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Sample records for spores confirmed mitosomal

  1. Immunolocalization of an alternative respiratory chain in Antonospora (Paranosema) locustae spores: mitosomes retain their role in microsporidial energy metabolism.

    Science.gov (United States)

    Dolgikh, Viacheslav V; Senderskiy, Igor V; Pavlova, Olga A; Naumov, Anton M; Beznoussenko, Galina V

    2011-04-01

    Microsporidia are a group of fungus-related intracellular parasites with severely reduced metabolic machinery. They lack canonical mitochondria, a Krebs cycle, and a respiratory chain but possess genes encoding glycolysis enzymes, a glycerol phosphate shuttle, and ATP/ADP carriers to import host ATP. The recent finding of alternative oxidase genes in two clades suggests that microsporidial mitosomes may retain an alternative respiratory pathway. We expressed the fragments of mitochondrial chaperone Hsp70 (mitHsp70), mitochondrial glycerol-3-phosphate dehydrogenase (mitG3PDH), and alternative oxidase (AOX) from the microsporidium Antonospora (Paranosema) locustae in Escherichia coli. Immunoblotting with antibodies against recombinant polypeptides demonstrated specific accumulation of both metabolic enzymes in A. locustae spores. At the same time comparable amounts of mitochondrial Hsp70 were found in spores and in stages of intracellular development as well. Immunoelectron microscopy of ultrathin cryosections of spores confirmed mitosomal localization of the studied proteins. Small amounts of enzymes of an alternative respiratory chain in merogonial and early sporogonial stages, alongside their accumulation in mature spores, suggest conspicuous changes in components and functions of mitosomes during the life cycle of microsporidia and the important role of these organelles in parasite energy metabolism, at least at the final stages of sporogenesis.

  2. Globulispora mitoportans n. g., n. sp., (Opisthosporidia: Microsporidia) a microsporidian parasite of daphnids with unusual spore organization and prominent mitosome-like vesicles

    Czech Academy of Sciences Publication Activity Database

    Vávra, Jiří; Hyliš, M.; Fiala, Ivan; Nebesářová, Jana

    2016-01-01

    Roč. 135, MAR (2016), s. 43-52 ISSN 0022-2011 Institutional support: RVO:60077344 Keywords : Microsporidia * Fungi * Daphnia * parasite * mitosome * phylogeny Subject RIV: EG - Zoology Impact factor: 2.379, year: 2016

  3. Screening and discovery of lineage-specific mitosomal membrane proteins in Entamoeba histolytica.

    Science.gov (United States)

    Santos, Herbert J; Imai, Kenichiro; Hanadate, Yuki; Fukasawa, Yoshinori; Oda, Toshiyuki; Mi-Ichi, Fumika; Nozaki, Tomoyoshi

    Entamoeba histolytica, an anaerobic intestinal parasite causing dysentery and extra-intestinal abscesses in humans, possesses highly reduced and divergent mitochondrion-related organelles (MROs) called mitosomes. This organelle lacks many features associated with canonical aerobic mitochondria and even other MROs such as hydrogenosomes. The Entamoeba mitosome has been found to have a compartmentalized sulfate activation pathway, which was recently implicated to have a role in amebic stage conversion. It also features a unique shuttle system via Tom60, which delivers proteins from the cytosol to the mitosome. In addition, only Entamoeba mitosomes possess a novel subclass of β-barrel outer membrane protein called MBOMP30. With the discoveries of such unique features of mitosomes of Entamoeba, there still remain a number of significant unanswered issues pertaining to this organelle. Particularly, the present understanding of the inner mitosomal membrane of Entamoeba is extremely limited. So far, only a few homologs for transporters of various substrates have been confirmed, while the components of the protein translocation complexes appear to be absent or are yet to be discovered. Employing a similar strategy as in our previous work, we collaborated to screen and discover mitosomal membrane proteins. Using a specialized prediction pipeline, we searched for proteins possessing α-helical transmembrane domains, which are unique to E. histolytica mitosomes. From the prediction algorithm, 25 proteins emerged as candidates, two of which were initially observed to be localized to the mitosomes. Further screening and analysis of the predicted proteins may provide clues to answer key questions on mitosomal evolution, biogenesis, dynamics, and biochemical processes. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  4. Mitosomes in Trophozoites and Cysts of the Reptilian Parasite Entamoeba invadens ▿

    Science.gov (United States)

    Siegesmund, Maria A.; Hehl, Adrian B.; van der Giezen, Mark

    2011-01-01

    Heat shock protein genes led to the discovery of mitosomes in Entamoeba histolytica, but mitosomes have not been described for any other Entamoeba species, nor have they been identified in the cyst stage. Here, we show that the distantly related reptilian pathogen Entamoeba invadens contains mitosomes, in both trophozoites and cysts, suggesting all Entamoeba species contain these organelles. PMID:21965513

  5. Mitosomes in trophozoites and cysts of the reptilian parasite Entamoeba invadens.

    Science.gov (United States)

    Siegesmund, Maria A; Hehl, Adrian B; van der Giezen, Mark

    2011-11-01

    Heat shock protein genes led to the discovery of mitosomes in Entamoeba histolytica, but mitosomes have not been described for any other Entamoeba species, nor have they been identified in the cyst stage. Here, we show that the distantly related reptilian pathogen Entamoeba invadens contains mitosomes, in both trophozoites and cysts, suggesting all Entamoeba species contain these organelles.

  6. Novel TPR-containing subunit of TOM complex functions as cytosolic receptor for Entamoeba mitosomal transport.

    Science.gov (United States)

    Makiuchi, Takashi; Mi-ichi, Fumika; Nakada-Tsukui, Kumiko; Nozaki, Tomoyoshi

    2013-01-01

    Under anaerobic environments, the mitochondria have undergone remarkable reduction and transformation into highly reduced structures, referred as mitochondrion-related organelles (MROs), which include mitosomes and hydrogenosomes. In agreement with the concept of reductive evolution, mitosomes of Entamoeba histolytica lack most of the components of the TOM (translocase of the outer mitochondrial membrane) complex, which is required for the targeting and membrane translocation of preproteins into the canonical aerobic mitochondria. Here we showed, in E. histolytica mitosomes, the presence of a 600-kDa TOM complex composed of Tom40, a conserved pore-forming subunit, and Tom60, a novel lineage-specific receptor protein. Tom60, containing multiple tetratricopeptide repeats, is localized to the mitosomal outer membrane and the cytosol, and serves as a receptor of both mitosomal matrix and membrane preproteins. Our data indicate that Entamoeba has invented a novel lineage-specific shuttle receptor of the TOM complex as a consequence of adaptation to an anaerobic environment.

  7. Novel TPR-containing subunit of TOM complex functions as cytosolic receptor for Entamoeba mitosomal transport

    Science.gov (United States)

    Makiuchi, Takashi; Mi-ichi, Fumika; Nakada-Tsukui, Kumiko; Nozaki, Tomoyoshi

    2013-01-01

    Under anaerobic environments, the mitochondria have undergone remarkable reduction and transformation into highly reduced structures, referred as mitochondrion-related organelles (MROs), which include mitosomes and hydrogenosomes. In agreement with the concept of reductive evolution, mitosomes of Entamoeba histolytica lack most of the components of the TOM (translocase of the outer mitochondrial membrane) complex, which is required for the targeting and membrane translocation of preproteins into the canonical aerobic mitochondria. Here we showed, in E. histolytica mitosomes, the presence of a 600-kDa TOM complex composed of Tom40, a conserved pore-forming subunit, and Tom60, a novel lineage-specific receptor protein. Tom60, containing multiple tetratricopeptide repeats, is localized to the mitosomal outer membrane and the cytosol, and serves as a receptor of both mitosomal matrix and membrane preproteins. Our data indicate that Entamoeba has invented a novel lineage-specific shuttle receptor of the TOM complex as a consequence of adaptation to an anaerobic environment. PMID:23350036

  8. An Interactome-Centered Protein Discovery Approach Reveals Novel Components Involved in Mitosome Function and Homeostasis in Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Samuel Rout

    2016-12-01

    Full Text Available Protozoan parasites of the genus Giardia are highly prevalent globally, and infect a wide range of vertebrate hosts including humans, with proliferation and pathology restricted to the small intestine. This narrow ecological specialization entailed extensive structural and functional adaptations during host-parasite co-evolution. An example is the streamlined mitosomal proteome with iron-sulphur protein maturation as the only biochemical pathway clearly associated with this organelle. Here, we applied techniques in microscopy and protein biochemistry to investigate the mitosomal membrane proteome in association to mitosome homeostasis. Live cell imaging revealed a highly immobilized array of 30-40 physically distinct mitosome organelles in trophozoites. We provide direct evidence for the single giardial dynamin-related protein as a contributor to mitosomal morphogenesis and homeostasis. To overcome inherent limitations that have hitherto severely hampered the characterization of these unique organelles we applied a novel interaction-based proteome discovery strategy using forward and reverse protein co-immunoprecipitation. This allowed generation of organelle proteome data strictly in a protein-protein interaction context. We built an initial Tom40-centered outer membrane interactome by co-immunoprecipitation experiments, identifying small GTPases, factors with dual mitosome and endoplasmic reticulum (ER distribution, as well as novel matrix proteins. Through iterative expansion of this protein-protein interaction network, we were able to i significantly extend this interaction-based mitosomal proteome to include other membrane-associated proteins with possible roles in mitosome morphogenesis and connection to other subcellular compartments, and ii identify novel matrix proteins which may shed light on mitosome-associated metabolic functions other than Fe-S cluster biogenesis. Functional analysis also revealed conceptual conservation of protein

  9. An Interactome-Centered Protein Discovery Approach Reveals Novel Components Involved in Mitosome Function and Homeostasis in Giardia lamblia.

    Science.gov (United States)

    Rout, Samuel; Zumthor, Jon Paulin; Schraner, Elisabeth M; Faso, Carmen; Hehl, Adrian B

    2016-12-01

    Protozoan parasites of the genus Giardia are highly prevalent globally, and infect a wide range of vertebrate hosts including humans, with proliferation and pathology restricted to the small intestine. This narrow ecological specialization entailed extensive structural and functional adaptations during host-parasite co-evolution. An example is the streamlined mitosomal proteome with iron-sulphur protein maturation as the only biochemical pathway clearly associated with this organelle. Here, we applied techniques in microscopy and protein biochemistry to investigate the mitosomal membrane proteome in association to mitosome homeostasis. Live cell imaging revealed a highly immobilized array of 30-40 physically distinct mitosome organelles in trophozoites. We provide direct evidence for the single giardial dynamin-related protein as a contributor to mitosomal morphogenesis and homeostasis. To overcome inherent limitations that have hitherto severely hampered the characterization of these unique organelles we applied a novel interaction-based proteome discovery strategy using forward and reverse protein co-immunoprecipitation. This allowed generation of organelle proteome data strictly in a protein-protein interaction context. We built an initial Tom40-centered outer membrane interactome by co-immunoprecipitation experiments, identifying small GTPases, factors with dual mitosome and endoplasmic reticulum (ER) distribution, as well as novel matrix proteins. Through iterative expansion of this protein-protein interaction network, we were able to i) significantly extend this interaction-based mitosomal proteome to include other membrane-associated proteins with possible roles in mitosome morphogenesis and connection to other subcellular compartments, and ii) identify novel matrix proteins which may shed light on mitosome-associated metabolic functions other than Fe-S cluster biogenesis. Functional analysis also revealed conceptual conservation of protein translocation

  10. Entamoeba mitosomes play an important role in encystation by association with cholesteryl sulfate synthesis.

    Science.gov (United States)

    Mi-ichi, Fumika; Miyamoto, Tomofumi; Takao, Shouko; Jeelani, Ghulam; Hashimoto, Tetsuo; Hara, Hiromitsu; Nozaki, Tomoyoshi; Yoshida, Hiroki

    2015-06-02

    Hydrogenosomes and mitosomes are mitochondrion-related organelles (MROs) that have highly reduced and divergent functions in anaerobic/microaerophilic eukaryotes. Entamoeba histolytica, a microaerophilic, parasitic amoebozoan species, which causes intestinal and extraintestinal amoebiasis in humans, possesses mitosomes, the existence and biological functions of which have been a longstanding enigma in the evolution of mitochondria. We previously demonstrated that sulfate activation, which is not generally compartmentalized to mitochondria, is a major function of E. histolytica mitosomes. However, because the final metabolites of sulfate activation remain unknown, the overall scheme of this metabolism and the role of mitosomes in Entamoeba have not been elucidated. In this study we purified and identified cholesteryl sulfate (CS) as a final metabolite of sulfate activation. We then identified the gene encoding the cholesteryl sulfotransferase responsible for synthesizing CS. Addition of CS to culture media increased the number of cysts, the dormant form that differentiates from proliferative trophozoites. Conversely, chlorate, a selective inhibitor of the first enzyme in the sulfate-activation pathway, inhibited cyst formation in a dose-dependent manner. These results indicate that CS plays an important role in differentiation, an essential process for the transmission of Entamoeba between hosts. Furthermore, we show that Mastigamoeba balamuthi, an anaerobic, free-living amoebozoan species, which is a close relative of E. histolytica, also has the sulfate-activation pathway in MROs but does not possess the capacity for CS production. Hence, we propose that a unique function of MROs in Entamoeba contributes to its adaptation to its parasitic life cycle.

  11. Evolution of mitosome metabolism and invasion-related proteins in Cryptosporidium.

    Science.gov (United States)

    Liu, Shiyou; Roellig, Dawn M; Guo, Yaqiong; Li, Na; Frace, Michael A; Tang, Kevin; Zhang, Longxian; Feng, Yaoyu; Xiao, Lihua

    2016-12-08

    The switch from photosynthetic or predatory to parasitic life strategies by apicomplexans is accompanied with a reductive evolution of genomes and losses of metabolic capabilities. Cryptosporidium is an extreme example of reductive evolution among apicomplexans, with losses of both the mitosome genome and many metabolic pathways. Previous observations on reductive evolution were largely based on comparative studies of various groups of apicomplexans. In this study, we sequenced two divergent Cryptosporidium species and conducted a comparative genomic analysis to infer the reductive evolution of metabolic pathways and differential evolution of invasion-related proteins within the Cryptosporidium lineage. In energy metabolism, Cryptosporidium species differ from each other mostly in mitosome metabolic pathways. Compared with C. parvum and C. hominis, C. andersoni possesses more aerobic metabolism and a conventional electron transport chain, whereas C. ubiquitum has further reductions in ubiquinone and polyisprenoid biosynthesis and has lost both the conventional and alternative electron transport systems. For invasion-associated proteins, similar to C. hominis, a reduction in the number of genes encoding secreted MEDLE and insulinase-like proteins in the subtelomeric regions of chromosomes 5 and 6 was also observed in C. ubiquitum and C. andersoni, whereas mucin-type glycoproteins are highly divergent between the gastric C. andersoni and intestinal Cryptosporidium species. Results of the study suggest that rapidly evolving mitosome metabolism and secreted invasion-related proteins could be involved in tissue tropism and host specificity in Cryptosporidium spp. The finding of progressive reduction in mitosome metabolism among Cryptosporidium species improves our knowledge of organelle evolution within apicomplexans.

  12. 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.

  13. 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...

  14. 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.

  15. 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.

  16. 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

  17. 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

  18. 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

  19. 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.

  20. 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.

  1. The distribution of Paenibacillus larvae spores in adult bees and honey and larval mortality, following the addition of American foulbrood diseased brood or spore-contaminated honey in honey bee (Apis mellifera) colonies.

    Science.gov (United States)

    Lindström, Anders; Korpela, Seppo; Fries, Ingemar

    2008-09-01

    Within colony transmission of Paenibacillus larvae spores was studied by giving spore-contaminated honey comb or comb containing 100 larvae killed by American foulbrood to five experimental colonies respectively. We registered the impact of the two treatments on P. larvae spore loads in adult bees and honey and on larval mortality by culturing for spores in samples of adult bees and honey, respectively, and by measuring larval survival. The results demonstrate a direct effect of treatment on spore levels in adult bees and honey as well as on larval mortality. Colonies treated with dead larvae showed immediate high spore levels in adult bee samples, while the colonies treated with contaminated honey showed a comparable spore load but the effect was delayed until the bees started to utilize the honey at the end of the flight season. During the winter there was a build up of spores in the adult bees, which may increase the risk for infection in spring. The results confirm that contaminated honey can act as an environmental reservoir of P. larvae spores and suggest that less spores may be needed in honey, compared to in diseased brood, to produce clinically diseased colonies. The spore load in adult bee samples was significantly related to larval mortality but the spore load of honey samples was not.

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

    Science.gov (United States)

    Lee, Geon Joon; Sim, Geon Bo; Choi, Eun Ha; Kwon, Young-Wan; Kim, Jun Young; Jang, Siun; Kim, Seong Hwan

    2015-01-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2015-01-14

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

  5. Seasonal and Diurnal Variation of Atmospheric Fungal Spore Concentrations in Hyderabad; Tandojam-Sindh and the Effects of Climatic Conditions

    International Nuclear Information System (INIS)

    Khan, M.; Parveen, A.; Qaisar, M.

    2016-01-01

    Airborne biological particles are present in every type of environment. Different types of geographical localities have different type of airspora, which affect human health. The current study is conducted for the first time to identify the airborne fungal spores from Hyderabad: Tando-Jam, Sindh. For this purpose, Burkard's 7-Days recording volumetric spore trap was used for a period of one year. A total of 68,183 spores/m/sup 3/ were recorded throughout the study period, belonging to 41 fungal spores types. The presented data revealed that Deuteromycetes spore type was predominant. Cladosporium sp. spores were detected in the highest concentration i.e., 50.83 percent, which was followed by Aspergillus sp. (18.63 percent) and Alternaria sp. (11.04 percent). The highest spore count was captured in the month of September-2008 (17,294 spores/m/sup 3/), while lowest spore count was found in the month of June-2009. Diurnal patterns of individual fungal spore types was observed to be mid-day to evening maxima for various species. Spearman rank correlation coefficient r was determined for correlation of fungal spore counts with climatic factors by using IBM software SPSS ver. 20. Results of the current study revealed that fungal spore concentration was increased in high humid weather while low count was found in hot and windy climate that was also confirmed by statistical analysis. The presented work demonstrated that various types of allergenic and phytopathogenic fungal spores were present in the atmosphere of Hyderabad: Tando-Jam. It was also observed that meteorological conditions had a significant impact on dispersal and concentration of fungal spores. (author)

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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 ...

  16. 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

  17. 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

  18. 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.

  19. 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.

  20. 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.

  1. CERN confirms LHC schedule

    CERN Multimedia

    2003-01-01

    The CERN Council held its 125th session on 20 June. Highlights of the meeting included confirmation that the LHC is on schedule for a 2007 start-up, and the announcement of a new organizational structure in 2004.

  2. Performance Confirmation Plan

    International Nuclear Information System (INIS)

    Lindner, E.N.

    2000-01-01

    As described, the purpose of the Performance Confirmation Plan is to specify monitoring, testing, and analysis activities for evaluating the accuracy and adequacy of the information used to determine that performance objectives for postclosure will be met. This plan defines a number of specific performance confirmation activities and associated test concepts in support of the MGR that will be implemented to fulfill this purpose. In doing so, the plan defines an approach to identify key factors and processes, predict performance, establish tolerances and test criteria, collect data (through monitoring, testing, and experiments), analyze these data, and recommend appropriate action. The process of defining which factors to address under performance confirmation incorporates input from several areas. In all cases, key performance confirmation factors are those factors which are: (1) important to safety, (2) measurable and predictable, and (3) relevant to the program (i.e., a factor that is affected by construction, emplacement, or is a time-dependent variable). For the present version of the plan, performance confirmation factors important to safety are identified using the principal factors from the RSS (CRWMS M and O 2000a) (which is derived from TSPA analyses) together with other available performance assessment analyses. With this basis, key performance confirmation factors have been identified, and test concepts and test descriptions have been developed in the plan. Other activities are also incorporated into the performance confirmation program outside of these key factors. Additional activities and tests have been incorporated when they are prescribed by requirements and regulations or are necessary to address data needs and model validation requirements relevant to postclosure safety. These other activities have been included with identified factors to construct the overall performance confirmation program

  3. Performance Confirmation Plan

    International Nuclear Information System (INIS)

    Lindner, E.N.

    2000-01-01

    As described, the purpose of the Performance Confirmation Plan is to specify monitoring, testing, and analysis activities for evaluating the accuracy and adequacy of the information used to determine that performance objectives for postclosure will be met. This plan defines a number of specific performance confirmation activities and associated test concepts in support of the MGR that will be implemented to fulfill this purpose. In doing so, the plan defines an approach to identify key factors and processes, predict performance, establish tolerances and test criteria, collect data (through monitoring, testing, and experiments), analyze these data, and recommend appropriate action. The process of defining which factors to address under performance confirmation incorporates input from several areas. In all cases, key performance confirmation factors are those factors which are: (1) important to safety, (2) measurable and predictable, and (3) relevant to the program (i.e., a factor that i s affected by construction, emplacement, or is a time-dependent variable). For the present version of the plan, performance confirmation factors important to safety are identified using the principal factors from the RSS (CRWMS M and O 2000a) (which is derived from TSPA analyses) together with other available performance assessment analyses. With this basis, key performance confirmation factors have been identified, and test concepts and test descriptions have been developed in the plan. Other activities are also incorporated into the performance confirmation program outside of these key factors. Additional activities and tests have been incorporated when they are prescribed by requirements and regulations or are necessary to address data needs and model validation requirements relevant to postclosure safety. These other activities have been included with identified factors to construct the overall performance confirmation program

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

    Science.gov (United States)

    LaDuc, Myron T.; Mohapatra, Bidyut

    2014-01-01

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

  5. 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

  6. 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.

  7. Repository performance confirmation

    International Nuclear Information System (INIS)

    Hansen, Francis D.

    2011-01-01

    Repository performance confirmation links the technical bases of repository science and societal acceptance. This paper explores the myriad aspects of what has been labeled performance confirmation in U.S. programs, which involves monitoring as a collection of distinct activities combining technical and social significance in radioactive waste management. This paper is divided into four parts: (1) A distinction is drawn between performance confirmation monitoring and other testing and monitoring objectives; (2) A case study illustrates confirmation activities integrated within a long-term testing and monitoring strategy for Yucca Mountain; (3) A case study reviews compliance monitoring developed and implemented for the Waste Isolation Pilot Plant; and (4) An approach for developing, evaluating and implementing the next generation of performance confirmation monitoring is presented. International interest in repository monitoring is exhibited by the European Commission Seventh Framework Programme 'Monitoring Developments for Safe Repository Operation and Staged Closure' (MoDeRn) Project. The MoDeRn partners are considering the role of monitoring in a phased approach to the geological disposal of radioactive waste. As repository plans advance in different countries, the need to consider monitoring strategies within a controlled framework has become more apparent. The MoDeRn project pulls together technical and societal experts to assimilate a common understanding of a process that could be followed to develop a monitoring program. A fundamental consideration is the differentiation of confirmation monitoring from the many other testing and monitoring activities. Recently, the license application for Yucca Mountain provided a case study including a technical process for meeting regulatory requirements to confirm repository performance as well as considerations related to the preservation of retrievability. The performance confirmation plan developed as part of the

  8. 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

  9. 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

  10. 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

  11. 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

  12. 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

  13. The relationships between air pollutants, meteorological parameters and concentration of airborne fungal spores

    International Nuclear Information System (INIS)

    Grinn-Gofron, Agnieszka; Strzelczak, Agnieszka; Wolski, Tomasz

    2011-01-01

    Fungal spores are an important component of bioaerosol and also considered to act as indicator of the level of atmospheric bio-pollution. Therefore, better understanding of these phenomena demands a detailed survey of airborne particles. The objective of this study was to examine the dependence of two the most important allergenic taxa of airborne fungi - Alternaria and Cladosporium - on meteorological parameters and air pollutant concentrations during three consecutive years (2006-2008). This study is also an attempt to create artificial neural network (ANN) forecasting models useful in the prediction of aeroallergen abundance. There were statistically significant relationships between spore concentration and environmental parameters as well as pollutants, confirmed by the Spearman's correlation rank analysis and high performance of the ANN models obtained. The concentrations of Cladosporium and Alternaria spores can be predicted with quite good accuracy from meteorological conditions and air pollution recorded three days earlier. - ANN models predict airspore contents from weather conditions and air pollutant.

  14. The relationships between air pollutants, meteorological parameters and concentration of airborne fungal spores

    Energy Technology Data Exchange (ETDEWEB)

    Grinn-Gofron, Agnieszka, E-mail: agofr@univ.szczecin.p [Department of Plant Taxonomy and Phytogeography, Faculty of Natural Science, University of Szczecin, Waska 13 Street, 71-415 Szczecin (Poland); Strzelczak, Agnieszka [Department of Food Process Engineering, Faculty of Food Science and Fisheries, West Pomeranian University of Technology, Szczecin (Poland); Wolski, Tomasz [Physical Oceanography Laboratory, University of Szczecin (Poland)

    2011-02-15

    Fungal spores are an important component of bioaerosol and also considered to act as indicator of the level of atmospheric bio-pollution. Therefore, better understanding of these phenomena demands a detailed survey of airborne particles. The objective of this study was to examine the dependence of two the most important allergenic taxa of airborne fungi - Alternaria and Cladosporium - on meteorological parameters and air pollutant concentrations during three consecutive years (2006-2008). This study is also an attempt to create artificial neural network (ANN) forecasting models useful in the prediction of aeroallergen abundance. There were statistically significant relationships between spore concentration and environmental parameters as well as pollutants, confirmed by the Spearman's correlation rank analysis and high performance of the ANN models obtained. The concentrations of Cladosporium and Alternaria spores can be predicted with quite good accuracy from meteorological conditions and air pollution recorded three days earlier. - ANN models predict airspore contents from weather conditions and air pollutant.

  15. 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.

  16. 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...

  17. Clostridium thermocellum Nitrilase Expression and Surface Display on Bacillus subtilis Spores.

    Science.gov (United States)

    Chen, Huayou; Zhang, Tianxi; Sun, Tengyun; Ni, Zhong; Le, Yilin; Tian, Rui; Chen, Zhi; Zhang, Chunxia

    2015-01-01

    Nitrilases are an important class of industrial enzymes. They require mild reaction conditions and are highly efficient and environmentally friendly, so they are used to catalyze the synthesis of carboxylic acid from nitrile, a process considered superior to conventional chemical syntheses. Nitrilases should be immobilized to overcome difficulties in recovery after the reaction and to stabilize the free enzyme. The nitrilase from Clostridium thermocellum was expressed, identified and displayed on the surface of Bacillus subtilis spores by using the spore coat protein G of B. subtilis as an anchoring motif. In a free state, the recombinant nitrilase catalyzed the conversion of 3-cyanopyridine to niacin and displayed maximum catalytic activity (8.22 units/mg protein) at 40 °C and pH 7.4. SDS-PAGE and Western blot were used to confirm nitrilase display. Compared with the free enzyme, the spore-immobilized nitrilase showed a higher tolerance for adverse environmental conditions. After the reaction, recombinant spores were recovered via centrifugation and reused 3 times to catalyze the conversion of 3-cyanopyridine with 75.3% nitrilase activity. This study demonstrates an effective means of nitrilase immobilization via spore surface display, which can be applied in biological processes or conversion. © 2015 S. Karger AG, Basel.

  18. 1-Octanol, a self-inhibitor of spore germination in Penicillium camemberti.

    Science.gov (United States)

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

    2016-08-01

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

  19. 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

  20. The impact of inducing germination of Bacillus anthracis and Bacillus thuringiensis spores on potential secondary decontamination strategies.

    Science.gov (United States)

    Omotade, T O; Bernhards, R C; Klimko, C P; Matthews, M E; Hill, A J; Hunter, M S; Webster, W M; Bozue, J A; Welkos, S L; Cote, C K

    2014-12-01

    Decontamination and remediation of a site contaminated by the accidental or intentional release of fully virulent Bacillus anthracis spores are difficult, costly and potentially damaging to the environment. Development of novel decontamination strategies that have minimal environmental impacts remains a high priority. Although ungerminated spores are amongst the most resilient organisms known, once exposed to germinants, the germinating spores, in some cases, become susceptible to antimicrobial environments. We evaluated the concept that once germinated, B. anthracis spores would be less hazardous and significantly easier to remediate than ungerminated dormant spores. Through in vitro germination and sensitivity assays, we demonstrated that upon germination, B. anthracis Ames spores and Bacillus thuringiensis Al Hakam spores (serving as a surrogate for B. anthracis) become susceptible to environmental stressors. The majority of these germinated B. anthracis and B. thuringiensis spores were nonviable after exposure to a defined minimal germination-inducing solution for prolonged periods of time. Additionally, we examined the impact of potential secondary disinfectant strategies including bleach, hydrogen peroxide, formaldehyde and artificial UV-A, UV-B and UV-C radiation, employed after a 60-min germination-induction step. Each secondary disinfectant employs a unique mechanism of killing; as a result, germination-induction strategies are better suited for some secondary disinfectants than others. These results provide evidence that the deployment of an optimal combination strategy of germination-induction/secondary disinfection may be a promising aspect of wide-area decontamination following a B. anthracis contamination event. By inducing spores to germinate, our data confirm that the resulting cells exhibit sensitivities that can be leveraged when paired with certain decontamination measures. This increased susceptibility could be exploited to devise more

  1. 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.)

  2. 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.).

  3. 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

  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. 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.

  6. 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

  7. 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

  8. 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 ...

  9. 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

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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

  19. Identification and Validation of Specific Markers of Bacillus anthracis Spores by Proteomics and Genomics Approaches*

    Science.gov (United States)

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

    2014-01-01

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

  20. [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.

  1. 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

  2. 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

  3. 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.

  4. Bacillus cereus spore formation, structure and germination

    NARCIS (Netherlands)

    Vries, de Y.P.

    2006-01-01

    Bacterial spores arespecializeddifferentiated celltypes for

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

    Science.gov (United States)

    From, Cecilie; van der Voort, Menno; Abee, Tjakko; Granum, Per Einar

    2012-06-01

    Bc1245 is a monocistronic chromosomal gene of Bacillus cereus ATCC 14579 encoding a putative protein of 143 amino acids identified in this study to have a spore-related function in B. cereus. Bc1245 is highly conserved in the genome of members of the B. cereus group, indicating an important function of the gene in this group of bacteria. Quantitative PCR revealed that bc1245 is transcribed late in sporulation (upon formation of phase-bright spores) and at the same time as the mother cell-specific transcription factor σ(K) . The σ(K) regulon includes structural components of the spore (such as coat proteins), and it is therefore plausible that bc1245 might encode a structural outer spore protein. This was confirmed by detection of BC1245 in exosporium extracts from B. cereus by immunoblotting against BC1245 antiserum. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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...

  13. A four-gene operon inBacillus cereusproduces two rare spore-decorating sugars.

    Science.gov (United States)

    Li, Zi; Mukherjee, Thiya; Bowler, Kyle; Namdari, Sholeh; Snow, Zachary; Prestridge, Sarah; Carlton, Alexandra; Bar-Peled, Maor

    2017-05-05

    Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. Accordingly, unraveling the sugar composition of bacterial cell surfaces can shed light on bacterial growth and pathogenesis. Here, we found that two rare sugars with a 3- C -methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM: C -methyltransferase, and NADPH-dependent CDP-3- C -methyl-6-deoxyhexose 4-reductase. The last enzyme predominantly yielded CDP-3- C -methyl-6-deoxygulose (CDP-cereose) and likely generated a 4-epimer CDP-3- C -methyl-6-deoxyallose (CDP-cillose). Some members of the B. cereus sensu lato group produce CDP-3- C -methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. Gene knockouts of the Bacillus C -methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. We also found that cereose represented 0.2-1% spore dry weight. Moreover, mutants lacking cereose germinated faster than the wild type, yet the mutants exhibited no changes in sporulation or spore resistance to heat. The findings reported here may provide new insights into the roles of the uncommon 3- C -methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus . © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Hexane abatement and spore emission control in a fungal biofilter-photoreactor hybrid unit

    Energy Technology Data Exchange (ETDEWEB)

    Saucedo-Lucero, J.O. [Department of Chemical Engineering and Environmental Technology, University of Valladolid, Dr. Mergelina s/n, 47011 Valladolid (Spain); IPICyT, Instituto Potosino de Investigación Científica y Tecnológica, División de Ciencias Ambientales, Camino a la Presa San José No. 2055, C.P., 78216 San Luis Potosí (Mexico); Quijano, G. [Department of Chemical Engineering and Environmental Technology, University of Valladolid, Dr. Mergelina s/n, 47011 Valladolid (Spain); Arriaga, S. [IPICyT, Instituto Potosino de Investigación Científica y Tecnológica, División de Ciencias Ambientales, Camino a la Presa San José No. 2055, C.P., 78216 San Luis Potosí (Mexico); Muñoz, R., E-mail: mutora@iq.uva.es [Department of Chemical Engineering and Environmental Technology, University of Valladolid, Dr. Mergelina s/n, 47011 Valladolid (Spain)

    2014-07-15

    Highlights: • A fungal biofilter/photoreactor was evaluated in terms of hexane and spore removal. • Biofilter supported elimination capacities of ≈35 g m{sup −3} h{sup −1} and CO{sub 2} yields of ≈75%. • The photocatalytic process slightly boosted the hexane abatement performance. • Biofilter emitted fungal spores at concentrations of 2.4 × 10{sup 3}–9.0 × 10{sup 4} CFU m{sup −3}. • Photo-assisted post-treatments resulted in spore deactivation efficiencies of 98%. - Abstract: The performance of a fungal perlite-based biofilter coupled to a post-treatment photoreactor was evaluated over 234 days in terms of n-hexane removal, emission and deactivation of fungal spores. The biofilter and photoreactor were operated at gas residence times of 1.20 and 0.14 min, respectively, and a hexane loading rate of 115 ± 5 g m{sup −3} h{sup −1}. Steady n-hexane elimination capacities of 30–40 g m{sup −3} h{sup −1} were achieved, concomitantly with pollutant mineralization efficiencies of 60–90%. No significant influence of biofilter irrigation frequency or irrigation nitrogen concentration on hexane abatement was recorded. Photolysis did not support an efficient hexane post-treatment likely due to the short EBRT applied in the photoreactor, while overall hexane removal and mineralization enhancements of 25% were recorded when the irradiated photoreactor was packed with ZnO-impregnated perlite. However, a rapid catalyst deactivation was observed, which required a periodic reactivation every 48 h. Biofilter irrigation every 3 days supported fungal spore emissions at concentrations ranging from 2.4 × 10{sup 3} to 9.0 × 10{sup 4} CFU m{sup −3}. Finally, spore deactivation efficiencies of ≈98% were recorded for the photolytic and photocatalytic post-treatment processes. This study confirmed the potential of photo-assisted post-treatment processes to mitigate the emission of hazardous fungal spores and boost the abatement performance of

  15. 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.

  16. 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.

  17. 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...

  18. 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.

  19. Comparative analysis of the concentration of fungal spores in the air of Lublin and Rzeszów (Eastern Poland

    Directory of Open Access Journals (Sweden)

    Idalia Kasprzyk

    2012-12-01

    Full Text Available Studies of the concentration of fungal spores were carried out in the cities of Lublin and Rzeszów simultaneously in 2002. At both sites the volumetric method of measurement was applied, using the Lanzoni VPPS 2000 trap. Only the allergenic taxa were analysed: Alternaria, Botrytis, Cladosporium, Drechslera type, Epicoccum, Torula, Stemphylium, Pithomyces, Polythrincium, and Ganoderma. The research showed considerable differences in the concentration and frequency of spores in the air at the sites compared. Higher mean concentrations of spores were usually observed in Lublin. Only for two taxa were the concentrations higher in Rzeszów. No significant differences were observed for the genus of Polythrincium and Torula. Also the lengths of periods of occurrence of the spores were determined using the 95% method. It was determined that the geobotanical conditions in Rzeszów have a positive effect on the lengthening of the presence of the spores in aeroplankton. The results of the observations were analysed statistically, which confirmed the occurrence of significant differences between the cities compared.

  20. 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.

  1. 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.

  2. 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.

  3. 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...

  4. 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

  5. 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.

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. 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

  11. 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

  12. 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 ...

  13. Newsletter Subscribe Confirmation | IDRC - International ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Thank you. Please check your email to confirm your subscription to the IDRC Bulletin. Please note: the link in your confirmation email is valid for 3 days, so please reply promptly. What we do · Funding · Resources · About IDRC. Knowledge. Innovation. Solutions. Careers · Contact Us · Site map. Sign up now for IDRC news ...

  14. 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.

  15. 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.

  16. 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.

  17. National validation study of a swab protocol for the recovery of Bacillus anthracis spores from surfaces.

    Science.gov (United States)

    Hodges, Lisa R; Rose, Laura J; O'Connell, Heather; Arduino, Matthew J

    2010-05-01

    Twelve Laboratory Response Network (LRN) affiliated laboratories participated in a validation study of a macrofoam swab protocol for the recovery, detection, and quantification of viable B. anthracis (BA) Sterne spores from steel surfaces. CDC personnel inoculated steel coupons (26cm(2)) with 1-4 log(10) BA spores and recovered them by sampling with pre-moistened macrofoam swabs. Phase 1 (P1) of the study evaluated swabs containing BA only, while dust and background organisms were added to swabs in Phase 2 (P2) to mimic environmental conditions. Laboratories processed swabs and enumerated spores by culturing eluted swab suspensions and counting colonies with morphology consistent with BA. Processed swabs were placed in enrichment broth, incubated 24h, and cultured by streaking for isolation. Real-time PCR was performed on selected colonies from P2 samples to confirm the identity of BA. Mean percent recovery (%R) of spores from the surface ranged from 15.8 to 31.0% (P1) and from 27.9 to 55.0% (P2). The highest mean percent recovery was 31.0% (sd 10.9%) for P1 (4 log(10) inoculum) and 55.0% (sd 27.6%) for P2 (1 log(10) inoculum). The overall %R was higher for P2 (44.6%) than P1 (24.1%), but the overall reproducibility (between-lab variability) was lower in P2 than in P1 (25.0 vs 16.5%CV, respectively). The overall precision (within-lab variability) was close to identical for P1 and P2 (44.0 and 44.1, respectively), but varied greatly between inoculum levels. The protocol demonstrated linearity in %R over the three inoculum levels and is able to detect between 26 and 5x10(6)spores/26cm(2). Sensitivity as determined by culture was >98.3% for both phases and all inocula, suggesting that the culture method maintains sensitivity in the presence of contaminants. The enrichment broth method alone was less sensitive for sampled swabs (66.4%) during P2, suggesting that the presence of background organisms inhibited growth or isolation of BA from the broth. The addition of

  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. Quantitative morphological descriptors confirm traditionally ...

    African Journals Online (AJOL)

    SARAH

    2015-09-30

    Sep 30, 2015 ... J. Appl. Biosci. 2015 Quantitative morphological descriptors confirm traditionally classified morphotypes of Pentadesma butyracea Sabine (clusiaceae). 8736. Quantitative morphological descriptors ...... Hijmans RJ, Cameron SE, Parra JL, Jones PG, Jarvis. A, 2004. The WorldClim interpolated global.

  20. 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.

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

    Science.gov (United States)

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

    2010-07-01

    Tempe, a Rhizopus ssp.-fermented soya bean food product, was investigated for bacteriostatic and/or bactericidal effects against cells and spores of the food-borne pathogen Bacillus cereus. Tempe extract showed a high antibacterial activity against B. cereus ATCC 14579 based on optical density and viable count measurements. This growth inhibition was manifested by a 4 log CFU ml(-1) reduction, within the first 15 min of exposure. Tempe extracts also rapidly inactivated B. cereus spores upon germination. Viability and membrane permeability assessments using fluorescence probes showed rapid inactivation and permeabilization of the cytoplasmic membrane confirming the bactericidal mode of action. Cooked beans and Rhizopus grown on different media did not show antibacterial activity, indicating the unique association of the antibacterial activity with tempe. Subsequent characterization of the antibacterial activity revealed that heat treatment and protease addition nullified the bactericidal effect, indicating the proteinaceous nature of the bioactive compound. During fermentation of soya beans with Rhizopus, compounds are released with extensive antibacterial activity against B. cereus cells and spores. The results show the potential of producing natural antibacterial compounds that could be used as ingredients in food preservation and pathogen control. © 2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology.

  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. 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.

  6. 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

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. A data analysis of the irradiation parameter D10 for bacteria and spores under various conditions.

    Science.gov (United States)

    van Gerwen, S J; Rombouts, F M; van't Riet, K; Zwietering, M H

    1999-09-01

    This paper provides approximate estimates for the irradiation parameter D10 to globally predict the effectiveness of any irradiation process. D10 is often reported to depend on many specific factors, implying that D10 cannot be estimated without exact knowledge of all factors involved. For specific questions these data can of course be useful but only if the conditions reported exactly match the specific question. Alternatively, this study determined the most relevant factors influencing D10, by quantitatively analyzing data from many references. The best first step appeared to be a classification of the data into vegetative bacteria and spores. As expected, spores were found to have significantly higher D10 values (average 2.48 kGy) than vegetative bacteria (average 0.762 kGy). Further analyses of the vegetative bacteria confirmed the expected extreme irradiation resistance of nonpathogenic Deinococcus radiodurans (average 10.4 kGy). Furthermore the analysis identified Enterococcus faecium, Alcaligenes spp., and several members of the Moraxella-Acinetobacter group as having very high resistance at very low temperatures (average 3.65 kGy). After exclusion of high- and low-resistance spores and some specific conditions showing relevant high or low D10 values, the average for spores was estimated to be 2.11 kGy. For vegetative bacteria this average was estimated to be 0.420 kGy. These approximate estimates are not definite, as they depend on the data used in the analyses. It is expected that inclusion of more data will not change the estimates to a great extent. The approximate estimates are therefore useful tools in designing and evaluating irradiation processes.

  13. Comparative analysis of immune effects in mice model: Clonorchis sinensis cysteine protease generated from recombinant Escherichia coli and Bacillus subtilis spores.

    Science.gov (United States)

    Wu, Zhanshuai; Tang, Zeli; Shang, Mei; Zhao, Lu; Zhou, Lina; Kong, Xiangzhan; Lin, Zhipeng; Sun, Hengchang; Chen, Tingjin; Xu, Jin; Li, Xuerong; Huang, Yan; Yu, Xinbing

    2017-07-01

    Clonorchiasis remains a nonnegligible public health problem in endemic areas. Cysteine protease of Clonorchis sinensis (CsCP) plays indispensable roles in the parasitic physiology and pathology, and has been exploited as a promising drug and vaccine candidate. In recent years, development of spore-based vaccines against multiple pathogens has attracted many investigators' interest. In previous studies, the recombinant Escherichia coli (BL21) and Bacillus subtilis spores expressing CsCP have been successfully constructed, respectively. In this study, the immune effects of CsCP protein purified from recombinant BL21 (rCsCP) and B. subtilis spores presenting CsCP (B.s-CsCP) in Balb/c mice model were conducted with comparative analysis. Levels of specific IgG, IgG1 and IgG2a were significantly increased in sera from both rCsCP and B.s-CsCP intraperitoneally immunized mice. Additionally, recombinant spores expressing abundant fusion CsCP (0.03125 pg/spore) could strongly enhance the immunogenicity of CsCP with significantly higher levels of IgG and isotypes. Compared with rCsCP alone, intraperitoneal administration of mice with spores expressing CsCP achieved a better effect of fighting against C. sinensis infection by slowing down the process of fibrosis. Our results demonstrated that a combination of Th1/Th2 immune responses could be elicited by rCsCP, while spores displaying CsCP prominently induced Th1-biased specific immune responses, and the complex cytokine network maybe mediates protective immune responses against C. sinensis. This work further confirmed that the usage of B. subtilis spores displaying CsCP is an effective way to against C. sinensis.

  14. Gender Confirmation Surgery: Guiding Principles.

    Science.gov (United States)

    Schechter, Loren S; D'Arpa, Salvatore; Cohen, Mimis N; Kocjancic, Ervin; Claes, Karel E Y; Monstrey, Stan

    2017-06-01

    At this time, no formal training or educational programs exist for surgeons or surgery residents interested in performing gender confirmation surgeries. To propose guiding principles designed to aid with the development of formal surgical training programs focused on gender confirmation surgery. We use expert opinion to provide a "first of its kind" framework for training surgeons to care for transgender and gender nonconforming individuals. We describe a multidisciplinary treatment model that describes an educational philosophy and the institution of quality parameters. This article represents the first step in the development of a structured educational program for surgical training in gender confirmation procedures. The World Professional Association for Transgender Health Board of Directors unanimously approved this article as the framework for surgical training. This article builds a framework for surgical training. It is designed to provide concepts that will likely be modified over time and based on additional data and evidence gathered through outcome measurements. We present an initial step in the formation of educational and technical guidelines for training surgeons in gender confirmation procedures. Schechter LS, D'Arpa S, Cohen MN, et al. Gender Confirmation Surgery: Guiding Principles. J Sex Med 2017;14:852-856. Copyright © 2017 International Society for Sexual Medicine. Published by Elsevier Inc. All rights reserved.

  15. 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.

  16. 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 ...

  17. 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...

  18. 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.

  19. 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...

  20. 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...

  1. 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

  2. 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

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. Constructing, Confirming, and Contesting Icons

    DEFF Research Database (Denmark)

    Mortensen, Mette

    2017-01-01

    to their reception as they help shape and delimit the publics and discourses surrounding visual icons. This article draws on existing research on visual icons and appropriations to develop a theoretical framework for how appropriations construct, confirm and contest icons and how personification constitutes the main......This article argues that appropriations are central to both the production and reception of visual icons. Appropriations are instrumental in iconization processes as they confirm and consolidate the iconic status by recycling the image in question. At the same time, appropriations are vital...... of these appropriations, two overall modes are singled out: the appropriations either decontextualize or recontextualize the figure of Kurdi. The two next analytical cases test the limits of decontextualization and recontextualization: Chinese artist Ai Weiwei decontextualizes the Kurdi imagery in a controversial...

  8. 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.

  9. 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.

  10. 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.

  11. Performance Confirmation Data Aquisition System

    Energy Technology Data Exchange (ETDEWEB)

    D.W. Markman

    2000-10-27

    The purpose of this analysis is to identify and analyze concepts for the acquisition of data in support of the Performance Confirmation (PC) program at the potential subsurface nuclear waste repository at Yucca Mountain. The scope and primary objectives of this analysis are to: (1) Review the criteria for design as presented in the Performance Confirmation Data Acquisition/Monitoring System Description Document, by way of the Input Transmittal, Performance Confirmation Input Criteria (CRWMS M&O 1999c). (2) Identify and describe existing and potential new trends in data acquisition system software and hardware that would support the PC plan. The data acquisition software and hardware will support the field instruments and equipment that will be installed for the observation and perimeter drift borehole monitoring, and in-situ monitoring within the emplacement drifts. The exhaust air monitoring requirements will be supported by a data communication network interface with the ventilation monitoring system database. (3) Identify the concepts and features that a data acquisition system should have in order to support the PC process and its activities. (4) Based on PC monitoring needs and available technologies, further develop concepts of a potential data acquisition system network in support of the PC program and the Site Recommendation and License Application.

  12. Opinion dynamics with confirmation bias.

    Directory of Open Access Journals (Sweden)

    Armen E Allahverdyan

    Full Text Available Confirmation bias is the tendency to acquire or evaluate new information in a way that is consistent with one's preexisting beliefs. It is omnipresent in psychology, economics, and even scientific practices. Prior theoretical research of this phenomenon has mainly focused on its economic implications possibly missing its potential connections with broader notions of cognitive science.We formulate a (non-Bayesian model for revising subjective probabilistic opinion of a confirmationally-biased agent in the light of a persuasive opinion. The revision rule ensures that the agent does not react to persuasion that is either far from his current opinion or coincides with it. We demonstrate that the model accounts for the basic phenomenology of the social judgment theory, and allows to study various phenomena such as cognitive dissonance and boomerang effect. The model also displays the order of presentation effect-when consecutively exposed to two opinions, the preference is given to the last opinion (recency or the first opinion (primacy -and relates recency to confirmation bias. Finally, we study the model in the case of repeated persuasion and analyze its convergence properties.The standard Bayesian approach to probabilistic opinion revision is inadequate for describing the observed phenomenology of persuasion process. The simple non-Bayesian model proposed here does agree with this phenomenology and is capable of reproducing a spectrum of effects observed in psychology: primacy-recency phenomenon, boomerang effect and cognitive dissonance. We point out several limitations of the model that should motivate its future development.

  13. Opinion dynamics with confirmation bias.

    Science.gov (United States)

    Allahverdyan, Armen E; Galstyan, Aram

    2014-01-01

    Confirmation bias is the tendency to acquire or evaluate new information in a way that is consistent with one's preexisting beliefs. It is omnipresent in psychology, economics, and even scientific practices. Prior theoretical research of this phenomenon has mainly focused on its economic implications possibly missing its potential connections with broader notions of cognitive science. We formulate a (non-Bayesian) model for revising subjective probabilistic opinion of a confirmationally-biased agent in the light of a persuasive opinion. The revision rule ensures that the agent does not react to persuasion that is either far from his current opinion or coincides with it. We demonstrate that the model accounts for the basic phenomenology of the social judgment theory, and allows to study various phenomena such as cognitive dissonance and boomerang effect. The model also displays the order of presentation effect-when consecutively exposed to two opinions, the preference is given to the last opinion (recency) or the first opinion (primacy) -and relates recency to confirmation bias. Finally, we study the model in the case of repeated persuasion and analyze its convergence properties. The standard Bayesian approach to probabilistic opinion revision is inadequate for describing the observed phenomenology of persuasion process. The simple non-Bayesian model proposed here does agree with this phenomenology and is capable of reproducing a spectrum of effects observed in psychology: primacy-recency phenomenon, boomerang effect and cognitive dissonance. We point out several limitations of the model that should motivate its future development.

  14. Performance Confirmation Data Acquisition System

    International Nuclear Information System (INIS)

    D.W. Markman

    2000-01-01

    The purpose of this analysis is to identify and analyze concepts for the acquisition of data in support of the Performance Confirmation (PC) program at the potential subsurface nuclear waste repository at Yucca Mountain. The scope and primary objectives of this analysis are to: (1) Review the criteria for design as presented in the Performance Confirmation Data Acquisition/Monitoring System Description Document, by way of the Input Transmittal, Performance Confirmation Input Criteria (CRWMS M and O 1999c). (2) Identify and describe existing and potential new trends in data acquisition system software and hardware that would support the PC plan. The data acquisition software and hardware will support the field instruments and equipment that will be installed for the observation and perimeter drift borehole monitoring, and in-situ monitoring within the emplacement drifts. The exhaust air monitoring requirements will be supported by a data communication network interface with the ventilation monitoring system database. (3) Identify the concepts and features that a data acquisition system should have in order to support the PC process and its activities. (4) Based on PC monitoring needs and available technologies, further develop concepts of a potential data acquisition system network in support of the PC program and the Site Recommendation and License Application

  15. 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...

  16. 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

  17. 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

  18. 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.

  19. Bacteriologically confirmed tuberculosis in children

    International Nuclear Information System (INIS)

    Ozere, I.; Sele, A.; Ozolina, A.

    2005-01-01

    Tuberculosis in children and adults is a growing problem with important public health implications. In Latvia the incidence of tuberculosis (TB) in children up to age 14 has increased from 7,1 per 100000 in 1992 to 28,8 per 100000 in 2003. The diagnosis of TB is confirmed by isolation and identification of M. tuberculosis (MT) from clinical specimen. Confirmation of the disease, however, is difficult in children due to poor bacilli excretion and even under the best circumstances only about 30-40% of pediatric TB cases are proved bacteriologically. Of the 370 pediatric TB cases diagnosed between January 1, 2001 and December 1, 2003 in Latvia, 53 (14,3%) were confirmed bacteriologically. The clinical, radiological, immunological and anamnestic features of confirmed TB can serve as cornerstones for diagnosing of TB, when culture is not available. Objective To evaluate the sensitivity of diagnostic criteria of TB, clinical and radiological manifestation of TB and drug susceptibility of MT isolated also. Methods All consecutive children (53 in total) up to age 14 diagnosed with bacteriologically confirmed TB during 01.01.2001. -01.12.2003. were prospectively evaluated for reasons mentioned above. Results Of the 53 children identified all but one had respiratory tract TB. 17(32,1 %) children were under 4 years of age, 9 (17%) children were 5-9 years old, but 27 (50,9%) patients were 10-14 years old. During evaluation data on TB source case were found in addition in 13 children and total TB contact history was positive in 37 (69,8%) patients. All clinical and radiographical forms of respiratory tract TB were diagnosed. The most common encountered forms were intrathoracic adenopathy in 10 (18,9%) cases and TB pneumonia in 6 (11,3%) cases in children aged 10-14 years. lnthrathoracic adenopathy associated with segmental parenchymal lesion was the most common form in children under 4 years of age -7 (13,2%) cases respectively. Conclusions 1. The clinical and radiological

  20. Model confirmation in climate economics.

    Science.gov (United States)

    Millner, Antony; McDermott, Thomas K J

    2016-08-02

    Benefit-cost integrated assessment models (BC-IAMs) inform climate policy debates by quantifying the trade-offs between alternative greenhouse gas abatement options. They achieve this by coupling simplified models of the climate system to models of the global economy and the costs and benefits of climate policy. Although these models have provided valuable qualitative insights into the sensitivity of policy trade-offs to different ethical and empirical assumptions, they are increasingly being used to inform the selection of policies in the real world. To the extent that BC-IAMs are used as inputs to policy selection, our confidence in their quantitative outputs must depend on the empirical validity of their modeling assumptions. We have a degree of confidence in climate models both because they have been tested on historical data in hindcasting experiments and because the physical principles they are based on have been empirically confirmed in closely related applications. By contrast, the economic components of BC-IAMs often rely on untestable scenarios, or on structural models that are comparatively untested on relevant time scales. Where possible, an approach to model confirmation similar to that used in climate science could help to build confidence in the economic components of BC-IAMs, or focus attention on which components might need refinement for policy applications. We illustrate the potential benefits of model confirmation exercises by performing a long-run hindcasting experiment with one of the leading BC-IAMs. We show that its model of long-run economic growth-one of its most important economic components-had questionable predictive power over the 20th century.

  1. Opinion Dynamics with Confirmation Bias

    Science.gov (United States)

    Allahverdyan, Armen E.; Galstyan, Aram

    2014-01-01

    Background Confirmation bias is the tendency to acquire or evaluate new information in a way that is consistent with one's preexisting beliefs. It is omnipresent in psychology, economics, and even scientific practices. Prior theoretical research of this phenomenon has mainly focused on its economic implications possibly missing its potential connections with broader notions of cognitive science. Methodology/Principal Findings We formulate a (non-Bayesian) model for revising subjective probabilistic opinion of a confirmationally-biased agent in the light of a persuasive opinion. The revision rule ensures that the agent does not react to persuasion that is either far from his current opinion or coincides with it. We demonstrate that the model accounts for the basic phenomenology of the social judgment theory, and allows to study various phenomena such as cognitive dissonance and boomerang effect. The model also displays the order of presentation effect–when consecutively exposed to two opinions, the preference is given to the last opinion (recency) or the first opinion (primacy) –and relates recency to confirmation bias. Finally, we study the model in the case of repeated persuasion and analyze its convergence properties. Conclusions The standard Bayesian approach to probabilistic opinion revision is inadequate for describing the observed phenomenology of persuasion process. The simple non-Bayesian model proposed here does agree with this phenomenology and is capable of reproducing a spectrum of effects observed in psychology: primacy-recency phenomenon, boomerang effect and cognitive dissonance. We point out several limitations of the model that should motivate its future development. PMID:25007078

  2. Confirmation of shutdown cooling effects

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Kotaro, E-mail: ksato@nelted.co.jp; Tabuchi, Masato; Sugimura, Naoki; Tatsumi, Masahiro [Nuclear Engineering, Limited, 1-3-7 Tosabori Nishi-ku, Osaka-shi, Osaka 550-0001 (Japan)

    2015-12-31

    After the Fukushima accidents, all nuclear power plants in Japan have gradually stopped their operations and have long periods of shutdown. During those periods, reactivity of fuels continues to change significantly especially for high-burnup UO{sub 2} fuels and MOX fuels due to radioactive decays. It is necessary to consider these isotopic changes precisely, to predict neutronics characteristics accurately. In this paper, shutdown cooling (SDC) effects of UO{sub 2} and MOX fuels that have unusual operation histories are confirmed by the advanced lattice code, AEGIS. The calculation results show that the effects need to be considered even after nuclear power plants come back to normal operation.

  3. Confirmation of shutdown cooling effects

    Science.gov (United States)

    Sato, Kotaro; Tabuchi, Masato; Sugimura, Naoki; Tatsumi, Masahiro

    2015-12-01

    After the Fukushima accidents, all nuclear power plants in Japan have gradually stopped their operations and have long periods of shutdown. During those periods, reactivity of fuels continues to change significantly especially for high-burnup UO2 fuels and MOX fuels due to radioactive decays. It is necessary to consider these isotopic changes precisely, to predict neutronics characteristics accurately. In this paper, shutdown cooling (SDC) effects of UO2 and MOX fuels that have unusual operation histories are confirmed by the advanced lattice code, AEGIS. The calculation results show that the effects need to be considered even after nuclear power plants come back to normal operation.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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.

  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. Stable transformation of ferns using spores as targets: Pteris vittata and Ceratopteris thalictroides.

    Science.gov (United States)

    Muthukumar, Balasubramaniam; Joyce, Blake L; Elless, Mark P; Stewart, C Neal

    2013-10-01

    Ferns (Pteridophyta) are very important members of the plant kingdom that lag behind other taxa with regards to our understanding of their genetics, genomics, and molecular biology. We report here, to our knowledge, the first instance of stable transformation of fern with recovery of transgenic sporophytes. Spores of the arsenic hyperaccumulating fern Pteris vittata and tetraploid 'C-fern Express' (Ceratopteris thalictroides) were stably transformed by Agrobacterium tumefaciens with constructs containing the P. vittata actin promoter driving a GUSPlus reporter gene. Reporter gene expression assays were performed on multiple tissues and growth stages of gametophytes and sporophytes. Southern-blot analysis confirmed stable transgene integration in recovered sporophytes and also confirmed that no plasmid from A. tumefaciens was present in the sporophyte tissues. We recovered seven independent transformants of P. vittata and four independent C. thalictroides transgenics. Inheritance analyses using β-glucuronidase (GUS) histochemical staining revealed that the GUS transgene was stably expressed in second generation C. thalictroides sporophytic tissues. In an independent experiment, the gusA gene that was driven by the 2× Cauliflower mosaic virus 35S promoter was bombarded into P. vittata spores using biolistics, in which putatively stable transgenic gametophytes were recovered. Transformation procedures required no tissue culture or selectable marker genes. However, we did attempt to use hygromycin selection, which was ineffective for recovering transgenic ferns. This simple stable transformation method should help facilitate functional genomics studies in ferns.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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...

  18. Spore Loads May Not be Used Alone as a Direct Indicator of the Severity of Nosema ceranae Infection in Honey Bees Apis mellifera (Hymenoptera:Apidae).

    Science.gov (United States)

    Zheng, Huo-Qing; Lin, Zhe-Guang; Huang, Shao-Kang; Sohr, Alex; Wu, Lyman; Chen, Yan Ping

    2014-12-01

    Nosema ceranae Fries et al., 1996, a microsporidian parasite recently transferred from Asian honey bees Apis cerana F., 1793, to European honey bees Apis mellifera L., 1758, has been suspected as one of the major culprits of the worldwide honey bee colony losses. Spore load is a commonly used criterion to describe the intensity of Nosema infection. In this study, by providing Nosema-infected bees with sterilized pollen, we confirmed that pollen feeding increased the spore loads of honey bees by several times either in the presence or absence of a queen. By changing the amount of pollen consumed by bees in cages, we showed that spore loads increased with an increase in pollen consumption. Nosema infections decrease honey bee longevity and transcription of vitellogenin, either with or without pollen feeding. However, the reduction of pollen consumption had a greater impact on honey bee longevity and vitellogenin level than the increase of spore counts caused by pollen feeding. These results indicate that spore loads may not be used alone as a direct indicator of the severity of N. ceranae infection in honey bees. © 2014 Entomological Society of America.

  19. Performance confirmation data acquisition system

    International Nuclear Information System (INIS)

    McAffee, D.A.; Raczka, N.T.

    1997-01-01

    As part of the Viability Assessment (VA) work, this QAP-3-9 document presents and evaluates a comprehensive set of viable concepts for collecting Performance Confirmation (PC) related data. The concepts include: monitoring subsurface repository air temperatures, humidity levels and gaseous emissions via the subsurface ventilation systems, and monitoring the repository geo-technical parameters and rock mass from bore-holes located along the perimeter main drifts and throughout a series of human-rated Observation Drifts to be located in a plane 25 meters above the plane of the emplacement drifts. A key element of this document is the development and analysis of a purposed multi-purpose Remote Inspection Gantry that would provide direct, real-time visual, thermal, and radiological monitoring of conditions inside operational emplacement drifts and close-up observations of in-situ Waste Packages. Preliminary finite-element analyses are presented that indicate the technological feasibility of operating an inspection gantry inside the operational emplacement drifts for short inspection missions lasting 2--3 hours. Overall reliability, availability, and maintainability of the PC data collection concepts are discussed. Preliminary concepts for PC data collection network are also provided

  20. Performance confirmation data acquisition system

    Energy Technology Data Exchange (ETDEWEB)

    McAffee, D.A.; Raczka, N.T. [Yucca Mountain Project, Las Vegas, NV (United States)

    1997-12-31

    As part of the Viability Assessment (VA) work, this QAP-3-9 document presents and evaluates a comprehensive set of viable concepts for collecting Performance Confirmation (PC) related data. The concepts include: monitoring subsurface repository air temperatures, humidity levels and gaseous emissions via the subsurface ventilation systems, and monitoring the repository geo-technical parameters and rock mass from bore-holes located along the perimeter main drifts and throughout a series of human-rated Observation Drifts to be located in a plane 25 meters above the plane of the emplacement drifts. A key element of this document is the development and analysis of a purposed multi-purpose Remote Inspection Gantry that would provide direct, real-time visual, thermal, and radiological monitoring of conditions inside operational emplacement drifts and close-up observations of in-situ Waste Packages. Preliminary finite-element analyses are presented that indicate the technological feasibility of operating an inspection gantry inside the operational emplacement drifts for short inspection missions lasting 2--3 hours. Overall reliability, availability, and maintainability of the PC data collection concepts are discussed. Preliminary concepts for PC data collection network are also provided.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. 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)

  7. Spore load and immune response of honey bees naturally infected by Nosema ceranae.

    Science.gov (United States)

    Li, Wenfeng; Evans, Jay D; Li, Jianghong; Su, Songkun; Hamilton, Michele; Chen, Yanping

    2017-12-01

    Nosema ceranae causes widespread infection in adult workers of European honey bees, Apis mellifera, and has often been linked to honey bee colony losses worldwide. Previous investigations of honey bee immune response to N. ceranae infection were largely based on laboratory experiment, however, little is known about the immune response of honey bees that are naturally infected by N. ceranae. Here, we compared the infection levels of N. ceranae in three different categories of adult bees (emergent bees, nurses, and foragers) and detected the host immune response to the N. ceranae infection under natural conditions. Our studies showed that the Nosema spore load and infection prevalence varied among the different types of adult workers, and both of them increased as honey bees aged: No infection was detected in emergent bees, nurses had a medium spore load and prevalence, while foragers were with the highest Nosema infection level and prevalence. Quantification of the mRNA levels of antimicrobial peptides (abaecin, apidaecin, defensin-1, defensin-2, and hymenoptaecin) and microbial recognition proteins (PGRP-S1, PGRP-S2, PGRP-S3, PGRP-LC, GNBP1-1, and GNBP1-2) confirmed the involvement of the Toll and/or Imd immune pathways in the host response to N. ceranae infection, and revealed an activation of host immune response by N. ceranae infection under natural conditions. Additionally, the levels of immune response were positively correlated with the Nosema spore loads in the infected bees. The information gained from this study will be relevant to the predictive modeling of honey bee disease dynamics for Nosema disease prevention and management.

  8. 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

  9. 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.

  10. 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

  11. 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

  12. 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...

  13. 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)

  14. 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...

  15. 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.

  16. 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

  17. 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.

  18. 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...

  19. 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

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. 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)

  10. 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.

  11. 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.

  12. 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...

  13. 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.

  14. 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

  15. 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.

  16. 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.

  17. 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

  18. 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

  19. 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

  20. 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 ...

  1. 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).

  2. 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

  3. 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 ...

  4. 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.

  5. 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

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

    Science.gov (United States)

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

    2018-04-25

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

  7. Improved eradication of Clostridium difficile spores from toilets of hospitalized patients using an accelerated hydrogen peroxide as the cleaning agent

    Directory of Open Access Journals (Sweden)

    Dueck Christine

    2010-09-01

    Full Text Available Abstract Background C. difficle spores in the environment of patients with C. difficile associated disease (CDAD are difficult to eliminate. Bleach (5000 ppm has been advocated as an effective disinfectant for the environmental surfaces of patients with CDAD. Few alternatives to bleach for non-outbreak conditions have been evaluated in controlled healthcare studies. Methods This study was a prospective clinical comparison during non-outbreak conditions of the efficacy of an accelerated hydrogen peroxide cleaner (0.5% AHP to the currently used stabilized hydrogen peroxide cleaner (0.05% SHP at manufacturer recommended use-dilution with respect to spore removal from toilets in a tertiary care facility. The toilets used by patients who had diarrhea with and without C. difficile associated disease (CDAD were cultured for C. difficile and were monitored using an ultraviolet mark (UVM to assess cleaning compliance on a daily basis 5 days per week. A total of 243 patients and 714 samples were analysed. The culture results were included in the analysis only if the UVM audit from the same day confirmed that the toilet had been cleaned. Results Our data demonstrated that the efficacy of spore killing is formulation specific and cannot be generalized. The OxivirTB® AHP formulation resulted in statistically significantly (p = 0.0023 lower levels of toxigenic C. difficile spores in toilets of patients with CDAD compared to the SHP formulation that was routinely being used (28% vs 45% culture positive. The background level of toxigenic C. difficile spores was 10% in toilets of patients with diarrhea not due to CDAD. The UVM audit indicated that despite the enhanced twice-daily cleaning protocol for CDAD patients cleaning was not achieved on approximately 30 - 40% of the days tested. Conclusion Our data indicate that the AHP formulation evaluated that has some sporicidal activity was significantly better than the currently used SHP formulation. This AHP

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

    Directory of Open Access Journals (Sweden)

    Pasco B. Avery

    2018-04-01

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

  9. 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

  10. 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)

  11. 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

  12. 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)

  13. 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

  14. 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.

  15. [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.

  16. 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

  17. 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

  18. 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)

  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. 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.)

  3. A real-time PCR-based strategy for the detection of Paenibacillus larvae vegetative cells and spores to improve the diagnosis and the screening of American foulbrood.

    Science.gov (United States)

    Martínez, J; Simon, V; Gonzalez, B; Conget, P

    2010-06-01

    To develop a real-time PCR-based strategy for the detection of Paenibacillus larvae vegetative cells and spores to improve the diagnosis and the screening of American foulbrood (AFB), the most harmful pathology of honeybee brood. A real-time PCR that allowed selective identification and quantification of P. larvae 16S rRNA sequence was developed. Using standard samples quantified by flow cytometry, detection limits of 37.5 vegetative cells ml(-1) and 10 spores ml(-1) were determined. Compared to spread plate method, this real-time PCR-based strategy allowed, in only 2 h, the detection of P. larvae in contaminated honeys. No false-positive results were obtained. Moreover, its detection limit was 100 times lower than that of the culture method (2 vs 200 spores g(-1) of honey). A rapid, selective, with low detection limit, sensitive and specific method to detect and quantify vegetative cells and spores of P. larvae is now available. In addition to honey samples, this real-time PCR-based strategy may be also applied to confirm AFB diagnosis in honeybee brood and to screen other apiary supplies and products (bees, pollen, wax), thus broadening the control of AFB spreading.

  4. 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.

  5. Muricholic acids inhibit Clostridium difficile spore germination and growth.

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    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.

  6. 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)

  7. 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.)

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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

  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. 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. 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.

  17. 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

  18. 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

  19. 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

  20. 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...

  1. 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...

  2. 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...

  3. 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) ...

  4. 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.

  5. Stable Transformation of Ferns Using Spores as Targets: Pteris vittata and Ceratopteris thalictroides1[W][OPEN

    Science.gov (United States)

    Muthukumar, Balasubramaniam; Joyce, Blake L.; Elless, Mark P.; Stewart, C. Neal

    2013-01-01

    Ferns (Pteridophyta) are very important members of the plant kingdom that lag behind other taxa with regards to our understanding of their genetics, genomics, and molecular biology. We report here, to our knowledge, the first instance of stable transformation of fern with recovery of transgenic sporophytes. Spores of the arsenic hyperaccumulating fern Pteris vittata and tetraploid ‘C-fern Express’ (Ceratopteris thalictroides) were stably transformed by Agrobacterium tumefaciens with constructs containing the P. vittata actin promoter driving a GUSPlus reporter gene. Reporter gene expression assays were performed on multiple tissues and growth stages of gametophytes and sporophytes. Southern-blot analysis confirmed stable transgene integration in recovered sporophytes and also confirmed that no plasmid from A. tumefaciens was present in the sporophyte tissues. We recovered seven independent transformants of P. vittata and four independent C. thalictroides transgenics. Inheritance analyses using β-glucuronidase (GUS) histochemical staining revealed that the GUS transgene was stably expressed in second generation C. thalictroides sporophytic tissues. In an independent experiment, the gusA gene that was driven by the 2× Cauliflower mosaic virus 35S promoter was bombarded into P. vittata spores using biolistics, in which putatively stable transgenic gametophytes were recovered. Transformation procedures required no tissue culture or selectable marker genes. However, we did attempt to use hygromycin selection, which was ineffective for recovering transgenic ferns. This simple stable transformation method should help facilitate functional genomics studies in ferns. PMID:23933990

  6. Calibration and Confirmation in Geophysical Models

    Science.gov (United States)

    Werndl, Charlotte

    2016-04-01

    For policy decisions the best geophysical models are needed. To evaluate geophysical models, it is essential that the best available methods for confirmation are used. A hotly debated issue on confirmation in climate science (as well as in philosophy) is the requirement of use-novelty (i.e. that data can only confirm models if they have not already been used before. This talk investigates the issue of use-novelty and double-counting for geophysical models. We will see that the conclusions depend on the framework of confirmation and that it is not clear that use-novelty is a valid requirement and that double-counting is illegitimate.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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...

  14. 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.

  15. 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...

  16. 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.

  17. 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

  18. Funding Alert Subscribe Confirmation | IDRC - International ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Thank you. Please check your email ( ) to confirm your subscription to the IDRC Funding Alerts. Please note: the link in your confirmation email is valid for 3 days, so please reply promptly. We fund researchers driving global change. Careers · Contact Us · Subscribe · Unsubscribe · Site map. Follow us; Facebook · Twitter ...

  19. 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

  20. 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.

  1. 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

  2. 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.

  3. Hourly predictive artificial neural network and multivariate regression tree models of Alternaria and Cladosporium spore concentrations in Szczecin (Poland)

    Science.gov (United States)

    Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka

    2009-11-01

    A study was made of the link between time of day, weather variables and the hourly content of certain fungal spores in the atmosphere of the city of Szczecin, Poland, in 2004-2007. Sampling was carried out with a Lanzoni 7-day-recording spore trap. The spores analysed belonged to the taxa Alternaria and Cladosporium. These spores were selected both for their allergenic capacity and for their high level presence in the atmosphere, particularly during summer. Spearman correlation coefficients between spore concentrations, meteorological parameters and time of day showed different indices depending on the taxon being analysed. Relative humidity (RH), air temperature, air pressure and clouds most strongly and significantly influenced the concentration of Alternaria spores. Cladosporium spores correlated less strongly and significantly than Alternaria. Multivariate regression tree analysis revealed that, at air pressures lower than 1,011 hPa the concentration of Alternaria spores was low. Under higher air pressure spore concentrations were higher, particularly when RH was lower than 36.5%. In the case of Cladosporium, under higher air pressure (>1,008 hPa), the spores analysed were more abundant, particularly after 0330 hours. In artificial neural networks, RH, air pressure and air temperature were the most important variables in the model for Alternaria spore concentration. For Cladosporium, clouds, time of day, air pressure, wind speed and dew point temperature were highly significant factors influencing spore concentration. The maximum abundance of Cladosporium spores in air fell between 1200 and 1700 hours.

  4. Role of visible light-activated photocatalyst on the reduction of anthrax spore-induced mortality in mice.

    Directory of Open Access Journals (Sweden)

    Jyh-Hwa Kau

    Full Text Available BACKGROUND: Photocatalysis of titanium dioxide (TiO(2 substrates is primarily induced by ultraviolet light irradiation. Anion-doped TiO(2 substrates were shown to exhibit photocatalytic activities under visible-light illumination, relative environmentally-friendly materials. Their anti-spore activity against Bacillus anthracis, however, remains to be investigated. We evaluated these visible-light activated photocatalysts on the reduction of anthrax spore-induced pathogenesis. METHODOLOGY/PRINCIPAL FINDINGS: Standard plating method was used to determine the inactivation of anthrax spore by visible light-induced photocatalysis. Mouse models were further employed to investigate the suppressive effects of the photocatalysis on anthrax toxin- and spore-mediated mortality. We found that anti-spore activities of visible light illuminated nitrogen- or carbon-doped titania thin films significantly reduced viability of anthrax spores. Even though the spore-killing efficiency is only approximately 25%, our data indicate that spores from photocatalyzed groups but not untreated groups have a less survival rate after macrophage clearance. In addition, the photocatalysis could directly inactivate lethal toxin, the major virulence factor of B. anthracis. In agreement with these results, we found that the photocatalyzed spores have tenfold less potency to induce mortality in mice. These data suggest that the photocatalysis might injury the spores through inactivating spore components. CONCLUSION/SIGNIFICANCE: Photocatalysis induced injuries of the spores might be more important than direct killing of spores to reduce pathogenicity in the host.

  5. Self-healing concrete by use of microencapsulated bacterial spores

    Energy Technology Data Exchange (ETDEWEB)

    Wang, J.Y. [Magnel Laboratory for Concrete Research, Faculty of Engineering and Architecture, Ghent University, TechnologieparkZwijnaarde 904, B-9052 Ghent (Belgium); Laboratory of Microbial Ecology and Technology (LabMET), Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent (Belgium); Soens, H. [Devan Chemicals NV, Klein Frankrijk 18, 9600 Ronse (Belgium); Verstraete, W. [Laboratory of Microbial Ecology and Technology (LabMET), Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent (Belgium); De Belie, N., E-mail: nele.debelie@ugent.be [Magnel Laboratory for Concrete Research, Faculty of Engineering and Architecture, Ghent University, TechnologieparkZwijnaarde 904, B-9052 Ghent (Belgium)

    2014-02-15

    Microcapsules were applied to encapsulate bacterial spores for self-healing concrete. The viability of encapsulated spores and the influence of microcapsules on mortar specimens were investigated first. Breakage of the microcapsules upon cracking was verified by Scanning Electron Microscopy. Self-healing capacity was evaluated by crack healing ratio and the water permeability. The results indicated that the healing ratio in the specimens with bio-microcapsules was higher (48%–80%) than in those without bacteria (18%–50%). The maximum crack width healed in the specimens of the bacteria series was 970 μm, about 4 times that of the non-bacteria series (max 250 μm). The overall water permeability in the bacteria series was about 10 times lower than that in non-bacteria series. Wet–dry cycles were found to stimulate self-healing in mortar specimens with encapsulated bacteria. No self-healing was observed in all specimens stored at 95%RH, indicating that the presence of liquid water is an essential component for self-healing.

  6. Spore attachment and extracellular mucilage of aquatic hyphomycetes.

    Science.gov (United States)

    Au, D W; Jones, E B; Moss, S T

    1996-01-01

    Stages in conidiun attachment to surfaces of Lemmoniera aquatica and Mycocentrospora filiformis (freshwater Hyphomycetes) were studied at the light microscope and scanning and transmission electron microscope levels. Sigmoid conidia of M. filiformis attach by pre-existing conidial mucilage at the spore pole and at a point along the conidial body. Tetraradiate conidia of L. aquatica attach by the thigmotropic release of mucilage at the tips of the three "arms";. Germination in both species is followed by the production of germ tubes, germ hyphae and appressoria. The chemical composition of the mucilage involved in attachment was determined by enzymatic studies and lectin-gold cytochemical studies. The major component was found to be acidic poly-saccharide, comprising mainly ß-1, 3-glucan, N-acetyl-D-glucosamine and N-acetyl-neuraminic acid. Variation in mucilage composition exists between the two species, among different structures of the same species, and on different regions of the same structure. This indicates that mucilage producton in the two species is a dynamic process.The ability to secure rapid spore attachment, often in turbulent condition, would be a competitive advantage to these saprobic fungi in the colonization of substrata.

  7. Identifying and quantifying Phakopsora pachyrhizi spores in rain.

    Science.gov (United States)

    Barnes, C W; Szabo, L J; Bowersox, V C

    2009-04-01

    In summers of 2005 and 2006, rain was collected weekly at over 100 selected National Atmospheric Deposition Program/National Trends Network sites across the soybean-growing region of the central and eastern United States. Rain samples were screened for Phakopsora pachyrhizi (causal agent of soybean rust) DNA using a nested real-time polymerase chain reaction assay. Over this time frame, P. pachyrhizi spores were detected in every state in the study, but more frequently in states along the Gulf and Atlantic coasts and along the Ohio River Valley westward to Kansas. A bimodal temporal distribution of samples testing positive for P. pachyrhizi was found in both years. However, there was a greater than threefold increase in the number of samples testing positive for P. pachyrhizi in 2006 compared with 2005, with the most significant increase in August. There was also an increase in the average number of spores per sample in 2006 relative to 2005. Sequence analysis of a subset of positive samples was used to validate the assay results. From the sequence analysis, two reliable polymorphic regions were found, resulting in six distinct genotypes. One genotype was found in 56% of the samples tested, whereas the other genotypes were found less frequently.

  8. Experience with confirmation measurement at Los Alamos

    International Nuclear Information System (INIS)

    Marshall, R.S.; Wagner, R.P.; Hsue, F.

    1985-01-01

    Confirmation measurements are used at Los Alamos in support of incoming and outgoing shipment accountibility and for support of both at 235 U and Pu inventories. Statistical data are presented to show the consistency of measurements on items of identical composition and on items measured at two facilitis using similar instruments. A description of confirmation measurement techniques used in support of 235 U and Pu inventories and a discussion on the ability of the measurements to identify items with misstated SNM are given

  9. Experience with confirmation measurement at Los Alamos

    International Nuclear Information System (INIS)

    Marshall, R.S.; Wagner, R.P.

    1985-01-01

    Confirmation measurements are used at Los Alamos in support of incoming and outgoing shipment accountability and for support of both 235 U and Pu inventories. Statistical data are presented to show the consistency of measurements on items of identical composition and on items measured at two facilities using similar instruments. A description of confirmation measurement techniques used in support of 235 U and Pu inventories and a discussion on the ability of the measurements to identify items with misstated SNM are given

  10. Assessment of mould spore exposure and relations to symptoms in wood trimmers

    NARCIS (Netherlands)

    Eduard, W.

    1993-01-01

    Relationships between exposure to mould spores, IgG antibodies against moulds and respiratory and febrile symptoms were studied among wood trimmers. A new method for quantitative assessment of mould spore exposure by scanning electron microscopy was developed. This method was validated by

  11. Distribution of mycorrhizal fungal spores in soils under agroforestry and monocultural coffee systems in Brazil

    NARCIS (Netherlands)

    Cardoso, I.M.; Boddington, C.L.; Janssen, B.H.; Oenema, O.; Kuyper, T.W.

    2003-01-01

    Deep-rooting trees in agroforestry systems may promote distribution of spores of arbuscular mycorrhizal fungi (AMF) at deeper soil levels. We investigated the vertical distribution of AMF spores in Oxisols under agroforestry and monocultural (unshaded) coffee systems in on-farm experiments (

  12. Bringing Evolution to a Technological Generation: A Case Study with the Video Game SPORE

    Science.gov (United States)

    Poli, DorothyBelle; Berenotto, Christopher; Blankenship, Sara; Piatkowski, Bryan; Bader, Geoffrey A.; Poore, Mark

    2012-01-01

    The video game SPORE was found to hold characteristics that stimulate higher-order thinking even though it rated poorly for accurate science. Interested in evaluating whether a scientifically inaccurate video game could be used effectively, we exposed students to SPORE during an evolution course. Students that played the game reported that they…

  13. Mutation Induction with UV- and X-radiations in spores and vegetative cells of Bacillus subtilis

    International Nuclear Information System (INIS)

    Tanooka, H.; Munakata, N.; Kitahara, S.

    1978-01-01

    Spores and vegetative cells of Bacillus subtilis strains with various defects in DNA-repair capacities (hcr - , ssp - , hcr - ssp - ) were irradiated with UV radiation or X-rays. Induced mutation frequency was determined from the observed frequency of prototrophic reversion of a suppressible auxotropic mutation. At equal physical dose, after either UV- or X-irradiation, spores were more resistant to mutations as well as to killing than were vegetative cells. However, quantitative comparison revealed that, at equally lethal doses, spores and vegetative cells were almost equally mutable by X-rays whereas spores were considerably less mutable by UV than were vegetative cells. Thus, as judged from their mutagenic efficiency relative to the lethality, X-ray-induced damage in the spore DNA and the vegetative DNA were equally mutagenic, while UV-induced DNA photoproducts in the spore were less mutagenic than those in vegetative cells. Post-treatment of UV-irradiated cells with caffeine decreased the survival and the induced mutation frequency for either spores or vegetative cells for all the strains. In X-irradiated spores however, a similar suppressing effect of caffeine was observed only for mutability of a strain lacking DNA polymerase I activity

  14. Removal of Bacillus anthracis sterne spore from commercial unpasteurized liquid egg white using crossflow microfiltration

    Science.gov (United States)

    Current pasteurization technology used by the egg industry is ineffective for destruction of spores such as those of Bacillus anthracis (BA). The validity of a cross-flow microfiltration (MF) process for separation of the attenuated strain of BA (Sterne) spores from commercial unpasteurized liquid ...

  15. Spore dispersal of fetid Lysurus mokusin by feces of mycophagous insects.

    Science.gov (United States)

    Chen, Gao; Zhang, Rui-Rui; Liu, Yang; Sun, Wei-Bang

    2014-08-01

    The ecological roles and biological mechanisms of zoochory in plants have long been foci in studies of co-evolutionary processes between plants and animals. However, the dispersal of fungal spores by animals has received comparatively little attention. In this study, the dispersal of spores of a selected fetid fungus, Lysurus mokusin, via feces of mycophagous insects was explored by: collecting volatiles emitted by the fungus using dynamic headspace extraction and analyzing them by GC-MS; testing the capacity of mycophagous insects to disperse its spores by counting spores in their feces; comparing the germinability of L. mokusin spores extracted from feces of nocturnal earwigs and natural gleba of the fungus; and assessing the ability of L. mokusin volatiles to attract insects in bioassays with synthetic scent mixtures. Numerous spores were detected in insects' feces, the bioassays indicated that L. mokusin odor (similar to that of decaying substances) attracts diverse generalist mycophagous insects, and passage through the gut of Anisolabis maritima earwigs significantly enhanced the germination rate of L. mokusin spores. Therefore, nocturnal earwigs and diurnal flies probably play important roles in dispersal of L. mokusin spores, and dispersal via feces may be an important common dispersal mechanism for fungal reproductive tissue.

  16. 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...

  17. 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.

  18. Felling Infected Oaks in Natural Stands Reduces Dissemination of Polyporus Hispidus Spores

    Science.gov (United States)

    E. R. Toole; F. I. McCracken

    1974-01-01

    Felling or girdling willow oaks which exhibited Polyporus hispidus cankers reduced sporophore production within 3 yr. Since spore dissemination was horizontal and downward, spores produced on felled trees did not spread as far as those from standing trees. Felling, therefore, is a more advantageous means of control.

  19. Analysis of germination and outgrowth of sorbic acid-stressed Bacillus cereus ATCC 14579 spores.

    NARCIS (Netherlands)

    Melis, van Clint; Nierop Groot, Masja; Tempelaars, Marcel; Moezelaar, Roy; Abee, Tjakko

    2010-01-01

    Sorbic acid (SA) is widely used as a preservative, but the effect of SA on spore germination and outgrowth has gained limited attention up to now. Therefore, the effect of sorbic acid on germination of spores of B. cereus strain ATCC 14579 was analyzed both at phenotype and transcriptome level.

  20. Changes in concentration of Alternaria and Cladosporium spores during summer storms

    Science.gov (United States)

    Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka

    2013-09-01

    Fungal spores are known to cause allergic sensitization. Recent studies reported a strong association between asthma symptoms and thunderstorms that could be explained by an increase in airborne fungal spore concentrations. Just before and during thunderstorms the values of meteorological parameters rapidly change. Therefore, the goal of this study was to create a predictive model for hourly concentrations of atmospheric Alternaria and Cladosporium spores on days with summer storms in Szczecin (Poland) based on meteorological conditions. For this study we have chosen all days of June, July and August (2004-2009) with convective thunderstorms. There were statistically significant relationships between spore concentration and meteorological parameters: positive for air temperature and ozone content while negative for relative humidity. In general, before a thunderstorm, air temperature and ozone concentration increased, which was accompanied by a considerable increase in spore concentration. During and after a storm, relative humidity increased while both air temperature ozone concentration along with spore concentrations decreased. Artificial neural networks (ANN) were used to assess forecasting possibilities. Good performance of ANN models in this study suggest that it is possible to predict spore concentrations from meteorological variables 2 h in advance and, thus, warn people with spore-related asthma symptoms about the increasing abundance of airborne fungi on days with storms.

  1. 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

  2. Evaluating the transport of bacillus subtilis spores as a potential surrogate for Cryptosporidium parvum Oocysts

    Science.gov (United States)

    The USEPA has recommended the use of aerobic spores as an indicator for Cryptosporidium oocysts when determining groundwater under the direct influence of surface water. Surface properties, interaction energies, transport, retention, and release behavior of B. subtilis spores were measured over a r...

  3. Germinant-enhanced decontamination of Bacillus spores adhered to iron and cement-mortar drinking water infrastructures.

    Science.gov (United States)

    Szabo, Jeffrey G; Muhammad, Nur; Heckman, Lee; Rice, Eugene W; Hall, John

    2012-04-01

    Germination was evaluated as an enhancement to decontamination methods for removing Bacillus spores from drinking water infrastructure. Germinating spores before chlorinating cement mortar or flushing corroded iron was more effective than chlorinating or flushing alone.

  4. DECONTAMINATION ASSESSMENT OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS, AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACTS USING A HYDROGEN PERIOXIDE GAS GENERATOR

    Science.gov (United States)

    Aims: To evaluate the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface materials using hydrogen peroxide gas. Methods and Results: B. anthracis, B. subtilis, and G. Stearothermophilus spores were dried on seven...

  5. Modelling the impact of fungal spore ice nuclei on clouds and precipitation

    International Nuclear Information System (INIS)

    Sesartic, Ana; Lohmann, Ulrike; Storelvmo, Trude

    2013-01-01

    Some fungal spore species have been found in laboratory studies to be very efficient ice nuclei. However, their potential impact on clouds and precipitation is not well known and needs to be investigated. Fungal spores as a new aerosol species were introduced into the global climate model (GCM) ECHAM5-HAM. The inclusion of fungal spores acting as ice nuclei in a GCM leads to only minor changes in cloud formation and precipitation on a global level; however, changes in the liquid water path and ice water path as well as stratiform precipitation can be observed in the boreal regions where tundra and forests act as sources of fungal spores. Although fungal spores contribute to heterogeneous freezing, their impact is reduced by their low numbers as compared to other heterogeneous ice nuclei. (letter)

  6. Recent progress in Bacillus subtilis spore-surface display: concept, progress, and future.

    Science.gov (United States)

    Wang, He; Wang, Yunxiang; Yang, Ruijin

    2017-02-01

    With the increased knowledge on spore structure and advances in biotechnology engineering, the newly developed spore-surface display system confers several inherent advantages over other microbial cell-surface display systems including enhanced stability and high safety. Bacillus subtilis is the most commonly used Bacillus species for spore-surface display. The expression of heterologous antigen or protein on the surface of B. subtilis spores has now been practiced for over a decade with noteworthy success. As an update and supplement to other previous reviews, we comprehensively summarize recent studies in the B. subtilis spore-surface display technique. We focus on its benefits as well as the critical factors affecting its display efficiency and offer suggestions for the future success of this field.

  7. Esterase activity as a novel parameter of spore germination in Bacillus anthracis

    International Nuclear Information System (INIS)

    Ferencko, Linda; Cote, Mindy A.; Rotman, Boris

    2004-01-01

    Spores of Bacillus anthracis were shown to produce esterase activity about 4 min after exposure to conventional germinants such as combinations of amino acids and purine ribosides. Neither amino acids nor ribosides alone induce germination and esterase activity. Expression of esterase activity was chloramphenicol resistant, and correlated with loss of spore refractivity, a traditional parameter of early germination. Based on these observations, we hypothesized that esterase activity could be used as a novel parameter for quantifying early events during spore germination. To test this hypothesis, we measured expression of esterase activity under a variety of germinating conditions. Using diacetyl fluorescein as fluorogenic substrate of esterases, we demonstrated that esterase activity was invariably induced whenever spores were triggered by known germinants. Moreover, D-alanine, an inhibitor of L-alanine-mediated germination, was found to significantly inhibit expression of esterase activity. In terms of molecular mechanisms, esterase expression could represent activation of proteases at the onset of spore germination

  8. Workshop on the Destruction of Bacterial Spores Held in Brussels, Belgium on May 1-3, 1985.

    Science.gov (United States)

    1985-05-03

    corrmunication. 0 We propose to evaluate the relevance to food processing of recent research ,. on resistance of bacterial spores to heat, and to reappraise the...spores from thermophiles is important in food spoilage . It also is an attribute useful in biological indicator systems • used to evaluate thermal... Bacterial spore injury - an update. 4 Food Prot. 44:776-786. Lewis, J.C., N.S. Snell, and G. Halderton. 1964. Dormancy and activation of :acterial spores, in

  9. Analysis of the Effects of a gerP Mutation on the Germination of Spores of Bacillus subtilis

    Science.gov (United States)

    2012-11-01

    Spores of Bacillus species are metabolically dormant and ex-tremely resistant to a wide variety of agents (38). As a conse- quence, these spores can...permeability barrier in dormant spores, the coat is a permeability barrier to large mole- cules (18, 20). Thus, it is possible that there are special...type and gerP spore germina- tion. Almost all bacteria have an alanine racemase activity essen- tial for the generation of the D-alanine needed for

  10. 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.

  11. Artificial neural network models of relationships between Alternaria spores and meteorological factors in Szczecin (Poland)

    Science.gov (United States)

    Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka

    2008-11-01

    Alternaria is an airborne fungal spore type known to trigger respiratory allergy symptoms in sensitive patients. Aiming to reduce the risk for allergic individuals, we constructed predictive models for the fungal spore circulation in Szczecin, Poland. Monthly forecasting models were developed for the airborne spore concentrations of Alternaria, which is one of the most abundant fungal taxa in the area. Aerobiological sampling was conducted over 2004-2007, using a Lanzoni trap. Simultaneously, the following meteorological parameters were recorded: daily level of precipitation; maximum and average wind speed; relative humidity; and maximum, minimum, average, and dew point temperature. The original factors as well as with lags (up to 3 days) were used as the explaining variables. Due to non-linearity and non-normality of the data set, the modelling technique applied was the artificial neural network (ANN) method. The final model was a split model with classification (spore presence or absence) followed by regression for spore seasons and log(x+1) transformed Alternaria spore concentration. All variables except maximum wind speed and precipitation were important factors in the overall classification model. In the regression model for spore seasons, close relationships were noted between Alternaria spore concentration and average and maximum temperature (on the same day and 3 days previously), humidity (with lag 1) and maximum wind speed 2 days previously. The most important variable was humidity recorded on the same day. Our study illustrates a novel approach to modelling of time series with short spore seasons, and indicates that the ANN method provides the possibility of forecasting Alternaria spore concentration with high accuracy.

  12. Fungal Spore Concentrations and Ergosterol Content in Aerosol Samples in the Caribbean During African Dust Events

    Science.gov (United States)

    Santos-Figueroa, G.; Bolaños-Rosero, B.; Mayol-Bracero, O. L.

    2015-12-01

    Fungal spores are a major component of primary biogenic aerosol particles that are emitted to the atmosphere, are ubiquitous, and play an important role in the chemistry and physics of the atmosphere, climate, and public health. Every year, during summer months, African dust (AD) particles are transported to the Caribbean region causing an increase in the concentrations of particulate matter in the atmosphere. AD is one of the most important natural sources of mineral particulate matter at the global scale, and many investigations suggest that it has the ability to transport dust-associated biological particles through long distances. The relationship between AD incursions and the concentration of fungal spores in the Caribbean region is poorly understood. In order to investigate the effects of AD incursions on fungal spore's emissions, fungal spore concentrations were monitored using a Burkard spore trap at the tropical montane cloud forest of Pico del Este at El Yunque National Forest, Puerto Rico. The presence of AD was supported with satellite images of aerosol optical thickness, and with the results from the air masses backward trajectories calculated with the NOAA HYSPLIT model. Basidiospores and Ascospores comprised the major components of the total spore's concentrations, up to a maximum of 98%, during both AD incursions and background days. A considerably decrease in the concentration of fungal spores during AD events was observed. Ergosterol, biomarker for measuring fungal biomass, concentrations were determined in aerosols that were sampled at a marine site, Cabezas de San Juan Nature Reserve, in Fajardo Puerto Rico, and at an urban site, Facundo Bueso building at the University of Puerto Rico. Additional efforts to understand the relationship between the arrival of AD to the Caribbean and a decrease in spore's concentrations are needed in order to investigate changes in local spore's vs the contribution of long-range spores transported within the AD.

  13. Mass production of spores of lactic acid-producing Rhizopus oryzae NBRC 5384 on agar plate.

    Science.gov (United States)

    Yamane, Tsuneo; Tanaka, Ryosuke

    2013-01-01

    Mass production of sporangiospores (spores) of Rhizopus oryzae NBRC 5384 (identical to NRRL 395 and ATCC 9363) on potato-dextrose-agar medium was studied aiming at starting its L(+)-lactic acid fermentation directly from spore inoculation. Various parameters including harvest time, sowed spore density, size of agar plate, height of air space, and incubation mode of plate (agar-on-bottom or agar-on-top) were studied. Ordinarily used shallow Petri dishes were found out to be unsuitable for the full growth of R. oryzae sporangiophores. In a very wide range of the sowed spore density, the smaller it was, the greater the number of the harvested spores was. It was also interesting to find out that R. oryzae grown downward vertically with a deep air space in an agar-on-top mode gave larger amount of spores than in an agar-on-bottom mode at 30°C for 7-day cultivation. Scale-up of the agar plate culture from 26.4 to 292 cm(2) was studied, resulting in the proportional relationship between the number of the harvested spores/plate and the plate area in the deep Petri dishes. The number of plates of 50 cm in diameter needed for 100 m(3) industrial submerged fermentation started directly from 2 × 10(5) spores/mL inoculum size was estimated as about 6, from which it was inferred that such a fermentation would be feasible. Designing a 50 cm plate and a method of spreading and collecting the spores were suggested. Bioprocess technological significance of the "full-scale industrial submerged fermentation started directly from spore inoculation omitting pre-culture" has been discussed. © 2013 American Institute of Chemical Engineers.

  14. A Gompertz regression model for fern spores germination

    Directory of Open Access Journals (Sweden)

    Gabriel y Galán, Jose María

    2015-06-01

    Full Text Available Germination is one of the most important biological processes for both seed and spore plants, also for fungi. At present, mathematical models of germination have been developed in fungi, bryophytes and several plant species. However, ferns are the only group whose germination has never been modelled. In this work we develop a regression model of the germination of fern spores. We have found that for Blechnum serrulatum, Blechnum yungense, Cheilanthes pilosa, Niphidium macbridei and Polypodium feuillei species the Gompertz growth model describe satisfactorily cumulative germination. An important result is that regression parameters are independent of fern species and the model is not affected by intraspecific variation. Our results show that the Gompertz curve represents a general germination model for all the non-green spore leptosporangiate ferns, including in the paper a discussion about the physiological and ecological meaning of the model.La germinación es uno de los procesos biológicos más relevantes tanto para las plantas con esporas, como para las plantas con semillas y los hongos. Hasta el momento, se han desarrollado modelos de germinación para hongos, briofitos y diversas especies de espermatófitos. Los helechos son el único grupo de plantas cuya germinación nunca ha sido modelizada. En este trabajo se desarrolla un modelo de regresión para explicar la germinación de las esporas de helechos. Observamos que para las especies Blechnum serrulatum, Blechnum yungense, Cheilanthes pilosa, Niphidium macbridei y Polypodium feuillei el modelo de crecimiento de Gompertz describe satisfactoriamente la germinación acumulativa. Un importante resultado es que los parámetros de la regresión son independientes de la especie y que el modelo no está afectado por variación intraespecífica. Por lo tanto, los resultados del trabajo muestran que la curva de Gompertz puede representar un modelo general para todos los helechos leptosporangiados

  15. Variation of desiccation tolerance and longevity in fern spores.

    Science.gov (United States)

    Ballesteros, Daniel; Hill, Lisa M; Walters, Christina

    2017-04-01

    This work contributes to the understanding of plant cell responses to extreme water stress when it is applied at different intensity and duration. Fern spores are used to explore survival at relative humidity (RH)moisture level. A RH of 10-25% corresponds well to sorption behavior parameters and is below the glass transition, measured using differential scanning calorimetry. Though response to RH was similar among species, the kinetics of deterioration varied considerably among species and this implies differences in the structure or mobility of molecules within the solidified cytoplasm. Our work suggests that desiccation damage occurs in desiccation tolerant cells, and that it is expressed as a time-dependent response, otherwise known as aging. Copyright © 2017 Elsevier GmbH. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2016-10-13

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

  17. Nonintrusive irradiated fuel inventory confirmation technique

    Energy Technology Data Exchange (ETDEWEB)

    Dowdy, E.J.; Nicholson, N.; Caldwell, J.T.

    1980-01-01

    Successful tests showing correlation between the intensity of the Cerenkov glow surrounding irradiated fuel assemblies in water-filled spent fuel storage ponds and the exposure and cooling times of assemblies have been concluded. Fieldable instruments used in subsequent tests confirmed that such measurements can be made easily and rapidly, without fuel assembly movement or the introduction of apparatus into the storage ponds.

  18. Alkaliphilic Bacillus species show potential application in concrete crack repair by virtue of rapid spore production and germination then extracellular calcite formation.

    Science.gov (United States)

    Sharma, T K; Alazhari, M; Heath, A; Paine, K; Cooper, R M

    2017-05-01

    Characterization of alkaliphilic Bacillus species for spore production and germination and calcite formation as a prelude to investigate their potential in microcrack remediation in concrete. Conditions, extent and timing of endospore production was determined by dark-field light microscopy; germination induction and kinetics were assessed by combining reduction in optical density with formation of refractile bodies by phase-contrast microscopy. Bacillus pseudofirmus was selected from several species as the most suitable isolate. Levels and timing of calcium carbonate precipitated in vitro by B. pseudofirmus were evaluated by atomic absorption spectroscopy and structural identity confirmed as calcite and aragonite by Raman spectroscopy and FTIR. The isolate produced copious spores that germinated rapidly in the presence of germinants l-alanine, inosine and NaCl. Bacterial cells produced CaCO 3 crystals in microcracks and the resulting occlusion markedly restricted water ingress. By virtue of rapid spore production and germination, calcium carbonate formation in vitro and in situ, leading to sealing of microcracks, B. pseudofirmus shows clear potential for remediation of concrete on a commercial scale. Microbial sealing of microcracks should become a practicable and sustainable means of increasing concrete durability. © 2017 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of The Society for Applied Microbiology.

  19. Quantifying the effect of water activity and storage temperature on single spore lag times of three moulds isolated from spoiled bakery products.

    Science.gov (United States)

    Dagnas, Stéphane; Gougouli, Maria; Onno, Bernard; Koutsoumanis, Konstantinos P; Membré, Jeanne-Marie

    2017-01-02

    The inhibitory effect of water activity (a w ) and storage temperature on single spore lag times of Aspergillus niger, Eurotium repens (Aspergillus pseudoglaucus) and Penicillium corylophilum strains isolated from spoiled bakery products, was quantified. A full factorial design was set up for each strain. Data were collected at levels of a w varying from 0.80 to 0.98 and temperature from 15 to 35°C. Experiments were performed on malt agar, at pH5.5. When growth was observed, ca 20 individual growth kinetics per condition were recorded up to 35days. Radius of the colony vs time was then fitted with the Buchanan primary model. For each experimental condition, a lag time variability was observed, it was characterized by its mean, standard deviation (sd) and 5 th percentile, after a Normal distribution fit. As the environmental conditions became stressful (e.g. storage temperature and a w lower), mean and sd of single spore lag time distribution increased, indicating longer lag times and higher variability. The relationship between mean and sd followed a monotonous but not linear pattern, identical whatever the species. Next, secondary models were deployed to estimate the cardinal values (minimal, optimal and maximal temperatures, minimal water activity where no growth is observed anymore) for the three species. That enabled to confirm the observation made based on raw data analysis: concerning the temperature effect, A. niger behaviour was significantly different from E. repens and P. corylophilum: T opt of 37.4°C (standard deviation 1.4°C) instead of 27.1°C (1.4°C) and 25.2°C (1.2°C), respectively. Concerning the a w effect, from the three mould species, E. repens was the species able to grow at the lowest a w (aw min estimated to 0.74 (0.02)). Finally, results obtained with single spores were compared to findings from a previous study carried out at the population level (Dagnas et al., 2014). For short lag times (≤5days), there was no difference between lag

  20. Cryogenic Irradiation of Bacillus Atrophaeus spores to understand microbial survival on Icy Bodies

    Science.gov (United States)

    Yerby, C. J.; Noell, A. C.; Hodyss, R. P.; Johnson, P. V.; Ponce, A.

    2017-12-01

    Bacterial Spores are useful indicator organisms for studying the survival of microbes and degradation of biomolecules on the surface of planetary icy bodies. To predict the limits of life's proliferation in space, specifically on icy bodies, it is essential to understand the ability of microbes to withstand photon and particle irradiation at cryogenic temperatures. Bacillus Atrophaeus spores were transferred onto stainless steel coupons by varied processes and subsequently frozen at Europan temperatures (16oK—273oK) in a vacuum at 8.7x10-8 Torr. An argon lamp bombarded the spore-containing coupons with a solar-like radiation spectra for a variety of times, and spores were removed from the coupons and enumerated in culture. To date, (n=43) coupons have been analyzed for spore kill-rates with regards to ice temperature and radiation exposure time. Results will be presented on the effect of cryogenic temperatures in improving radiation resistance of bacterial spores. This works also details methodology improvements by comparing different spore deposition and recovery methods before and after cryogenic irradiation.

  1. 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.

  2. Mushroom's spore size and time of fruiting are strongly related: is moisture important?

    Science.gov (United States)

    Kauserud, Håvard; Heegaard, Einar; Halvorsen, Rune; Boddy, Lynne; Høiland, Klaus; Stenseth, Nils Chr

    2011-04-23

    Most basidiomycete fungi produce annual short-lived sexual fruit bodies from which billions of microscopic spores are spread into the air during a short time period. However, little is known about the selective forces that have resulted in some species fruiting early and others later in the fruiting season. This study of relationships between morphological and ecological characteristics, climate factors and time of fruiting are based upon thorough statistical analyses of 66 520 mapped records from Norway, representing 271 species of autumnal fruiting mushroom species. We found a strong relationship between spore size and time of fruiting; on average, a doubling of spore size (volume) corresponded to 3 days earlier fruiting. Small-spored species dominate in the oceanic parts of Norway, whereas large-spored species are typical of more continental parts. In separate analyses, significant relationships were observed between spore size and climate factors. We hypothesize that these relationships are owing to water balance optimization, driven by water storage in spores as a critical factor for successful germination of primary mycelia in the drier micro-environments found earlier in the fruiting season and/or in continental climates.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2011-06-01

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

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

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    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.

  5. 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.

  6. Ultrasonography for confirmation of gastric tube placement.

    Science.gov (United States)

    Tsujimoto, Hiraku; Tsujimoto, Yasushi; Nakata, Yukihiko; Akazawa, Mai; Kataoka, Yuki

    2017-04-17

    Gastric tubes are commonly used for the administration of drugs and tube feeding for people who are unable to swallow. Feeding via a tube misplaced in the trachea can result in severe pneumonia. Therefore, the confirmation of tube placement in the stomach after tube insertion is important. Recent studies have reported that ultrasonography provides good diagnostic accuracy estimates in the confirmation of appropriate tube placement. Hence, ultrasound could provide a promising alternative to X-rays in the confirmation of tube placement, especially in settings where X-ray facilities are unavailable or difficult to access. To assess the diagnostic accuracy of ultrasound for gastric tube placement confirmation. We searched the Cochrane Library (2016, Issue 3), MEDLINE (to March 2016), Embase (to March 2016), National Institute for Health Research (NIHR) PROSPERO Register (to May 2016), Aggressive Research Intelligence Facility Databases (to May 2016), ClinicalTrials.gov (to May 2016), ISRCTN registry (May 2016), World Health Organization International Clinical Trials Registry Platform (to May 2016) and reference lists of articles, and contacted study authors. We included studies that evaluated the diagnostic accuracy of naso- and orogastric tube placement confirmed by ultrasound visualization using X-ray visualization as the reference standard. We included cross-sectional studies, and case-control studies. We excluded case series or case reports. Studies were excluded if X-ray visualization was not the reference standard or if the tube being placed was a gastrostomy or enteric tube. Two review authors independently assessed the risk of bias and extracted data from each of the included studies. We contacted authors of the included studies to obtain missing data. We identified 10 studies (545 participants and 560 tube insertions) which met our inclusion criteria.No study was assigned low risk of bias or low concern in every QUADAS-2 domain. We judged only three (30

  7. Small, acid-soluble proteins bound to DNA protect Bacillus subtilis spores from killing by dry heat.

    OpenAIRE

    Setlow, B; Setlow, P

    1995-01-01

    Dry Bacillus subtilis spores lacking their two major DNA-binding proteins (small, acid-soluble proteins [SASP] alpha and beta) were much more sensitive to dry heat than were wild-type spores. Survivors of dry heat treatment of both wild-type and mutant spores exhibited a high frequency of mutations, and the DNA from the heated spores had increased numbers of single-strand breaks. These data indicate that SASP alpha and beta provide significant protection to spore DNA against the damaging effe...

  8. Effect of plasterboard composition on Stachybotrys chartarum growth and biological activity of spores.

    Science.gov (United States)

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

    2003-07-01

    The effects of plasterboard composition on the growth and sporulation of Stachybotrys chartarum as well as on the inflammatory potential of the spores were studied. S. chartarum was grown on 13 modified plasterboards under saturated humidity conditions. The biomass was estimated by measuring the ergosterol content of the S. chartarum culture while the spore-induced cytotoxicity and production of nitric oxide (NO), tumor necrosis factor alpha (TNF-alpha), and interleukin-6 in mouse macrophages was used to illustrate the bioactivity of spores. The ergosterol content of S. chartarum correlated with the number of spores collected from plasterboards. The growth and sporulation decreased compared to that of the reference board in those cases where (i) the liner was treated with biocide, (ii) starch was removed from the plasterboard, or (iii) desulfurization gypsum was used in the core. Spores collected from all the plasterboards were toxic to the macrophages. The biocide added to the core did not reduce the growth; in fact, the spores collected from that board evoked the highest cytotoxicity. The conventional additives used in the core had inhibitory effects on growth. Recycled plasterboards used in the core and the board lacking the starch triggered spore-induced TNF-alpha production in macrophages. In summary, this study shows that the growth of a strain of S. chartarum on plasterboard and the subsequent bioactivity of spores were affected by minor changes to the composition of the core or liners, but it could not be totally prevented without resorting to the use of biocides. However, incomplete prevention of microbial growth by biocides even increased the cytotoxic potential of the spores.

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

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    M. Lauren Donnelly

    2017-01-01

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

  10. Evaluation of the effects of fragmented steam exposure cycles on the survival of bacterial spores.

    Science.gov (United States)

    Shirtz, J T; Soli, T C; Allen, W E; Stellwag, E J; McConnell, T J

    1999-01-01

    The purpose of this study was to examine the population and resistance characteristics of bacterial spores which have been exposed to an abbreviated steam sterilization cycle. The philosophy of many pharmaceutical manufacturers is to require a second complete terminal sterilization cycle in the event of an unplanned interruption during the terminal sterilization of a production batch. The impact of abbreviated steam sterilization cycles was examined for their effect on the survivability and resistance of bacterial spores following an inadequate sterilization cycle. Steam sterilization cycles of two minutes and four minutes were performed on separate groups of Biological Indicator spore strips. These groups were then held at room temperature and re-exposed to a range of sterilization conditions after 24, 48, and 72 hours, i.e., start cycle, abort, hold, start cycle, abort. Spore survivor curves were calculated and resistance estimations were determined. The results of the study indicated that the log level of the surviving spores remained fairly constant, but variability within groups increased as sterilization time increased. The resistance of these surviving spores, as measured by D value, also remained relatively constant throughout the holding period. Abbreviated cycles were similarly conducted on ampules containing a spore suspension, and the spore populations and moist heat resistances were determined over time. Contrary to the spore strip, the population of the subject ampules was less stable showing a gradual decline over the same observation period. The study also included a comparison of the surviving population of short and long fragmented cycles. The results of this study demonstrate that a second complete sterilization cycle is unnecessary to assure the absence of living matter in the sterilized units.

  11. Intestinal calcium and bile salts facilitate germination of Clostridium difficile spores.

    Directory of Open Access Journals (Sweden)

    Travis J Kochan

    2017-07-01

    Full Text Available Clostridium difficile (C. difficile is an anaerobic gram-positive pathogen that is the leading cause of nosocomial bacterial infection globally. C. difficile infection (CDI typically occurs after ingestion of infectious spores by a patient that has been treated with broad-spectrum antibiotics. While CDI is a toxin-mediated disease, transmission and pathogenesis are dependent on the ability to produce viable spores. These spores must become metabolically active (germinate in order to cause disease. C. difficile spore germination occurs when spores encounter bile salts and other co-germinants within the small intestine, however, the germination signaling cascade is unclear. Here we describe a signaling role for Ca2+ during C. difficile spore germination and provide direct evidence that intestinal Ca2+ coordinates with bile salts to stimulate germination. Endogenous Ca2+ (released from within the spore and a putative AAA+ ATPase, encoded by Cd630_32980, are both essential for taurocholate-glycine induced germination in the absence of exogenous Ca2+. However, environmental Ca2+ replaces glycine as a co-germinant and circumvents the need for endogenous Ca2+ fluxes. Cd630_32980 is dispensable for colonization in a murine model of C. difficile infection and ex vivo germination in mouse ileal contents. Calcium-depletion of the ileal contents prevented mutant spore germination and reduced WT spore germination by 90%, indicating that Ca2+ present within the gastrointestinal tract plays a critical role in C. difficile germination, colonization, and pathogenesis. These data provide a biological mechanism that may explain why individuals with inefficient intestinal calcium absorption (e.g., vitamin D deficiency, proton pump inhibitor use are more prone to CDI and suggest that modulating free intestinal calcium is a potential strategy to curb the incidence of CDI.

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

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    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.

  13. Decolorization of indigo carmine by laccase displayed on Bacillus subtilis spores.

    Science.gov (United States)

    Cho, Eun-Ah; Seo, Jiyoung; Lee, Dong-Woo; Pan, Jae-Gu

    2011-06-10

    Blue multicopper oxidases, laccases displayed on the surface of Bacillus spores were used to decolorize a widely used textile dyestuff, indigo carmine. The laccase-encoding gene of Bacillus subtilis, cotA, was cloned and expressed in B. subtilis DB104, and the expressed enzyme was spontaneously localized on Bacillus spores. B. subtilis spores expressing laccase exhibited maximal activity for the oxidation of 2,2'-azino-bis (3-ethylthiazoline-6-sulfonate) (ABTS) at pH 4.0 and 80°C, and for the decolorization of indigo carmine at pH 8.0 and 60°C. The displayed enzyme retained 80% of its original activity after pre-treatment with organic solvents such as 50% acetonitrile and n-hexane for 2h at 37°C. The apparent K(m) of the enzyme displayed on spores was 443±124 μM for ABTS with a V(max) of 150 ± 16 U/mg spores. Notably, 1mg of spores displaying B. subtilis laccase (3.4 × 10(2)U for ABTS as a substrate) decolorized 44.6 μg indigo carmine in 2h. The spore reactor (0.5 g of spores corresponding to 1.7×10(5)U in 50 mL) in a consecutive batch recycling mode decolorized 223 mg indigo carmine/L to completion within 42 h at pH 8.0 and 60°C. These results suggest that laccase displayed on B. subtilis spores can serve as a powerful environmental tool for the treatment of textile dye effluent. Copyright © 2011 Elsevier Inc. All rights reserved.

  14. The fastest flights in nature: high-speed spore discharge mechanisms among fungi.

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    Levi Yafetto

    Full Text Available BACKGROUND: A variety of spore discharge processes have evolved among the fungi. Those with the longest ranges are powered by hydrostatic pressure and include "squirt guns" that are most common in the Ascomycota and Zygomycota. In these fungi, fluid-filled stalks that support single spores or spore-filled sporangia, or cells called asci that contain multiple spores, are pressurized by osmosis. Because spores are discharged at such high speeds, most of the information on launch processes from previous studies has been inferred from mathematical models and is subject to a number of errors. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we have used ultra-high-speed video cameras running at maximum frame rates of 250,000 fps to analyze the entire launch process in four species of fungi that grow on the dung of herbivores. For the first time we have direct measurements of launch speeds and empirical estimates of acceleration in these fungi. Launch speeds ranged from 2 to 25 m s(-1 and corresponding accelerations of 20,000 to 180,000 g propelled spores over distances of up to 2.5 meters. In addition, quantitative spectroscopic methods were used to identify the organic and inorganic osmolytes responsible for generating the turgor pressures that drive spore discharge. CONCLUSIONS/SIGNIFICANCE: The new video data allowed us to test different models for the effect of viscous drag and identify errors in the previous approaches to modeling spore motion. The spectroscopic data show that high speed spore discharge mechanisms in fungi are powered by the same levels of turgor pressure that are characteristic of fungal hyphae and do not require any special mechanisms of osmolyte accumulation.

  15. Effect of relative humidity on the aerodynamic diameter and respiratory deposition of fungal spores

    Science.gov (United States)

    Reponen, Tiina; Willeke, Klaus; Ulevicius, Vidmantas; Reponen, Auvo; Grinshpun, Sergey A.

    Exposure to airborne fungal spores may cause respiratory symptoms. The hygroscopicity of airborne spores may significantly affect their aerodynamic diameter, and thus change their deposition pattern in the human respiratory tract. We have investigated the change in aerodynamic diameter of five different fungal species as a function of relative humidity. Liquid and dry dispersion methods were explored for the aerosolization of the fungal spores. A new system that produces non-aggregated spore aerosol directly from a moldy surface was designed and found suitable for this study. The spores were aerosolized from a mold growth on agar by ducting dry air over the surface, and spore chains in the flow were broken up by passing the entire flow through a critical orifice. Size-spectrometric measurements with an Aerodynamic Particle Sizer showed that the aerodynamic diameter of the tested fungal spores does not change significantly when the relative humidity increases from 30% to 90%. A more distinct spore size increase was found at a relative humidity of ˜ 100%. The highest change of the aerodynamic diameter was found with Cladosporium cladosporioides: it increased from 1.8 μm to 2.3 μm when the relative humidity increased from 30% to ˜ 100%. The size increase corresponds to an approximate doubling of the particle volume. In order to estimate the effect of hygroscopic growth on the respiratory deposition of spores, the mean depositions in the human respiratory tract were calculated for fungal spores with various size changes due to hygroscopic growth. A recently developed model of the International Commission of Radiological Protection was used for the respiratory deposition calculations. We found that the 27% increase in Cladosporium size results in a 20-30% increase in the respiratory deposition of these spores. We conclude that most fungal spores are only slightly hygroscopic and the hygroscopic increase does not significantly affect their respiratory deposition. Our

  16. 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

  17. The immunological characteristics and probiotic function of recombinant Bacillus subtilis spore expressing Clonorchis sinensis cysteine protease.

    Science.gov (United States)

    Tang, Zeli; Shang, Mei; Chen, Tingjin; Ren, Pengli; Sun, Hengchang; Qu, Hongling; Lin, Zhipeng; Zhou, Lina; Yu, Jinyun; Jiang, Hongye; Zhou, Xinyi; Li, Xuerong; Huang, Yan; Xu, Jin; Yu, Xinbing

    2016-12-19

    Clonorchiasis, a food-borne zoonosis, is caused by Clonorchis sinensis. The intestinal tract and bile ducts are crucial places for C. sinensis metacercariae to develop into adult worms. The endospore of Bacillus subtilis is an ideal oral immunization vehicle for delivery of heterologous antigens to intestine. Cysteine protease of C. sinensis (CsCP) is an endogenous key component in the excystment of metacercariae and other physiological or pathological processes. We constructed a fusion gene of CotC (a coat protein)-CsCP and obtained B. subtilis spores with recombinant plasmid of pEB03-CotC-CsCP (B.s-CotC-CsCP). CotC-CsCP expressed on spores' surface was detected by Western blotting and immunofluorescence. Immunological characteristics of recombinant spore coat protein were evaluated in a mouse model. The levels of CsCP-specific antibodies were detected by ELISA. Effects of recombinant spores on mouse intestine were evaluated by histological staining. The activities of biochemical enzymes in serum were assayed by microplate. Liver sections of infected mice were evaluated by Ishak score after Masson's trichrome. The B.s-CotC-CsCP spores displayed CsCP on their coat. Specific IgG and isotypes were significantly induced by coat proteins of B.s-CotC-CsCP spores after subcutaneous immunization. IgA levels in intestinal mucus and bile of B.s-CotC-CsCP orally treated mice significantly increased. Additionally, more IgA-secreting cells were observed in enteraden and lamina propria regions of the mouse jejunum, and an increased amount of acidic mucins in intestines were also observed. There were no significant differences in enzyme levels of serum among groups. No inflammatory injury was observed in the intestinal tissues of each group. The degree of liver fibrosis was significantly reduced after oral immunization with B.s-CotC-CsCP spores. Bacillus subtilis spores maintained the original excellent immunogenicity of CsCP expressed on their surface. Both local and systemic

  18. 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.

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

    Science.gov (United States)

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

    2018-04-01

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

  20. Photoinduction of Spore Germination in Marchantia polylmorpha L.is Mediated by Photosynthesis

    OpenAIRE

    Tadashi, Nakazato; Akeo, Kadota; Masamitsu, Wada; Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University; Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University; Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University

    1999-01-01

    The effects of light on spore germination (protrusion of protonemata) in the liverwort Marchantia polymorpha L.were examined. Spore germination was found to be light dependent and light irradiation for 10h or longer was necessary. Test using specific wavelengths showed that the entire spectrum from near UV to red light was effective, red light being the most effective. Spore germination could be induced by intermittent irradiation with 15-min red light pulses given every 1 or 2 h for 24 h. Th...

  1. 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

    Full Text Available Lisa J Bazzle,1 Marc A Cubeta,2 Steven L Marks,1 David C Dorman3 1Department of Clinical Sciences, College of Veterinary Medicine, 2Department of Plant Pathology, College of Agriculture and Life Sciences, Center for Integrated Fungal Research, 3Department of Molecular and Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Purpose: Mushroom poisoning is a recurring and challenging problem in veterinary medicine. Diagnosis of mushroom exposure in animals is hampered by the lack of rapid diagnostic tests. Our study evaluated the feasibility of using flotation concentration and microscopic evaluation of spores for mushroom identification. Evaluation of this method in living animals exposed to toxigenic mushrooms is limited by ethical constraints; therefore, we relied upon the use of an in vitro model that mimics the oral and gastric phases of digestion. Methods: In our study, mycologist-identified toxigenic (poisonous and nontoxigenic fresh mushrooms were collected in North Carolina, USA. In phase 1, quantitative spore recovery rates were determined following magnesium sulfate, modified Sheather's sugar solution, and zinc sulfate flotation (n=16 fungal species. In phase 2, mushrooms (n=40 fungal species were macerated and digested for up to 2 hours in a salivary and gastric juice simulant. The partially digested material was acid neutralized, filtered, and spores concentrated using zinc sulfate flotation followed by microscopic evaluation of spore morphology. Results: Mean spore recovery rates for the three flotation fluids ranged from 32.5% to 41.0% (P=0.82. Mean (± standard error of the mean Amanita spp. spore recovery rates were 38.1%±3.4%, 36.9%±8.6%, and 74.5%±1.6% (P=0.0012 for the magnesium sulfate, Sheather's sugar, and zinc sulfate solutions, respectively. Zinc sulfate flotation following in vitro acid digestion (phase 2 yielded spore numbers adequate for microscopic visualization in

  2. PERFORMANCE CONFIRMATION IN-SITU INSTRUMENTATION

    International Nuclear Information System (INIS)

    N.T. Raczka

    2000-01-01

    The purpose of this document is to identify and analyze the types of in-situ instruments and methods that could be used in support of the data acquisition portion of the Performance Confirmation (PC) program at the potential nuclear waste repository at Yucca Mountain. The PC program will require geomechanical , geophysical, thermal, and hydrologic instrumentation of several kinds. This analysis is being prepared to document the technical issues associated with each type of measurement during the PC period. This analysis utilizes the ''Performance Confirmation Input Criteria'' (CRWMS M andO 1999a) as its starting point. The scope of this analysis is primarily on the period after the start of waste package emplacement and before permanent closure of the repository, a period lasting between 15 and 300 years after last package emplacement (Stroupe 2000, Attachment 1, p. 1). The primary objectives of this analysis are to: (1) Review the design criteria as presented in the ''Performance Confirmation Input Criteria'' (CRWMS M andO 1999a). The scope of this analysis will be limited to the instrumentation related to parameters that require continuous monitoring of the conditions underground. (2) Preliminary identification and listing of the data requirements and parameters as related to the current repository layout in support of PC monitoring. (3) Preliminary identification of methods and instrumentation for the acquisition of the required data. Although the ''Performance Confirmation Input Criteria'' (CRWMS M andO 1999a) defines a broad range of data that must be obtained from a variety of methods, the focus of this analysis is on instrumentation related to the performance of the rock mass and the formation of water in the repository environment, that is obtainable from in-situ observation, testing, and monitoring

  3. Heat Flux Inhibition by Whistlers: Experimental Confirmation

    International Nuclear Information System (INIS)

    Eichler, D.

    2002-01-01

    Heat flux in weakly magnetized collisionless plasma is, according to theoretical predictions, limited by whistler turbulence that is generated by heat flux instabilities near threshold. Observations of solar wind electrons by Gary and coworkers appear to confirm the limit on heat flux as being roughly the product of the magnetic energy density and the electron thermal velocity, in agreement with prediction (Pistinner and Eichler 1998)

  4. The effects of meteorological factors on airborne fungal spore concentration in two areas differing in urbanisation level

    Science.gov (United States)

    Oliveira, M.; Ribeiro, H.; Delgado, J. L.; Abreu, I.

    2009-01-01

    Although fungal spores are an ever-present component of the atmosphere throughout the year, their concentration oscillates widely. This work aims to establish correlations between fungal spore concentrations in Porto and Amares and meteorological data. The seasonal distribution of fungal spores was studied continuously (2005-2007) using volumetric spore traps. To determine the effect of meteorological factors (temperature, relative humidity and rainfall) on spore concentration, the Spearman rank correlation test was used. In both locations, the most abundant fungal spores were Cladosporium, Agaricus, Agrocybe, Alternaria and Aspergillus/Penicillium, the highest concentrations being found during summer and autumn. In the present study, with the exception of Coprinus and Pleospora, spore concentrations were higher in the rural area than in the urban location. Among the selected spore types, spring-autumn spores ( Coprinus, Didymella, Leptosphaeria and Pleospora) exhibited negative correlations with temperature and positive correlations both with relative humidity and rainfall level. On the contrary, late spring-early summer (Smuts) and summer spores ( Alternaria, Cladosporium, Epicoccum, Ganoderma, Stemphylium and Ustilago) exhibited positive correlations with temperature and negative correlations both with relative humidity and rainfall level. Rust, a frequent spore type during summer, had a positive correlation with temperature. Aspergillus/Penicillium, showed no correlation with the meteorological factors analysed. This knowledge can be useful for agriculture, allowing more efficient and reliable application of pesticides, and for human health, by improving the diagnosis and treatment of respiratory allergic disease.

  5. Effects of meteorological factors on airborne bracken ( Pteridium aquilinum (L.) Kuhn.) spores in Salamanca (middle-west Spain)

    Science.gov (United States)

    Rodríguez de La Cruz, David; Sánchez Reyes, Estefanía; Sánchez Sánchez, José

    2009-05-01

    Temporal variation of airborne bracken ( Pteridium aquilinum) spores concentration in Salamanca during 10 years from January 1998 to December 2007 were studied by using a Burkard spore trap, and correlations with some meteorological parameters were analyzed. The number of spores that were counted was very low, due probably to the distance between the spore trap and the main bracken populations which were located 70 km away from the city. Long-range transport caused by winds coming from the Second Quadrant (IIQ) is supposed to be responsible for the appearance of bracken spores in Salamanca. The season period from August to late October shows the most intense spore dispersal process, with an early morning distribution along the day. Years 2002 and 2007 with a low quantity of airborne spores were also characterized by low mean temperatures, always under 18°C from May to June. Daily spore concentration shows positive correlation with temperature and sun hours but negative with IVQ winds and with relative humidity. No correlation between daily spore concentration and rainfall was found. Also, a positive correlation between number of spores and IIQ winds was observed during the main spore season (MSS) and prepeak period (PRE).

  6. 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

  7. Roles of DacB and spm proteins in clostridium perfringens spore resistance to moist heat, chemicals, and UV radiation.

    Science.gov (United States)

    Paredes-Sabja, Daniel; Sarker, Nahid; Setlow, Barbara; Setlow, Peter; Sarker, Mahfuzur R

    2008-06-01

    Clostridium perfringens food poisoning is caused mainly by enterotoxigenic type A isolates that typically possess high spore heat resistance. Previous studies have shown that alpha/beta-type small, acid-soluble proteins (SASP) play a major role in the resistance of Bacillus subtilis and C. perfringens spores to moist heat, UV radiation, and some chemicals. Additional major factors in B. subtilis spore resistance are the spore's core water content and cortex peptidoglycan (PG) structure, with the latter properties modulated by the spm and dacB gene products and the sporulation temperature. In the current work, we have shown that the spm and dacB genes are expressed only during C. perfringens sporulation and have examined the effects of spm and dacB mutations and sporulation temperature on spore core water content and spore resistance to moist heat, UV radiation, and a number of chemicals. The results of these analyses indicate that for C. perfringens SM101 (i) core water content and, probably, cortex PG structure have little if any role in spore resistance to UV and formaldehyde, presumably because these spores' DNA is saturated with alpha/beta-type SASP; (ii) spore resistance to moist heat and nitrous acid is determined to a large extent by core water content and, probably, cortex structure; (iii) core water content and cortex PG cross-linking play little or no role in spore resistance to hydrogen peroxide; (iv) spore core water content decreases with higher sporulation temperatures, resulting in spores that are more resistant to moist heat; and (v) factors in addition to SpmAB, DacB, and sporulation temperature play roles in determining spore core water content and thus, spore resistance to moist heat.

  8. Universal nucleic acids sample preparation method for cells, spores and their mixture

    Science.gov (United States)

    Bavykin, Sergei [Darien, IL

    2011-01-18

    The present invention relates to a method for extracting nucleic acids from biological samples. More specifically the invention relates to a universal method for extracting nucleic acids from unidentified biological samples. An advantage of the presently invented method is its ability to effectively and efficiently extract nucleic acids from a variety of different cell types including but not limited to prokaryotic or eukaryotic cells and/or recalcitrant organisms (i.e. spores). Unlike prior art methods which are focused on extracting nucleic acids from vegetative cell or spores, the present invention effectively extracts nucleic acids from spores, multiple cell types or mixtures thereof using a single method. Important that the invented method has demonstrated an ability to extract nucleic acids from spores and vegetative bacterial cells with similar levels effectiveness. The invented method employs a multi-step protocol which erodes the cell structure of the biological sample, isolates, labels, fragments nucleic acids and purifies labeled samples from the excess of dye.

  9. Rupturing of Biological Spores As a Source of Secondary Particles in Amazonia.

    Science.gov (United States)

    China, Swarup; Wang, Bingbing; Weis, Johannes; Rizzo, Luciana; Brito, Joel; Cirino, Glauber G; Kovarik, Libor; Artaxo, Paulo; Gilles, Mary K; Laskin, Alexander

    2016-11-15

    Airborne biological particles, such as fungal spores and pollen, are ubiquitous in the Earth's atmosphere and may influence the atmospheric environment and climate, impacting air quality, cloud formation, and the Earth's radiation budget. The atmospheric transformations of airborne biological spores at elevated relative humidity remain poorly understood and their climatic role is uncertain. Using an environmental scanning electron microscope (ESEM), we observed rupturing of Amazonian fungal spores and subsequent release of submicrometer size fragments after exposure to high humidity. We find that fungal fragments contain elements of inorganic salts (e.g., Na and Cl). They are hygroscopic in nature with a growth factor up to 2.3 at 96% relative humidity, thus they may potentially influence cloud formation. Due to their hygroscopic growth, light scattering cross sections of the fragments are enhanced by up to a factor of 10. Furthermore, rupturing of fungal spores at high humidity may explain the bursting events of new particle formation in Amazonia.

  10. The Ultraviolet Photochemistry and Photobiology of Vegetative Cells and Spores of Bacillus megaterium

    Science.gov (United States)

    Donnellan, J. E.; Stafford, R. S.

    1968-01-01

    The ultraviolet (UV) photochemistry and photobiology of spores and vegetative cells of Bacillus megaterium have been studied. The response of vegetative cells of B. megaterium appears qualitatively similar to those of Escherichia coli, Micrococcus radiodurans, and Bacillus subtilis with respect to photoproduct formation and repair mechanisms. UV irradiation, however, does not produce cyclobutane-type thymine dimers in the DNA of spores, although other thymine photo-products are produced. The photoproducts do not disappear after photoreactivation, but they are eliminated from the DNA by a dark-repair mechanism different from that found for dimers in vegetative cells. Irradiations performed at three wavelengths produce the same amounts of spore photoproduct and give the same survival curves. Variation of the sporulation medium before irradiation results in comparable alterations in the rate of spore photoproduct production and in survival. PMID:4966691

  11. Characterization of the Surface Morphology of Bacillus Spores by Atomic Force Microscopy

    National Research Council Canada - National Science Library

    Zolock, Ruth

    2002-01-01

    .... kurstaki, Bacillus cereus strain 569, and Bacillus globigii var. niger. The spores were separated from a nutrient agar culture by filtering and centrifugation, suspended in deionized water, and immobilized on a graphite substrate by spin-coating...

  12. Lethality of Bacillus Anthracis Spores Due to Short Duration Heating Measured Using Infrared Spectroscopy

    National Research Council Canada - National Science Library

    Goetz, Kristina M

    2005-01-01

    In this research, Bacillus anthracis spores were subjected to bursts of heat lasting on the order of one second in duration using a laser system to simulate the explosive environment from an agent defeat weapon...

  13. Characterization of the Surface Morphology of Bacillus Spores by Atomic Force Microscopy

    National Research Council Canada - National Science Library

    Zolock, Ruth

    2002-01-01

    The surface morphology of Bacillus spores was resolved by atomic force microscopy in order to determine if characteristic surface features could be used to distinguish between closely related species...

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

    CSIR Research Space (South Africa)

    Vismer, HF

    1995-08-01

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

  15. 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

  16. Annual Pollen and Spore Sedimentation Record off South Java in Indian Ocean

    Science.gov (United States)

    Poliakova, Anastasia; Rixen, Tim; Behling, Hermann

    2013-04-01

    Marine sediments are excellent archives recording environmental changes in the ocean as well as on land. In particular pollen and spores preserved in marine sediments could provide crucial information on land use and climate changes in the past. However, in order to better understand and interpret sedimentary records studies on modern pollen and spore transportation and sedimentation is needed. Therefore a sediment trap was deployed for about one year (December 2001 - November 2002) off South Java in the Indian Ocean at a water-depth of about 2000 m. Abundance and taxa composition of pollen and spores collected by the sediment trap reflect climatic (monsoon conditions and ocean currents) as well as biological (flowering periods, migration ability of pollen) factors controlling their sedimentation. Pollen and, at a lower rate, pteridophyta spore concentration tends to increase during non-monsoon period.

  17. 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.

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

    Digital Repository Service at National Institute of Oceanography (India)

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

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

  19. 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...

  20. Troubleshooting Requests e-mail Confirmation

    CERN Document Server

    TS Department

    2004-01-01

    In an ongoing effort to improve quality of the repair requests, a new e-mail confirmation automatic system will be implemented starting from the 21st October. All repair requests transmitted to the TCR (72201) or the FM Helpdesk (77777) will be confirmed in an e-mail to the requestor, provided that the latter has a valid e-mail address in the HR database. The e-mail will contain a reference number, a brief description of the problem, the location and a contact where more information can be obtained. A second e-mail will be sent when the processing of the repair request is finished. We hope that this initiative will improve the transparency and quality of our service. Helpdesk Troubleshooting Requests (reminder) We remind you that all the repair requests and other communication concerning the CERN machine buildings have to be transmitted to the TCR via 72201, whereas the ones concerning tertiary buildings are handled directly by the FM helpdesk under the phone number 77777, i.e. problems on systems and equ...

  1. [Cutaneous gnathostomiasis, first confirmed case in Colombia].

    Science.gov (United States)

    Jurado, Leonardo F; Palacios, Diana M; López, Rocío; Baldión, Margarita; Matijasevic, Eugenio

    2015-01-01

    Gnathostomiasis is a parasitic zoonosis caused by some species of helminthes belonging to the genus Gnathostoma . It has a wide clinical presentation and its diagnosis is a challenge. Tropical and subtropical countries are endemic, and its transmission is associated with eating raw or undercooked meat from fresh water animals. Increasing global tourism and consuming exotic foods have produced a noticeable rise in cases of the disease in the last decades. However, in our country, there has not been any confirmed case of gnathostomiasis previously reported. We present the case of a 63-year-old Colombian man with an international travel history, who presented with gastrointestinal symptoms. During the hospital stay, he developed a cutaneous lesion on the upper right abdominal quadrant, where later, a larva was found. A morphological study allowed us to identify it as Gnathostoma spinigerum . As such, this is the first report of an imported case of gnathostomiasis confirmed in Colombia. This article describes the principles, etiology, pathogenic cycle and treatment of this disease with special considerations to our patient´s particular features.

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

    Science.gov (United States)

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

    2016-04-01

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

  3. Ganoderin A, a novel 9,11-secosterol from Ganoderma lucidum spores oil.

    Science.gov (United States)

    Ge, Fa-Huan; Duan, Ming-Hui; Li, Jing; Shi, Qing-Long

    2017-12-01

    In this study, four sterols were isolated from the Ganoderma lucidum spores oil obtained via supercritical CO 2 extraction. Four chemical constituents were ganoderin A (1), chaxine B (2), ergosterol, (3) and stellasterol (4). All the separated ingredients were characterized using spectral data interpretation and by comparing with reported data. Noticeably, stellasterol and chaxine B were both firstly isolated from Ganoderma lucidum spores oil and ganoderin A was shown to bear an unprecedented skeleton.

  4. 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

  5. [Analysis of bacterial colonization associated with Gigaspora margarita spores by green fluorescence protein (GFP) marked technology].

    Science.gov (United States)

    Long, Liangkun; Yao, Qing; Ai, Yuncan; Zhu, Honghui

    2009-05-01

    We analyzed bacterial colonization associated with spores of arbuscular mycorrhizal fungi (AMF) Gigaspora margarita, to indicate their ecological niche, and to provide information for further researches on their populations or functions. Six bacteria strains (Peanibacillus sp. M060106-1, Peanibacillus sp. M061122-2, Peanibacillus sp. M061122-6, Bacillus sp. M061122-4, Bacillus sp. M061122-10 and Brevibacillus sp. M061122-12) isolated from G. margarita spores were tagged with green fluorescence protein (GFP) using the carrier plasmid pNF8 (gfp-mut1). We analyzed the ecological niche and population dynamics of tagged strains on G. margarita under different conditions by using fluorescent microscope and/or plate counts. Four strains (M060106-1, M061122-6, M061122-10 and M061122-12) were tagged with GFP, showing high plasmid stability. These tagged strains possessed the basic characteristics identical to their original strains and, hence, were fit for short-term study of environmental colonization. All four GFP-tagged strains colonized the spore wall of G. margarita, and M061122-6 and M061122-12 further colonized the fungal hyphae. Under different pH conditions,the population dynamic of each GFP-tagged strain on the spores showed the same trend, i.e. first increased and then decreased, and the effects on the population size varied with different pH value. GFP-tagged strains colonized the spores of low viability more easily than those of high viability, and the population dynamic on the spores of high viability was different for each tagged strain. The isolated bacteria associated with G. margarita spores can re-colonize the fungal spores, whereas their colonizing ability depends on their characteristics and environmental factors. These data contributes to the further understanding of populations and functions of AMF-associated bacteria.

  6. Mechanism and site of inhibition of Bacillus cereus spore outgrowth by nitrosothiols

    International Nuclear Information System (INIS)

    Morris, S.L.

    1982-01-01

    Structure vs. activity studies demonstrate that nitrosothiols inhibit outgrowth of B. cereus spores by reversible covalent bond formation with sensitive spore components. Kinetic studies of the binding of nitrosothiols and iodoacetate, a known sulfhydryl reagent, show that they complete for the same spore sites. Since two other nitrite derivatives, the Perigo factor and the transferrin inhibitor, interfere with iodoacetate label uptake in a kinetically similar fashion, all of these compounds may inhibit spore outgrowth by interacting with the same spore thiol groups. Disruption of spores which have been inhibited by radioactive iodoacetate demonstrates that much of the label is incorporated into a membrane-rich fraction that sediments as a single peak on a sucrose density gradient. SDS gel electrophoresis and autofluorography allows the identification of four intensely labelled proteins with molecular weights of 13,000, 28,000, 29,000, and 30,000. If the iodoacetate labelling is carried out in the presence of nitrosothiol, incorporation is greatly reduced into all components. When germinating spores are labelled with succinate or the lactose analog, o-nitrophenylgalactopyranoside, a significant reduction in the amount of label bound is also observed suggesting that two iodoacetate-reactive sites may be the succinate and lactose permease systems. Severe decreases in the transport of succinate and lactose into iodoacetate and nitrosothiol inhibited spores further implicates a nitrosothiol (iodoacetate) permease interaction. Iodoacetate and nitrosothiols therefore may exert their inhibitory effects by interfering with critical membrane protein sulfhydryl groups, possibly by a a covalent modification mechanism. Some of these sensitive thiols may be involved in active transport processes

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2016-09-01

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

  8. The Role of Aquaporins in pH-Dependent Germination of Rhizopus delemar Spores.

    Directory of Open Access Journals (Sweden)

    Tidhar Turgeman

    Full Text Available Rhizopus delemar and associated species attack a wide range of fruit and vegetables after harvest. Host nutrients and acidic pH are required for optimal germination of R. delemar, and we studied how this process is triggered. Glucose induced spore swelling in an acidic environment, expressed by an up to 3-fold increase in spore diameter, whereas spore diameter was smaller in a neutral environment. When suspended in an acidic environment, the spores started to float, indicating a change in their density. Treatment of the spores with HgCl2, an aquaporin blocker, prevented floating and inhibited spore swelling and germ-tube emergence, indicating the importance of water uptake at the early stages of germination. Two putative candidate aquaporin-encoding genes-RdAQP1 and RdAQP2-were identified in the R. delemar genome. Both presented the conserved NPA motif and six-transmembrane domain topology. Expressing RdAQP1 and RdAQP2 in Arabidopsis protoplasts increased the cells' osmotic water permeability coefficient (Pf compared to controls, indicating their role as water channels. A decrease in R. delemar aquaporin activity with increasing external pH suggested pH regulation of these proteins. Substitution of two histidine (His residues, positioned on two loops facing the outer side of the cell, with alanine eliminated the pH sensing resulting in similar Pf values under acidic and basic conditions. Since hydration is critical for spore switching from the resting to activate state, we suggest that pH regulation of the aquaporins can regulate the initial phase of R. delemar spore germination, followed by germ-tube elongation and host-tissue infection.

  9. 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.

  10. Comparison of fungal spores concentrations measured with wideband integrated bioaerosol sensor and Hirst methodology

    Science.gov (United States)

    Fernández-Rodríguez, S.; Tormo-Molina, R.; Lemonis, N.; Clot, B.; O'Connor, D. J.; Sodeau, John R.

    2018-02-01

    The aim of this work was to provide both a comparison of traditional and novel methodologies for airborne spores detection (i.e. the Hirst Burkard trap and WIBS-4) and the first quantitative study of airborne fungal concentrations in Payerne (Western Switzerland) as well as their relation to meteorological parameters. From the traditional method -Hirst trap and microscope analysis-, sixty-three propagule types (spores, sporangia and hyphae) were identified and the average spore concentrations measured over the full period amounted to 4145 ± 263.0 spores/m3. Maximum values were reached on July 19th and on August 6th. Twenty-six spore types reached average levels above 10 spores/m3. Airborne fungal propagules in Payerne showed a clear seasonal pattern, increasing from low values in early spring to maxima in summer. Daily average concentrations above 5000 spores/m3 were almost constant in summer from mid-June onwards. Weather parameters showed a relevant role for determining the observed spore concentrations. Coniferous forest, dominant in the surroundings, may be a relevant source for airborne fungal propagules as their distribution and predominant wind directions are consistent with the origin. The comparison between the two methodologies used in this campaign showed remarkably consistent patterns throughout the campaign. A correlation coefficient of 0.9 (CI 0.76-0.96) was seen between the two over the time period for daily resolutions (Hirst trap and WIBS-4). This apparent co-linearity was seen to fall away once increased resolution was employed. However at higher resolutions upon removal of Cladosporium species from the total fungal concentrations (Hirst trap), an increased correlation coefficient was again noted between the two instruments (R = 0.81 with confidence intervals of 0.74 and 0.86).

  11. Measurements of DNA Damage and Repair in Bacillus anthracis Sterne Spores by UV Radiation

    Science.gov (United States)

    2014-09-18

    contains an antibiotic resistance gene, which gives the cell an advantage when it keeps the plasmid. Studies will be conducted on Ba spores and...are extremely resistant to the environment and can survive in soil for decades [2]. Since the spore contains few energy compounds such as ATP and...glucose, and 10mM MgCl2. The solution was incubated for 1 hour so expression of the antibiotic resistance could occur prior to plating on selective

  12. The contribution of endogenous and exogenous effects to radiation-induced damage in the bacterial spore

    International Nuclear Information System (INIS)

    Jacobs, G.P.; Samuni, A.; Czapski, G.

    1985-01-01

    Radical scavengers such as polyethylene glycol 400 and 4000 and bovine albumin have been used to define the contribution of exogenous and endogenous effects to the gamma-radiation-induced damage in aqueous buffered suspensions of Bacillus pumilus spores. The results indicate that this damage in the bacterial spore is predominantly endogenous both in the presence of 1 atmosphere of oxygen, and in anoxia. (author)

  13. 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

  14. 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)

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

    Science.gov (United States)

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

    2016-04-01

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

  16. The Role of Aquaporins in pH-Dependent Germination of Rhizopus delemar Spores.

    Science.gov (United States)

    Turgeman, Tidhar; Shatil-Cohen, Arava; Moshelion, Menachem; Teper-Bamnolker, Paula; Skory, Christopher D; Lichter, Amnon; Eshel, Dani

    2016-01-01

    Rhizopus delemar and associated species attack a wide range of fruit and vegetables after harvest. Host nutrients and acidic pH are required for optimal germination of R. delemar, and we studied how this process is triggered. Glucose induced spore swelling in an acidic environment, expressed by an up to 3-fold increase in spore diameter, whereas spore diameter was smaller in a neutral environment. When suspended in an acidic environment, the spores started to float, indicating a change in their density. Treatment of the spores with HgCl2, an aquaporin blocker, prevented floating and inhibited spore swelling and germ-tube emergence, indicating the importance of water uptake at the early stages of germination. Two putative candidate aquaporin-encoding genes-RdAQP1 and RdAQP2-were identified in the R. delemar genome. Both presented the conserved NPA motif and six-transmembrane domain topology. Expressing RdAQP1 and RdAQP2 in Arabidopsis protoplasts increased the cells' osmotic water permeability coefficient (Pf) compared to controls, indicating their role as water channels. A decrease in R. delemar aquaporin activity with increasing external pH suggested pH regulation of these proteins. Substitution of two histidine (His) residues, positioned on two loops facing the outer side of the cell, with alanine eliminated the pH sensing resulting in similar Pf values under acidic and basic conditions. Since hydration is critical for spore switching from the resting to activate state, we suggest that pH regulation of the aquaporins can regulate the initial phase of R. delemar spore germination, followed by germ-tube elongation and host-tissue infection.

  17. The Role of Aquaporins in pH-Dependent Germination of Rhizopus delemar Spores

    Science.gov (United States)

    Turgeman, Tidhar; Shatil-Cohen, Arava; Moshelion, Menachem; Teper-Bamnolker, Paula; Skory, Christopher D.; Lichter, Amnon; Eshel, Dani

    2016-01-01

    Rhizopus delemar and associated species attack a wide range of fruit and vegetables after harvest. Host nutrients and acidic pH are required for optimal germination of R. delemar, and we studied how this process is triggered. Glucose induced spore swelling in an acidic environment, expressed by an up to 3-fold increase in spore diameter, whereas spore diameter was smaller in a neutral environment. When suspended in an acidic environment, the spores started to float, indicating a change in their density. Treatment of the spores with HgCl2, an aquaporin blocker, prevented floating and inhibited spore swelling and germ-tube emergence, indicating the importance of water uptake at the early stages of germination. Two putative candidate aquaporin-encoding genes—RdAQP1 and RdAQP2—were identified in the R. delemar genome. Both presented the conserved NPA motif and six-transmembrane domain topology. Expressing RdAQP1 and RdAQP2 in Arabidopsis protoplasts increased the cells' osmotic water permeability coefficient (Pf) compared to controls, indicating their role as water channels. A decrease in R. delemar aquaporin activity with increasing external pH suggested pH regulation of these proteins. Substitution of two histidine (His) residues, positioned on two loops facing the outer side of the cell, with alanine eliminated the pH sensing resulting in similar Pf values under acidic and basic conditions. Since hydration is critical for spore switching from the resting to activate state, we suggest that pH regulation of the aquaporins can regulate the initial phase of R. delemar spore germination, followed by germ-tube elongation and host-tissue infection. PMID:26959825

  18. Spores of Clostridium difficile clinical isolates display a diverse germination response to bile salts.

    Directory of Open Access Journals (Sweden)

    Daniela Heeg

    Full Text Available Clostridium difficile spores play a pivotal role in the transmission of infectious diarrhoea, but in order to cause disease spores must complete germination and return to vegetative cell growth. While the mechanisms of spore germination are well understood in Bacillus, knowledge of C. difficile germination remains limited. Previous studies have shown that bile salts and amino acids play an important role in regulating the germination response of C. difficile spores. Taurocholate, in combination with glycine, can stimulate germination, whereas chenodeoxycholate has been shown to inhibit spore germination in a C. difficile clinical isolate. Our recent studies of C. difficile sporulation characteristics have since pointed to substantial diversity among different clinical isolates. Consequently, in this study we investigated how the germination characteristics of different C. difficile isolates vary in response to bile salts. By analysing 29 isolates, including 16 belonging to the BI/NAP1/027 type, we show that considerable diversity exists in both the rate and extent of C. difficile germination in response to rich medium containing both taurocholate and glycine. Strikingly, we also show that although a potent inhibitor of germination for some isolates, chenodeoxycholate does not inhibit the germination, or outgrowth, of all C. difficile strains. Finally, we provide evidence that components of rich media may induce the germination of C. difficile spores, even in the absence of taurocholate. Taken together, these data suggest that the mechanisms of C. difficile spore germination in response to bile salts are complex and require further study. Furthermore, we stress the importance of studying multiple isolates in the future when analysing the nutrients or chemicals that either stimulate or inhibit C. difficile spore germination.

  19. Mutation induction in spores of Bacillus subtilis by accelerated very heavy ions

    International Nuclear Information System (INIS)

    Baltschukat, K.; Horneck, G.; Buecker, H.; Facius, R.; Schaefer, M.

    1986-01-01

    Mutation induction (resistance to sodium azide) in spores of Bacillus subtilis was investigated after irradiation with heavy ions from Neon to Uranium with specific particle energies between 0.17 and 18.6 MeV/u. A strong dependence of the mutation induction cross section on particle charge and energy was observed. From the results it was concluded that mutation induction in bacterial spores by very heavy ions is mainly caused by secondary electrons. (orig.)

  20. 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.

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

    Science.gov (United States)

    Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka

    2011-03-01

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

  2. [Study the rudimentary immunoregulatory mechanisms of Ganoderma Spore oil on immunocompromized mice].

    Science.gov (United States)

    Yi, Youjin; Hu, Shun; Xiong, Xingyao; Liu, Dongbo; Zhong, Yingli

    2012-09-01

    To study the rudimentary immunoregulatory mechanisms of Ganoderma spore oil on immunocompromized mice model. Thrity KM mice were randomly selected and assigned into three groups (ten animals per group): the model control group, Ganoderma Lucidum spores oil group and the normal control group. The model control group and Ganoderma Lucidum spores oil group were injected intraperitoneally with cyclophosphamide at 40 mg x kg(-1) d to generate a immunocompromized mice model. The normal control group were administered with 0.9% NaCl solution 0.1 ml/10 g BW as placebo. All agents were given orally once a day, given for consecutive 30 days, Ganoderma Lucidum spores oil group 150 mg/kg, the others given maize 0.1 ml/10 g BW. The serum TNF-alpha , IFN-gamma content of the mice through ELISA kit and the expression levels of IL-2, IL-10, IL-12, IL-4, IFN-gamma, TNF-alpha mRNA in mouse spleen and thymus were examined by RT-PCR to rudimentary study its immunoregulatory mechanisms. Ganoderma spore oil can significantly increased the content of TNF-alpha and IFN-gamma in the serum and the expression levels of IL-2, IL-10, IL-12, IL-4, IFN-gamma, TNF-alpha mRNA in spleen and thymus, with obvious difference from the model control (P Ganoderma spore oil can be able to improve the above cytokine ion expression to immunoregulate the immunocompromized mice.

  3. 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.

  4. Detection limit of Clostridium botulinum spores in dried mushroom samples sourced from China.

    Science.gov (United States)

    Malakar, Pradeep K; Plowman, June; Aldus, Clare F; Xing, Zengtao; Zhao, Yong; Peck, Michael W

    2013-08-16

    A survey of dried mushrooms (Lentinula edodes (Shiitake) and Auricularia auricula (Wood Ear)) sourced from China was carried out to determine the natural contamination of these mushrooms with spores of proteolytic Clostridium botulinum and non-proteolytic C. botulinum. The mushrooms were collected from supermarkets and retailers in 21 cities in China during October 2008. Spore loads of C. botulinum in mushrooms have a degree of uncertainty and variability and this study contributes valuable data for determining prevalence of spores of C. botulinum in mushrooms. An optimized detection protocol that combined selective enrichment culture with multiplex PCR was used to test for spores of proteolytic and non-proteolytic C. botulinum. Detection limits were calculated, using a maximum likelihood protocol, from mushroom samples inoculated with defined numbers of spores of proteolytic C. botulinum or non-proteolytic C. botulinum. Based on the maximum likelihood detection limit, it is estimated that dried mushroom A. auricula contained botulinum, and botulinum. Dried L. edodes contained botulinum and it was not possible to determine reliable detection limits for spores of non-proteolytic C. botulinum using the current detection protocol. © 2013 Elsevier B.V. All rights reserved.

  5. Detection of Bacillus anthracis spores from environmental water using bioluminescent reporter phage.

    Science.gov (United States)

    Nguyen, C; Makkar, R; Sharp, N J; Page, M A; Molineux, I J; Schofield, D A

    2017-11-01

    We investigated the ability of a temperate Bacillus anthracis reporter phage (Wβ::luxAB-2), which transduces bioluminescence to infected cells, to detect viable spores from deliberately contaminated environmental water samples. Environmental water was inoculated with spores and assayed with Wβ::luxAB-2. Bioluminescent signals directly correlated with input phage and spore concentrations. A limit of detection of 10 1 and 10 2 CFU per ml within 8 h was achieved from pond and lake water, respectively. Detection was greatly simplified by minimizing sample processing steps without spore extraction. The complex endogenous microbial flora and salt content of brackish water challenged the assay, extending the detection time to 12 h for a sensitivity of 10 2 CFU per ml. Phage-mediated bioluminescence was strictly dependent on bacterial physiology, being significantly reduced in mid/late log phase cells. This was shown to be due to an inability of the phage to adsorb. The reporter phage Wβ::luxAB-2 displays potential for simplified detection of viable spores from contaminated water samples within 12 h. A deliberate aerosol release of spores could lead to widespread contamination, leaving large areas uninhabitable until remediation. An essential requirement of this restoration process is the development of simplified detection assays in different environmental matrices. © 2017 The Society for Applied Microbiology.

  6. Starch in plasterboard sustains Streptomyces californicus growth and bioactivity of spores.

    Science.gov (United States)

    Murtoniemi, T; Keinänen, M M; Nevalainen, A; Hirvonen, M-R

    2003-01-01

    The effects of plasterboard composition on Streptomyces californicus growth and bioactivity of spores were studied. Streptomyces californicus was grown on 13 modified plasterboards under saturated humidity conditions. The total content of fatty acid methyl esters was used for quantifying S. californicus biomass, while the spore-induced cytotoxicity and production of nitric oxide (NO), tumour necrosis factor-alpha, and interleukine-6 (IL-6) in mouse macrophages was used to assess the bioactivity of spores. Removal of starch completely from the plasterboard or only from the core reduced significantly the biomass production and the biological activity of spores in comparison with reference board. The biocide added into the core or on the liner decreased the growth markedly and inhibited the sporulation totally. The biomass production correlated positively with the spore number, cytotoxicity, and production of NO and IL-6. Streptomyces californicus grew under nutrient limitation on all studied plasterboards. The starch is the major factor enabling S. californicus to grow and to produce biologically active metabolites on plasterboard. The composition of building material has an impact on microbial growth and bioactivity of spores which may be involved in complex mechanisms leading to respiratory symptoms in the occupants in moisture damaged buildings.

  7. [Irradiation effect of 60Co gamma ray on Aspergillus flavus spores in normal saline and maize].

    Science.gov (United States)

    Zhong, Kai; Gao, Xiang; Ji, Rong

    2011-05-01

    To observe irradiation effect of gamma ray on Aspergilllus flavus spores. Aspergillus flavus spores of different concentration were prepared on two media and were exposed to gamma ray. D10 values and theoretical minimal sterilizing doses were calculated by colony counting results. Actual minimal sterilizing doses were verified according to theoretical dose. D10 value and minimal sterilizing dose of high concentration spore in normal saline were 0.39 kGy and 2.5 kGy. Those of low concentration spore in normal saline were 0.47 kGy and 1.5 kGy. Dl0 value and minimal sterilizing dose of high concentration spore in maize were 0.67 kGy and 3.0 kGy. Those of low concentration spore in maize were 0.72 kGy and 2.0 kGy. 3.0 Gy may be suitable for grain processing to sterilize Aspergillus flavus.

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    Alba Adriana Vallejo-Cardona

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

  10. Evaluation of the allergenicity of tropical pollen and airborne spores in Singapore.

    Science.gov (United States)

    Chew, F T; Lim, S H; Shang, H S; Dahlia, M D; Goh, D Y; Lee, B W; Tan, H T; Tan, T K

    2000-04-01

    Sensitization to pollen and spores of the Southeast Asian tropical region is not well documented. This study evaluated the allergenicity of the tropical airspora in Singapore. On the basis of the results of an aerobiologic survey of the airspora profile of Singapore, crude extracts of 23 main spore (fungal and fern) and pollen types were prepared. A total of 231 patients with asthma and/or allergic rhinitis and 76 healthy controls were evaluated by skin prick test (SPT). Total and specific IgE levels were also quantified by the fluorescence allergosorbent test (FAST). All 23 allergenic extracts tested elicited positive SPT responses. Among the patients with atopic diseases, extracts of oil-palm pollen (Elaeis guineensis) were observed to have the highest frequency of positive reactions (40%), followed by extracts of resam-fern spores (Dicranopteris linearis) (34%) and sea-teak pollen (Podocarpus polystachyus) (33.8%). Fungal spores with the highest SPT responses were Curvularia spp. (26-32%) and Drechslera-like spores (31%). Positive responses to these extracts correlated with total serum IgE levels of the subjects and were significantly associated with the presence of atopic disease. We have documented sensitization to tropical pollen and spores in our population. Its association with atopy suggests that it has a role in allergic diseases in the tropics.

  11. Progress in Bacillus subtilis Spore Surface Display Technology towards Environment, Vaccine Development, and Biocatalysis.

    Science.gov (United States)

    Chen, Huayou; Ullah, Jawad; Jia, Jinru

    2017-01-01

    Spore surface display is the most desirable with enhanced effects, low cost, less time consuming and the most promising technology for environmental, medical, and industrial development. Spores have various applications in industry due to their ability to survive in harsh industrial processes including heat resistance, alkaline tolerance, chemical tolerance, easy recovery, and reusability. Yeast and bacteria, including gram-positive and -negative, are the most frequently used organisms for the display of various proteins (eukaryotic and prokaryotic), but unlike spores, they can rupture easily due to nutritive properties, susceptibility to heat, pH, and chemicals. Hence, spores are the best choice to avoid these problems, and they have various applications over nonspore formers due to amenability for laboratory purposes. Various strains of Clostridium and Bacillus are spore formers, but the most suitable choice for display is Bacillus subtilis because, according to the WHO, it is safe to humans and considered as "GRAS" (generally recognized as safe). This review focuses on the application of spore surface display towards industries, vaccine development, the environment, and peptide library construction, with cell surface display for enhanced protein expression and high enzymatic activity. Different vectors, coat proteins, and statistical analyses can be used for linker selection to obtain greater expression and high activity of the displayed protein. © 2017 S. Karger AG, Basel.

  12. Inactivation of dried bacteria and bacterial spores by means of gamma irradiation at high temperatures.

    Science.gov (United States)

    Emborg, C

    1974-05-01

    Dried preparations with Streptococcus faecium, strain A(2)1, and spores of Bacillus sphaericus, strain C(I)A, normally used for control of the microbiological efficiency of radiation sterilization plants and preparations with spores of Bacillus subtilis, normally used for control of sterilization by dry heat, formalin, and ethylene oxide, as well as similar preparations with Micrococcus radiodurans, strain R(1), and spores of Bacillus globigii (B. subtilis, var. niger) were gamma irradiated with dose rates from 16 to 70 krad/h at temperatures from 60 to 100 C. At 80 C the radiation response of the spore preparations was the same as at room temperature, whereas the radiation resistance of the preparations with the two vegetative strains was reduced. At 100 C the radiation response of preparations with spores of B. subtilis was unaffected by the high temperature, whereas at 16 and and 25 krad/h the radiation resistance of the radiation-resistant sporeformer B. sphaericus, strain C(I)A, was reduced to the level of radiation resistance of preparations with spores of B. subtilis. It is concluded that combinations of heat and gamma irradiation at the temperatures and dose rates tested may have very few practical applications in sterilization of medical equipment.

  13. The SpmA/B and DacF proteins of Clostridium perfringens play important roles in spore heat resistance.

    Science.gov (United States)

    Orsburn, Benjamin; Sucre, Katie; Popham, David L; Melville, Stephen B

    2009-02-01

    Strains of Clostridium perfringens that cause acute food poisoning have been shown to produce spores that are significantly more heat resistant than those of other strains. Previous studies demonstrated that the spore core density and the ratio of spore cortex peptidoglycan relative to the germ cell wall were factors that correlated with the heat resistance of a C. perfringens spore. To further evaluate these relationships, mutant strains of C. perfringens SM101 were constructed with null mutations in dacF, encoding a D,D-carboxypeptidase, and in the spmA-spmB operon, which is involved in spore core dehydration. The dacF mutant was shown to produce less spore cortex peptidoglycan and had a corresponding decrease in spore heat resistance. The spmA-spmB strain produced highly unstable spores with significantly lower core densities and increased heat sensitivity, which were easily destroyed during treatments affecting the spore coat layers. These results support the previous assertion that a threshold core density as well as a high ratio of cortex peptidoglycan relative to the germ cell wall contribute to the formation of a more heat-resistant spore in this species.

  14. Re-aerosolization of Bacillus thuringiensis spores from concrete and turf.

    Science.gov (United States)

    Bishop, A H; O'Sullivan, C M; Lane, A; Butler Ellis, M C; Sellors, W J

    2017-05-01

    Spores of Bacillus anthracis deposited on surfaces can become airborne again as a result of air currents and mechanical forces. As such, they are a potential source of infection by inhalation. Spores of Bacillus thuringiensis were used to quantify this phenomenon in a simulation of outdoor conditions. Concrete and turf surfaces were inoculated by aerosol to produce high spore densities (greater than 1 × 10 9  CFU per m 2 ) which were then subjected to the passage of air at 10 ms -1 with and without simulated walking. Re-aerosolized spores were sampled by wetted wall cyclone air samplers. The mean total re-aerosolization rate from concrete (m -2  min -1 ) was 1·16 × 10 -3 for wind alone and 3·2 × 10 -3 for wind and simulated walking while for turf the respective values were 2·7 × 10 -4 and 6·7 × 10 -4 . Following the malicious and/or accidental release of an aerosol of Bacillus anthracis spores, the immediate risk of human inhalation would decrease as the spores were deposited on surfaces or diluted by wind flow. There is, however, a concern that the deposited spores could become re-aerosolized and so present an ongoing hazard. Using an accurate simulant for B. anthracis spores a method is reported here that allowed the enumeration of re-aerosolized spores from concrete and turf by wind flow and footfall. Under the conditions used, the rates of re-aerosolization were low. These findings will need to be verified under real outdoor conditions before the true significance in terms of secondary exposure to pathogenic spores can be assessed. © 2017 Crown copyright. Letter of Applied Microbiology © 2017 The Society for Applied Microbiology. This article is published with the permission of the Controller of HMSO and the Queen's Printer for Scotland.

  15. 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

  16. Small acid-soluble spore proteins of Clostridium acetobutylicum are able to protect DNA in vitro and are specifically cleaved by germination protease GPR and spore protease YyaC.

    Science.gov (United States)

    Wetzel, Daniela; Fischer, Ralf-Jörg

    2015-11-01

    Small acid-soluble proteins (SASPs) play an important role in protection of DNA in dormant bacterial endospores against damage by heat, UV radiation or enzymic degradation. In the genome of the strict anaerobe Clostridium acetobutylicum, five genes encoding SASPs have been annotated and here a further sixth candidate is suggested. The ssp genes are expressed in parallel dependent upon Spo0A, a master regulator of sporulation. Analysis of the transcription start points revealed a σG or a σF consensus promoter upstream of each ssp gene, confirming a forespore-specific gene expression. SASPs were termed SspA (Cac2365), SspB (Cac1522), SspD (Cac1620), SspF (Cac2372), SspH (Cac1663) and Tlp (Cac1487). Here it is shown that with the exception of Tlp, every purified recombinant SASP is able to bind DNA in vitro thereby protecting it against enzymic degradation by DNase I. Moreover, SspB and SspD were specifically cleaved by the two germination-specific proteases GPR (Cac1275) and YyaC (Cac2857), which were overexpressed in Escherichia coli and activated by an autocleavage reaction. Thus, for the first time to our knowledge, GPR-like activity and SASP specificity could be demonstrated for a YyaC-like protein. Collectively, the results assign SspA, SspB, SspD, SspF and SspH of C. acetobutylicum as members of α/β-type SASPs, whereas Tlp seems to be a non-DNA-binding spore protein of unknown function. In acetic acid-extracted proteins of dormant spores of C. acetobutylicum, SspA was identified almost exclusively, indicating its dominant biological role as a major α/β-type SASP in vivo.

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

    Directory of Open Access Journals (Sweden)

    Sirec Teja

    2012-08-01

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

  18. On the origin of heterogeneity in (preservation) resistance of Bacillus spores: Input for a ‘systems’ analysis approach of bacterial spore outgrowth

    NARCIS (Netherlands)

    Hornstra, L.M.; ter Beek, A.; Smelt, J.P.; Kallemeijn, W.W.; Brul, S.

    2009-01-01

    Bacterial spores are the ultimate (stress) ‘survival capsules’. They allow strains from the Bacillus and Clostridium species to survive harsh environmental conditions. In addition to the decision to enter sporulation the decision to do the reverse (germinate) is also a decisive event after which

  19. Synthesis of a Bacillus subtilis small, acid-soluble spore protein in Escherichia coli causes cell DNA to assume some characteristics of spore DNA

    International Nuclear Information System (INIS)

    Setlow, B.; Hand, A.R.; Setlow, P.

    1991-01-01

    Small, acid-soluble proteins (SASP) of the alpha/beta-type are associated with DNA in spores of Bacillus subtilis. Induction of synthesis of alpha/beta-type SASP in Escherichia coli resulted in rapid cessation of DNA synthesis, followed by a halt in RNA and then protein accumulation, although significant mRNA and protein synthesis continued. There was a significant loss in viability associated with SASP synthesis in E. coli: recA+ cells became extremely long filaments, whereas recA mutant cells became less filamentous. The nucleoids of cells with alpha/beta-type SASP were extremely condensed, as viewed in both light and electron microscopes, and immunoelectron microscopy showed that the alpha/beta-type SASP were associated with the cell DNA. Induction of alpha/beta-type SASP synthesis in E. coli increased the negative superhelical density of plasmid DNA by approximately 20%; UV irradiation of E. coli with alpha/beta-type SASP gave reduced yields of thymine dimers but significant amounts of the spore photoproduct. These changes in E. coli DNA topology and photochemistry due to alpha/beta-type SASP are similar to the effects of alpha/beta-type SASP on the DNA in Bacillus spores, further suggesting that alpha/beta-type SASP are a major factor determining DNA properties in bacterial spores

  20. Confirming a case of ball lightening production

    Energy Technology Data Exchange (ETDEWEB)

    Balyberdin, V.V.

    1975-01-01

    Model confirmation of ball lightening production was carried out on a three-cascade unit having a working voltage of 150 kV in an air atmosphere at a pressure of 730 to 750 mg Hg std. with C-shaped bending of the lightening drawoff. The electrical discharge parameters were held constant as follows: maximum current amplitude of 7 kA, oscillatory period of the quenched discharge of 6.5 x 10/sup -6/ sec, and energy used up in the unit approximately 2.2 k joules. Moving pictures are presented of the generation of one long luminous bundle displaced from a solid with a velocity up to 240 m/sec, whose luminosity was observed over a period of approximately 3 x 10/sup -3/ sec. Analysis of the experimental results showed that the appearance of plasma bundles is associated with the development of turbulent flows of a low-temperature plasma near the tip of a solid. (SJR)

  1. Confirmation of Kepler Planet Candidates around Giants

    Science.gov (United States)

    Sato, Bun-ei

    2014-01-01

    Detecting planetary transits is extremely valuable not only because it makes an independent confirmation of a planet from Doppler method but also because the photometric transits yield unambiguous information on planet masses and radii, and thus mean density and interior structure, and obliquity of planetary orbits. Planets around giants and subgiants have been intensively surveyed over the decade by precise radial velocity (RV) measurements, mainly from the viewpoint of planet searches around intermediate-mass (1.5-5M⊙) stars. The planets show remarkable properties in their masses and orbital parameters. Unfortunately, however, it is quite difficult to detect transiting ones around such evolved stars because of the large sizes of the host stars. Therefore, our understanding of properties of planets around intermediate-mass stars are still far behind from those around solar-like stars. In S13B, we made the first RV follow-up observations with HDS for the transiting planet candidates around giants found by Kepler space telescope, and identified promising candidates of true transiting planets. Here we propose to take additional RV data for them with HDS in order to make sure that the RV periodicity is the same as the transit one.

  2. Web-Based Honorarium Confirmation System Prototype

    Science.gov (United States)

    Wisswani, N. W.; Catur Bawa, I. G. N. B.

    2018-01-01

    Improving services in academic environment can be applied by regulating salary payment process for all employees. As a form of control to maintain financial transparency, employees should have information concerning salary payment process. Currently, notification process of committee honorarium will be accepted by the employees in a manual manner. The salary will be received by the employee bank account and to know its details, they should go to the accounting unit to find out further information. Though there are some employees entering the accounting unit, they still find difficulty to obtain information about detailed honor information that they received in their accounts. This can be caused by many data collected and to be managed. Based on this issue, this research will design a prototype of web-based system for accounting unit system in order to provide detailed financial transaction confirmation to employee bank accounts that have been informed through mobile banking system. This prototype will be developed with Waterfall method through testing on final users after it is developed through PHP program with MySQL as DBMS

  3. Confirmation of ETI: initial organizational response

    Science.gov (United States)

    Harrison, Albert A.

    2003-08-01

    Perhaps the most crucial responses to the confirmation of extraterrestrial intelligence will come from organizations, rather than from individual people. Among the key organizations that will help shape humanity's response are political institutions such as the US Congress, administrative bodies such as the US Department of State, security agencies, the military, professional societies, and the media. Although popular culture and individual beliefs will affect organizational performance, organizational reactions will depend also on organizational cultures and traditions, administrative structures, communication patterns, decision-making processes, and the actions of other organizations. Prompt and effective responses may be blocked by sociopolitical constraints, jurisdictional disputes, cumbersome structures and procedures, stresses that frequently slow and distort information processing, and potentially counterproductive efforts to maintain positive organizational images. Efforts undertaken by governmental agencies will be hampered by public perceptions of low credibility. Foresight and advance preparation are among the steps that organizations may take to prepare for contact, but conservative values, skepticism towards SETI, and competing organizational priorities make serious preparation unlikely.

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

    DEFF Research Database (Denmark)

    Skottrup, Peter; Hearty, Stephen; Frøkiær, Hanne

    This study describes a biosensing principle for detection of fungal spores using surface plasmon resonance (SPR). The approach involves the use of a monoclonal antibody (mab) and a SPR sensor for label-free detection of the model organism Puccinia striiformis f.sp. tritici (Pst) a biotrophic fungus...... causing wheat yellow rust. We have developed mabs towards intact whole spores and used a subtractive inhibition format for detection of spores in solution. The antibody was incubated with different spore concentrations and the remaining free antibody was quantified using a BIAcore® 3000 sensor. Decreasing...... binding of mab to the sensor surface was observed as the Pst urediniospore concentration was increased. The detection range for the assay was 1.7 x 106 – 5.3 x 104 spores/ml. This study describes the first use of SPR for detection of fungal spores and the generic principle has the potential to be used...

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

    DEFF Research Database (Denmark)

    Skottrup, Peter Durand; Hearty, Stephen; Frøkiær, Hanne

    This study describes a biosensing principle for detection of fungal spores using surface plasmon resonance (SPR). The approach involves the use of a monoclonal antibody (mab) and a SPR sensor for label-free detection of the model organism Puccinia striiformis f.sp. tritici (Pst) a biotrophic fungus...... causing wheat yellow rust. We have developed mabs towards intact whole spores and used a subtractive inhibition format for detection of spores in solution. The antibody was incubated with different spore concentrations and the remaining free antibody was quantified using a BIAcore® 3000 sensor....... Decreasing binding of mab to the sensor surface was observed as the Pst urediniospore concentration was increased. The detection range for the assay was 1.7 x 106 – 5.3 x 104 spores/ml. This study describes the first use of SPR for detection of fungal spores and the generic principle has the potential...

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

    DEFF Research Database (Denmark)

    Skottrup, Peter; Hearty, Stephen; Frøkiær, Hanne

    2005-01-01

    This study describes a biosensing principle for detection of fungal spores using surface plasmon resonance (SPR). The approach involves the use of a monoclonal antibody (mab) and a SPR sensor for label-free detection of the model organism Puccinia striiformis f.sp. tritici (Pst) a biotrophic fungus...... causing wheat yellow rust. We have developed mabs towards intact whole spores and used a subtractive inhibition format for detection of spores in solution. The antibody was incubated with different spore concentrations and the remaining free antibody was quantified using a BIAcore® 3000 sensor. Decreasing...... binding of mab to the sensor surface was observed as the Pst urediniospore concentration was increased. The detection range for the assay was 1.7 x 106 – 5.3 x 104 spores/ml. This study describes the first use of SPR for detection of fungal spores and the generic principle has the potential to be used...

  7. Most Probable Number Rapid Viability PCR Method to Detect Viable Spores of Bacillus anthracis in Swab Samples

    Energy Technology Data Exchange (ETDEWEB)

    Letant, S E; Kane, S R; Murphy, G A; Alfaro, T M; Hodges, L; Rose, L; Raber, E

    2008-05-30

    This note presents a comparison of Most-Probable-Number Rapid Viability (MPN-RV) PCR and traditional culture methods for the quantification of Bacillus anthracis Sterne spores in macrofoam swabs generated by the Centers for Disease Control and Prevention (CDC) for a multi-center validation study aimed at testing environmental swab processing methods for recovery, detection, and quantification of viable B. anthracis spores from surfaces. Results show that spore numbers provided by the MPN RV-PCR method were in statistical agreement with the CDC conventional culture method for all three levels of spores tested (10{sup 4}, 10{sup 2}, and 10 spores) even in the presence of dirt. In addition to detecting low levels of spores in environmental conditions, the MPN RV-PCR method is specific, and compatible with automated high-throughput sample processing and analysis protocols.

  8. 'Omics' for microbial food stability: Proteomics for the development of predictive models for bacterial spore stress survival and outgrowth.

    Science.gov (United States)

    Abhyankar, Wishwas; Stelder, Sacha; de Koning, Leo; de Koster, Chris; Brul, Stanley

    2017-01-02

    Bacterial spores are ubiquitous in nature. They are stress resistant entities that are a concern to microbiological food stability due to their environmental stress resistance. In addition germinating and outgrowing spores at undesired times and places pose a significant health burden. The challenge is amplified due to the heterogeneous germination and outgrowth behaviour of isogenic spore populations. We discuss the role of different 'omics' techniques, proteomics in particular, to study spore biology in detail. With examples, the use of label-based and label-free quantitative proteomics approaches in understanding the spore physiology is demonstrated. Also the need of genomics, single cell analyses and analysis of cellular physiology is discussed briefly. Certainly accurate comprehensive data obtained from omics methods and molecular physiology will underpin the development of robust molecular models of bacterial spore germination and outgrowth. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Changes in ultraviolet resistance and photoproduct formation as early events in spore germination of Bacillus cereus T

    International Nuclear Information System (INIS)

    Irie, R.

    1978-01-01

    In order to determine the timing of the change in the state of DNA in bacterial spores during the course of germination, L-alanine-induced germination of Bacillus cereus spores was interrupted by 0.3M CaCl 2 as an inhibitor, and the resulting semi-refractive spores (spores at the end of the first phase of germination) were examined for UV-resistance and photoproduct formation. Upon UV-irradiation, these spores, still having a semi-refractile core as observed under a phase-contrast microscope, gave rise to mainly the cyclobutane-type thymine dimer. It was concluded that change in the stats of the spore DNA occurs early in the process of germination, i.e. before the refractility of the core is lost. It was also found that CaCl 2 markedly prolonged the duration of the transient UV-resistant stage. (author)

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2016-09-30

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

  11. Inhibition of Bacillus licheniformis spore growth in milk by nisin, monolaurin, and pH combinations.

    Science.gov (United States)

    Mansour, M; Amri, D; Bouttefroy, A; Linder, M; Milliere, J B

    1999-02-01

    The effects of nisin and monolaurin, alone and in combination, were investigated on Bacillus licheniformis spores in milk at 37 degrees C. In the absence of inhibitors, germinated spores developed into growing vegetative cells and started sporulation at the end of the exponential phase. In the presence of nisin (25 IU ml-1), spore outgrowth was inhibited (4 log10 reduction at 10 h). Regrowth appeared between 10 and 24 h and reached a high population level (1.25 x 10(8) cfu ml-1) after 7 d. Monolaurin (250 micrograms ml-1) had a bacteriostatic effect during the first 10 h but thereafter, regrowth occurred slowly with a population level after 7 d (4 x 10(5) cfu ml-1) lower than that of nisin. Different combined effects of nisin (between 0 and 42 IU ml-1), monolaurin (ranging from 0 to 300 micrograms ml-1), pH values (between 5.0 and 7.0) and spore loads (10(3), 10(4), 10(5) spores ml-1) were investigated using a Doehlert matrix in order to study the main effects of these factors and the different interactions. Results were analysed using the Response Surface Methodology (RSM) and indicated that nisin and monolaurin had no action on spores before germination; only pH values had a significant effect (P monolaurin (100 micrograms ml-1) in combination acted synergistically on outgrown spores and vegetative cells, showing total inhibition at pH 6.0, without regrowth, within 7 d at 37 degrees C.

  12. 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.

  13. High fungal spore burden with predominance of Aspergillus in hospital air of a tertiary care hospital in Chandigarh

    Directory of Open Access Journals (Sweden)

    S M Rudramurthy

    2016-01-01

    Full Text Available The prevalence of fungal spores in the hospital air is essential to understand the hospital-acquired fungal infections. Air conditioners (ACs used in hospitals may either reduce spores in air or be colonised by fungi and aid in its dissemination. The present study was conducted to assess the fungal spore burden in AC and non-AC areas. We found a high fungal spore count in air irrespective of whether the area was AC or non-AC. The most predominant species isolated were Aspergillus flavus and Aspergillus fumigatus. Such high concentrations of pathogenic fungi in air may predispose individuals to develop disease.

  14. Spatial and temporal distribution of Alternaria spores in the Iberian Peninsula atmosphere, and meteorological relationships: 1993-2009

    Science.gov (United States)

    Aira, María-Jesús; Rodríguez-Rajo, Francisco-Javier; Fernández-González, María; Seijo, Carmen; Elvira-Rendueles, Belén; Abreu, Ilda; Gutiérrez-Bustillo, Montserrat; Pérez-Sánchez, Elena; Oliveira, Manuela; Recio, Marta; Tormo, Rafael; Morales, Julia

    2013-03-01

    This paper provides an updated of airborne Alternaria spore spatial and temporal distribution patterns in the Iberian Peninsula, using a common non-viable volumetric sampling method. The highest mean annual spore counts were recorded in Sevilla (39,418 spores), Mérida (33,744) and Málaga (12,947), while other sampling stations never exceeded 5,000. The same cities also recorded the highest mean daily spore counts (Sevilla 109 spores m-3; Mérida 53 spores m-3 and Málaga 35 spores m-3) and the highest number of days on which counts exceeded the threshold levels required to trigger allergy symptoms (Sevilla 38 % and Mérida 30 % of days). Analysis of annual spore distribution patterns revealed either one or two peaks, depending on the location and prevailing climate of sampling stations. For all stations, average temperature was the weather parameter displaying the strongest positive correlation with airborne spore counts, whilst negative correlations were found for rainfall and relative humidity.

  15. Analysis of the predicting variables for daily and weekly fluctuations of two airborne fungal spores: Alternaria and Cladosporium

    Science.gov (United States)

    Recio, Marta; del Mar Trigo, María; Docampo, Silvia; Melgar, Marta; García-Sánchez, José; Bootello, Lourdes; Cabezudo, Baltasar

    2012-11-01

    Alternaria and Cladosporium are two fungal taxa whose spores (conidia) are included frequently in aerobiological studies of outdoor environments. Both spore types are present in the atmosphere of Malaga (Spain) throughout almost the entire year, although they reach their highest concentrations during spring and autumn. To establish predicting variables for daily and weekly fluctuations, Spearman's correlations and stepwise multiple regressions between spore concentrations (measured using a volumetric 7-day recorder) and meteorological variables were made with results obtained for both spore types in 1996 and 1997. Correlations and regressions were also made between the different taxa and their concentrations in different years. Significant and positive correlation coefficients were always obtained between spore concentrations of both taxa, followed by temperature, their concentrations in different years, sunshine hours and relative humidity (this last in a negative sense). For the two spore types we obtained higher correlation and regression coefficients using weekly data. We showed different regression models using weekly values. From the results and a practical point of view, it was concluded that weekly values of the atmospheric concentration of Alternaria spores can be predicted from the maximum temperature expected and its concentrations in the years sampled. As regards the atmospheric concentration of Cladoposrium spores, the weekly values can be predicted based on the concentration of Alternaria spores, thus saving the time and effort that would otherwise be employed in counting them by optical microscopy.

  16. Manned in Situ Confirmation of Lunar Ice

    Science.gov (United States)

    Gerené, S. P. B.; Hummeling, R. W. J.; Ockels, W. J.

    A study is performed to investigate the feasibility of a manned expedition to the Moon using the European Ariane-5 launcher. The primary objective of this lunar mission is to confirm the presence of water at the South-Pole craters. It is believed that these permanently shadowed craters contain water in the form of ice. Secondary objective is to perform lunar surface science and making a first step towards a lunar outpost. Early results show that a minimum of two Ariane-5 launches is required. In this `two Ariane' scenario the first launch will bring a Lunar Landing Vehicle (LLV) into low lunar orbit. The second will launch two astronauts in a Crew Transfer Vehicle into a rendez- vous trajectory with the LLV. Arrived at the Moon, the astronauts will enter the LLV, undock from the CTV and land at the designated site located near the rim of the South-Pole Shackleton crater. The transfer strategy for both spacecraft will be the so-called direct transfer, taking about four days. At arrival the LLV will start mapping the landing site at a ground resolution of one meter. As a consequence of the polar orbit, the CTV has to arrive fourteen days later and surface operations can take about twelve days, accumulating in a total mission-duration of 36 days. 32 days for the CTV and 22 days for the LLV. In case a `two Ariane' flight does not posses sufficient capabilities also a `three Ariane' scenario is developed, in which the LLV is split-up into two stages and launched separately. These two will dock at the Moon forming a descent stage and an ascent stage. The third launch will be a CTV. During surface operations, astronauts will set up a solar power unit, install the sample retrieval system and carry out surface science. Samples of the crater floor will be retrieved by means of a probe or robot guided along a cable suspended over the crater rim. Also, this paper shows the way in which European astronauts can be brought to the Moon for other future missions, like the

  17. Development of method for evaluating cell hardness and correlation between bacterial spore hardness and durability

    Science.gov (United States)

    2012-01-01

    Background Despite the availability of conventional devices for making single-cell manipulations, determining the hardness of a single cell remains difficult. Here, we consider the cell to be a linear elastic body and apply Young’s modulus (modulus of elasticity), which is defined as the ratio of the repulsive force (stress) in response to the applied strain. In this new method, a scanning probe microscope (SPM) is operated with a cantilever in the “contact-and-push” mode, and the cantilever is applied to the cell surface over a set distance (applied strain). Results We determined the hardness of the following bacterial cells: Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and five Bacillus spp. In log phase, these strains had a similar Young’s modulus, but Bacillus spp. spores were significantly harder than the corresponding vegetative cells. There was a positive, linear correlation between the hardness of bacterial spores and heat or ultraviolet (UV) resistance. Conclusions Using this technique, the hardness of a single vegetative bacterial cell or spore could be determined based on Young’s modulus. As an application of this technique, we demonstrated that the hardness of individual bacterial spores was directly proportional to heat and UV resistance, which are the conventional measures of physical durability. This technique allows the rapid and direct determination of spore durability and provides a valuable and innovative method for the evaluation of physical properties in the field of microbiology. PMID:22676476

  18. Isolation and analysis of bacteria associated with spores of Gigaspora margarita.

    Science.gov (United States)

    Cruz, A F; Horii, S; Ochiai, S; Yasuda, A; Ishii, T

    2008-06-01

    The aim of this work was to observe bacteria associated with the spores of Gigaspora margarita, an arbuscular mycorrhizal fungus (AMF). First, a direct analysis of DNA from sterilized spores indicated the bacteria belonging to the genus Janthinobacterium. In the second assay, two bacterial strains were isolated by osmosis from protoplasts, which were derived from spores by using two particular enzymes: lysing enzymes and yatalase. After isolation, cultivation and identification by their DNA as performed in the first experiment, the species with the closest relation were Janthinobacterium lividum (KCIGM01) and Paenibacillus polymyxa (KCIGM04) isolated with lysing enzymes and yatalase respectively. Morphologically, J. lividum was Gram negative and oval, while P. polymyxa was also oval, but Gram positive. Both strains had antagonistic effects to the pathogenic fungi Rosellimia necatrix, Pythium ultimum, Fusarium oxysporum and Rhizoctonia solani. In particular, J. lividum was much stronger in this role. However, in phosphorus (P) solubilization P. polymyxa functioned better than J. lividum. This experiment had revealed two new bacteria species (P. polymyxa and J. lividum), associated with AMF spores, which functioned to suppress diseases and to solubilize P. AMF spores could be a useful source for bacterial antagonists to soil-borne diseases and P solubilization.

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

    Directory of Open Access Journals (Sweden)

    Novaković Milana S.

    2013-01-01

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

  20. 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.

  1. Inactivation of Bacillus subtilis spores by combined pulsed light and thermal treatments.

    Science.gov (United States)

    Artíguez, Mari Luz; Martínez de Marañón, Iñigo

    2015-12-02

    The combined effect of pulsed light (PL) and heat processing was evaluated on the inactivation of Bacillus subtilis spores. Those processes were applied separately and the time between both treatments was modified to evaluate whether the effect of the first treatment is maintained for a long time. B. subtilis spores subjected to sublethal pre-treatments were more sensitive to subsequent treatments (PL or thermal treatments) than untreated spores. Heating followed by PL was the most effective combination in reducing B. subtilis counts. Bacterial spores remained sensitized to subsequent treatment for at least 24 h of storage in water, whatever the temperature was (4 or 30°C). Sensitivity of B. subtilis cells to PL or heat processing increased after germination in a nutrient broth, being equally sensitive from 3 to 24 h. Vegetative cells maintained their enhanced sensitivity to subsequent processing after spore germination. The results of this work demonstrate that the combination of heating and PL treatment is a promising preservation method for microbial inactivation. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. The relation between growth of four microbes on six different plasterboards and biological activity of spores.

    Science.gov (United States)

    Murtoniemi, T; Hirvonen, M-R; Nevalainen, A; Suutari, M

    2003-03-01

    Microbial growth on water-damaged building materials is commonly associated with adverse health effects in the occupants. We examined the growth of Stachybotrys chartarum, Aspergillus versicolor, Penicillium spinulosum, and Streptomyces californicus, isolated from water-damaged buildings, on six different brands of plasterboards. The microbial growth was compared with the biological activity of the spores, that is the potential to induce cytotoxicity and proinflammatory mediators in RAW264.7 macrophages. These results showed that the microbial growth on plasterboard depended on both the microbial strain and the brand of plasterboard used. The biological activity of spores appeared to be regulated by different growth conditions on plasterboards so that good microbial growth was associated with a low bioactivity of the spores, whereas the spores collected from plasterboard supporting only weak growth usually were biologically active. Cytotoxicity of either S. chartarum or A. versicolor did not correlate with any particular growth conditions or induced inflammatory responses. Instead, there were positive correlations between cytotoxicity and levels of induced proinflammatory cytokines for P. spinulosum and S. californicus. These data suggest that both the microbial growth on plasterboard and the resulting bioactivity of spores vary and might be affected by changing the growth conditions provided by the plasterboards.

  3. Development of method for evaluating cell hardness and correlation between bacterial spore hardness and durability

    Directory of Open Access Journals (Sweden)

    Nakanishi Koichi

    2012-06-01

    Full Text Available Abstract Background Despite the availability of conventional devices for making single-cell manipulations, determining the hardness of a single cell remains difficult. Here, we consider the cell to be a linear elastic body and apply Young’s modulus (modulus of elasticity, which is defined as the ratio of the repulsive force (stress in response to the applied strain. In this new method, a scanning probe microscope (SPM is operated with a cantilever in the “contact-and-push” mode, and the cantilever is applied to the cell surface over a set distance (applied strain. Results We determined the hardness of the following bacterial cells: Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and five Bacillus spp. In log phase, these strains had a similar Young’s modulus, but Bacillus spp. spores were significantly harder than the corresponding vegetative cells. There was a positive, linear correlation between the hardness of bacterial spores and heat or ultraviolet (UV resistance. Conclusions Using this technique, the hardness of a single vegetative bacterial cell or spore could be determined based on Young’s modulus. As an application of this technique, we demonstrated that the hardness of individual bacterial spores was directly proportional to heat and UV resistance, which are the conventional measures of physical durability. This technique allows the rapid and direct determination of spore durability and provides a valuable and innovative method for the evaluation of physical properties in the field of microbiology.

  4. 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.

  5. Crop harvest in Central Europe causes episodes of high airborne Alternaria spore concentrations in Copenhagen

    Science.gov (United States)

    Skjøth, C. A.; Sommer, J.; Frederiksen, L.; Gosewinkel Karlson, U.

    2012-06-01

    This study tests the hypothesis that Danish agricultural areas are the main source to airborne Alternaria spores in Copenhagen, Denmark. We suggest that the source to the overall load is mainly local, but with intermittent Long Distance Transport (LDT) from more remote agricultural areas. This hypothesis is supported by investigating a 10 yr bi-hourly record of Alternaria spores in the air from Copenhagen. This record shows 232 clinically relevant episodes with a distinct daily profile. The data analysis also revealed potential LDT episodes almost every year. A source map and analysis of atmospheric transport suggest that LDT always originates from the main agricultural areas in Central Europe. A dedicated emission study in cereal crops under harvest during 2010 also supports our hypothesis. The emission study showed that although the fields had been treated against fungal infections, harvesting still produced large amounts of airborne fungal spores. It is likely that such harvesting periods can cause clinically relevant levels of fungal spores in the atmosphere. Our findings suggest that crop harvest in Central Europe causes episodes of high airborne Alternaria spore concentrations in Copenhagen as well as other urban areas in this region. It is likely that such episodes could be simulated using atmospheric transport models.

  6. 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.

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

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    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.

  8. Development of a new technique using glass beads for dry dispersion of airborne fungal spores.

    Science.gov (United States)

    Shimasaki, Noriko; Okaue, Akira; Kikuno, Ritsuko; Okuda, Shunji; Abe, Keiko

    2015-01-01

    To evaluate the removal of airborne microbes by air cleaners, a technique for generating airborne fungal spores in the dry state in a test chamber (dry dispersion) become necessary. The Society of Indoor Environment Japan (SIEJ) published SIEJ Standard Method No. 20110001 (SIEJ standard),in which an aerial ultrasonic oscillator was used as the device for dry dispersion. However, a more versatile apparatus is also necessary from a practical point of view. Therefore, we developed a new device using glass beads for the dispersion. Glass beads and a fungal sheet containing spores of Wallemia sebi were set in a midget impinger, which was connected to a compressor and a compact test chamber (1 m(3)). Air was blown into the impinger from the compressor. The spores on the fungal sheet were released by impingement of the glass beads when the beads were induced to float by the air blown into the impinger, and the spores were introduced to the chamber by the airflow. This newly developed technique can be used in a compact chamber system and could be applicable as an improved method for generating airborne fungal spores in the dry state in the SIEJ standard.

  9. Mucorales spores induce a proinflammatory cytokine response in human mononuclear phagocytes and harbor no rodlet hydrophobins.

    Science.gov (United States)

    Wurster, Sebastian; Thielen, Vanessa; Weis, Philipp; Walther, Paul; Elias, Johannes; Waaga-Gasser, Ana Maria; Dragan, Mariola; Dandekar, Thomas; Einsele, Hermann; Löffler, Jürgen; Ullmann, Andrew J

    2017-11-17

    Mucormycoses are life-threatening infections in immunocompromised patients. This study characterizes the response of human mononuclear cells to different Mucorales and Ascomycota. PBMC, monocytes, and monocyte derived dendritic cells (moDCs) from healthy donors were stimulated with resting and germinated stages of Mucorales and Ascomycota. Cytokine response and expression of activation markers were studied. Both inactivated germ tubes and resting spores of Rhizopus arrhizus and other human pathogenic Mucorales species significantly stimulated mRNA synthesis and secretion of proinflammatory cytokines. Moreover, R. arrhizus spores induced the upregulation of co-stimulatory molecules on moDCs and a specific T-helper cell response. Removal of rodlet hydrophobins by hydrofluoric acid treatment of A. fumigatus conidia resulted in enhanced immunogenicity, whereas the cytokine response of PBMCs to dormant R. arrhizus spores was not influenced by hydrofluoric acid. Scanning electron micrographs of Mucorales spores did not exhibit any morphological correlates of rodlet hydrophobins. Taken together, this study revealed striking differences in the response of human mononuclear cells to resting stages of Ascomycota and Mucorales, which may be explained by absence of an immunoprotective hydrophobin layer in Mucorales spores.

  10. 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.

  11. Computer-assisted image processing to detect spores from the fungus Pandora neoaphidis.

    Science.gov (United States)

    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 slides, where autonomous image recording precedes computerised image analysis. The purpose of the present image analysis is to support human visual inspection of imagery data - not to replace it. The workflow has three components:•Preparation of slides for microscopy.•Image recording.•Computerised image processing where the initial part is, as usual, segmentation depending on the actual data product. Then comes identification of blobs, calculation of principal axes of blobs, symmetry operations and projection on a three parameter egg shape space.

  12. 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)

  13. Influence of industrial smoke on the germination spores of certain lichens

    Energy Technology Data Exchange (ETDEWEB)

    Kofler, L.; Jacquard, F.; Martin, J.F.

    1968-01-01

    Dust particles produced by calcium carbide and iron-alloy factories strongly inhibit the germination of the spores of Physcia pulverulenia. The spores of Xanthoria parietina and above ali of Lecanora hageni are much more resistant. This agrees with the distribution of these Lichens around the factories. The three species show the same scale of resistance with respect to town dust. The basicity of the particles plays certainly a part in the inhibition of spores, but this can not be the only active factor. Up to now attention has been drawn upon the damages caused by gaseous pollutants to Lichen gonidia. The present work shows that solid particles can sometimes be responsible for Lichen scarcity and that the Lichen fungus is also sensitive to certain pollutants. 9 references, 1 figure, 5 tables.

  14. Species Specific Bacterial Spore Detection Using Lateral-Flow Immunoassay with DPA-Triggered Tb Luminescence

    Science.gov (United States)

    Ponce, Adrian

    2003-01-01

    A method of detecting bacterial spores incorporates (1) A method of lateral-flow immunoassay in combination with (2) A method based on the luminescence of Tb3+ ions to which molecules of dipicolinic acid (DPA) released from the spores have become bound. The present combination of lateral-flow immunoassay and DPA-triggered Tb luminescence was developed as a superior alternative to a prior lateral-flow immunoassay method in which detection involves the visual observation and/or measurement of red light scattered from colloidal gold nanoparticles. The advantage of the present combination method is that it affords both (1) High selectivity for spores of the species of bacteria that one seeks to detect (a characteristic of lateral-flow immunoassay in general) and (2) Detection sensitivity much greater (by virtue of the use of DPA-triggered Tb luminescence instead of gold nanoparticles) than that of the prior lateral-flow immunoassay method

  15. Comparison of the interleukin-1β-inducing potency of allergenic spores from higher fungi (Basidiomycetes) in a cryopreserved human whole blood system

    Science.gov (United States)

    Rivera-Mariani, Félix E.; Vysyaraju, Kranthi; Negherbon, Jesse; Levetin, Estelle; Horner, W. Elliot; Hartung, Thomas; Breysse, Patrick N.

    2014-01-01

    Background Spores from basidiomycete fungi (basidiospores) are highly prevalent in the atmosphere of urban and rural settings. Studies have confirmed their potential to affect human health as allergens. Less is known about their potential to serve as stimuli of the innate immune system and induce pro-inflammatory reactions. Methods In this study, we evaluated the pro-inflammatory potential of spores from 11 allergenic gilled (Pleurotus ostreatus, Oudemansiella radicata, Armillaria tabescens, Coprinus micaceus, Pluteus cervinus, Chlorophyllum molybdites) and non-gilled (Pisolithus arhizus, Merulius tremullosus, Calvatia cyathiformis, Lycoperdon pyriforme, Boletus bicolor) basidiomycetes fungi based on their potency to induce the release of the pro-inflammatory cytokine interleukin (IL)-1β in a cryopreserved human whole blood system. In addition, the role of morphological features of the spores (surface area, shape, and pigmentation) were examined for their role in the spores’ interleukin (IL)-1β-including potency. Peripheral blood from healthy volunteers was collected, pooled, and cryopreserved. After stimulating the cryopreserved pooled blood with 106 to 103 basidiospores/ml, the concentration of IL-1β in culture supernatants was determined with ELISA. Results Basidiospores manifested concentration-dependent IL-1β-inducing potency, which was more noteworthy among basidiospores from gilled basidiomycetes. At higher concentrations of basidiospores, the IL-1β-inducing potency was able to be differentiated in the cryopreserved human whole blood system. Morphological features did not correlate with the IL-1β-inducing potency of the basidiospores, suggesting that non-morphological properties modulate the IL-1β-inducing potency. Conclusion Our data provides evidence of the pro-inflammatory potential of basidiospores, and the utility of cryopreserved human whole blood as a human-based in-vitro system to study the immune reactivity of allergenic basidiospores. PMID

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

    Science.gov (United States)

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

    2018-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Z. I. Antoniolli

    2002-09-01

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

  18. Effects of microbial cocultivation on inflammatory and cytotoxic potential of spores.

    Science.gov (United States)

    Murtoniemi, Timo; Penttinen, Piia; Nevalainen, Aino; Hirvonen, Maija-Riitta

    2005-11-01

    Microbial growth on moisture-damaged building materials is commonly associated with adverse health effects in the occupants. In moisture damage situations, the environmental conditions as well as the dominant microbial species will vary, leading to a diversity of microbes and continual changes in the different microbial populations. Currently, very little is known about the effects of microbial cocultures on the potential harmfulness of the microbial population. In this study we have investigated the effects of cocultivation of certain indoor air microbes on the inflammatory and cytotoxic potential of their spores. We grew various microbial combinations made from strains of Streptomyces californicus, Stachybotrys chartarum, Aspergillus versicolor, and Penicillium spinulosum on wetted plasterboard. After 5 or 10 wk of growth, the spores were collected from the plasterboards, mouse RAW264.7 macrophages were exposed to the spores, and after 24 h the induced inflammatory and cytotoxic responses were analyzed. Among all the tested microbes and their combinations, the spores of Str. californicus proved to be the most potent inducer of cytotoxicity and inflammatory responses. These results indicate also that microbial coculture may support the growth of certain microbes with high immunotoxic potency such as Str.californicus. Furthermore, coculture containing S. chartarum and A. versicolor caused a synergistic increase in cytotoxicity compared to the sum response induced by the pure cultures, but no effect on inflammatory responses was detected. Generally, spore-induced cytotoxicity and production of inflammatory markers increased during the growth period from 5 to 10 wk, suggesting that the immunotoxic potency of spores increases with time.

  19. Evaluation of some physical and chemical treatments for inactivating microsporidian spores isolated from fish.

    Science.gov (United States)

    Leiro, José M; Piazzon, Carla; Domínguez, Berta; Mallo, Natalia; Lamas, Jesús

    2012-05-15

    Microsporidia are a large diverse group of intracellular parasites now considered as fungi. They are particularly prevalent in fish and are recognized as important opportunistic parasites in humans. Although the mode of transmission of microsporidia has not been fully clarified, the consumption and manipulation of infected fish may be a risk factor for humans. Comparative analysis of rDNA sequence revealed that the microsporidians used in the present study had 99-100% identity with anglerfish microsporidians of the genus Spraguea and very low identity with microsporidians that infect humans. Microsporidian spores were exposed to different physical and chemical treatments: freezing at -20°C for 24-78 h, heating at 60°C for 5-15 min, microwaving at 700 W, 2.45 GHz for 15-60s, and treatment with ethanol at concentrations of between 1 and 70% for 15 min. The viability of the spores after each treatment was evaluated by two methods: a) haemocytometer counts, measuring the extrusion of the polar filament in control and treated spores, and b) a fluorometric method, testing the membrane integrity by propidium iodide exclusion. The results of both methods were concordant. Spores were inactivated by freezing at -20°C for more than 48 h, by heating to 60°C for 10 min and by microwaving at 750 W, for 20s. Exposure to 70% ethanol for 15 min also inactivated microsporidian spores. The results suggest that both freezing and heating are effective treatments for destroying microsporidian spores in European white anglerfish, and that 70% ethanol could be used by fish processors to disinfect their hands and the utensils used in processing fish. The fluorometric method can be used as an alternative to haemocytometer counts in disinfection studies aimed at establishing strategies for inactivating and reducing the viability and the potential infectivity of microsporidians present in fish or in the environment. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. 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.

  1. 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

  2. Purine and its analogues and radiation damage in Bacillus megaterium spores

    Energy Technology Data Exchange (ETDEWEB)

    Powers, E.L.

    1986-12-01

    As an extension of results obtained from radiation studies on caffeine both in other laboratories and more recently in this laboratory using the bacterial spore as the test system, six compounds with chemical structures closely resembling that of caffeine were tested as radiation modifiers. Of these compounds, purine, adenine and hypoxanthine resembled caffeine in sensitizing spores to radiation, while theobromine, xanthine and theophylline did not. These responses are discussed in relation to the electron sequestration hypothesis of cellular sensitization to high-energy radiation.

  3. High Pressure Germination of Bacillus subtilis Spores with Alterations in Levels and Types of Germination Proteins

    Science.gov (United States)

    2014-01-01

    Microbiol 102, 65 76. Butzin, X.Y., Troiano, A.J., Coleman , W.H., Griffiths , K.K., Doona, C.J., Feeherry, F.E., Wang, G., Li, Y. Q. et al. (2012...germination, possibly because it is essential for organization of GRs in a complex in spores’ inner membrane ( Griffiths et aL 2011). Journal of... Griffiths , K.K., Zhang, J., Cowan, A.E., Yu, J. and Setlow, P. (2011) Germination proteins in the inner membrane of dormant Bacillus subtilis spores

  4. Proposed role of lactate in germination of hypochlorite-treated Clostridium botulinum spores.

    OpenAIRE

    Foegeding, P M; Busta, F F

    1983-01-01

    Clostridium botulinum 12885A spores treated with hypochlorite required added DL-calcium lactate for L-alanine germination. Lactate was the active component of calcium lactate. Equimolar concentrations of L-malate, but not of DL-propionate, could replace lactate, suggesting that the alpha-hydroxy acid structure is important. Neither lactate nor malate was an effective germinant for buffer-treated or hypochlorite-treated spores. If the L-alanine concentration was increased 100-fold (to 450 mM),...

  5. Solving the aerodynamics of fungal flight: how air viscosity slows spore motion.

    Science.gov (United States)

    Fischer, Mark W F; Stolze-Rybczynski, Jessica L; Davis, Diana J; Cui, Yunluan; Money, Nicholas P

    2010-01-01

    Viscous drag causes the rapid deceleration of fungal spores after high-speed launches and limits discharge distance. Stokes' law posits a linear relationship between drag force and velocity. It provides an excellent fit to experimental measurements of the terminal velocity of free-falling spores and other instances of low Reynolds number motion (Re10 m s(-1) and travel as far as 2.5m (Re>100). Copyright © 2010 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  6. 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

  7. Ascospores of large-spored Metschnikowia species are genuine meiotic products of these yeasts

    DEFF Research Database (Denmark)

    Marinoni, G.; Piskur, Jure; Lachance, M.A.

    2003-01-01

    The asci of Metschnikowia species normally contain two ascospores (never more), raising the question of whether these spores are true meiotic products. We investigated this problem by crossing genetically-marked strains of the haploid, heterothallic taxa, Metschnikowia hawaiiensis, Metschnikowia...... continentalis var. continentalis, and M. continentalis var. borealis. Asci were dissected and the segregation patterns for various phenotypes analyzed. In all cases (n = 47) both mating types (h(+) and h(-)) were recovered in pairs of sister spores, casting further uncertainty as to whether normal meiosis takes...

  8. 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)

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

    Science.gov (United States)

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

    2015-01-01

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

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

    NARCIS (Netherlands)

    Voort, van der M.; Abee, T.

    2013-01-01

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

  11. Novel Sample Preparation Method for Safe and Rapid Detection of Bacillus anthracis Spores in Environmental Powders and Nasal Swabs

    OpenAIRE

    Luna, Vicki A.; King, Debra; Davis, Carisa; Rycerz, Tony; Ewert, Matthew; Cannons, Andrew; Amuso, Philip; Cattani, Jacqueline

    2003-01-01

    Bacillus anthracis spores have been used as a biological weapon in the United States. We wanted to develop a safe, rapid method of sample preparation that provided safe DNA for the detection of spores in environmental and clinical specimens. Our method reproducibly detects B. anthracis in samples containing

  12. Estimating the abundance of airborne pollen and fungal spores at variable elevations using an aircraft: how high can they fly?

    Science.gov (United States)

    Damialis, Athanasios; Kaimakamis, Evangelos; Konoglou, Maria; Akritidis, Ioannis; Traidl-Hoffmann, Claudia; Gioulekas, Dimitrios

    2017-03-16

    Airborne pollen and fungal spores are monitored mainly in highly populated, urban environments, for allergy prevention purposes. However, their sources can frequently be located outside cities' fringes with more vegetation. So as to shed light to this paradox, we investigated the diversity and abundance of airborne pollen and fungal spores at various environmental regimes. We monitored pollen and spores using an aircraft and a car, at elevations from sea level to 2,000 m above ground, in the region of Thesssaloniki, Greece. We found a total of 24 pollen types and more than 15 spore types. Pollen and spores were detected throughout the elevational transect. Lower elevations exhibited higher pollen concentrations in only half of plant taxa and higher fungal spore concentrations in only Ustilago. Pinaceae and Quercus pollen were the most abundant recorded by airplane (>54% of the total). Poaceae pollen were the most abundant via car measurements (>77% of the total). Cladosporium and Alternaria spores were the most abundant in all cases (aircraft: >69% and >17%, car: >45% and >27%, respectively). We conclude that pollen and fungal spores can be diverse and abundant even outside the main source area, evidently because of long-distance transport incidents.

  13. Characterization of germination and outgrowth of sorbic acid-stressed Bacillus cereus ATCC 14579 spores: Phenotype and transcriptome analysis

    NARCIS (Netherlands)

    Melis, van C.C.J.; Nierop Groot, M.N.; Tempelaars, M.H.; Moezelaar, R.; Abee, T.

    2011-01-01

    Sorbic acid (SA) is widely used as a preservative, but the effect of SA on spore germination and outgrowth has gained limited attention up to now. Therefore, the effect of sorbic acid on germination of spores of Bacillus cereus strain ATCC 14579 was analyzed both at phenotype and transcriptome

  14. Recovery of heat treated Bacillus cereus spores is affected by matrix composition and factors with putative functions in damage repair

    NARCIS (Netherlands)

    Warda, A.K.; Tempelaars, M.H.; Abee, T.; Nierop Groot, M.N.

    2016-01-01

    The ability of spores to recover and grow out after food processing is affected by cellular factors and by the outgrowth conditions. In the current communication we studied the recovery and outgrowth of individually sorted spores in BHI and rice broth media and on agar plates using flow cytometry.

  15. Plate assay for determining the time of production of protease, cellulase, and pectinases by germinating fungal spores.

    Science.gov (United States)

    Hagerman, A E; Blau, D M; McClure, A L

    1985-12-01

    A new method for detecting enzymes produced by fungal spores during germination is described here. With this method, the production of enzymes such as protease, cellulase, or pectinase can be correlated with the extent of spore germination. Germination is studied in vitro on agar-based media containing protein, cellulose, or pectin. The spores are immobilized on a permeable membrane mounted on the substrate-containing medium. At various times after inoculation the membrane-bound spores are removed and the medium is stained. The extent of germination is assessed by microscopic examination of the spores and the presence of active hydrolytic enzymes is revealed by the staining. The staining methods are sensitive; detection limits are 1 X 10(-3) unit of cellulase; 2 X 10(-4) unit of protease; 3 X 10(-3) unit of pectin lyase; 3.5 units of polygalacturonase; 2 X 10(-3) unit of pectin methyl esterase. The method has been demonstrated by studying the production of enzymes by germinating conidia of Botrytis cinerea. Cellulase and protease were present before any spores germinated. Pectin lyase was first observed when at least 80% of the spores had germinated. Pectin methyl esterase and polygalacturonase were not produced by the spores.

  16. Contribution of endogenous and exogenous damage to the total radiation-induced damage in the bacterial spore

    International Nuclear Information System (INIS)

    Jacobs, G.P.; Samuni, A.; Czapski, G.

    1980-01-01

    Radical scavengers such as polyethylene glycol 4000 and bovine albumin have been used to define the contribution of exogenous and endogenous damage to the total radiation-induced damage in aqueous buffered suspensions of Bacillus pumilus spores. The results indicate that this damage in the bacterial spore is predominantly endogenous

  17. Estimating the abundance of airborne pollen and fungal spores at variable elevations using an aircraft: how high can they fly?

    Science.gov (United States)

    Damialis, Athanasios; Kaimakamis, Evangelos; Konoglou, Maria; Akritidis, Ioannis; Traidl-Hoffmann, Claudia; Gioulekas, Dimitrios

    2017-03-01

    Airborne pollen and fungal spores are monitored mainly in highly populated, urban environments, for allergy prevention purposes. However, their sources can frequently be located outside cities’ fringes with more vegetation. So as to shed light to this paradox, we investigated the diversity and abundance of airborne pollen and fungal spores at various environmental regimes. We monitored pollen and spores using an aircraft and a car, at elevations from sea level to 2,000 m above ground, in the region of Thesssaloniki, Greece. We found a total of 24 pollen types and more than 15 spore types. Pollen and spores were detected throughout the elevational transect. Lower elevations exhibited higher pollen concentrations in only half of plant taxa and higher fungal spore concentrations in only Ustilago. Pinaceae and Quercus pollen were the most abundant recorded by airplane (>54% of the total). Poaceae pollen were the most abundant via car measurements (>77% of the total). Cladosporium and Alternaria spores were the most abundant in all cases (aircraft: >69% and >17%, car: >45% and >27%, respectively). We conclude that pollen and fungal spores can be diverse and abundant even outside the main source area, evidently because of long-distance transport incidents.

  18. Relationships among North American and Japanese Laetiporus isolates inferred from molecular phylogenetics and single-spore incompatibility reactions

    Science.gov (United States)

    Mark T. Banik; Daniel L. Lindner; Yuko Ota; Tsutomu. Hattori

    2010-01-01

    Relationships were investigated among North American and Japanese isolates of Laetiporus using phylogenetic analysis of ITS sequences and single-spore isolate incompatibility. Single-spore isolate pairings revealed no significant compatibility between North American and Japanese isolates. ITS analysis revealed 12 clades within the core ...

  19. 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.

  20. Effect of Gamma Radiation on Spore Germination and Mycelial Growth of Penicillium Expansum, Post harvest Disease of Apple Fruit

    International Nuclear Information System (INIS)

    Mostafavi, H. A.; Mirmajlessi, S. M.; Mirjalili, S. M.; Fathollahi, H.; Mansouripour, S. M.; Babaei, M.

    2012-01-01

    Blue mold caused by Penicillium expansum causes most of the losses during the storage period in the world. In this study, the inhibition effect of different doses of gamma radiation on spore germination and mycelial growth of Penicillium expansum was investigated. As a result, the Penicillium expansum was recovered from infected apple fruits. In order to evaluate the gamma radiation effect on the spore germination, spore suspension (10 4 spore/ml) exposed to 0, 100, 300 and 600 grey, using Co-60 gamma cell with a dose rate of 0.2 Gy/Sec. Also, a disk of mycelium (0.5 cm 2 ) was removed from the edge of a three-days colony and transferred to PDA plates and irradiated to 0, 1500, 2000, 2500, 3000 and 3500 Gy. The results showed that, the irradiation has completely inhibited the spore germination at 600 Gy. While, a dose of 3000 Gy completely inhibited the mycelial growth of Penicillium expansum.

  1. Friction and Adhesion Forces of Bacillus thuringiensis Spores on Planar Surfaces in Atmospheric Systems

    Energy Technology Data Exchange (ETDEWEB)

    Kweon, Hyojin [Georgia Inst. of Technology, Atlanta, GA (United States); Yiacoumi, Sotira [Georgia Inst. of Technology, Atlanta, GA (United States); Tsouris, Costas [Georgia Inst. of Technology, Atlanta, GA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2011-11-07

    The kinetic friction force and the adhesion force of Bacillus thuringiensis spores on planar surfaces in atmospheric systems were studied using atomic force microscopy. The influence of relative humidity (RH) on these forces varied for different surface properties including hydrophobicity, roughness, and surface charge. The friction force of the spore was greater on a rougher surface than on mica, which is atomically flat. As RH increases, the friction force of the spores decreases on mica whereas it increases on rough surfaces. The influence of RH on the interaction forces between hydrophobic surfaces is not as strong as for hydrophilic surfaces. The friction force of the spore is linear to the sum of the adhesion force and normal load on the hydrophobic surface. In conclusion, the poorly defined surface structure of the spore and the adsorption of contaminants from the surrounding atmosphere are believed to cause a discrepancy between the calculated and measured adhesion forces.

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

    Directory of Open Access Journals (Sweden)

    R. Iannone

    2011-02-01

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

  3. Evaluation of anti-microbial activity of spore powder of Ganoderma lucidum on clinical isolates of Prevotella intermedia: A pilot study

    Directory of Open Access Journals (Sweden)

    Ranganath N Nayak

    2015-01-01

    Full Text Available Aim: This study aimed at evaluating the anti-microbial activity of spore powder of Ganoderma lucidum on Prevotella intermedia isolated from subgingival plaque from chronic periodontitis patients. Settings and Design: Written informed consent was obtained from each subject enrolled in the study. The Institutional Ethics Committee granted the ethical clearance for the study. Materials and Methods: This study included 20 patients diagnosed with chronic periodontitis. Pooled subgingival plaque samples were collected using sterile curettes from the deepest sites of periodontal pockets. The collected samples were then transported in 1 mL of reduced transport fluid. The organisms were cultured and confirmed. These organisms were then used for minimum inhibitory concentration (MIC procedure. Statistical Analysis: Mean of the MIC value obtained was calculated. Results: Thirteen out of the 20 clinical samples were tested that showed sensitivity at various concentrations. Five samples showed sensitivity at all concentrations. Twelve samples showed sensitivity at 8 mcg/ml. Eleven samples showed sensitivity at 4 mcg/ml, 8 samples showed sensitivity at 2 mcg/ml, and 5 samples showed sensitivity even at 1 mcg/ml. Mean MIC value of G. lucidum spore powder for P. intermedia obtained was 3.62 mcg/ml. Conclusion: G. lucidum with its multipotential bioactivity could be used as an anti-microbial, in conjunction with conventional therapy in periodontal disease.

  4. Identification of a protein interacting with the spore wall protein SWP26 of Nosema bombycis in a cultured BmN cell line of silkworm.

    Science.gov (United States)

    Zhu, Feng; Shen, Zhongyuan; Hou, Jiange; Zhang, Jiao; Geng, Tao; Tang, Xudong; Xu, Li; Guo, Xijie

    2013-07-01

    Nosema bombycis is a silkworm parasite that causes severe economic damage to sericulture worldwide. It is the first microsporidia to be described in the literature, and to date, very little molecular information is available regarding microsporidian physiology and their relationships with their hosts. Therefore, the interaction between the microsporidia N. bombycis and its host silkworm, Bombyx mori, was analyzed in this study. The microsporidian spore wall proteins (SWPs) play a specific role in spore adherence to host cells and recognition by the host during invasion. In this study, SWP26 fused with enhanced green fluorescence protein (EGFP) was expressed in BmN cells by using a Bac-to-Bac expression system. Subsequently, the turtle-like protein of B. mori (BmTLP) was determined to interact with SWP26 via the use of anti-EGFP microbeads. This interaction was then confirmed by yeast two-hybrid analysis. The BmTLP cDNA encodes a polypeptide of 447 amino acids that includes a putative signal peptide of 27 amino acid residues. In addition, the BmTLP protein contains 2 immunoglobulin (IG) domains and 2 IGc2-type domains, which is the typical domain structure of IG proteins. The results of this study indicated that SWP26 interacts with the IG-like protein BmTLP, which contributes to the infectivity of N. bombycis to its host silkworm. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Evaluation of anti-microbial activity of spore powder of Ganoderma lucidum on clinical isolates of Prevotella intermedia: A pilot study.

    Science.gov (United States)

    Nayak, Ranganath N; Dixitraj, P T; Nayak, Aarati; Bhat, Kishore

    2015-09-01

    This study aimed at evaluating the anti-microbial activity of spore powder of Ganoderma lucidum on Prevotella intermedia isolated from subgingival plaque from chronic periodontitis patients. Written informed consent was obtained from each subject enrolled in the study. The Institutional Ethics Committee granted the ethical clearance for the study. This study included 20 patients diagnosed with chronic periodontitis. Pooled subgingival plaque samples were collected using sterile curettes from the deepest sites of periodontal pockets. The collected samples were then transported in 1 mL of reduced transport fluid. The organisms were cultured and confirmed. These organisms were then used for minimum inhibitory concentration (MIC) procedure. Mean of the MIC value obtained was calculated. Thirteen out of the 20 clinical samples were tested that showed sensitivity at various concentrations. Five samples showed sensitivity at all concentrations. Twelve samples showed sensitivity at 8 mcg/ml. Eleven samples showed sensitivity at 4 mcg/ml, 8 samples showed sensitivity at 2 mcg/ml, and 5 samples showed sensitivity even at 1 mcg/ml. Mean MIC value of G. lucidum spore powder for P. intermedia obtained was 3.62 mcg/ml. G. lucidum with its multipotential bioactivity could be used as an anti-microbial, in conjunction with conventional therapy in periodontal disease.

  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. Interaction of airway epithelial cells (A549) with spores and mycelium of Aspergillus fumigatus

    NARCIS (Netherlands)

    Zhang, ZH; Liu, RY; Noordhoek, JA; Kauffman, HF

    2005-01-01

    Objective. To study the interaction of airway epithelial cell line A549 with fragments of mycelium, spores of Aspergitlus fumigatus in vitro and to determine if toll-like receptors (TLRs) are involved in the process. Methods. A549 cells were exposed to fragments of A. fumigatus mycelium, zymosan and

  8. 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

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

    NARCIS (Netherlands)

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

    2007-01-01

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

  10. Beetroot-Pigment-Derived Colorimetric Sensor for Detection of Calcium Dipicolinate in Bacterial Spores

    Science.gov (United States)

    Gonçalves, Letícia Christina Pires; Da Silva, Sandra Maria; DeRose, Paul C.; Ando, Rômulo Augusto; Bastos, Erick Leite

    2013-01-01

    In this proof-of-concept study, we describe the use of the main red beet pigment betanin for the quantification of calcium dipicolinate in bacterial spores, including Bacillus anthracis. In the presence of europium(III) ions, betanin is converted to a water-soluble, non-luminescent orange 1∶1 complex with a stability constant of 1.4×105 L mol–1. The addition of calcium dipicolinate, largely found in bacterial spores, changes the color of the aqueous solution of [Eu(Bn)+] from orange to magenta. The limit of detection (LOD) of calcium dipicolinate is around 2.0×10–6 mol L–1 and the LOD determined for both spores, B. cereus and B. anthracis, is (1.1±0.3)×106 spores mL–1. This simple, green, fast and low cost colorimetric assay was selective for calcium dipicolinate when compared to several analogous compounds. The importance of this work relies on the potential use of betalains, raw natural pigments, as colorimetric sensors for biological applications. PMID:24019934

  11. Vertical transmission of endobacteria in the arbuscular mycorrhizal fungus Gigaspora margarita through generation of vegetative spores.

    Science.gov (United States)

    Bianciotto, V; Genre, A; Jargeat, P; Lumini, E; Bécard, G; Bonfante, P

    2004-06-01

    Arbuscular mycorrhizal (AM) fungi living in symbiotic association with the roots of vascular plants have also been shown to host endocellular rod-shaped bacteria. Based on their ribosomal sequences, these endobacteria have recently been identified as a new taxon, Candidatus Glomeribacter gigasporarum. In order to investigate the cytoplasmic stability of the endobacteria in their fungal host and their transmission during AM fungal reproduction (asexual), a system based on transformed carrot roots and single-spore inocula of Gigaspora margarita was used. Under these in vitro sterile conditions, with no risk of horizontal contamination, the propagation of endobacteria could be monitored, and it was shown, by using primers designed for both 16S and 23S ribosomal DNAs, to occur through several vegetative spore generations (SG0 to SG4). A method of confocal microscopy for quantifying the density of endobacteria in spore cytoplasm was designed and applied; endobacteria were consistently found in all of the spore generations, although their number rapidly decreased from SG0 to SG4. The study demonstrates that a vertical transmission of endobacteria takes place through the fungal vegetative generations (sporulation) of an AM fungus, indicating that active bacterial proliferation occurs in the coenocytic mycelium of the fungus, and suggests that these bacteria are obligate endocellular components of their AM fungal host.

  12. Detection of a novel bacterium associated with spores of the arbuscular mycorrhizal fungus Gigaspora margarita.

    Science.gov (United States)

    Long, Liangkun; Yao, Qing; Ai, Yuncan; Deng, Mingrong; Zhu, Honghui

    2009-06-01

    With PCR-denaturing gradient gel electrophoresis analysis, two bacterial 16S rRNA gene V3 region sequences, 7A and 7B, were detected in association with the crushed spores of the arbuscular mycorrhizal fungus Gigaspora margarita W.N. Becker & I.R. Hall 1976 MAFF520054. DNA sequencing and phylogenetic analysis revealed that 7B was mostly related to the documented cytoplasm endosymbiotic bacterium Candidatus Glomeribacter gigasporarum of G. margarita, but 7A could not be confidently assigned to a known taxon. Further characterization of 7A was conducted by obtaining its almost complete 16S rRNA gene sequence via PCR amplification and sequencing. BLAST search indicates that the 16S rRNA gene sequence did not match any identified species sequences in the GenBank database. Further detection revealed that 7A was also associated with the clean G. margarita MAFF520054 spores that were obtained by the surface-sterilized method or dual culture with Ri T-DNA transformed carrot roots. Many ellipse-shaped or egg-shaped bacterium-like organisms were clustered in layer 3 of the fungal spore wall by transmission electron microscopy observation. Our results indicate that 7A represents a novel bacterial population associated with G. margarita MAFF520054 spores, and its doubtless location (wall or cytoplasm) remains unclear based on the present data.

  13. Ascospores of large-spored Metschnikowia species are genuine meiotic products of these yeasts

    DEFF Research Database (Denmark)

    Marinoni, G.; Piskur, Jure; Lachance, M.A.

    2003-01-01

    continentalis var. continentalis, and M. continentalis var. borealis. Asci were dissected and the segregation patterns for various phenotypes analyzed. In all cases (n = 47) both mating types (h(+) and h(-)) were recovered in pairs of sister spores, casting further uncertainty as to whether normal meiosis takes...

  14. 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...

  15. Storage Effects on Sample Integrity of Environmental Surface Sampling Specimens with Bacillus anthracis Spores.

    Science.gov (United States)

    Perry, K Allison; O'Connell, Heather A; Rose, Laura J; Noble-Wang, Judith A; Arduino, Matthew J

    The effect of packaging, shipping temperatures and storage times on recovery of Bacillus anthracis . Sterne spores from swabs was investigated. Macrofoam swabs were pre-moistened, inoculated with Bacillus anthracis spores, and packaged in primary containment or secondary containment before storage at -15°C, 5°C, 21°C, or 35°C for 0-7 days. Swabs were processed according to validated Centers for Disease Control/Laboratory Response Network culture protocols, and the percent recovery relative to a reference sample (T 0 ) was determined for each variable. No differences were observed in recovery between swabs held at -15° and 5°C, (p ≥ 0.23). These two temperatures provided significantly better recovery than swabs held at 21°C or 35°C (all 7 days pooled, p ≤ 0.04). The percent recovery at 5°C was not significantly different if processed on days 1, 2 or 4, but was significantly lower on day 7 (day 2 vs. 7, 5°C, 10 2 , p=0.03). Secondary containment provided significantly better percent recovery than primary containment, regardless of storage time (5°C data, p ≤ 0.008). The integrity of environmental swab samples containing Bacillus anthracis spores shipped in secondary containment was maintained when stored at -15°C or 5°C and processed within 4 days to yield the optimum percent recovery of spores.

  16. Reduced spore germination explains sensitivity of reef-building algae to climate change stressors.

    Directory of Open Access Journals (Sweden)

    Alexandra Ordoñez

    Full Text Available Reduced seawater pH and changes in carbonate chemistry associated with ocean acidification (OA decrease the recruitment of crustose coralline algae (CCAcf., an important coral-reef builder. However, it is unclear whether the observed decline in recruitment is driven by impairment of spore germination, or post-settlement processes (e.g. space competition. To address this, we conducted an experiment using a dominant CCA, Porolithon cf. onkodes to test the independent and combined effects of OA, warming, and irradiance on its germination success and early development. Elevated CO2 negatively affected several processes of spore germination, including formation of the germination disc, initial growth, and germling survival. The magnitude of these effects varied depending on the levels of temperature and irradiance. For example, the combination of high CO2 and high temperature reduced formation of the germination disc, but this effect was independent of irradiance levels, while spore abnormalities increased under high CO2 and high temperature particularly in combination with low irradiance intensity. This study demonstrates that spore germination of CCA is impacted by the independent and interactive effects of OA, increasing seawater temperature and irradiance intensity. For the first time, this provides a mechanism for how the sensitivity of critical early life history processes to global change may drive declines of adult populations of key marine calcifiers.

  17. 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

  18. Spore production in Paecilomyces lilacinus (Thom.) samson strains on agro-industrial residues

    OpenAIRE

    Robl, Diogo; Sung, Letizia B.; Novakovich, Jo?o Henrique; Marangoni, Paulo R.D.; Zawadneak, Maria Aparecida C.; Dalzoto, Patricia R.; Gabardo, Juarez; Pimentel, Ida Chapaval

    2009-01-01

    Paecilomyces lilacinus has potential for pests control. We aimed to analyze mycelial growth and spore production in P. lilacinus strains in several agro-industrial residues and commercial media. This study suggests alternative nutrient sources for fungi production and that the biotechnological potential of agro-industrial refuses could be employed in byproducts development.

  19. Characterization of small-spored Alternaria from Argentinean crops through a polyphasic approach

    DEFF Research Database (Denmark)

    da Cruz Cabral, Lucía; Rodriguero, Marcela; Stenglein, Sebastián

    2017-01-01

    exporter of agricultural products, so it is essential to thoroughly understand the physiological behaviour of this pathogen in a food safety context. Thus, the objective of this work was to characterize small-spored Alternaria spp. obtained from tomato fruits, pepper fruits, wheat grains and blueberries...

  20. Optimization of Verticillium lecanii spore production in solid-state fermentation on sugarcane bagasse

    NARCIS (Netherlands)

    Shi, Y.; Xu, X.; Zhu, Y.

    2009-01-01

    Verticillium lecanii is an entomopathogen with high potential in biological control of pests. We developed a solid-state fermentation with sugarcane bagasse as carrier absorbing liquid medium to propagate V. lecanii spores. Using statistical experimental design, we optimized the medium composition

  1. Simple, Inexpensive, and Rapid Way to Produce Bacillus subtilis Spores for the Guthrie Bioassay

    Science.gov (United States)

    Franklin, Martha L.; Clark, William A.

    1981-01-01

    Esculin agar has been found to be a simple, inexpensive, rapid, and reliable means to promote production of spores of inhibitor-sensitive clones of Bacillus subtilis strains ATCC 6051 and 6633 for use in the Guthrie bioassay screening tests for genetic metabolic disorders. Images PMID:6790564

  2. 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...

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

    DEFF Research Database (Denmark)

    Hepper, C.M.; Jakobsen, Iver

    1983-01-01

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

  4. 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

  5. In situ reticulate sphenophyllalean spores, Pennsylvanian (Bolsovian) of the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Bek, Jiří; Libertín, M.

    2010-01-01

    Roč. 159, 1/2 (2010), s. 56-61 ISSN 0034-6667 R&D Projects: GA AV ČR IAA301110701 Institutional research plan: CEZ:AV0Z30130516 Keywords : sphenophylls * Bowmanites * in situ spores * Pennsylvanian Subject RIV: DB - Geology ; Mineralogy Impact factor: 1.985, year: 2010

  6. 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

  7. A rapid colorimetric assay for mold spore germination using XTT tetrazolium salt

    Science.gov (United States)

    Carol A. Clausen; Vina W. Yang

    2011-01-01

    Current laboratory test methods to measure efficacy of new mold inhibitors are time consuming, some require specialized test equipment and ratings are subjective. Rapid, simple quantitative assays to measure the efficacy of mold inhibitors are needed. A quantitative, colorimetric microassay was developed using XTT tetrazolium salt to metabolically assess mold spore...

  8. Airborne pollen and spore survey in relation to allergy and plant ...

    African Journals Online (AJOL)

    Airborne bio-particles of allergic significance were recorded at a height of 15m in Nsukka during September 1999 – February 2000. Spores of fungi and pollen grains, which are important part of the exposure that may lead to allergic discomfort and plant diseases, dominated the particles. Other primary sources of the allergic ...

  9. Rupturing of Biological Spores As a Source of Secondary Particles in Amazonia

    Energy Technology Data Exchange (ETDEWEB)

    China, Swarup; Wang, Bingbing; Weis, Johannes; Rizzo, Luciana; Brito, Joel; Cirino, Glauber G.; Kovarik, Libor; Artaxo, Paulo; Gilles, Mary K.; Laskin, Alexander

    2016-11-15

    Airborne biological particles, such as fungal spores and pollen, are ubiquitous in the Earth’s atmosphere and play an important role in the atmospheric environment and climate, impacting air quality, cloud formation, and the Earth’s radiation budget. The atmospheric transformations of airborne biological spores at elevated relative humidity remain poorly understood and their climatic role is uncertain. Using an environmental scanning electron microscope (ESEM), we observed rupturing of Amazonian fungal spores and subsequent release of nanometer to submicron size fragments after exposure to high humidity. We find that fungal fragments contain elements of inorganic salts (e.g., Na and Cl). They are hygroscopic in nature with a growth factor up to 2.3 at 96% relative humidity, thus they may potentially influence cloud formation. Due to their hygroscopic growth, light scattering cross sections of the fragments are enhanced by up to a factor of 10. Furthermore, rupturing of fungal spores at high humidity may explain the bursting events of nanoparticles and may provide insight into new particle formation in Amazonia.

  10. Abiotic and microbiotic factors controlling biofilm formation of thermophilic spore formers.

    NARCIS (Netherlands)

    Zhao, Y.; Caspers, M.P.; Metselaar, K.I.; de Boer, W.P.H.; Roeselers, G.; Moezelaar, R.; Nierop Groot, M.N.; Montijn, R.; Abee, T.; Kort, R.

    2013-01-01

    One of the major concerns in the production of dairy concentrates is the risk of contamination by heat-resistant spores from thermophilic bacteria. In order to acquire more insight in the composition of microbial communities occurring in the dairy concentrate industry, a bar-coded 16S amplicon

  11. Autofluorescence of fossil pollen and spores with special reference to age determination and coalification

    NARCIS (Netherlands)

    Gijzel, van Pieter

    1967-01-01

    In the present study the primary fluorescence phenomena of fossil pollen and spores are described. This new method in palynology is based on a large number of fluorescence microscopical observations and spectrophotometrical determinations of palynomorphs from deposits of various type and age. It

  12. Trichoderma sp. spores and Kluyveromyces marxianus cells magnetic separation: Immobilization on chitosan-coated magnetic nanoparticles.

    Science.gov (United States)

    Palacios-Ponce, Sócrates; Ramos-González, Rodolfo; Ruiz, Héctor A; Aguilar, Miguel A; Martínez-Hernández, José L; Segura-Ceniceros, Elda P; Aguilar, Cristóbal N; Michelena, Georgina; Ilyina, Anna

    2017-07-03

    In the present study, the interactions between chitosan-coated magnetic nanoparticles (C-MNP) and Trichoderma sp. spores as well as Kluyveromyces marxianus cells were studied. By Plackett-Burman design, it was demonstrated that factors which directly influenced on yeast cell immobilization and magnetic separation were inoculum and C-MNP quantity, stirring speed, interaction time, and volume of medium, while in the case of fungal spores, the temperature also was disclosed as an influencing factor. Langmuir and Freundlich models were applied for the mathematical analysis of adsorption isotherms at 30°C. For Trichoderma sp. spore adsorption isotherm, the highest correlation coefficient was observed for lineal function of Langmuir model with a maximum adsorption capacity at 5.00E + 09 spores (C-MNP g -1 ). Adsorption isotherm of K. marxianus cells was better adjusted to Freundlich model with a constant (K f ) estimated as 2.05E + 08 cells (C-MNP g -1 ). Both systems may have a novel application in fermentation processes assisted with magnetic separation of biomass.

  13. Heat-induced oxidative injury contributes to inhibition of Botrytis cinerea spore germination and growth

    Science.gov (United States)

    The inhibitory effect of a heat treatment (HT) on Botrytis cinerea, a major postharvest fungal pathogen, and the possible mode of action were investigated. Spore germination and germ tube elongation of B. cinerea were both increasingly and significantly inhibited by a HT (43 degrees C) for 10, 20 o...

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

    NARCIS (Netherlands)

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

    2010-01-01

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

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

    NARCIS (Netherlands)

    Farenhorst, Marit; Knols, Bart G. J.

    2010-01-01

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

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

    NARCIS (Netherlands)

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

    2007-01-01

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

  17. Carbonization measurements of pollen-grains and spores and their application

    NARCIS (Netherlands)

    Gutjahr, C.C.M.

    1966-01-01

    This study presents the equipment and technique used in the author’s palynological carbonization studies. The equipment consists mainly of modified Zeiss microphotographie equipment. A photoelectric cell fitted with a field restrictor is used to measure the light absorption of a spore or pollen

  18. Regurgitated pellets of Merops apiaster as fomites of infective Nosema ceranae (Microsporidia) spores.

    Science.gov (United States)

    Higes, Mariano; Martín-Hernández, Raquel; Garrido-Bailón, Encarna; Botías, Cristina; García-Palencia, Pilar; Meana, Aránzazu

    2008-05-01

    The importance of transmission factor identification is of great epidemiological significance. The bee-eater (Merops apiaster) is a widely distributed insectivorous bird, locally abundant mainly in arid and semi-arid areas of southern Europe, northern Africa and western Asia but recently has been seen breeding in central Europe and Great Britain. Bee-eaters predominantly eat insects, especially bees, wasps and hornets. On the other hand, Nosema ceranae is a Microsporidia recently described as a parasite in Apis mellifera honeybees in Europe. Due to the short time since its description scarce epidemiological data are available. In this study we investigate the role of the regurgitated pellets of the European bee-eater as fomites of infective spores of N. ceranae. Spore detection in regurgitated pellets of M. apiaster is described [phase-contrast microscopy (PCM) and polymerase chain reaction (PCR) methods]. Eighteen days after collection N. ceranae spores still remain viable and their infectivity is shown after artificial infection of Nosema-free 8-day-old adult bees. The epidemiological consequences of the presence of Nosema spores in this fomites are discussed.

  19. Fluorescence-Activated Cell Sorting of Live Versus Dead Bacterial Cells and Spores

    Science.gov (United States)

    Bernardini, James N.; LaDuc, Myron T.; Diamond, Rochelle; Verceles, Josh

    2012-01-01

    This innovation is a coupled fluorescence-activated cell sorting (FACS) and fluorescent staining technology for purifying (removing cells from sampling matrices), separating (based on size, density, morphology, and live versus dead), and concentrating cells (spores, prokaryotic, eukaryotic) from an environmental sample.

  20. 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.