, chemical and biological characteristics of soil; climatic factors, equipment used, method of application, method of storage, handling and disposal of waste, site characteristics (proximity of ground and underground waters, biodiversity and sensitivity of the environment. Microorganisms play an important role in pesticide degradation as they are able to utilize the biogenic elements from those compounds, as well as energy for their physiological processes. On the other hand, pesticides are more or less toxic substances that can have adverse effect on populations of microorganisms and prevent their development, reduce their abundance, deplete their taxonomic complexity and create communities with a lower level of diversity and reduced physiological activity.The article discusses complex interactions between pesticides and microorganisms in soil immediately after application and over the ensuing period. Data on changes in the abundance of some systematic and physiological groups of microorganisms, their microbial biomass and enzymatic activity caused under pesticide activity are discussed as indicators of these processes.

  17. Functional Properties of Microorganisms in Fermented Foods

    OpenAIRE

    Jyoti Prakash Tamang; Dong Hwa Shin; Soo Wan eChae; Su-Jin eJung

    2016-01-01

    Fermented foods have unique functional properties imparting some health benefits to consumers due to presence of functional microorganisms, which possess probiotics properties, antimicrobial, antioxidant, peptide production, etc. Health benefits of some global fermented foods are synthesis of nutrients, prevention of cardiovascular disease, prevention of cancer, gastrointestinal disorders, allergic reactions, diabetes, among others. The present paper is aimed to review the information on some...

  18. Antibiotic Sensitivity Pattern of Microorganisms Isolated from ...

    African Journals Online (AJOL)

    Antibiotic sensitivity pattern of microorganisms isolated from smoked and frozen fishes sold in Benin and Warri metropolis were investigated. Adopting microbiological standard techniques, the results of the bacterial counts and fungal counts ranged from 5.4 x 106 (Ekpan market) to 25.1 x 106 (Ekpan market) and 1.1 x 105 ...

  19. Ecophysiology of microorganisms in microbial elctrolysis cells

    NARCIS (Netherlands)

    Croese, E.

    2012-01-01

    One of the main challenges for improvement of the microbial electrolysis cell (MEC) has been the reduction of the cost of the cathode catalyst. As catalyst at the cathode, microorganisms offer great possibilities. Previous research has shown the principle possibilities for the biocathode for H2

  20. Tapping uncultured microorganisms through metagenomics for drug ...

    African Journals Online (AJOL)

    Natural products have been an important historical source of therapeutic agents. Microorganisms are major source of bioactive natural products, and several microbial products including antibiotics, anti-inflammatory, anti-tumour, immunosuppressants and others are currently used as therapeutic agents for human and ...

  1. Microorganisms may play role in waste management

    International Nuclear Information System (INIS)

    Anon.

    1990-01-01

    ORNL researchers are studying microorganisms that remove metals from water for possible use in managing radioactive and hazardous wastes in Oak Ridge and elsewhere. These organisms including bacteria, fungi and algae, selectively absorb metals from water. Thus, in high concentrations, they could be used to treat wastewater containing radioactive or toxic metals

  2. Microorganisms in the aetiology of atherosclerosis

    NARCIS (Netherlands)

    Morré, S. A.; Stooker, W.; Lagrand, W. K.; van den Brule, A. J.; Niessen, H. W.

    2000-01-01

    Recent publications have suggested that infective pathogens might play an important role in the pathogenesis of atherosclerosis. This review focuses on these microorganisms in the process of atherosclerosis. The results of in vitro studies, animal studies, tissue studies, and serological studies

  3. Microorganisms as Indicators of Soil Health

    DEFF Research Database (Denmark)

    Nielsen, M. N.; Winding, A.; Binnerup, S.

    Microorganisms are an essential part of living soil and of outmost importance for soil health. As such they can be used as indicators of soil health. This report reviews the current and potential future use of microbial indicators of soil health and recommends specific microbial indicators for soil...... indicators into soil monitoring programmes as they become applicable....

  4. The influence of selected nanomaterials on microorganisms

    Czech Academy of Sciences Publication Activity Database

    Brandeburová, P.; Birošová, L.; Vojs, M.; Kromka, Alexander; Gál, M.; Tichý, J.; Híveš, J.; Mackul´ak, T.

    2017-01-01

    Roč. 148, č. 3 (2017), s. 525-530 ISSN 0026-9247 R&D Projects: GA ČR GA15-01687S Institutional support: RVO:68378271 Keywords : nanomaterials * nanotechnologies * microorganisms * toxicity Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 1.282, year: 2016

  5. Estimation of Available Phosphorus in Soil Using the Population of Arbuscular Mycorrhizal Fungi Spores

    Directory of Open Access Journals (Sweden)

    Machfud Effendy

    2011-09-01

    Full Text Available Soil microbes, such as arbuscular mycorrhizal fungi (AMF have the ability to dissolve unavailable phosphorus (P and they can be used as an indicator of the P availability in soil. The study was conducted on upland soil in East Java. The soil was sampled twice, before and after planting at the harvesting time. The population of AMF spores and soil P availability were observed. The AMF spores were isolated using wet sieving method, decanting, and followed by sucrose density gradient centrifugation. The available P was observed using the Olsen extraction. The numbers of AMF spore was corelated with available P, moreover the numbers of AMF spore was compared to the availabality of P. The results showed that the total number of AMF spores at six sites were ranged from a little to midle, and the available P ranged from low to high level. All soil site samples had a linear corelation between numbers of AMF spore and available P in soil. The greater the number of AMF spore, the higher the available P in soil. It was likely that the availability of P in soil can be predicted by the population of AMF spores in soil. Therefore, the number of AMF spore can be need as a biological method to predict the available P in soil and to make a recommendation the use of P fertilizer.

  6. Non-Seasonal Variation of Airborne Aspergillus Spore Concentration in a Hospital Building

    Directory of Open Access Journals (Sweden)

    Michael Oberle

    2015-10-01

    Full Text Available Nosocomial fungal infections are gaining increased attention from infectiologists. An adequate investigation into the levels of airborne Aspergillus and other fungal spores in hospital settings, under normal conditions, is largely unknown. We monitored airborne spore contamination in a Swiss hospital building in order to establish a seasonally-dependent base-line level. Air was sampled using an impaction technique, twice weekly, at six different locations over one year. Specimens were seeded in duplicate on Sabouraud agar plates. Grown colonies were identified to genus levels. The airborne Aspergillus spore concentration was constantly low throughout the whole year, at a median level of 2 spores/m3 (inter-quartile range = IQR 1–4, and displayed no seasonal dependency. The median concentration of other fungal spores was higher and showed a distinct seasonal variability with the ambient temperature change during the different seasons: 82 spores/m3 (IQR 26–126 in summer and 9 spores/m3 (IQR 6–15 in winter. The spore concentration varied considerably between the six sampling sites in the building (10 to 26 spores/m3. This variability may explain the variability of study results in the literature.

  7. Bacillus spores and their relevant chemicals studied by terahertz time domain spectroscopy

    Science.gov (United States)

    Tang, Jianhua; Yang, Bin; Llewellyn, Ian; Cutler, Ronald R.; Donnan, Robert S.

    2014-01-01

    Terahertz time domain spectroscopy has been used to investigate 0.2-2.2 THz transmission responses of Bacillus spores and their related chemical components. Whilst no THz signatures could be clearly associated with either sporulated cells or their chief chemical components, differing degrees of signal attenuation and frequency-dependent light scattering were observed depending on spore composition and culture media. The observed monotonic increase in absorption by spores over this THz spectral domain is mainly from Mie scattering and also from remnant water bound to the spores.

  8. Effects of meteorological factors on the levels of Alternaria spores on a potato crop

    Science.gov (United States)

    Escuredo, Olga; Seijo, Maria Carmen; Fernández-González, Maria; Iglesias, Isabel

    2011-03-01

    Alternaria solani Soraeur produces early blight in Solanum tuberosum L., leading to significant agricultural losses. The current study was carried out on the extensive potato crop situated in north-western of Spain during 2007, 2008 and 2009. In this area potato crops are the most important source of income. In this work we used a Hirst-type volumetric spore-trap for the aerobiological monitoring of Alternaria spores. The highest spore concentrations were recorded during the 2009 cycle (10,555 spores), and the lowest concentrations were recorded during the 2008 cycle (5,471 spores). Over the 3 years of study, the highest concentrations were registered during the last stage of the crop. The aim of the study was to observe the influence of meteorological factors on the concentration of Alternaria spores, which can lead to serious infection and early blight. Prediction of the stages during which a crop is particularly vulnerable to infection allows for adjustment of the application of fungicide and is of environmental and agricultural importance. For this reason, we tested three models (P-Days, DD and IWP) to predict the first treatment and decrease the negative effect that these spores have on potato crops. The parameter that showed the most significant correlation with spore concentrations was minimum temperature. We used ARIMA (autoregressive integrated model of running mean) time-series models to determine the forecast. We considered weather data as predictor variables and the concentration of spores on the previous day as the fixed variable.

  9. Decontamination of B. globigii spores from drinking water infrastructure using disinfectants

    Data.gov (United States)

    U.S. Environmental Protection Agency — Decontamination of Bacillus spores adhered to common drinking water infrastructure surfaces was evaluated using a variety of disinfectants. Corroded iron and...

  10. Two distinct groups within the Bacillus subtilis group display significantly different spore heat resistance properties.

    Science.gov (United States)

    Berendsen, Erwin M; Zwietering, Marcel H; Kuipers, Oscar P; Wells-Bennik, Marjon H J

    2015-02-01

    The survival of bacterial spores after heat treatment and the subsequent germination and outgrowth in a food product can lead to spoilage of the food product and economical losses. Prediction of time-temperature conditions that lead to sufficient inactivation requires access to detailed spore thermal inactivation kinetics of relevant model strains. In this study, the thermal inactivation kinetics of spores of fourteen strains belonging to the Bacillus subtilis group were determined in detail, using both batch heating in capillary tubes and continuous flow heating in a micro heater. The inactivation data were fitted using a log linear model. Based on the spore heat resistance data, two distinct groups (p subtilis group could be identified. One group of strains had spores with an average D120 °C of 0.33 s, while the spores of the other group displayed significantly higher heat resistances, with an average D120 °C of 45.7 s. When comparing spore inactivation data obtained using batch- and continuous flow heating, the z-values were significantly different, hence extrapolation from one system to the other was not justified. This study clearly shows that heat resistances of spores from different strains in the B. subtilis group can vary greatly. Strains can be separated into two groups, to which different spore heat inactivation kinetics apply. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Bacillus subtilis spores PROTECT experiment Space-exposed and Mars-exposed vs. Earth-control

    Data.gov (United States)

    National Aeronautics and Space Administration — Because of their ubiquity and resistance to spacecraft decontamination bacterial spores are considered likely potential forward contaminants on robotic missions to...

  12. A genetic algorithm-Bayesian network approach for the analysis of metabolomics and spectroscopic data: application to the rapid identification of Bacillus spores and classification of Bacillus species.

    Science.gov (United States)

    Correa, Elon; Goodacre, Royston

    2011-01-26

    The rapid identification of Bacillus spores and bacterial identification are paramount because of their implications in food poisoning, pathogenesis and their use as potential biowarfare agents. Many automated analytical techniques such as Curie-point pyrolysis mass spectrometry (Py-MS) have been used to identify bacterial spores giving use to large amounts of analytical data. This high number of features makes interpretation of the data extremely difficult We analysed Py-MS data from 36 different strains of aerobic endospore-forming bacteria encompassing seven different species. These bacteria were grown axenically on nutrient agar and vegetative biomass and spores were analyzed by Curie-point Py-MS. We develop a novel genetic algorithm-Bayesian network algorithm that accurately identifies sand selects a small subset of key relevant mass spectra (biomarkers) to be further analysed. Once identified, this subset of relevant biomarkers was then used to identify Bacillus spores successfully and to identify Bacillus species via a Bayesian network model specifically built for this reduced set of features. This final compact Bayesian network classification model is parsimonious, computationally fast to run and its graphical visualization allows easy interpretation of the probabilistic relationships among selected biomarkers. In addition, we compare the features selected by the genetic algorithm-Bayesian network approach with the features selected by partial least squares-discriminant analysis (PLS-DA). The classification accuracy results show that the set of features selected by the GA-BN is far superior to PLS-DA.

  13. Characteristics and determinants of ambient fungal spores in Hualien, Taiwan

    Science.gov (United States)

    Ho, Hsiao-Man; Rao, Carol Y.; Hsu, Hsiao-Hsien; Chiu, Yueh-Hsiu; Liu, Chi-Ming; Chao, H. Jasmine

    Characteristics and determinants of ambient aeroallergens are of much concern in recent years because of the apparent health impacts of allergens. Yet relatively little is known about the complex behaviors of ambient aeroallergens. To address this issue, we monitored ambient fungal spores in Hualien, Taiwan from 1993-1996 to examine the compositions and temporal variations of fungi, and to evaluate possible determinants. We used a Burkard seven-day volumetric spore trap to collect daily fungal spores. Air pollutants, meteorological factors, and Asian dust events were included in the statistical analyses to predict fungal levels. We found that the most dominant fungal categories were ascospores, followed by Cladosporium and Aspergillus/Penicillium. The majority of the fungal categories had significant diurnal and seasonal variations. Total fungi, Cladosporium, Ganoderma, Arthrinium/Papularia, Cercospora, Periconia, Alternaria, Botrytis, and PM 10 had significantly higher concentrations ( p<0.05) during the period affected by Asian dust events. In multiple regression models, we found that temperature was consistently and positively associated with fungal concentrations. Other factors correlated with fungal concentrations included ozone, particulate matters with an aerodynamic diameter less than 10 μm (PM 10), relative humidity, rainfall, atmospheric pressure, total hydrocarbons, carbon monoxide, nitrogen dioxide, and sulfur dioxide. Most of the fungal categories had higher levels in 1994 than in 1995-96, probably due to urbanization of the study area. In this study, we demonstrated complicated interrelationships between fungi and air pollution/meteorological factors. In addition, long-range transport of air pollutants contributed significantly to local aeroallergen levels. Future studies should examine the health impacts of aeroallergens, as well as the synergistic/antagonistic effects of weather, and local and global-scale air pollutions.

  14. Fern spore longevity in saline water: can sea bottom sediments maintain a viable spore bank?

    NARCIS (Netherlands)

    Groot, de G.A.; During, H.

    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 interand intraspecific diversity, and are increasingly exploited to facilitate diversity

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

    Science.gov (United States)

    Warda, Alicja K; den Besten, Heidy M W; Sha, Na; Abee, Tjakko; Nierop Groot, Masja N

    2015-05-18

    Spoilage of heat treated foods can be caused by the presence of surviving spore-formers. It is virtually impossible to prevent contamination at the primary production level as spores are ubiquitous present in the environment and can contaminate raw products. As a result spore inactivation treatments are widely used by food producing industries to reduce the microbial spore loads. However consumers prefer mildly processed products that have less impact on its quality and this trend steers industry towards milder preservation treatments. Such treatments may result in damaged instead of inactivated spores, and these spores may germinate, repair, and grow out, possibly leading to quality and safety issues. The ability to repair and grow out is influenced by the properties of the food matrix. In the current communication we studied the outgrowth from heat damaged Bacillus cereus ATCC 14579 spores on Anopore membrane, which allowed following outgrowth heterogeneity of individual spores on broccoli and rice-based media as well as standard and mildly acidified (pH 5.5) meat-based BHI. Rice, broccoli and BHI pH 5.5 media resulted in delayed outgrowth from untreated spores, and increased heterogeneity compared to BHI pH 7.4, with the most pronounced effect in rice media. Exposure to wet heat for 1 min at 95 °C caused 2 log inactivation and approximately 95% of the spores in the surviving fraction were damaged resulting in substantial delay in outgrowth based on the time required to reach a maximum microcolony size of 256 cells. The delay was most pronounced for heat-treated spores on broccoli medium followed by spores on rice media (both untreated and treated). Interestingly, the increase in outgrowth heterogeneity of heat treated spores on BHI pH 7.4 was more pronounced than on rice, broccoli and BHI pH 5.5 conceivably reflecting that conditions in BHI pH 7.4 better support spore damage repair. This study compares the effects of three main factors, namely heat treatment, p

  16. Cytological and proteomic analyses of horsetail (Equisetum arvense L. spore germination

    Directory of Open Access Journals (Sweden)

    Qi eZhao

    2015-06-01

    Full Text Available Spermatophyte pollen tubes and root hairs have been used as single-cell-type model systems to understand the molecular processes underlying polar growth of plant cells. Horsetail (Equisetum arvense L. is a perennial herb species in Equisetopsida, which creates separately growing spring and summer stems in its life cycle. The mature chlorophyllous spores produced from spring stems can germinate without dormancy. Here we report the cellular features and protein expression patterns in five stages of horsetail spore germination (mature spores, rehydrated spores, double-celled spores, germinated spores, and spores with protonemal cells. Using 2-DE combined with mass spectrometry, 80 proteins were found to be abundance changed upon spore germination. Among them, proteins involved in photosynthesis, protein turnover, and energy supply were over-represented. Thirteen proteins appeared as proteoforms on the gels, indicating the potential importance of post-translational modification. In addition, the dynamic changes of ascorbate peroxidase, peroxiredoxin, and dehydroascorbate reductase implied that reactive oxygen species homeostasis is critical in regulating cell division and tip-growth. The diverse expression patterns of proteins in photosynthesis, energy supply, lipid and amino acid metabolism indicated that heterotrophic and autotrophic metabolism were necessary in light-dependent germination of the spores. Twenty-six proteins were involved in protein synthesis and fate, indicating that protein turnover is vital to spore germination. Furthermore, the altered abundance of small G protein Ran, 14-3-3 protein, actin, and Caffeoyl-CoA O-methyltransferase revealed that signaling transduction, vesicle trafficking, cytoskeleton dynamics, and cell wall modulation were critical to cell division and polar growth. These findings lay a foundation toward understanding the molecular mechanisms underlying fern spore asymmetric division and rhizoid polar growth.

  17. Induced Sporicidal Activity of Chlorhexidine against Clostridium difficile Spores under Altered Physical and Chemical Conditions

    Science.gov (United States)

    Nerandzic, Michelle M.; Donskey, Curtis J.

    2015-01-01

    Background Chlorhexidine is a broad-spectrum antimicrobial commonly used to disinfect the skin of patients to reduce the risk of healthcare-associated infections. Because chlorhexidine is not sporicidal, it is not anticipated that it would have an impact on skin contamination with Clostridium difficile, the most important cause of healthcare-associated diarrhea. However, although chlorhexidine is not sporicidal as it is used in healthcare settings, it has been reported to kill spores of Bacillus species under altered physical and chemical conditions that disrupt the spore’s protective barriers (e.g., heat, ultrasonication, alcohol, or elevated pH). Here, we tested the hypothesis that similarly altered physical and chemical conditions result in enhanced sporicidal activity of chlorhexidine against C. difficile spores. Principal Findings C. difficile spores became susceptible to heat killing at 80°C within 15 minutes in the presence of chlorhexidine, as opposed to spores suspended in water which remained viable. The extent to which the spores were reduced was directly proportional to the concentration of chlorhexidine in solution, with no viable spores recovered after 15 minutes of incubation in 0.04%–0.0004% w/v chlorhexidine solutions at 80°C. Reduction of spores exposed to 4% w/v chlorhexidine solutions at moderate temperatures (37°C and 55°C) was enhanced by the presence of 70% ethanol. However, complete elimination of spores was not achieved until 3 hours of incubation at 55°C. Elevating the pH to ≥9.5 significantly enhanced the killing of spores in either aqueous or alcoholic chlorhexidine solutions. Conclusions Physical and chemical conditions that alter the protective barriers of C. difficile spores convey sporicidal activity to chlorhexidine. Further studies are necessary to identify additional agents that may allow chlorhexidine to reach its target within the spore. PMID:25861057

  18. Bacterial spore inactivation by atmospheric-pressure plasmas in the presence or absence of UV photons as obtained with the same gas mixture

    International Nuclear Information System (INIS)

    Boudam, M K; Moisan, M; Saoudi, B; Popovici, C; Gherardi, N; Massines, F

    2006-01-01

    This paper comprises two main parts: a review of the literature on atmospheric-pressure discharges used for micro-organism inactivation, focused on the inactivation mechanisms, and a presentation of our research results showing, in particular, that UV photons can be the dominant species in the inactivation process. The possibility of achieving spore inactivation through UV radiation using an atmospheric-pressure discharge or its flowing afterglow is the object of a continuing controversy. In fact, the review of the literature that we present shows that a majority of researchers have come to the conclusion that, at atmospheric pressure, chemically reactive species such as free radicals, metastable atoms and molecules always control the inactivation process, while UV photons play only a minor role or no role at all. In contrast, only a few articles suggest or claim that UV photons coming from atmospheric-pressure discharges can, in some cases, inactivate micro-organisms, but the experimental data presented and the supporting arguments brought forward in that respect are relatively incomplete. Using a dielectric-barrier discharge operated at atmospheric pressure in an N 2 -N 2 O mixture, we present, for the first time, experiments where micro-organisms are subjected to plasma conditions such that, on the one hand, UV radiation is strong or, on the other hand, there is no UV radiation, the two different situations being obtained with the same experimental arrangement, including the same gas mixture, N 2 -N 2 O. To achieve maximum UV radiation, the concentration of the oxidant molecule (N 2 O) added to N 2 needs to be tuned carefully, resulting then in the fastest inactivation rate. The concentration range of the oxidant molecule in the mixture for which the UV intensity is significant is extremely narrow, a fact that possibly explains why such a mode of plasma sterilization was not readily observed. The survival curves obtained under dominant UV radiation conditions

  19. [The study of mycolytic properties of aerobic spore-forming bacteria producing extracellular chitinases].

    Science.gov (United States)

    Aktuganov, G E; Melent'ev, A I; Galimzianova, N F; Shirokov, A V

    2008-01-01

    The mycolytic activity of 27 strains of antagonistic bacilli belonging to two taxonomic groups (18 strains of Bacillus subtilis and 9 strains of Paenibacillus ehimensis) capable of induced synthesis of chitinolytic enzymes was studied. Most of the B. subtilis strains neither displayed visible mycolytic effects on the phytopathogenic fungus Bipolaris sorokiniana in vitro, nor produced chitinases in the presence of an auto-claved mycelium. On the contrary, P. ehimensis strains grown under conditions favorable for induction of chitinases and other hydrolases exhibited a pronounced lytic effect on B. sorokiniana and actively grew by utilizing mycelium as the sole source of carbon and nitrogen. Comparison of the mycolytic activities of extracellular hydrolases in the studied strains demonstrated low correlation between chitinase production and the ability of the strains to degrade the cell walls of B. sorokiniana. Characterization of enzyme profiles in the studied strains revealed that beta-1,3-glucanase was a more significant factor than chitinase for determining the mycolytic potential of bacteria and their ability to utilize the mycelium of phytopathogenic fungi as a growth substrate.

  20. Safety assesment of Bacillus clausii UBBC07, a spore forming probiotic

    Directory of Open Access Journals (Sweden)

    Suvarna G. Lakshmi

    Full Text Available Probiotics are vital bacteria that colonize the intestine and modify its microflora with benefits for the host. Very few members of the Bacillus group are recognized as safe for use and hence only a few strains are available as commercial preparations for application in humans and animals. Acute and subacute studies in rats were conducted to establish safety of Bacillus clausii (B. clausii UBBC07. In the acute toxicity study, the oral LD50 for B. clausii UBBC07 was found to be >5000 mg/kg (630 billion cfu/kg body weight. The NOAEL for B. clausii UBBC07 was found to be 1000 (126 billion cfu mg/kg body weight/day by oral route in the subacute toxicity study. There were no significant differences between control and treated groups in any of the endpoints assessed using an OECD443 or OECD407 protocol.B. clausii UBBC07 was found to be resistant to three antibiotics −clindamycin, erythromycin and chloramphenicol. Analysis of the whole genome sequence of B. clausii UBBC07 revealed that the antibiotic resistance genes are present in chromosomal DNA which is intrinsic and not transferable. Toxin genes were also found to be absent. These results suggest consumption of B. clausii UBBC07 is safe for humans. Keywords: Acute toxicity, Subacute toxicity, NOAEL, Bacillus clausii UBBC07, Whole genome