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Sample records for acid-soluble spore proteins

  1. 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.

  2. Roles of Small, Acid-Soluble Spore Proteins and Core Water Content in Survival of Bacillus subtilis Spores Exposed to Environmental Solar UV Radiation▿

    OpenAIRE

    Moeller, Ralf; Setlow, Peter; Reitz, Günther; Nicholson, Wayne L.

    2009-01-01

    Spores of Bacillus subtilis contain a number of small, acid-soluble spore proteins (SASP) which comprise up to 20% of total spore core protein. The multiple α/β-type SASP have been shown to confer resistance to UV radiation, heat, peroxides, and other sporicidal treatments. In this study, SASP-defective mutants of B. subtilis and spores deficient in dacB, a mutation leading to an increased core water content, were used to study the relative contributions of SASP and increased core water conte...

  3. Influence of acid-soluble proteins from bivalve Siliqua radiata ligaments on calcium carbonate crystal growth

    Science.gov (United States)

    Huang, Zeng-Qiong; Zhang, Gang-Sheng

    2016-08-01

    In vitro biomimetic synthesis of calcium carbonate (CaCO3) in the presence of shell proteins is a heavily researched topic in biomineralization. However, little is known regarding the function of bivalve ligament proteins in the growth of CaCO3 crystals. In this study, using fibrous protein K58 from Siliqua radiata ligaments or coverslips as substrates, we report the results of our study of CaCO3 precipitation in the presence or absence of acid-soluble proteins (ASP) from inner ligament layers. ASP can disturb the controlling function of K58 or a coverslip on the crystalline phase, resulting in the formation of aragonite, calcite, and vaterite. In addition, we identified the following four primary components from ASP by mass spectroscopy: alkaline phosphatase (ALP), ABC transporter, keratin type II cytoskeletal 1 (KRT 1), and phosphate ABC transporter, phosphate-binding protein (PstS). Further analysis revealed that the first three proteins and especially ALP, which is important in bone mineralisation, could affect the polymorphism and morphology of CaCO3 crystals by trapping calcium ions in their domains. Our results indicate that ALP may play an important role in the formation of aragonite in S. radiata ligaments. This paper may facilitate our understanding of the biomineralization process.

  4. Protective Role of Spore Structural Components in Determining Bacillus subtilis Spore Resistance to Simulated Mars Surface Conditions

    OpenAIRE

    Moeller, Ralf; Schuerger, Andrew C.; Reitz, Günther; Nicholson, Wayne L.

    2012-01-01

    Spores of wild-type and mutant Bacillus subtilis strains lacking various structural components were exposed to simulated Martian atmospheric and UV irradiation conditions. Spore survival and mutagenesis were strongly dependent on the functionality of all of the structural components, with small acid-soluble spore proteins, coat layers, and dipicolinic acid as key protectants.

  5. Bacillus anthracis Spore Surface Protein BclA Mediates Complement Factor H Binding to Spores and Promotes Spore Persistence.

    Science.gov (United States)

    Wang, Yanyu; Jenkins, Sarah A; Gu, Chunfang; Shree, Ankita; Martinez-Moczygemba, Margarita; Herold, Jennifer; Botto, Marina; Wetsel, Rick A; Xu, Yi

    2016-06-01

    Spores of Bacillus anthracis, the causative agent of anthrax, are known to persist in the host lungs for prolonged periods of time, however the underlying mechanism is poorly understood. In this study, we demonstrated that BclA, a major surface protein of B. anthracis spores, mediated direct binding of complement factor H (CFH) to spores. The surface bound CFH retained its regulatory cofactor activity resulting in C3 degradation and inhibition of downstream complement activation. By comparing results from wild type C57BL/6 mice and complement deficient mice, we further showed that BclA significantly contributed to spore persistence in the mouse lungs and dampened antibody responses to spores in a complement C3-dependent manner. In addition, prior exposure to BclA deletion spores (ΔbclA) provided significant protection against lethal challenges by B. anthracis, whereas the isogenic parent spores did not, indicating that BclA may also impair protective immunity. These results describe for the first time an immune inhibition mechanism of B. anthracis mediated by BclA and CFH that promotes spore persistence in vivo. The findings also suggested an important role of complement in persistent infections and thus have broad implications. PMID:27304426

  6. Bacillus anthracis Spore Surface Protein BclA Mediates Complement Factor H Binding to Spores and Promotes Spore Persistence.

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    Yanyu Wang

    2016-06-01

    Full Text Available Spores of Bacillus anthracis, the causative agent of anthrax, are known to persist in the host lungs for prolonged periods of time, however the underlying mechanism is poorly understood. In this study, we demonstrated that BclA, a major surface protein of B. anthracis spores, mediated direct binding of complement factor H (CFH to spores. The surface bound CFH retained its regulatory cofactor activity resulting in C3 degradation and inhibition of downstream complement activation. By comparing results from wild type C57BL/6 mice and complement deficient mice, we further showed that BclA significantly contributed to spore persistence in the mouse lungs and dampened antibody responses to spores in a complement C3-dependent manner. In addition, prior exposure to BclA deletion spores (ΔbclA provided significant protection against lethal challenges by B. anthracis, whereas the isogenic parent spores did not, indicating that BclA may also impair protective immunity. These results describe for the first time an immune inhibition mechanism of B. anthracis mediated by BclA and CFH that promotes spore persistence in vivo. The findings also suggested an important role of complement in persistent infections and thus have broad implications.

  7. SWP5, a Spore Wall Protein, Interacts with Polar Tube Proteins in the Parasitic Microsporidian Nosema bombycis

    OpenAIRE

    Li, Zhi; Pan, Guoqing; Li, Tian; Wei HUANG; Chen, Jie; Geng, Lina; Yang, Donglin; Wang, Linling; Zhou, Zeyang

    2012-01-01

    Microsporidia are a group of eukaryotic intracellular parasites that infect almost all vertebrates and invertebrates. The microsporidian invasion process involves the extrusion of a unique polar tube into host cells. Both the spore wall and the polar tube play an important role in microsporidian pathogenesis. So far, five spore wall proteins (SWP1, SWP2, Enp1, Enp2, and EcCDA) from Encephalitozoon intestinalis and Encephalitozoon cuniculi and five spore wall proteins (SWP32, SWP30, SWP26, SWP...

  8. Improvement on Acid-solubility of Soy Protein Isolate with Enzymatic Method%酶法提高大豆分离蛋白酸溶性的研究

    Institute of Scientific and Technical Information of China (English)

    黄橙子; 王静; 高红亮; 崔红亮; 常忠义

    2013-01-01

    通过单因素试验研究了植酸酶添加量、酶解时间、pH、温度和料液浓度对大豆蛋白酸溶度、透光率和料液粘度的影响,在单因素试验的基础上对pH、温度、料液浓度和时间进行四因素三水平的正交试验,结果表明,当植酸酶添加量为0.6%时,酶解的最佳条件组合为pH3.0,温度40℃,料液浓度10%,酶解时间45 min,此时大豆分离蛋白的酸溶度为45.79%,透光率为68.5%,与优化前相比,分别提高了30.65%和29.4%.%Soy protein isolate(SPI)has been used in the food industry for decades,but its application in acidic food products was Limited because of its low solubility in the acidic condition. The objective of the experiments was to improve the solubility of SPI under acidic conditions by hydrolyzing the phytic acid with phytase and to find the optimum conditions for the enzymatic hydrolyzing process. During the experiments, acid-soluble properties,including acid-solubility,transmittance,and viscosity of the processed material have been observed. Single factor and orthogonal experiments revealed that when the solid content of the curd slurry was 10% ,the best phytase adding amount was 0. 6% ,the ideal condition for the enzymatic reaction was pH3.0, 40℃,45 min. After the enzymatic treatment,acid-solubility and transmittance of SPI reached 45.79% and 68. 5% ,increased by 30.65% and 29.4% ,respectively.

  9. Identification of the immunogenic spore and vegetative proteins of Bacillus anthracis vaccine strain A16R.

    Directory of Open Access Journals (Sweden)

    Xiankai Liu

    Full Text Available Immunoproteomics was used to screen the immunogenic spore and vegetative proteins of Bacillus anthracis vaccine strain A16R. The spore and vegetative proteins were separated by 2D gel electrophoresis and transferred to polyvinylidene difluoride membranes, and then western blotting was performed with rabbit immune serum against B.anthracis live spores. Immunogenic spots were cut and digested by trypsin. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed to identify the proteins. As a result, 11 and 45 immunogenic proteins were identified in the spores and vegetative cells, respectively; 26 of which have not been reported previously. To verify their immunogenicity, 12 of the identified proteins were selected to be expressed, and the immune sera from the mice vaccinated by the 12 expressed proteins, except BA0887, had a specific western blot band with the A16R whole cellular lytic proteins. Some of these immunogenic proteins might be used as novel vaccine candidates themselves or for enhancing the protective efficacy of a protective-antigen-based vaccine.

  10. Improved Proteomic Analysis Following Trichloroacetic Acid Extraction of Bacillus anthracis Spore Proteins

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

    2015-08-07

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Proteomic analysis is dependent upon efficient extraction of proteins from bacterial samples without introducing bias toward extraction of particular protein classes. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrich for certain classes of proteins. The method presented here is technically simple and does not require specialized equipment such as a mechanical disrupter. Our data reveal that for particularly challenging samples, such as B. anthracis Sterne spores, trichloroacetic acid extraction improved the number of proteins identified within a sample compared to bead beating (714 vs 660, respectively). Further, TCA extraction enriched for 103 known spore specific proteins whereas bead beating resulted in 49 unique proteins. Analysis of C. botulinum samples grown to 5 days, composed of vegetative biomass and spores, showed a similar trend with improved protein yields and identification using our method compared to bead beating. Interestingly, easily lysed samples, such as B. anthracis vegetative cells, were equally as effectively processed via TCA and bead beating, but TCA extraction remains the easiest and most cost effective option. As with all assays, supplemental methods such as implementation of an alternative preparation method may provide additional insight to the protein biology of the bacteria being studied.

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

    NARCIS (Netherlands)

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

    2012-01-01

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

  12. Identification of a Protein Subset of the Anthrax Spore Immunome in Humans Immunized with the Anthrax Vaccine Adsorbed Preparation

    OpenAIRE

    Kudva, Indira T.; Griffin, Robert W.; Garren, Jeonifer M.; Calderwood, Stephen B.; John, Manohar

    2005-01-01

    We identified spore targets of Anthrax Vaccine Adsorbed (AVA)-induced immunity in humans by screening recombinant clones of a previously generated, limited genomic Bacillus anthracis Sterne (pXO1+, pXO2−) expression library of putative spore surface (spore-associated [SA]) proteins with pooled sera from human adults immunized with AVA (immune sera), the anthrax vaccine currently approved for use by humans in the United States. We identified 69 clones that reacted specifically with pooled immu...

  13. Spore Resistance Properties.

    Science.gov (United States)

    Setlow, Peter

    2014-10-01

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

  14. Comparative Study on the Infectivity and Spore Surface Protein of Nosema bombycis and Its Morphological Variant Strain

    Institute of Scientific and Technical Information of China (English)

    HUANG Shao-kang; LU Xing-meng

    2005-01-01

    A new morphological variant strain of microsporidium was produced by infecting the mulberry looper, Hemerophila atrilineata [Phthonandria atrilineata], with Nosema bombycis successively for 24 times, and named 24Nbh. Comparative studies on morphology, infectivity and spore surface protein were conducted. 24Nbh was short and wide, and had a significant difference (P<0.01) over the Nb spores. The infectivity tests conducted on second instar silkworm larvae showed that IC50 of 24Nbh was 1.98× 104 spores mL-1 and of Nb was 1.72× 103 spores mL-1, thus indicating that the infectivity of Nb decreased 11.5 times after multiplying in mulberry looper for 24 times. The IC50 of spores from silkworm infected with 24 Nbh was 6.9 times less than Nb, showing that the infectivity of 24Nbh spores rejuvenated very fast when reinfected to silkworms, further more, the length and width of such spore was larger than 24Nbh (P<0.01) and smaller than Nb (P<0.05).The SDS-PAGE profiles of Nb and 24Nbh were generally the same, 4 distinct proteins of 12, 17, 30, 33 kDa were obtained with difference in quantity. When 120 μg of protein was applied for 2D-PAGE, five suspected different proteins with difference in quantity were observed. These results demonstrate that these differential proteins maybe associated with variation in infectivity of the spores.

  15. A gene encoding a vicilin-like protein is specifically expressed in fern spores. Evolutionary pathway of seed storage globulins.

    Science.gov (United States)

    Shutov, A D; Braun, H; Chesnokov, Y V; Bäumlein, H

    1998-02-15

    The isolation and characterisation of a cDNA coding for a vicilin-like protein of the fern Matteuccia struthiopteris is described. The corresponding gene is specifically expressed during late stages of spore development. Extensive sequence comparisons suggest that the fern protein can be considered as a molecular missing link between single-domain germin/spherulin-like proteins and two-domain seed storage globulins of gymnosperms and angiosperms. Further, evidence is provided for the existence of a superfamily of structurally related, functionally different proteins which includes storage globulins of the vicilin and legumin families, a membrane-associated sucrose-binding protein of soybean, a Forssman antigen-binding lectin of velvet bean, the precursor of the vacuolar membrane bound proteins MP27/MP32 of pumpkin, the embryogenesis-specific protein Gea8 of carrot, the fern-spore-specific protein described here as well as the functionally diverse family of germins/germin-like proteins and the spherulins of myxomycetes. We propose that seed storage globulins of spermatophytes evolved from desiccation-related single-domain proteins of prokaryotes via a duplicated two-domain ancestor that is best represented by the extant fern spore-specific vicilin-like protein.

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

    OpenAIRE

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

    2015-01-01

    The germination of spore-forming bacteria in high-salinity environments is of applied interest for food microbiology and soil ecology. It has previously been shown that high salt concentrations detrimentally affect Bacillus subtilis spore germination, rendering this process slower and less efficient. The mechanistic details of these salt effects, however, remained obscure. Since initiation of nutrient germination first requires germinant passage through the spores' protective integuments, the...

  17. Structural and Functional Analysis of the GerD Spore Germination Protein of Bacillus Species

    OpenAIRE

    Li, Yunfeng; Jin, Kai; Ghosh, Sonali; Devarakonda, Parvathimadhavi; Carlson, Kristina; Davis, Andrew; Stewart, Kerry-Ann V.; Cammett, Elizabeth; Rossi, Patricia Pelczar; Setlow, Barbara; Lu, Min; Setlow, Peter; Hao, Bing

    2014-01-01

    Spore germination in Bacillus species represents an excellent model system with which to study the molecular mechanisms underlying the nutritional control of growth and development. Binding of specific chemical nutrients to their cognate receptors located in the spore inner membrane triggers the germination process that leads to a resumption of metabolism in spore outgrowth. Recent studies suggest that the inner membrane GerD lipoprotein plays a critical role in the receptor-mediated activati...

  18. Interaction and assembly of two novel proteins in the spore wall of the microsporidian species Nosema bombycis and their roles in adherence to and infection of host cells.

    Science.gov (United States)

    Yang, Donglin; Pan, Guoqing; Dang, Xiaoqun; Shi, Yawei; Li, Chunfeng; Peng, Pai; Luo, Bo; Bian, Maofei; Song, Yue; Ma, Cheng; Chen, Jie; Ma, Zhengang; Geng, Lina; Li, Zhi; Tian, Rui; Wei, Cuifang; Zhou, Zeyang

    2015-04-01

    Microsporidia are obligate intracellular parasites with rigid spore walls that protect against various environmental pressures. Despite an extensive description of the spore wall, little is known regarding the mechanism by which it is deposited or the role it plays in cell adhesion and infection. In this study, we report the identification and characterization of two novel spore wall proteins, SWP7 and SWP9, in the microsporidian species Nosema bombycis. SWP7 and SWP9 are mainly localized to the exospore and endospore of mature spores and the cytoplasm of sporonts, respectively. In addition, a portion of SWP9 is targeted to the spore wall of sporoblasts earlier than SWP7 is. Both SWP7 and SWP9 are specifically colocalized to the spore wall in mature spores. Furthermore, immunoprecipitation, far-Western blotting, unreduced SDS-PAGE, and yeast two-hybrid data demonstrated that SWP7 interacted with SWP9. The chitin binding assay showed that, within the total spore protein, SWP9 and SWP7 can bind to the deproteinated chitin spore coats (DCSCs) of N. bombycis. However, binding of the recombinant protein rSWP7-His to the DCSCs is dependent on the combination of rSWP9-glutathione S-transferase (GST) with the DCSCs. Finally, rSWP9-GST, anti-SWP9, and anti-SWP7 antibodies decreased spore adhesion and infection of the host cell. In conclusion, SWP7 and SWP9 may have important structural capacities and play significant roles in modulating host cell adherence and infection in vitro. A possible major function of SWP9 is as a scaffolding protein that supports other proteins (such as SWP7) that form the integrated spore wall of N. bombycis. PMID:25605761

  19. Characterization of the spore surface and exosporium proteins of Clostridium sporogenes; implications for Clostridium botulinum group I strains.

    Science.gov (United States)

    Janganan, Thamarai K; Mullin, Nic; Tzokov, Svetomir B; Stringer, Sandra; Fagan, Robert P; Hobbs, Jamie K; Moir, Anne; Bullough, Per A

    2016-10-01

    Clostridium sporogenes is a non-pathogenic close relative and surrogate for Group I (proteolytic) neurotoxin-producing Clostridium botulinum strains. The exosporium, the sac-like outermost layer of spores of these species, is likely to contribute to adhesion, dissemination, and virulence. A paracrystalline array, hairy nap, and several appendages were detected in the exosporium of C. sporogenes strain NCIMB 701792 by EM and AFM. The protein composition of purified exosporium was explored by LC-MS/MS of tryptic peptides from major individual SDS-PAGE-separated protein bands, and from bulk exosporium. Two high molecular weight protein bands both contained the same protein with a collagen-like repeat domain, the probable constituent of the hairy nap, as well as cysteine-rich proteins CsxA and CsxB. A third cysteine-rich protein (CsxC) was also identified. These three proteins are also encoded in C. botulinum Prevot 594, and homologues (75-100% amino acid identity) are encoded in many other Group I strains. This work provides the first insight into the likely composition and organization of the exosporium of Group I C. botulinum spores. PMID:27375261

  20. Bacterial spore survival after exposure to HZE particle bombardment -implication for the lithopanspermia hypothesis.

    Science.gov (United States)

    Moeller, Ralf; Berger, Thomas; Matthiä, Daniel; Okayasu, Ryuichi; Kitamura, H.; Reitz, Guenther

    transcriptional response during spore germination" (Moeller et al., 2008 [3]). To simulate the interplanetary journey of a meteorite, stacks of spore-samples on gabbro slides in different depths were exposed. Spore survival and the rate of the induced mutations (i.e., sporulation-deficiency (Spo-)) depended on the LET of the applied species of ions as well as on the location (and depth) of the irradiated spores in the artificial meteorite. The exposure to high LET iron ions led to a low level of spore survival and increased frequency of mutation to Spo-compared to low-energy charged particles compared to the low LET helium ions. In order to obtain insights on the role of DNA repair by nonhomologous end joining (NHEJ), homologous recombination (HR) and apurinic/apyrimidinic (AP) endonucleases in B. subtilis spore resistance to high-energy charged particles has been studied in parallel. Spores deficient in NHEJ and AP endonucleases were significantly more sensitive to HZE particle bombardment than were the HR-mutant and wild-type spores, indicating that NHEJ and AP endonucleases provide DNA break repair pathways during spore germination. ((References: [1] Arrhenius, S. 1903. Die Verbreitung des Lebens im Weltenraum. Umschau 7:481-485.; [2] Nicholson, W. L. 2009. Ancient micronauts: interplanetary transport of microbes by cosmic impacts. Trends Mi-crobiol. 17:243-250.; [3] Moeller, R., P. Setlow, G. Horneck, T. Berger, G. Reitz, P. Rettberg, A. J. Doherty, R. Okayasu, and W. L. Nicholson. 2008. Roles of the major, small, acid-soluble spore proteins and spore-specific and universal DNA repair mechanisms in resistance of Bacillus subtilis spores to ionizing radiation from X-rays and high-energy charged-particle bombardment. J. Bacteriol. 190:1134-1140.))

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

    NARCIS (Netherlands)

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

    2011-01-01

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

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

    NARCIS (Netherlands)

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

    2013-01-01

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

  3. Yeast spore germination: a requirement for Ras protein activity during re-entry into the cell cycle.

    OpenAIRE

    Herman, P K; Rine, J.

    1997-01-01

    Saccharomyces cerevisiae spore germination is a process in which quiescent, non-dividing spores become competent for mitotic cell division. Using a novel assay for spore uncoating, we found that spore germination was a multi-step process whose nutritional requirements differed from those for mitotic division. Although both processes were controlled by nutrient availability, efficient spore germination occurred in conditions that did not support cell division. In addition, germination did not ...

  4. Purification and characterization of the acid soluble 26-kDa polypeptide from soybean seeds.

    Science.gov (United States)

    Momma, M; Haraguchi, K; Saito, M; Chikuni, K; Harada, K

    1997-08-01

    Whey proteins from soybean seeds of Japanese varieties were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Among 11 varieties of soybean, three green and one black soybeans lacked a 26-kDa band that was found in all yellow soybeans. In this paper, the 26-kDa protein was named AS26k (acid soluble 26-kDa protein) temporarily. The AS26k protein was purified from Glycine max cv. Nattosyoryu, which is yellow soybean, through four purification steps: 30-35% saturated ammonium sulfate fractionation, ion exchange chromatography on S Sepharose Fast Flow, gel filtration on Sephadex G-100, and hydrophobic chromatography on phenyl Sepharose CL-4B. Purified AS26k was cleaved with V8 proteinase from Staphylococcus aureus or CNBr. The cleaved polypeptide contained two typical dehydrin motif sequences: DEYGNPV and (M)DKIKEKLPG, and a 19 amino acids sequence similar to a pea dehydrin. Native AS26k had a molecular mass of 32 kDa on gel filtration and a pl of 7.2 on two-dimensional PAGE. Similarly to other dehydrins and late embryogenesis abundant (LEA) proteins, AS26k was rich in hydrophilic amino acids, and highly heat stable. These results showed that AS26k was a dehydrin, a group II LEA protein in soybean seeds. PMID:9301109

  5. Separation of Protein Crystals from Spores of Bacillus thuringiensis by Ludox Gradient Centrifugation

    OpenAIRE

    Zhu, Yu Sheng; Brookes, Allan; Carlson, Ken; Filner, Philip

    1989-01-01

    A method is described for the purification of Bacillus thuringiensis protein crystals by Ludox gradient centrifugation. This method is simple, inexpensive, fast, and efficient compared with other techniques. It has been successfully used to purify and characterize the protein crystals from several B. thuringiensis strains.

  6. REMOVAL OF ACID-SOLUBLE LIGNIN FROM BIOMASS EXTRACTS USING AMBERLITE XAD-4 RESIN

    Directory of Open Access Journals (Sweden)

    Thomas James Schwartz

    2010-09-01

    Full Text Available This paper describes a method for the removal of acid-soluble lignin from acid hydrolyzed hemicelluloses extracted from a mixture of northern hardwood chips, by using Amberlite XAD-4 resin, which was shown to remove 100% of furan derivatives and 90% of acid-soluble lignin. Subsequent fermentation of the resin treated hydrolyzates gave ethanol yields as high as 97% of theoretical and showed a marked increase in fermentation rate. Regeneration of resin performed with 75% acetone was 85% efficient with respect to acid soluble lignin.

  7. A Novel Immunogenic Spore Coat-Associated Protein in Bacillus Anthracis: Characterization via Proteomics Approaches and a Vector-Based Vaccine System

    OpenAIRE

    Liu, Yu-Tsueng; Lin, Shwu-Bin; Huang, Cheng-Po; Huang, Chun-Ming

    2007-01-01

    New generation anthrax vaccines have been actively explored with the aim of enhancing efficacies and decreasing undesirable side effects that could be caused by licensed vaccines. Targeting novel antigens and/or eliminating the requirements for multiple needle injections and adjuvants are major objectives in the development of new anthrax vaccines. Using proteomics approaches, we identified a spore coat-associated protein (SCAP) in Bacillus anthracis. An E. coli vector-based vaccine system wa...

  8. Physical interaction and assembly of Bacillus subtilis spore coat proteins CotE and CotZ studied by atomic force microscopy.

    Science.gov (United States)

    Liu, Huiqing; Qiao, Haiyan; Krajcikova, Daniela; Zhang, Zhe; Wang, Hongda; Barak, Imrich; Tang, Jilin

    2016-08-01

    The spore of Bacillus subtilis, a dormant type of cell, is surrounded by a complex multilayered protein structure known as the coat. It is composed of over 70 proteins and essential for the spore to withstand extreme environmental conditions and allow germination under favorable conditions. However, understanding how the properties of the coat arise from the interactions among all these proteins is an important challenge. Moreover, many specific protein-protein interactions among the coat proteins are crucial for coat assembly. In this study, atomic force microscopy (AFM) based single molecule force spectroscopy (SMFS) was applied to investigate the interaction as a dynamic process between two morphogenetic coat proteins, CotE and CotZ. The unbinding force and kinetic parameters characterizing the interaction between CotE and CotZ were obtained. It is found that there is a strong affinity between CotE and CotZ. Furthermore, the assembly behaviors of CotE and CotZ, individually or in combination, were studied by AFM at solid-liquid interfaces. Our results revealed that CotE-CotZ assembly is dependent on their molar ratios and the interaction between CotE and CotZ involves in the CotE-CotZ assembly. PMID:27320701

  9. APPLICATIONS OF RAMAN SPECTROSCOPY FOR CHEMICAL CHARACTERIZATION AND PROTEIN CONFORMATION OF AGARICUS BISPORUS (LANGE) IMBACH. (AGARICOMYCETIDAE) SPORES

    OpenAIRE

    Lamrood PY, Ralegankar SD* and Harpale VM

    2014-01-01

    A Raman spectra was obtained for a spore suspension of an edible mushroom Agaricus bisporus using iRaman 2013 with a variable laser power (max. upto 300mW), near infrared 785 nm diode laser and resolution time 3 cm-1. The spore sample was exposed for 140s with excitation laser power 30mW to obtain optimum peaks. The spectra revealed Raman frequencies range from 242 to 2690 cm-1. Raman spectroscopy shows a strong S-S stretching vibrational band in the region of 500-550 cm-1 suggesting that the...

  10. 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.

  11. A new chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica affects Soybean Asian rust (Phakopsora pachyrhizi spore germination

    Directory of Open Access Journals (Sweden)

    Mehta Angela

    2011-02-01

    Full Text Available Abstract Background Asian rust (Phakopsora pachyrhizi is a common disease in Brazilian soybean fields and it is difficult to control. To identify a biochemical candidate with potential to combat this disease, a new chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica (CaclXIP leaves was cloned into the pGAPZα-B vector for expression in Pichia pastoris. Results A cDNA encoding a chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica (CaclXIP, was isolated from leaves. The amino acid sequence predicts a (β/α8 topology common to Class III Chitinases (glycoside hydrolase family 18 proteins; GH18, and shares similarity with other GH18 members, although it lacks the glutamic acid residue essential for catalysis, which is replaced by glutamine. CaclXIP was expressed as a recombinant protein in Pichia pastoris. Enzymatic assay showed that purified recombinant CaclXIP had only residual chitinolytic activity. However, it inhibited xylanases from Acrophialophora nainiana by approx. 60% when present at 12:1 (w/w enzyme:inhibitor ratio. Additionally, CaclXIP at 1.5 μg/μL inhibited the germination of spores of Phakopsora pachyrhizi by 45%. Conclusions Our data suggests that CaclXIP belongs to a class of naturally inactive chitinases that have evolved to act in plant cell defence as xylanase inhibitors. Its role on inhibiting germination of fungal spores makes it an eligible candidate gene for the control of Asian rust.

  12. Characterization of Acid Soluble Collagen from Redbelly Yellowtail Fusilier Fish Skin (Caesio cuning

    Directory of Open Access Journals (Sweden)

    Ika Astiana

    2016-04-01

    Full Text Available Fish skin can be used as raw material for producing collagen. The collagen can be extracted by chemicalor combination of chemical and enzymatic processes. Extraction of collagen chemically can do with theacid process that produces acid soluble collagen (ASC. This study aimed to determine the optimumconcentration and time of pretreatment and extraction, also to determine the characteristics of the acidsoluble collagen from the skin of yellow tail fish. Extraction of collagen done by pretreatment using NaOH atthe concentration of 0.05; 0.1; and 0.15 M and extraction using acetic acid at the concentration of 0.3; 0.5; and0.7 M. Pretreatment NaOH with concentration 0.05 M and soaking time of 8 hours is the best combinationfor eliminating non collagen protein. Combination treatment of acetic acid at the concentration of 0.3 Mfor 3 days obtained the best solubility. The yield of collagen ASC was 18.4±1.49% (db and 5.79±0.47%(wb. Amino acid composition that is dominant in the ASC collagen was glycine (25.09±0.003%, alanine(13.71±0.075%, and proline (12.15±0.132%. Collagen from yellow tail fish skin has α1, α2, β and γprotein structure with the molecular weight of 125, 113, 170-181, and 208 KDa. The transition and meltingtemperatures of collagen were 67.69oC and 144.4oC. The surface structure of collagen by analysis of SEM hasfibers on the surface.Keywords: cholesterol, fatty acids, meat tissue, proximate, red snapper (L. argentimaculatus

  13. An ancient and conserved function for Armadillo-related proteins in the control of spore and seed germination by abscisic acid.

    Science.gov (United States)

    Moody, Laura A; Saidi, Younousse; Gibbs, Daniel J; Choudhary, Anushree; Holloway, Daniel; Vesty, Eleanor F; Bansal, Kiran Kaur; Bradshaw, Susan J; Coates, Juliet C

    2016-08-01

    Armadillo-related proteins regulate development throughout eukaryotic kingdoms. In the flowering plant Arabidopsis thaliana, Armadillo-related ARABIDILLO proteins promote multicellular root branching. ARABIDILLO homologues exist throughout land plants, including early-diverging species lacking true roots, suggesting that early-evolving ARABIDILLOs had additional biological roles. Here we investigated, using molecular genetics, the conservation and diversification of ARABIDILLO protein function in plants separated by c. 450 million years of evolution. We demonstrate that ARABIDILLO homologues in the moss Physcomitrella patens regulate a previously undiscovered inhibitory effect of abscisic acid (ABA) on spore germination. Furthermore, we show that A. thaliana ARABIDILLOs function similarly during seed germination. Early-diverging ARABIDILLO homologues from both P. patens and the lycophyte Selaginella moellendorffii can substitute for ARABIDILLO function during A. thaliana root development and seed germination. We conclude that (1) ABA was co-opted early in plant evolution to regulate functionally analogous processes in spore- and seed-producing plants and (2) plant ARABIDILLO germination functions were co-opted early into both gametophyte and sporophyte, with a specific rooting function evolving later in the land plant lineage. PMID:27040616

  14. Method for the manufacture of nitric acid soluble mixed oxide fuel pellets

    International Nuclear Information System (INIS)

    For the manufacture of nitric acid-soluble mixed oxide fuel pellets with adjustable proportions, the starting powder is ground down to a primary grain size of < 2 μm together with a halogen-free grinding aid and subsequently mixed. The change is then granulated in a rotating chamber, pressed into pellet form and sintered. (orig.)

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

    Science.gov (United States)

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

    2015-01-01

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

  16. Cryopreservation of fern spores

    Science.gov (United States)

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

  17. 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.

  18. ADAM Family Protein Mde10 Is Essential for Development of Spore Envelopes in the Fission Yeast Schizosaccharomyces pombe

    OpenAIRE

    Nakamura, Tomohiro; Abe, Hiroko; Hirata, Aiko; Shimoda, Chikashi

    2004-01-01

    We report the identification of Schizosaccharomyces pombe mde10+ as a gene possessing a FLEX element, which forms a binding site for the meiosis-specific transcription factor Mei4. In fact, mde10+ is transcribed only in diploid cells that are induced to meiosis in a Mei4-dependent manner. Western blot analysis indicated that the epitope-tagged Mde10 protein accumulates transiently during meiosis and then rapidly decreases. Mde10 is a multidomain protein containing a metalloprotease catalytic ...

  19. 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.

  20. Science hub spore data

    Data.gov (United States)

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

  1. 利用枯草芽孢衣壳蛋白表面展示β-半乳糖苷酶%Functional Display of β-galactosidase on the Spore Surface of Bacillus subtilis Using Spore Coat Protein as Anchor Motif

    Institute of Scientific and Technical Information of China (English)

    王贺; 杨瑞金; 华霄; 赵伟; 张文斌

    2012-01-01

    分别将枯草芽孢杆菌(Bacillussubtilis 168)芽孢衣壳蛋白CotB、CotC、CotG和CotX的启动子和编码序列与来自嗜热脂肪芽孢杆菌(BacillusstearothermophilusIAMll001)的β-半乳糖苷酶基因bgaB进行重组,构建融合表达cotB—bgaB、eotC—bgaB、eotG—bgaB和eotX—bgaB的整合型重组质粒。将4种重组质粒分别转入枯草芽孢杆菌Bacillussubtilis168(trp。),获得了能在芽孢表面展示的重组菌株PB701、PB702、PB703和PB704。经Westernblot检测,4种重组菌株均表达了预期分子量的融合蛋白,初步表明β-半乳糖苷酶被锚定在重组菌株的芽孢表面。以oNPG为底物测定4种重组菌株芽孢表面展示β-半乳糖苷酶的水解能力,得到的酶活分别为0.14、0.06、0.22和0.20U/mL。%In this work, we developed an efficient spore display system that a model protein β-galactosidase was anchored on the spore surface of Bacillus subtilis 168 based on the use of spore coat proteins. The PCR-amplifying cotB, cotC, cotG and cotX were ligated with pMD-19T and digested with XbaI and KpnI, and then subcloned into vector pJS700a previously digested with the same two restriction enzymes, finally resulted in the plasmids pJSB, pJSC, pJSG and pJSX. To construct the gene fusions, the bgaB from Bacillus stearothermophilus IAMll001 was cloned into the KpnI and EcoRI sites of plasmid pJSB, pJSC, pJS G and pJSX to generate generating the plasmids pJSBB, pJSCB, pJSGB and pJSXB,respectively After linearization with BgllI restriction endonuclease, the four re- combinant integrative plasmids were transformed into B. subtilis 168 to yield the recombinant strain PB701, PB702, PB703 and PB704,respectively. Results from Western blot analysis showed that the fusion protein was immobilized on the spore surface. Using oNPG as substrate, the enzyme activity of spore-displaying β-galactosidase was assayed and they were 0.14, 0.06, 0.22 and 0.20 U/mL for PB701, PB702, PB

  2. 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.

  3. Acid-soluble breakdown of homologous deoxyribonucleic acid adsorbed by Haemophilus influenzae: its biological significance

    Energy Technology Data Exchange (ETDEWEB)

    Stuy, J.H.

    1974-11-01

    Competent bacteria of Haemophilus influenzae strain Rd were exposed to various kinds of radioactive deoxyribonucleic acid (DNA) for short periods of time and at relatively low temperature. The fate of phage HP1 DNA was studied most extensively. Adsorbed DNA was partially acid solubilized by lysogens and by nonlysogens with very similar kinetics. The biological activity of the DNA decreased extensively in both lysogenic and nonlysogenic recipients. 2,4-Dinitrophenol had no effect on the acid solubilization but largely abolished the biological inactivation. Inactivation kinetics for three different markers and for the triple combination were roughly the same. The presence of 2,4-dinitrophenol in the medium, or the HP1 prophage in the chromosome, did not alter this observation. This suggests that acid solubilization involves the destruction of whole DNA molecules. In view of the absence of DNA homology between phage and host, it is concluded that acid-soluble breakdown of adsorbed transforming DNA is not an integral part of the donor DNA integration process. Behavior of mutant bacteria indicates that neither exonuclease III nor exonuclease V is involved.

  4. Anthrax Spores under a microscope

    Science.gov (United States)

    2003-01-01

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

  5. Spore: Spawning Evolutionary Misconceptions?

    Science.gov (United States)

    Bean, Thomas E.; Sinatra, Gale M.; Schrader, P. G.

    2010-10-01

    The use of computer simulations as educational tools may afford the means to develop understanding of evolution as a natural, emergent, and decentralized process. However, special consideration of developmental constraints on learning may be necessary when using these technologies. Specifically, the essentialist (biological forms possess an immutable essence), teleological (assignment of purpose to living things and/or parts of living things that may not be purposeful), and intentionality (assumption that events are caused by an intelligent agent) biases may be reinforced through the use of computer simulations, rather than addressed with instruction. We examine the video game Spore for its depiction of evolutionary content and its potential to reinforce these cognitive biases. In particular, we discuss three pedagogical strategies to mitigate weaknesses of Spore and other computer simulations: directly targeting misconceptions through refutational approaches, targeting specific principles of scientific inquiry, and directly addressing issues related to models as cognitive tools.

  6. Binding Affinity of Glycoconjugates to BACILLUS Spores and Toxins

    Science.gov (United States)

    Rasol, Aveen; Eassa, Souzan; Tarasenko, Olga

    2010-04-01

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

  7. New Rapid Spore Assay

    Science.gov (United States)

    Kminek, Gerhard; Conley, Catharine

    2012-07-01

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

  8. MECHANISM OF FUSARIUM TRICINCTUM (CORDA SACC. SPORE INACTIVATION BY CHLORINE DIOXIDE

    Directory of Open Access Journals (Sweden)

    Zhao Chen

    2015-06-01

    Full Text Available The mechanism of Fusarium tricinctum (Corda Sacc. spore inactivation by chlorine dioxide (ClO2 was investigated. During F. tricinctum spore inactivation by ClO2, protein, DNA, and metal ion leakage, enzyme activity, and cell ultrastructure were examined. Protein and DNA leakages were not detected, while there were metal ion leakages of K+, Ca2+, and Mg2+, which were well-correlated with the inactivation rate. The enzyme activities of glucose-6-phosphate dehydrogenase, citrate synthase, and phosphofructokinase were inhibited and were also well-correlated with the inactivation rate. Electron micrographs showed the ultrastructural modifications of spores and demonstrated that spores were heavily distorted and collapsed from their regular structure. Spore surface damage and disruption in inner components was also severe. The metal ion leakage, the inhibition of enzyme activities, and the damage of spore structure were significant in F. tricinctum spore inactivation by ClO2.

  9. The dynamics of acid-soluble phosphorus compounds in the course of winter and spring wheat germination under various thermic conditions. Part II. Labile phosphorus after hydrolysis of the acid-soluble fraction

    Directory of Open Access Journals (Sweden)

    A. Barbaro

    2015-06-01

    Full Text Available The changes in labile phosphorus compounds content during germination of wheat were investigated. These compounds were determined in acid-soluble germ extracts separated into fractions according to the solubility of their barium salts. Low germination temperature was found to raise the labile phosphorus content in the fraction of insoluble barium salts. If we assume that labile P of this fraction consisted mainly of adenosinedi- and triphosphates, it would seem that the rise, in the ATP and ADP level under the influence of low temperature may be essential for initiating flowering in winter varieties.

  10. Spore formation and toxin production in Clostridium difficile biofilms.

    Directory of Open Access Journals (Sweden)

    Ekaterina G Semenyuk

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2005-02-18

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

  12. Ultrastructure and properties of Paecilomyces lilacinus spores

    Energy Technology Data Exchange (ETDEWEB)

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

    2002-10-01

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

  13. Ultrastructure and properties of Paecilomyces lilacinus spores.

    Science.gov (United States)

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

    2002-10-01

    Strains of the filamentous soil fungus Paecilomyces lilacinus are currently being developed for use as biological control agents against root-knot, cyst, and other plant-parasitic nematodes. The inoculum applied in the field consists mainly of spores. This study was undertaken to examine the size, ultrastructure, and rodlet layers of P. lilacinus spores and the effect of the culture method on structural and functional spore properties. A rodlet layer was identified on aerial spores only. Other differences noted between aerial spores and those produced in submerged culture included the size and appearance of spores and thickness of spore coat layers when examined with transmission electron microscopy. The two spore types differed in UV tolerance, with aerial spores being less sensitive to environmentally relevant UV radiation. Also, viability after drying and storage was better with the aerial spores. Both spore types exhibited similar nematophagous ability. PMID:12489777

  14. Vacuum-induced Mutations In Bacillus Subtilis Spores

    Science.gov (United States)

    Munakata, N.; Maeda, M.; Hieda, K.

    During irradiation experiments with vacuum-UV radiation using synchrotron sources, we made unexpected observation that Bacillus subtilis spores of several recombination-deficient strains lost colony-forming ability by the exposure to high vacuum alone. Since this suggested the possible injury in spore DNA, we looked for mutation induction using the spores of strains HA101 (wild-type repair capability) and TKJ6312 (excision and spore repair deficient) that did not lose survivability. It was found that the frequency of nalidixic-acid resistant mutation increased several times in both of these strains by the exposure to high vacuum (10e-4 Pa after 24 hours). The analysis of sequence changes in gyrA gene showed that the majority of mutations carried a unique allele (gyrA12) of tandem double-base substitutions from CA to TT. The observation has been extended to rifampicin resistant mutations, the majority of that carried substitutions from CA to TT or AT in rpoB gene. On the other hand, when the spores of strains PS578 and PS2319 (obtained from P. Setlow) that are defective in a group of small acidic proteins (alpha/beta-type SASP) were similarly treated, none of the mutants analyzed carried such changes. This suggests that the unique mutations might be induced by the interaction of small acidic proteins with spore DNA under forced dehydration. The results indicate that extreme vacuum causes severe damage in spore DNA, and provide additional constraint to the long-term survival of bacterial spores in the space environment.

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

    Science.gov (United States)

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

    2016-06-20

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

  16. Hydrazine vapor inactivates Bacillus spores

    Science.gov (United States)

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

    2016-05-01

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

  17. Spore and the sociocultural moment

    Science.gov (United States)

    Meyer, W. Max

    2012-12-01

    Analyses of the game Spore have centered on the important issues of accuracy of evolution content and engendering interest in science. This paper suggests that examination of the degree of scaffolding necessary to use the game in pedagogy is a missing part of the discussion, and then questions the longevity of the Spore discussion relative to the general dissatisfaction with the science presented in the game. The paper proposes that analysis of Spore and other technological tools in science education may be embedded in an historical moment which directs the discussion towards satisfying sociocultural and organizational needs and away from pedagogical ones.

  18. 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.

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

    International Nuclear Information System (INIS)

    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)

  20. Translating physics to microbiology: spore resistance to terrestrial and extraterrestrial extremes

    OpenAIRE

    Moeller, Ralf; Raguse, Marina; Nagler, Katja; Fuchs, Felix M.

    2016-01-01

    Spore-forming bacteria are of particular concern in the context of planetary protection because their tough endospores are capable of withstanding certain sterilization procedures as well as harsh environments. Spores of Bacillus subtilis have been shown to be suitable dosimeters for probing extreme terrestrial and extraterrestrial environmental conditions in astrobiological and environmental studies. During dormancy spores are metabolically inactive; thus substantial DNA, protein, tRNA and r...

  1. 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.

  2. Isolation and characterization of acid-soluble collagen from the scales of marine fishes from Japan and Vietnam.

    Science.gov (United States)

    Minh Thuy, Le Thi; Okazaki, Emiko; Osako, Kazufumi

    2014-04-15

    Acid-soluble collagen (ASC) was successfully extracted from the scales of lizard fish (Saurida spp.) and horse mackerel (Trachurus japonicus) from Japan and Vietnam and grey mullet (Mugil cephalis), flying fish (Cypselurus melanurus) and yellowback seabream (Dentex tumifrons) from Japan. ASC yields were about 0.43-1.5% (on a dry weight basis), depending on the species. The SDS-PAGE profile showed that the ASCs were type I collagens, and consisted of two different α chains, α1 and α2, as well as a β component. ASC of horse mackerel from Vietnam contained a higher imino acid level than that from Japan. ASC denaturation temperature (Td) ranged from 26 to 29 °C, depending on fish species and imino acid content (pcollagens was observed at pHs 1-3. Collagen solubility decreased sharply at NaCl concentrations >0.4M, regardless of fish type. PMID:24295705

  3. The Role of Bacterial Spores in Metal Cycling and Their Potential Application in Metal Contaminant Bioremediation.

    Science.gov (United States)

    Butterfield, Cristina N; Lee, Sung-Woo; Tebo, Bradley M

    2016-04-01

    Bacteria are one of the premier biological forces that, in combination with chemical and physical forces, drive metal availability in the environment. Bacterial spores, when found in the environment, are often considered to be dormant and metabolically inactive, in a resting state waiting for favorable conditions for them to germinate. However, this is a highly oversimplified view of spores in the environment. The surface of bacterial spores represents a potential site for chemical reactions to occur. Additionally, proteins in the outer layers (spore coats or exosporium) may also have more specific catalytic activity. As a consequence, bacterial spores can play a role in geochemical processes and may indeed find uses in various biotechnological applications. The aim of this review is to introduce the role of bacteria and bacterial spores in biogeochemical cycles and their potential use as toxic metal bioremediation agents. PMID:27227313

  4. Spore populations among bulk tank raw milk and dairy powders are significantly different.

    Science.gov (United States)

    Miller, Rachel A; Kent, David J; Watterson, Matthew J; Boor, Kathryn J; Martin, Nicole H; Wiedmann, Martin

    2015-12-01

    To accommodate stringent spore limits mandated for the export of dairy powders, a more thorough understanding of the spore species present will be necessary to develop prospective strategies to identify and reduce sources (i.e., raw materials or in-plant) of contamination. We characterized 1,523 spore isolates obtained from bulk tank raw milk (n=33 farms) and samples collected from 4 different dairy powder-processing plants producing acid whey, nonfat dry milk, sweet whey, or whey protein concentrate 80. The spores isolated comprised 12 genera, at least 44 species, and 216 rpoB allelic types. Bacillus and Geobacillus represented the most commonly isolated spore genera (approximately 68.9 and 12.1%, respectively, of all spore isolates). Whereas Bacillus licheniformis was isolated from samples collected from all plants and farms, Geobacillus spp. were isolated from samples from 3 out of 4 plants and just 1 out of 33 farms. We found significant differences between the spore population isolated from bulk tank raw milk and those isolated from dairy powder plant samples, except samples from the plant producing acid whey. A comparison of spore species isolated from raw materials and finished powders showed that although certain species, such as B. licheniformis, were found in both raw and finished product samples, other species, such as Geobacillus spp. and Anoxybacillus spp., were more frequently isolated from finished powders. Importantly, we found that 8 out of 12 genera were isolated from at least 2 different spore count methods, suggesting that some spore count methods may provide redundant information if used in parallel. Together, our results suggest that (1) Bacillus and Geobacillus are the predominant spore contaminants in a variety of dairy powders, implying that future research efforts targeted at elucidating approaches to reduce levels of spores in dairy powders should focus on controlling levels of spore isolates from these genera; and (2) the spore

  5. Effects of meteorological conditions on spore plumes

    Science.gov (United States)

    Burch, M.; Levetin, E.

    2002-05-01

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

  6. Glycoprotein and protein markers for strain differentiation and growth environment or media attribution

    Energy Technology Data Exchange (ETDEWEB)

    Wunschel, David S.; Fox, Alvin; Wahl, Karen L.

    2012-01-01

    Recent experience with Bacillus spore characterization has demonstrated that protein markers can provide potentially vital identifying and bioforensic information. The masses of constitutively expressed proteins and their peptide fragments can be used to identify bacterial isolates. Protein marker mass variation information reflects the underlying amino acid sequence variation to provide complementary information to genetic sequence analysis. Protein markers (identified by mass or sequence) that are conserved or variable can be readily selected. In contrast, genetic primers, as used in PCR, target conserved genetic regions. Furthermore, protein markers are relatively stable compared to nucleic acids and may remain in samples for longer periods of time. This is important to consider when the source, age and condition of samples may vary in a forensic investigation. Examples of constitutively expressed proteins that have been extensively characterized include the exosporium BclA and BclB proteins and small acid soluble proteins (SASPs). Finally, gene expression (usually assessed at the mRNA level) can vary in response to different environmental conditions. As a result, the profile of protein markers of the organism also reflects the culture environment. Mass spectrometric tools can be used to access the same information on culture-related protein expression variation. However, unlike genetic methods, with proteomic methodology there is the potential to define exactly which medium was employed for organism growth. This potential could provide additional clues for forensic attribution

  7. 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...

  8. Triple fixation of Bacillus subtilis dormant spores.

    OpenAIRE

    Kozuka, S; Tochikubo, K

    1983-01-01

    A triple-fixation method with a sequential application of 5% glutaraldehyde, 1% osmium tetroxide, and 2% potassium permanganate gave superior preservation of the ultrastructure of Bacillus subtilis dormant spores with a thick spore coat.

  9. Perchloric acid-soluble proteins from goat liver inhibit chemical carcinogenesis of Syrian hamster cheek-pouch carcinoma

    OpenAIRE

    Ghezzo, F; G.N. Berta; Bussolati, B; Bosio, A; Corvetti, G.; Di Carlo, F.; Bussolati, G; Guglielmone, R; Bartorelli, A.

    1999-01-01

    Chemically induced Syrian hamster cheek-pouch squamous cell carcinoma is very similar to the corresponding human tumour. This paper describes a blind study in which inhibition of dimethylbenzanthracene-induced cheek-pouch tumours by a goat liver extract denominated UK101 was investigated. Less than 40% of animals treated with UK101 developed tumours compared with 100% of the controls. Intermediate results (80%) were noted in a positive control group treated with Calmette–Guérin bacillus. Immu...

  10. 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.

  11. 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.

  12. Natamycin and the germinating spore

    NARCIS (Netherlands)

    van Leeuwen, M.R.

    2009-01-01

    Fungi cause enormous food losses worldwide due to crop infection and food spoilage. Contamination by fungi often starts with dispersal vehicles (spores or conidia) that are dispersed either by air and water. A crucial step in fungal contamination is the process of germination, which is followed by m

  13. The Gonzo Scientist. Flunking Spore.

    Science.gov (United States)

    Bohannon, John

    2008-10-24

    The blockbuster video game Spore is being marketed as a science-based adventure that brings evolution, cell biology, and even astrophysics to the masses. But after grading the game's science with a team of researchers, the Gonzo Scientist has some bad news. PMID:18948523

  14. Influence of certain essential oils on carcinogen-metabolizing enzymes and acid-soluble sulfhydryls in mouse liver.

    Science.gov (United States)

    Banerjee, S; Sharma, R; Kale, R K; Rao, A R

    1994-01-01

    The influence of essential oils from naturally occurring plant dietary items such as cardamom, celery seed, cumin seed, coriander, ginger, nutmeg, and zanthoxylum on the activities of hepatic carcinogen-metabolizing enzymes (cytochrome P450, aryl hydrocarbon hydroxylase, and glutathione S-transferase) and acid-soluble sulfhydryl level was investigated in Swiss albino mice. Each oil was fed by gavage at 10 microliters/day for 14 days, and then the animals were sacrificed and their hepatic enzyme activities and sulfhydryl levels were evaluated. Only nutmeg and zanthoxylum oils induced cytochrome P450 level significantly (p oil caused a significant reduction in its activity (p oil (p nutmeg oil caused a significant reduction in its activity (p oils did not significantly alter the level of cytochrome P450 and aryl hydrocarbon hydroxylase activity. Glutathione S-transferase activity was significantly elevated in all experimental groups (p oils of cardamom (p nutmeg (p oils affects the host enzymes associated with activation and detoxication of xenobiotic compounds, including chemical carcinogens and mutagens. PMID:8072879

  15. MECHANISM OF FUSARIUM TRICINCTUM (CORDA) SACC. SPORE INACTIVATION BY CHLORINE DIOXIDE

    OpenAIRE

    Zhao Chen

    2015-01-01

    The mechanism of Fusarium tricinctum (Corda) Sacc. spore inactivation by chlorine dioxide (ClO2) was investigated. During F. tricinctum spore inactivation by ClO2, protein, DNA, and metal ion leakage, enzyme activity, and cell ultrastructure were examined. Protein and DNA leakages were not detected, while there were metal ion leakages of K+, Ca2+, and Mg2+, which were well-correlated with the inactivation rate. The enzyme activities of glucose-6-phosphate dehydrogenase, citrate synthase, and ...

  16. Isolating and Purifying Clostridium difficile Spores

    Science.gov (United States)

    Edwards, Adrianne N.; McBride, Shonna M.

    2016-01-01

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

  17. A Serum Response Factor homolog is required for spore differentiation in Dictyostelium.

    Science.gov (United States)

    Escalante, R; Sastre, L

    1998-10-01

    A homolog of the Serum Response Factor (SRF) has been isolated from Dictyostelium discoideum and its function studied by analyzing the consequences of its gene disruption. The MADS-box region of Dictyostelium SRF (DdSRF) is highly conserved with those of the human, Drosophila and yeast homologs. srfA is a developmentally regulated gene expressed in prespore and spore cells. This gene plays an essential role in sporulation as its disruption leads to abnormal spore morphology and loss of viability. The mutant spores were round and cellulose deposition seemed to be partially affected. Initial prestalk and prespore cell differentiation did not seem to be compromised in the mutant since the expression of several cell-type-specific markers were found to be unaffected. However, the mRNA level of the spore marker spiA was greatly reduced. Activation of the cAMP-dependent protein kinase (PKA) by 8-Br-cAMP was not able to fully bypass the morphological defects of srfA- mutant spores, although this treatment induced spiA mRNA expression. Our results suggest that DdSRF is required for full maturation of spores and participates in the regulation of the expression of the spore-coat marker spiA and probably other maturation genes necessary for proper spore cell differentiation. PMID:9729488

  18. Bacillus subtilis spores as vaccine adjuvants: further insights into the mechanisms of action.

    Directory of Open Access Journals (Sweden)

    Renata Damásio de Souza

    Full Text Available Bacillus subtilis spores have received growing attention regarding potential biotechnological applications, including the use as probiotics and in vaccine formulations. B. subtilis spores have also been shown to behave as particulate vaccine adjuvants, promoting the increase of antibody responses after co-administration with antigens either admixed or adsorbed on the spore surface. In this study, we further evaluated the immune modulatory properties of B. subtilis spores using a recombinant HIV gag p24 protein as a model antigen. The adjuvant effects of B. subtilis spores were not affected by the genetic background of the mouse lineage and did not induce significant inflammatory or deleterious effects after parenteral administration. Our results demonstrated that co-administration, but not adsorption to the spore surface, enhanced the immunogenicity of that target antigen after subcutaneous administration to BALB/c and C57BL/6 mice. Spores promoted activation of antigen presenting cells as demonstrated by the upregulation of MHC and CD40 molecules and enhanced secretion of pro-inflammatory cytokines by murine dendritic cells. In addition, in vivo studies indicated a direct role of the innate immunity on the immunomodulatory properties of B. subtilis spores, as demonstrated by the lack of adjuvant effects on MyD88 and TLR2 knockout mouse strains.

  19. Fatty Acid Profiles for Differentiating Growth Medium Formulations Used to Culture Bacillus cereus T-strain Spores.

    Science.gov (United States)

    Ehrhardt, Christopher J; Murphy, Devonie L; Robertson, James M; Bannan, Jason D

    2015-07-01

    Microbial biomarkers that indicate aspects of an organism's growth conditions are important targets of forensic research. In this study, we examined fatty acid composition as a signature for the types of complex nutrients in the culturing medium. Bacillus cereus T-strain spores were grown in medium formulations supplemented with one of the following: peptone (meat protein), tryptone (casein protein), soy protein, and brain-heart infusion. Cellular biomass was profiled with fatty acid methyl ester (FAME) analysis. Results showed peptone cultures produced spores enriched in straight-chained lipids. Tryptone cultures produced spores enriched in branched-odd lipids when compared with peptone, soy, and brain-heart formulations. The observed FAME variation was used to construct a set of discriminant functions that could help identify the nutrients in a culturing recipe for an unknown spore sample. Blinded classification tests were most successful for spores grown on media containing peptone and tryptone, showing 88% and 100% correct identification, respectively. PMID:25854710

  20. 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

  1. [Comparison of susceptibility of spores of Bacillus subtilis and Czech strains of Clostridium difficile to disinfectants].

    Science.gov (United States)

    Votava, M; Slitrová, B

    2009-02-01

    An important factor in the prevention of nosocomial outbreaks caused by Clostridium difficile ribotype 027 is the disinfection of a patient environment by reliable sporicidal disinfectants. Sporicidal activity of particular agents is tested on spores of Bacillus subtilis. Questions are brought up if the disinfectant which works on B. subtilis spores will be equally effective on the spores of C. difficile. Therefore we have compared the effects of five disinfectants available on the Czech market on the spores of collection strains of both microbes and on the spores of ten C. difficile field strains isolated from feces of hospitalized patients. The effective substances were: disinfectant No. 1 chloramine B, No. 2 chlorine dioxide, No. 3 formaldehyde and ethan-2-dion, No. 4 peracetic and acetic acids and hydrogen peroxide, No. 5 ethanol and propan-2-ol. The testing was performed using the dilution neutralization method according to (SN EN 13704, the agent reducing the number of spores by more than 3 orders was considered sporicidal. In addition to the standard time 60 min a 15-minutes exposition was used and the effect was tested also under the protein burden. Disinfectant No. 1 showed better effect on the C. difficile than B. subtilis spores, even in lower (1%) concentration. Similarly, the sensitivity of the C. difficile spores to disinfectants No. 2 and 3 was somewhat higher. The sporicidity of the disinfectant No. 4 was so high that it reduced the number of spores of all strains within 15 minutes by more than 4 orders; possible difference in the susceptibility of spores was not observed. Whereas the disinfectant No. 5 was not reliably effective on the spores of B. subtilis, surprisingly it showed the sporicidal effect on the spores of field C. difficile strains. We conclude that spores of field C. difficile strains in particular turned out to be more sensitive to disinfectants than the spores of the collection strain ofB. subtilis. Therefore B. subtilis remains

  2. Kinetics of germination of individual spores of Geobacillus stearothermophilus as measured by raman spectroscopy and differential interference contrast microscopy.

    Directory of Open Access Journals (Sweden)

    Tingting Zhou

    Full Text Available Geobacillus stearothermophilus is a gram-positive, thermophilic bacterium, spores of which are very heat resistant. Raman spectroscopy and differential interference contrast microscopy were used to monitor the kinetics of germination of individual spores of G. stearothermophilus at different temperatures, and major conclusions from this work were as follows. 1 The CaDPA level of individual G. stearothermophilus spores was similar to that of Bacillus spores. However, the Raman spectra of protein amide bands suggested there are differences in protein structure in spores of G. stearothermophilus and Bacillus species. 2 During nutrient germination of G. stearothermophilus spores, CaDPA was released beginning after a lag time (T(lag between addition of nutrient germinants and initiation of CaDPA release. CaDPA release was complete at T(release, and DT(release (T(release - T(lag was 1-2 min. 3 Activation by heat or sodium nitrite was essential for efficient nutrient germination of G. stearothermophilus spores, primarily by decreasing T(lag values. 4 Values of T(lag and T(release were heterogeneous among individual spores, but DT(release values were relatively constant. 5 Temperature had major effects on nutrient germination of G. stearothermophilus spores, as at temperatures below 65°C, average T(lag values increased significantly. 6 G. stearothermophilus spore germination with exogenous CaDPA or dodecylamine was fastest at 65°C, with longer T(lag values at lower temperatures. 7 Decoating of G. stearothermophilus spores slowed nutrient germination slightly and CaDPA germination significantly, but increased dodecylamine germination markedly. These results indicate that the dynamics and heterogeneity of the germination of individual G. stearothermophilus spores are generally similar to that of Bacillus species.

  3. Reticulate spore ornamentation in Strobilomyces

    Institute of Scientific and Technical Information of China (English)

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

    2011-01-01

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

  4. 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.

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

    OpenAIRE

    Cote, Christopher K.; Susan L. Welkos

    2015-01-01

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

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

    Science.gov (United States)

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

    2016-06-20

    Bacillus subtilis spores represent a suitable platform for the adsorption of proteins, enzymes and viral particles at physiological conditions. In the present work, we demonstrate that purified spores can also adsorb DNA on their surface after treatment with cationic molecules. In addition, we demonstrate that DNA-coated B. subtilis spores can be used as particulate carriers and act as an alternative to gold microparticles for the biolistic (gene gun) administration of plasmid DNA in mice. Gene gun delivery of spores pre-treated with DODAB (dioctadecyldimethylammonium bromide) allowed efficient plasmid DNA absorption and induced protein expression levels similar to those obtained with gold microparticles. More importantly, we demonstrated that a DNA vaccine adsorbed on spores can be loaded into biolistic cartridges and efficiently delivered into mice, which induced specific cellular and antibody responses. Altogether, these data indicate that B. subtilis spores represent a simple and low cost alternative for the in vivo delivery of DNA vaccines by the gene gun technology. PMID:27130499

  7. Ptaquiloside in Bracken Spores from Britain

    DEFF Research Database (Denmark)

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

    2013-01-01

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

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

    OpenAIRE

    Phillips, A. P.; Martin, K L

    1983-01-01

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

  9. 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

  10. 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.

  11. 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 metabolism

  12. 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)

    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

  13. Sensitive, Rapid Detection of Bacterial Spores

    Science.gov (United States)

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

    2009-01-01

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

  14. 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.

  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. Ptaquiloside in bracken spores from Britain

    DEFF Research Database (Denmark)

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

    2013-01-01

    Secondary metabolites from bracken fern (Pteridium aquilinum (L.) Kuhn) are suspected of causing cancer in humans. The main carcinogen is the highly water-soluble norsesquiterpene glucoside ptaquiloside, which may be ingested by humans through food, e.g. via contaminated water, meat or milk. It has...... in a collection of spores from Britain. Ptaquiloside was present in all samples, with a maximum of 29μgg−1, which is very low compared to other parts of the fern. Considering the low abundance of spores in breathing air under normal conditions, this exposure route is likely to be secondary to milk or drinking...

  17. 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

  18. CotC-CotU Heterodimerization during Assembly of the Bacillus subtilis Spore Coat▿

    OpenAIRE

    Isticato, Rachele; Pelosi, Assunta; Zilhão, Rita, 1959-; Baccigalupi, Loredana; Henriques, Adriano O.; De Felice, Maurilio; Ricca, Ezio

    2007-01-01

    We report evidence that CotC and CotU, two previously identified components of the Bacillus subtilis spore coat, are produced concurrently in the mother cell chamber of the sporulating cell under the control of σK and GerE and immediately assembled around the forming spore. In the coat, the two proteins interact to form a coat component of 23 kDa. The CotU-CotC interaction was not detected in two heterologous hosts, suggesting that it occurs only in B. subtilis. Monomeric forms of both CotU a...

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

    CERN Document Server

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

    2009-01-01

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

  20. Recovery of Heat Treated Bacillus cereus Spores Is Affected by Matrix Composition and Factors with Putative Functions in Damage Repair.

    Science.gov (United States)

    Warda, Alicja K; Tempelaars, Marcel H; Abee, Tjakko; Nierop Groot, Masja 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. We show that recovery of wet heat treated Bacillus cereus ATCC 14579 spores is affected by matrix composition with highest recovery in BHI broth or on rice agar plates, compared to BHI agar plates and rice broth. Data show that not only media composition but also its liquid or solid state affect the recovery of heat treated spores. To determine the impact of factors with putative roles in recovery of heat treated spores, specific genes previously shown to be highly expressed in outgrowing heat-treated spores were selected for mutant construction. Spores of nine B. cereus ATCC 14579 deletion mutants were obtained and their recovery from wet heat treatment was evaluated using BHI and rice broth and agar plates. Deletion mutant spores showed different capacity to recover from heat treatment compared to wild type with the most pronounced effect for a mutant lacking BC5242, a gene encoding a membrane protein with C2C2 zinc finger which resulted in over 95% reduction in recovery compared to the wild type in BHI broth. Notably, similar relative performance of wild type and mutants was observed using the other recovery conditions. We obtained insights on the impact of matrix composition and state on recovery of individually sorted heat treated spores and identified cellular factors with putative roles in this process. These results may provide leads for future developments in design of more efficient combined preservation treatments. PMID:27486443

  1. 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.

  2. 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

  3. 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 c

  4. 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.

  5. Effects of Citral on Aspergillus flavus Spores by Quasi-elastic Light Scattering and Multiplex Microanalysis Techniques

    Institute of Scientific and Technical Information of China (English)

    Man LUO; Li-Ke JIANG; Yao-Xiong HUANG; Ming XIAO; Bo LI; Guo-Lin ZOU

    2004-01-01

    Citral refined from Litsea cubeba oil has been found to have a strong influence on fungi,especially Aspergillus flavus. Multiplex microanalysis and quasi-elastic light scattering techniques were applied to study the effects of citral on Aspergillus flavus spores from the levels of membrane, organelle and intracellular macromolecule. It was found that citral injured the wall and the membrane of A. flavus spore,resulting in decrease of its elasticity. After entering the cell, citral not only influenced the genetic expression of mitochondrion reduplication and its morphology, but also changed the aggregation of protein-like macromolecules. As a result, cells, organelles and macromolecules lost their normal structures and functions,eventually leading to the loss of germination ability of A. flavus spores. Since Litsea cubeba oil as food additive and antifungal agent is safe and less poisonous, it is important to elucidate the inhibitory mechanisms of Litsea cubeba oil on the germination ability ofA. flavus spore.

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

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Malkin, A J

    2008-06-02

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

  7. Main airborne Ascomycota spores: characterization by culture, spore morphology, ribosomal DNA sequences and enzymatic analysis.

    Science.gov (United States)

    Oliveira, Manuela; Amorim, M Isabel; Ferreira, Elsa; Delgado, Luís; Abreu, Ilda

    2010-04-01

    The aim of this work was to identify the main allergy-related Ascomycetes fungal spores present in the atmosphere of Porto, using different and complementary techniques. The atmospheric sampling, performed in the atmosphere of Porto (Portugal) from August 2006 to July 2008, indicated Cladosporium, Penicillium, Aspergillus and Alternaria as the main fungal spore taxa. Alternaria and Cladosporium peaks were registered during summer. Aspergillus and Penicillium highest values were registered from late winter to early spring. Additionally, the Andersen sampler allowed the culture and isolation of the collected viable spores subsequently used for different identification approaches. The internal-transcribed spacer region of the nuclear ribosomal repeat unit sequences of airborne Ascomycetes fungi isolates revealed 11 taxonomically related fungal species. Among the identified taxa, Penicillum and Aspergillus presented the highest diversity, while only one species of Cladosporium and Alternaria, respectively, were identified. All selected fungal spore taxa possessed phosphatase, esterase, leucine arylamidase and beta-glucosidase enzymatic activity, while none had lipase, cystine arylamidase, trypsin or beta-glucuronidase activity. The association between the spore cell wall morphology, DNA-based techniques and enzymatic activity approaches allowed a more reliable identification procedure of the airborne Ascomycota fungal spores. PMID:20143229

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2001-07-01

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

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

    Science.gov (United States)

    Górny, Rafał L.; Reponen, Tiina; Grinshpun, Sergey A.; Willeke, Klaus

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

  10. Mushroom spore dispersal by convectively-driven winds

    CERN Document Server

    Dressaire, Emilie; Song, Boya; Roper, Marcus

    2015-01-01

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

  11. Spore development and nuclear inheritance in arbuscular mycorrhizal fungi

    Directory of Open Access Journals (Sweden)

    Hijri Mohamed

    2011-02-01

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

  12. Effect of individual or combined treatment by γ-irradiation or temperature (high or low) on bacillus subtilis spores and its application for sterilization of ground beef

    International Nuclear Information System (INIS)

    The combination of two lethal agents such as irradiation and temperature (high or sub zero) resulted in synergistic death or B. subtilis spores (as indicated by decrease in the thermal D-value). The extent of this synergism in killing a spore population depended mainly on the sequence on application of the two physical agents. Irradiation-temperature (high or sub zero) sequence killed more but injured less B. subtilis spores than temperature irradiation sequence or irradiation and temperature applied separately. Storage at -200C killed more spores than storage at -20C if carried after irradiation, while the reverse was true of storage was prior irradiation. An irradiation dose of 8 KGY followed by thermal exposure to 700C for 1 hr is suggested for the sterilization of ground beef. Irradiation induced certain quantitative changes on the amino-N, protein-N, RNA and DNA of the first subcultures of irradiated spores with stimulatory effect at low irradiation doses and inhibitory effect at the high irradiation doses. This might explain the increased sensitivity of irradiated spores to subsequent exposure to unfavourable temperature (high or sub zero). Exposure of B. subtilis spore to 700C induced a stimulation in the amino- and protein-N of the resulting cells while exposure to 800C resulted in a significant decrease in the amino-N. The protein-N remained more or less the same

  13. Thirty-four identifiable airborne fungal spores in Havana, Cuba.

    Science.gov (United States)

    Almaguer, Michel; Aira, María-Jesús; Rodríguez-Rajo, F Javier; Fernandez-Gonzalez, Maria; Rojas-Flores, Teresa I

    2015-01-01

    The airborne fungal spore content in Havana, Cuba, collected by means a non-viable volumetric methodology, was studied from November 2010 - October 2011. The study, from a qualitative point of view, allowed the characterization of 29 genera and 5 fungal types, described following the Saccardo´s morphotypes, as well as their morphobiometrical characteristics. In the amerospores morphotype, the conidia of 7 genera (with ascospores, basidiospores and uredospores) and 5 fungal types were included. Among phragmospores morphotype, the ascospores and conidia of 12 different genera were identified. The dictyospores morphotype only included conidial forms from 6 genera. Finally, the less frequent morphotypes were staurospores, didymospores and distosepted spores. In general, the main worldwide spread mitosporic fungi also predominated in the Havana atmosphere, accompanied by some ascospores and basidiospores. Cladosporium cladosporioides type was the most abundant with a total of 148,717 spores, followed by Leptosphaeria, Coprinus and the Aspergillus-Penicillium type spores, all of them with total values ranging from 20,591 - 16,392 spores. The higher monthly concentrations were registered in January (31,663 spores) and the lowest in December (7,314 spores). Generally, the average quantity of spores recorded during the months of the dry season (20,599 spores) was higher compared with that observed during the rainy season (17,460 spores).

  14. Airborne Spore Analysis of Karabük Atmosphere

    Directory of Open Access Journals (Sweden)

    Ayşe Kaplan

    2014-05-01

    Full Text Available In order to identify types and amounts of airborne allergenic spore dispersal in the atmosphere of Karabük by gravimetric method in 2006 and 2007, two Durham samplers were situated on roof and garden of Technical Education Faculty of Karabük University between the dates January 1, 2006 and December 31, 2007. As a result of the analysis a total of 2822.2±625.01 spore/cm2 spore quantity belonging to 21 types was identified. Of this total, 1106±250.33 spore/cm² was observed in 2006 and 1716±374.68 spore/cm² was observed in 2007. Spore concentrations revealed no statistically differences between two samplers (t=0.1527-1.1355, p>0.05. The relationship between spore concentrations and meteorological factors was displayed by Spearman Correlation analysis. The highest quantity of fungal spores and Myxomycetes were determined in June and July. Cladosporium, Alternaria, Ustilago, Myxomycetes and unidentified Ascomycetes spores were recorded as dominant. In the end of this study, a two-year spore calendar was prepared.

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

    Science.gov (United States)

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

    2016-08-01

    Clostridium species and Bacillus spp. are spore-forming bacteria that cause hospital infections. The spores from these bacteria are transmitted from patient to patient via healthcare workers' hands. Although alcohol-based hand rubbing is an important hand hygiene practice, it is ineffective against bacterial spores. Therefore, healthcare workers should wash their hands with soap when they are contaminated with spores. However, the extent of health care worker hand contamination remains unclear. The aim of this study is to determine the level of bacterial spore contamination on healthcare workers' hands. The hands of 71 healthcare workers were evaluated for bacterial spore contamination. Spores attached to subject's hands were quantitatively examined after 9 working hours. The relationship between bacterial spore contamination and hand hygiene behaviors was also analyzed. Bacterial spores were detected on the hands of 54 subjects (76.1%). The mean number of spores detected was 468.3 CFU/hand (maximum: 3300 CFU/hand). Thirty-seven (52.1%) and 36 (50.7%) subjects were contaminated with Bacillus subtilis and Bacillus cereus, respectively. Nineteen subjects (26.8%) were contaminated with both Bacillus species. Clostridium difficile was detected on only one subject's hands. There was a significant negative correlation between the hand contamination level and the frequency of handwashing (r = -0.44, P time since last handwashing (r = 0.34, P handwashing during daily patient care. PMID:27236515

  16. Correction of axial chromatic aberrations in confocal Raman microspectroscopic measurements of a single microbial spore.

    Science.gov (United States)

    Lasch, Peter; Hermelink, Antje; Naumann, Dieter

    2009-06-01

    Herein we describe a strategy for correcting the longitudinal or axial component of chromatic aberration in confocal Raman microspectroscopy. The method is based on measuring a vertical series of confocal Raman sections of samples by a high numerical aperture Raman microscope. Using the known characteristics of the wavelength-dependent focal shift of the optical system, the Raman intensities can be corrected to allow the rearrangement of Raman data from different focal planes. In the present study the computational correction routine was applied to an experimental data set of 4-dimensional (xyz spatial and the spectral dimension) confocal Raman spectra collected from single spores of Bacillus cereus. After correcting the axial component of the chromatic aberration, univariate and multivariate spectral parameters were obtained and used in the following for 3D segmentation and volume rendering on the basis of the structural and compositional information contained in the Raman spectra of the spore. Using univariate Raman intensities from defined functional group frequencies or k-means cluster membership values as a multivariate parameter for volume rendering, we demonstrate a high degree of correlation between confocal Raman microspectroscopy and the spores' morphology. In this paper we will also present cluster mean spectra which will be discussed in light of the presence of proteins and Ca-DPA, a calcium chelate of dipicolinic acid in the spore. PMID:19475143

  17. Assay for Spore Wall Integrity Using a Yeast Predator.

    Science.gov (United States)

    Okada, Hiroki; Neiman, Aaron M; Ohya, Yoshikazu

    2016-01-01

    During the budding yeast life cycle, a starved diploid cell undergoes meiosis followed by production of four haploid spores, each surrounded by a spore wall. The wall allows the spores to survive in harsh environments until conditions improve. Spores are also more resistant than vegetative cells to treatments such as ether vapor, glucanases, heat shock, high salt concentrations, and exposure to high or low pH, but the relevance of these treatments to natural environmental stresses remains unclear. This protocol describes a method for assaying the yeast spore wall under natural environmental conditions by quantifying the survival of yeast spores that have passed through the digestive system of a yeast predator, the fruit fly. PMID:27480715

  18. Live-imaging of Bacillus subtilis spore germination and outgrowth

    OpenAIRE

    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 eliminate or inactivate these bacterial spores in foods. In this regard food industry uses different preservation methods such as thermal-treatment, weak acids, antimicrobial compounds etc. Complete therm...

  19. Surface tension propulsion of fungal spores by use of microdroplets

    CERN Document Server

    Noblin, Xavier; Dumais, Jacques

    2010-01-01

    Many edible mushrooms eject their spores (about 10 microns in size) at high speed (about 1 m/s) using surface tension forces in a few microseconds. Basically the coalescence of a droplet with the spore generates the necessary momentum to eject the spore. We have detailed this mechanism in \\cite{noblin2}. In this article, we give some details about the high speed movies (up to 250000 fps) of mushrooms' spores ejection attached to this submission. This video was submitted as part of the Gallery of Fluid Motion 2010 which is showcase of fluid dynamics videos.

  20. Application of gaseous ozone for inactivation of Bacillus subtilis spores.

    Science.gov (United States)

    Aydogan, Ahmet; Gurol, Mirat D

    2006-02-01

    The effectiveness of gaseous ozone (O3) as a disinfectant was tested on Bacillus subtilis spores, which share the same physiological characteristics as Bacillus anthracis spores that cause the anthrax disease. Spores dried on surfaces of different carrier material were exposed to O3 gas in the range of 500-5000 ppm and at relative humidity (RH) of 70-95%. Gaseous O3 was found to be very effective against the B. subtilis spores, and at O3 concentrations as low as 3 mg/L (1500 ppm), approximately 3-log inactivation was obtained within 4 hr of exposure. The inactivation curves consisted of a short lag phase followed by an exponential decrease in the number of surviving spores. Prehydration of the bacterial spores has eliminated the initial lag phase. The inactivation rate increased with increasing O3 concentration but not >3 mg/L. The inactivation rate also increased with increase in RH. Different survival curves were obtained for various surfaces used to carry spores. Inactivation rates of spores on glass, a vinyl floor tile, and office paper were nearly the same. Whereas cut pile carpet and hardwood flooring surfaces resulted in much lower inactivation rates, another type of carpet (loop pile) showed significant enhancement in the inactivation of the spores. PMID:16568801

  1. Adhesion of Colletotrichum lindemuthianum Spores to Phaseolus vulgaris Hypocotyls and to Polystyrene

    OpenAIRE

    Young, David H.; Kauss, Heinrich

    1984-01-01

    Adhesion of Colletotrichum lindemuthianum spores to Phaseolus vulgaris hypocotyls and to polystyrene was inhibited by the respiratory inhibitors sodium azide and antimycin A, indicating a requirement for metabolic activity in adhesion. Various commercial proteins and Tween 80 also reduced adhesion to both surfaces. Binding was enhanced by the presence of salts: sodium, potassium, calcium, and magnesium chlorides were equally effective. The removal of surface wax from hypocotyls by chloroform ...

  2. MALDI-based intact spore mass spectrometry of downy and powdery mildews.

    Science.gov (United States)

    Chalupová, Jana; Sedlářová, Michaela; Helmel, Michaela; Rehulka, Pavel; Marchetti-Deschmann, Martina; Allmaier, Günter; Sebela, Marek

    2012-08-01

    Fast and easy identification of fungal phytopathogens is of great importance in agriculture. In this context, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as a powerful tool for analyzing microorganisms. This study deals with a methodology for MALDI-TOF MS-based identification of downy and powdery mildews representing obligate biotrophic parasites of crop plants. Experimental approaches for the MS analyses were optimized using Bremia lactucae, cause of lettuce downy mildew, and Oidium neolycopersici, cause of tomato powdery mildew. This involved determining a suitable concentration of spores in the sample, selection of a proper MALDI matrix, looking for the optimal solvent composition, and evaluation of different sample preparation methods. Furthermore, using different MALDI target materials and surfaces (stainless steel vs polymer-based) and applying various conditions for sample exposure to the acidic MALDI matrix system were investigated. The dried droplet method involving solvent evaporation at room temperature was found to be the most suitable for the deposition of spores and MALDI matrix on the target and the subsequent crystallization. The concentration of spore suspension was optimal between 2 and 5 × 10(9) spores per ml. The best peptide/protein profiles (in terms of signal-to-noise ratio and number of peaks) were obtained by combining ferulic and sinapinic acids as a mixed MALDI matrix. A pretreatment of the spore cell wall with hydrolases was successfully introduced prior to MS measurements to obtain more pronounced signals. Finally, a novel procedure was developed for direct mass spectra acquisition from infected plant leaves. PMID:22899506

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

    Science.gov (United States)

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

    2016-08-01

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

  4. Model simulations of fungal spore distribution over the Indian region

    Science.gov (United States)

    Ansari, Tabish U.; Valsan, Aswathy E.; Ojha, N.; Ravikrishna, R.; Narasimhan, Balaji; Gunthe, Sachin S.

    2015-12-01

    Fungal spores play important role in the health of humans, animals, and plants by constituting a class of the primary biological aerosol particles (PBAPs). Additionally, these could mediate the hydrological cycle by acting as nuclei for ice and cloud formation (IN and CCN respectively). Various processes in the biosphere and the variations in the meteorological conditions control the releasing mechanism of spores through active wet and dry discharge. In the present paper, we simulate the concentration of fungal spores over the Indian region during three distinct meteorological seasons by combining a numerical model (WRF-Chem) with the fungal spore emissions based on land-use type. Maiden high-resolution regional simulations revealed large spatial gradient and strong seasonal dependence in the concentration of fungal spores over the Indian region. The fungal spore concentrations are found to be the highest during winter (0-70 μg m-3 in December), moderately higher during summer (0-35 μg m-3 in May) and lowest during the monsoon (0-25 μg m-3 in July). The elevated concentrations during winter are attributed to the shallower boundary layer trapping the emitted fungal spores in smaller volume. In contrast, the deeper boundary layer mixing in May and stronger monsoonal-convection in July distribute the fungal spores throughout the lower troposphere (∼5 km). We suggest that the higher fungal spore concentrations during winter could have potential health impacts. While, stronger vertical mixing could enable fungal spores to influence the cloud formation during summer and monsoon. Our study provides the first information about the distribution and seasonal variation of fungal spores over the densely populated and observationally sparse Indian region.

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

    DEFF Research Database (Denmark)

    Holtgrewe-Stukenbrock, Eva; Rosendahl, Søren

    2005-01-01

    A nested multiplex PCR (polymerase chain reaction) approach was used for multilocus genotyping of arbuscular mycorrhizal fungal populations. This method allowed us to amplify multiple loci from Glomus single spores in a single PCR amplification. Variable introns in the two protein coding genes Gm...

  6. Live-imaging of Bacillus subtilis spore germination and outgrowth

    NARCIS (Netherlands)

    R. Pandey

    2014-01-01

    Spores of Gram-positive bacteria such as Bacillus and Clostridium cause huge economic losses to the food industry. In food products, spores survive under food preservation conditions and subsequent germination and outgrowth eventually causes food spoilage. Therefore efforts are being made to elimina

  7. 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.

  8. Chenodeoxycholate Is an Inhibitor of Clostridium difficile Spore Germination▿

    OpenAIRE

    Sorg, Joseph A.; Sonenshein, Abraham L.

    2008-01-01

    Some cholate derivatives that are normal components of bile can act with glycine to induce the germination of Clostridium difficile spores, but at least one bile component, chenodeoxycholate, does not induce germination. Here we show that chenodeoxycholate inhibits the germination of C. difficile spores in response to cholate and taurocholate.

  9. Comparison of hand hygiene procedures for removing Bacillus cereus spores.

    Science.gov (United States)

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

    2014-01-01

    Bacillus cereus is a spore-forming bacterium. B. cereus occasionally causes nosocomial infections, in which hand contamination with the spores plays an important role. Therefore, hand hygiene is the most important practice for controlling nosocomial B. cereus infections. This study aimed to determine the appropriate hand hygiene procedure for removing B. cereus spores. Thirty volunteers' hands were experimentally contaminated with B. cereus spores, after which they performed 6 different hand hygiene procedures. We compared the efficacy of the procedures in removing the spores from hands. The alcohol-based hand-rubbing procedures scarcely removed them. The soap washing procedures reduced the number of spores by more than 2 log10. Extending the washing time increased the spore-removing efficacy of the washing procedures. There was no significant difference in efficacy between the use of plain soap and antiseptic soap. Handwashing with soap is appropriate for removing B. cereus spores from hands. Alcohol-based hand-rubbing is not effective. PMID:25252644

  10. Factors influencing Saprolegnia spp. spore numbers in Norwegian salmon hatcheries.

    Science.gov (United States)

    Thoen, E; Evensen, Ø; Skaar, I

    2016-06-01

    A quantitative survey of Saprolegnia spp. in the water systems of Norwegian salmon hatcheries was performed. Water samples from 14 salmon hatcheries distributed along the Norwegian coastline were collected during final incubation in the hatcheries. Samples of inlet and effluent water were analyzed to estimate Saprolegnia propagule numbers. Saprolegnia spores were found in all samples at variable abundance. Number of spores retrieved varied from 50 to 3200 L(-1) in inlet water and from 30 to >5000 L(-1) in effluent water. A significant elevation of spore levels in effluent water compared to inlet water was detected. The estimated spore levels were related to recorded managerial and environmental parameters, and the number of spores in inlet water and temperature was the factor having most influence on the spore concentration in the incubation units (effluent water). Further, the relative impact of spore concentration on hatching rates was investigated by correlation analysis. From this was found that even high spore counts did not impact significantly on hatching success. PMID:26123005

  11. 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.

  12. Characterizing aeroallergens by infrared spectroscopy of fungal spores and pollen.

    Directory of Open Access Journals (Sweden)

    Boris Zimmermann

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

  13. 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.

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

    Science.gov (United States)

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

    2015-01-01

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

  15. Rugged single domain antibody detection elements for Bacillus anthracis spores and vegetative cells.

    Directory of Open Access Journals (Sweden)

    Scott A Walper

    Full Text Available Significant efforts to develop both laboratory and field-based detection assays for an array of potential biological threats started well before the anthrax attacks of 2001 and have continued with renewed urgency following. While numerous assays and methods have been explored that are suitable for laboratory utilization, detection in the field is often complicated by requirements for functionality in austere environments, where limited cold-chain facilities exist. In an effort to overcome these assay limitations for Bacillus anthracis, one of the most recognizable threats, a series of single domain antibodies (sdAbs were isolated from a phage display library prepared from immunized llamas. Characterization of target specificity, affinity, and thermal stability was conducted for six sdAb families isolated from rounds of selection against the bacterial spore. The protein target for all six sdAb families was determined to be the S-layer protein EA1, which is present in both vegetative cells and bacterial spores. All of the sdAbs examined exhibited a high degree of specificity for the target bacterium and its spore, with affinities in the nanomolar range, and the ability to refold into functional antigen-binding molecules following several rounds of thermal denaturation and refolding. This research demonstrates the capabilities of these sdAbs and their potential for integration into current and developing assays and biosensors.

  16. Methods for neutralizing anthrax or anthrax spores

    Energy Technology Data Exchange (ETDEWEB)

    Sloan, Mark A; Vivekandanda, Jeevalatha; Holwitt, Eric A; Kiel, Johnathan L

    2013-02-26

    The present invention concerns methods, compositions and apparatus for neutralizing bioagents, wherein bioagents comprise biowarfare agents, biohazardous agents, biological agents and/or infectious agents. The methods comprise exposing the bioagent to an organic semiconductor and exposing the bioagent and organic semiconductor to a source of energy. Although any source of energy is contemplated, in some embodiments the energy comprises visible light, ultraviolet, infrared, radiofrequency, microwave, laser radiation, pulsed corona discharge or electron beam radiation. Exemplary organic semiconductors include DAT and DALM. In certain embodiments, the organic semiconductor may be attached to one or more binding moieties, such as an antibody, antibody fragment, or nucleic acid ligand. Preferably, the binding moiety has a binding affinity for one or more bioagents to be neutralized. Other embodiments concern an apparatus comprising an organic semiconductor and an energy source. In preferred embodiments, the methods, compositions and apparatus are used for neutralizing anthrax spores.

  17. Novel Secretion Apparatus Maintains Spore Integrity and Developmental Gene Expression in Bacillus subtilis

    Science.gov (United States)

    Meisner, Jeffrey; Serrano, Monica; Henriques, Adriano O.; Moran, Charles P.; Rudner, David Z.

    2009-01-01

    Sporulation in Bacillus subtilis involves two cells that follow separate but coordinately regulated developmental programs. Late in sporulation, the developing spore (the forespore) resides within a mother cell. The regulation of the forespore transcription factor σG that acts at this stage has remained enigmatic. σG activity requires eight mother-cell proteins encoded in the spoIIIA operon and the forespore protein SpoIIQ. Several of the SpoIIIA proteins share similarity with components of specialized secretion systems. One of them resembles a secretion ATPase and we demonstrate that the ATPase motifs are required for σG activity. We further show that the SpoIIIA proteins and SpoIIQ reside in a multimeric complex that spans the two membranes surrounding the forespore. Finally, we have discovered that these proteins are all required to maintain forespore integrity. In their absence, the forespore develops large invaginations and collapses. Importantly, maintenance of forespore integrity does not require σG. These results support a model in which the SpoIIIA-SpoIIQ proteins form a novel secretion apparatus that allows the mother cell to nurture the forespore, thereby maintaining forespore physiology and σG activity during spore maturation. PMID:19609349

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

    Science.gov (United States)

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

    2015-11-01

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

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

    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

  20. 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

  1. 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

  2. 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

  3. Micromotors to capture and destroy anthrax simulant spores.

    Science.gov (United States)

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

    2015-03-01

    Towards addressing the need for detecting and eliminating biothreats, we describe a micromotor-based approach for screening, capturing, isolating and destroying anthrax simulant spores in a simple and rapid manner with minimal sample processing. The B. globilli antibody-functionalized micromotors can recognize, capture and transport B. globigii spores in environmental matrices, while showing non-interactions with excess of non-target bacteria. Efficient destruction of the anthrax simulant spores is demonstrated via the micromotor-induced mixing of a mild oxidizing solution. The new micromotor-based approach paves a way to dynamic multifunctional systems that rapidly recognize, isolate, capture and destroy biological threats. PMID:25622851

  4. [Sporogenesis, sporoderm and mature spore ornamentation in Lycopodiaceae].

    Science.gov (United States)

    Rincon Baron, Edgar Javier; Rolleri, Cristina Hilda; Passarelli, Lilian M; Espinosa Matías, Silvia; Torres, Alba Marina

    2014-09-01

    Studies on reproductive aspects, spore morphology and ultrastructure of Lycopodiaceae are not very common in the scientific literature, and constitute essential information to support taxonomic and systematic relationships among the group. In order to complete existing information, adding new and broader contributions on these topics, a comparative analysis of the sporogenesis ultrastructure, with emphasis on cytological aspects of the sporocyte coat development, tapetum, monoplastidic and polyplastidic meiosis, sporoderm ontogeny and ornamentation of the mature spores, was carried out in 43 taxa of eight genera of the Lycopodiaceae: Austrolycopodium, Diphasium, Diphasiastrum, Huperzia (including Phlegmariurus), Lycopodium, Lycopodiella, Palhinhaea and Pseudolycopodiella growing in the Andes of Colombia and the Neotropics. For this study, the transmission elec- tron microscopy (TEM) samples were collected in Cauca and Valle del Cauca Departments, while most of the spores for scanning electron microscopy (SEM) analysis were obtained from herbarium samples. We followed standard preparation procedures for spore observation by TEM and SEM. Results showed that the sporocyte coat is largely composed by primary wall components; the sporocyte develop much of their metabolic activity in the production of their coat, which is retained until the spores release; protective functions for the diploid cells undergoing meiosis is postulated here for this layer. The abundance of dictyosomes in the sporocyte cytoplasm was related to the formation and development of the sporocyte coat. Besides microtubule activity, the membrane of sporocyte folds, associated with electrodense material, and would early determine the final patterns of spore ornamentation. Monoplastidic condition is common in Lycopodium s.l., whereas polyplastidic condition was observed in species of Huperzia and Lycopodiella s. l. In monoplastidic species, the tapetum presents abun- dant multivesicular bodies, while in

  5. Identification of Major Sporulation Proteins of Myxococcus xanthus Using a Proteomic Approach▿

    OpenAIRE

    Dahl, John L.; Tengra, Farah K; Dutton, David; Yan, Jinyuan; Andacht, Tracy M.; Coyne, Lia; Windell, Veronica; Garza, Anthony G.

    2007-01-01

    Myxococcus xanthus is a soil-dwelling, gram-negative bacterium that during nutrient deprivation is capable of undergoing morphogenesis from a vegetative rod to a spherical, stress-resistant spore inside a domed-shaped, multicellular fruiting body. To identify proteins required for building stress-resistant M. xanthus spores, we compared the proteome of liquid-grown vegetative cells with the proteome of mature fruiting body spores. Two proteins, protein S and protein S1, were differentially ex...

  6. Investigating synergism during sequential inactivation of Bacillus subtilis spores with several disinfectants.

    Science.gov (United States)

    Cho, Min; Kim, Jae-Hong; Yoon, Jeyong

    2006-08-01

    The sequential application of ozone, chlorine dioxide, or UV followed by free chlorine was performed to investigate the synergistic inactivation of Bacillus subtilis spores. The greatest synergism was observed when chlorine dioxide was used as a primary disinfectant followed by secondary disinfection with free chlorine. A lesser synergistic effect was observed when ozone was used as the primary disinfectant, but no synergism was observed when UV was used as the primary disinfectant. When free chlorine was used as the primary disinfectant (i.e., sequential application in the reverse order), the synergistic effect was shown only when chlorine dioxide was applied as the secondary disinfectant. The synergistic effect observed could be related to damage to the spore coat during primary disinfection, suggested by the loss of proteins from spores during disinfectant treatment. The greatest synergism observed by the chlorine dioxide/free chlorine pair suggested that common reaction sites might exist for these disinfectants. The concept of percent synergistic effect was introduced to quantitatively compare the extent of synergistic effects in the sequential disinfection processes.

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

    DEFF Research Database (Denmark)

    Zimmerman, Kolea; Levitis, Daniel; Pringle, Anne

    2014-01-01

    In this 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....... In this fungus, pigmented spores are viable and unpigmented spores are inviable. These results show that while both parents influence all these traits, maternal influence is strongest on both fertility and mortality traits until the spores are physiologically independent of the maternal cytoplasm....

  8. Effects of temperature and desiccation on ex situ conservation of nongreen fern spores

    Science.gov (United States)

    Conservation of the genetic diversity of ferns is limited by the paucity of ex situ spore banks. Conflicting reports of fern spore response to low temperature and moisture impedes establishment of fern spore banks. There is little information available to evaluate longevity of fern spores under dif...

  9. Oxidation mechanism of Penicillium digitatum spores through neutral oxygen radicals

    Science.gov (United States)

    Hashizume, Hiroshi; Ohta, Takayuki; Takeda, Keigo; Ishikawa, Kenji; Hori, Masaru; Ito, Masafumi

    2014-01-01

    To investigate the inactivation process of Penicillium digitatum spores through neutral oxygen species, the spores were treated with an atmospheric-pressure oxygen radical source and observed in-situ using a fluorescent confocal-laser microscope. The treated spores were stained with two fluorescent dyes, 1,1‧-dioctadecyl-3,3,Y,3‧-tetramethylindocarbocyanine perchlorate (DiI) and diphenyl-1-pyrenylphosphine (DPPP). The intracellular organelles as well as the cell membranes in the spores treated with the oxygen radical source were stained with DiI without a major morphological change of the membranes. DPPP staining revealed that the organelles were oxidized by the oxygen radical treatment. These results suggest that neutral oxygen species, especially atomic oxygen, induce a minor structural change or functional inhibition of cell membranes, which leads to the oxidation of the intracellular organelles through the penetration of reactive oxygen species into the cell.

  10. Small Probes for Orbital Return of Experiments (SPORE) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Analogous to the CubeSat standardization of micro-satellites, the SPORE flight system architecture will utilize a modular design approach to provide low-cost...

  11. Surface Bacterial-Spore Assay Using Tb3+/DPA Luminescence

    Science.gov (United States)

    Ponce, Adrian

    2007-01-01

    Equipment and a method for rapidly assaying solid surfaces for contamination by bacterial spores are undergoing development. The method would yield a total (nonviable plus viable) spore count of a surface within minutes and a viable-spore count in about one hour. In this method, spores would be collected from a surface by use of a transparent polymeric tape coated on one side with a polymeric adhesive that would be permeated with one or more reagent(s) for detection of spores by use of visible luminescence. The sticky side of the tape would be pressed against a surface to be assayed, then the tape with captured spores would be placed in a reader that illuminates the sample with ultraviolet light and counts the green luminescence spots under a microscope to quantify the number of bacterial spores per unit area. The visible luminescence spots seen through the microscope would be counted to determine the concentration of spores on the surface. This method is based on the chemical and physical principles of methods described in several prior NASA Tech Briefs articles, including Live/Dead Spore Assay Using DPA-Triggered Tb Luminescence (NPO-30444), Vol. 27, No. 3 (March 2003), page 7a. To recapitulate: The basic idea is to exploit the observations that (1) dipicolinic acid (DPA) is present naturally only in bacterial spores; and (2) when bound to Tb3+ ions, DPA triggers intense green luminescence of the ions under ultraviolet excitation; (3) DPA can be released from the viable spores by using L-alanine to make them germinate; and (4) by autoclaving, microwaving, or sonicating the sample, one can cause all the spores (non-viable as well as viable) to release their DPA. One candidate material for use as the adhesive in the present method is polydimethysiloxane (PDMS). In one variant of the method for obtaining counts of all (viable and nonviable) spores the PDMS would be doped with TbCl3. After collection of a sample, the spores immobilized on the sticky tape surface

  12. Anthrax Spores Make an Essential Contribution to Vaccine Efficacy

    OpenAIRE

    Brossier, Fabien; Levy, Martine; Mock, Michèle

    2002-01-01

    Anthrax is caused by Bacillus anthracis, a gram-positive spore-forming bacterium. Septicemia and toxemia rapidly lead to death in infected mammal hosts. Currently used acellular vaccines against anthrax consist of protective antigen (PA), one of the anthrax toxin components. However, in experimental animals such vaccines are less protective than live attenuated strains. Here we demonstrate that the addition of formaldehyde-inactivated spores (FIS) of B. anthracis to PA elicits total protectio...

  13. 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.

  14. Natural Dissemination of Bacillus anthracis Spores in Northern Canada

    OpenAIRE

    Dragon, D C; Bader, D. E.; Mitchell, J.; Woollen, N.

    2005-01-01

    Soil samples were collected from around fresh and year-old bison carcasses and areas not associated with known carcasses in Wood Buffalo National Park during an active anthrax outbreak in the summer of 2001. Sample selection with a grid provided the most complete coverage of a site. Soil samples were screened for viable Bacillus anthracis spores via selective culture, phenotypic analysis, and PCR. Bacillus anthracis spores were isolated from 28.4% of the samples. The highest concentrations of...

  15. Lipids stimulate spore germination in the entomopathogenic ascomycete Ascosphaera aggregata.

    Science.gov (United States)

    James, R R; Buckner, J S

    2004-10-01

    The alfalfa leafcutting bee (Megachile rotundata) is solitary and managed on a large scale for pollination of alfalfa seed crops. The bees nest in holes drilled in wood or polystyrene blocks, and their larvae are highly prone to a fungal disease called chalkbrood. The most prevalent form of chalkbrood is caused by Ascosphaera aggregata, but this ascomycete is difficult to culture. Hyphae will grow on standard fungal media, but spore germination is difficult to achieve and highly variable. We found that germination can be enhanced with oils. Lipids derived from plants and bee larvae increased germination from 50% (without oil) to 75-85% (with oil). Percent germination was significantly greater in the presence of lipids but germination was not significantly different when different oils, including mineral oil, were used. A. aggregata spores oriented along the oil-aqueous interface in the broth in a polar fashion, with swelling and germ tube formation always occurring into the aqueous portion of the broth. The other half of the spore tended to attach to a lipid droplet, where it remained, without swelling, during germ tube formation. The physical attachment of spores to the oil-aqueous interface is what most probably stimulates spore germination, as opposed to some nutritional stimulation. However, further research is needed to determine if and where the spores encounter such an interface when germinating in the host gut, where germination normally occurs. PMID:15645171

  16. Availability of websites offering to sell psilocybin spores and psilocybin.

    Science.gov (United States)

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

    2009-09-01

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

  17. The Luna stain, an improved selective stain for detection of microsporidian spores in histologic sections

    OpenAIRE

    Peterson, Tracy S.; Spitsbergen, Jan M.; Feist, Stephen W.; Kent, Michael L

    2011-01-01

    Microsporidia in histologic sections are most often diagnosed by observing spores in host tissues. Spores are easy to identify if they occur in large aggregates or xenomas when sections are stained with hematoxylin and eosin (H&E). However, individual spores are not frequently detected in host tissues with conventional H&E staining, particularly if spores are scattered within the tissues, areas of inflammation or small spores in nuclei (i.e., Nucleospora salmonis). Hence, a variety of selecti...

  18. Quantitative Analysis of Spatial-Temporal Correlations during Germination of Spores of Bacillus Species ▿

    OpenAIRE

    Zhang, JinQiao; Garner, Will; Setlow, Peter; Yu, Ji

    2011-01-01

    Bacteria of Bacillus species sporulate upon starvation, and the resultant dormant spores germinate when the environment appears likely to allow the resumption of vegetative growth. Normally, the rates of germination of individual spores in populations are very heterogeneous, and the current work has investigated whether spore-to-spore communication enhances the synchronicity of germination. In order to do this work, time-lapse optical images of thousands of individual spores were captured dur...

  19. UV-Photobiology of bacterial spores in space

    Science.gov (United States)

    Horneck, Gerda; Douki, Thierry; Cadet, Jean; Panitz, Corinna; Rabbow, Elke; Moeller, Ralf; Rettberg, Petra

    The vast, cold and radiation filled regimes of outer space present on one hand an environmental challenge for any form of terrestrial life; on the other hand they constitute a unique platform for astrobiology research. Major environmental parameters of space that are of interest to astrobiology are (i) space vacuum, (ii) solar electromagnetic radiation, above all the high energy UV radiation, (iii) galactic cosmic radiation, (iv) extreme temperature fluctuations, and (v) microgravity. Exposure facilities on board of Earth orbiting satellites and the International Space Station (ISS) have provided unique opportunities to study biological and chemical processes in response to those parameters directly in space. Endospores of Bacillus spp., especially B. subtilis, characterized by an extreme resistance to environmental insults and an incredible longevity have served as experimental models in studies on (i) the role of the ozone layer in protecting our biosphere; (ii) the likelihood of the interplanetary transfer of life via meteorites, i.e. the hypothesis of lithopanspermia; (iii) the habitability of Mars; (iv) the need for planetary protection measures; and (v) the molecular mechanisms underlying the extreme lethality of solar extraterrestrial UV-radiation. Role of the ozone layer in protecting our biosphere: Using solar extraterrestrial UV radiation and a set of optical filters, the terrestrial UV radiation climate at different ozone concentration was simulated and the biologically effective irradiance was measured with B. subtilis spores immobilized in a biofilm. With decreasing (simulated) ozone concentrations the biologically effective solar irradiance strongly increased by nearly 1000-fold for early Earth conditions before the ozone layer was built up. Likelihood of lithopanspermia: In an impact-driven scenario of lithopanspermia, rock-dwelling microorganisms - after being ejected from a planet - may wander through space for extended periods of time before being

  20. Investigating the Inactivation Mechanism of Bacillus subtilis Spores by High Pressure CO2.

    Science.gov (United States)

    Rao, Lei; Zhao, Feng; Wang, Yongtao; Chen, Fang; Hu, Xiaosong; Liao, Xiaojun

    2016-01-01

    The objective of this study was to investigate the inactivation mechanism of Bacillus subtilis spores by high pressure CO2 (HPCD) processing. The spores of B. subtilis were subjected to heat at 0.1 MPa or HPCD at 6.5-20 MPa, and 64-86°C for 0-120 min. The germination, the permeability of inner membrane (IM) and cortex, the release of pyridine-2, 6-dicarboxylic acid (DPA), and changes in the morphological and internal structures of spores were investigated. The HPCD-treated spores did not lose heat resistance and their DPA release was lower than the inactivation, suggesting that spores did not germinate during HPCD. The flow cytometry analysis suggested that the permeability of the IM and cortex of HPCD-treated spores was increased. Furthermore, the DPA of the HPCD-treated spores were released in parallel with their inactivation and the fluorescence photomicrographs showed that these treated spores were stained by propidium iodide, ensuring that the permeability of IM of spores was increased by HPCD. The scanning electron microscopy photomicrographs showed that spores were crushed into debris or exhibited a hollowness on the surface, and the transmission electron microscopy photomicrographs exhibited an enlarged core, ruptured and indistinguishable IM and a loss of core materials in the HPCD-treated spores, indicating that HPCD damaged the structures of the spores. These findings suggested that HPCD inactivated B. subtilis spores by directly damaging the structure of the spores, rather than inducing germination of the spores. PMID:27656175

  1. Effect of Acid Soluble Aluminium on Inclusions in Non-Oriented Electrical Steel%无取向电工钢中酸溶铝含量对夹杂物的影响

    Institute of Scientific and Technical Information of China (English)

    夏文勇; 刘静静; 秦哲; 侯泽旺; 仇圣桃; 王建军

    2012-01-01

    为提高无取向电工钢成品性能及优化生产,采用SEM-EDS等分析方法分别对比分析了无取向电工钢生产时,不同酸溶铝含量的生产工艺对钢中夹杂物的影响.研究表明,当钢中酸溶铝含量w(Als)为0.2%~0.4%时,精炼过程所生成的夹杂物为高熔点的Al2O3和球状的MgO-Al2O3类夹杂物;当钢中w(A1s)小于0.005%时,精炼过程所生成的夹杂物主要为塑性硅铝锰酸盐类夹杂;各工位夹杂物平均数量呈递减的趋势,且w(A1s)为0.2%~0.4%的高碱度渣系控制生成的夹杂物总量低于后者.%In order to improve the properties of non-oriented electrical steel and optimize the production, the effect of different content of acid soluble aluminium on inclusions in non-oriented electrical steel was investigated by the method of SEM-EDS. The results show that when acid-soluble aluminium content in steel is 0. 2%-0. 4%, inclusions in the refining stage are AI2 O3 with high melting point and globular MgO-Al2 O3. While acid-soluble aluminium content in steel less than 0. 005%, inclusions in the refining stage are Si-Al mixed manganese salts. The average size distribution of inclusion in each station is followed by a decreasing trend.

  2. 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

    2016-04-01

    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. PMID:26801572

  3. Fungal Spores Viability on the International Space Station

    Science.gov (United States)

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

    2016-11-01

    In this study we investigated the security of a spaceflight experiment from two points of view: spreading of dried fungal spores placed on the different wafers and their viability during short and long term missions on the International Space Station (ISS). Microscopic characteristics of spores from dried spores samples were investigated, as well as the morphology of the colonies obtained from spores that survived during mission. The selected fungal species were: Aspergillus niger, Cladosporium herbarum, Ulocladium chartarum, and Basipetospora halophila. They have been chosen mainly based on their involvement in the biodeterioration of different substrate in the ISS as well as their presence as possible contaminants of the ISS. From biological point of view, three of the selected species are black fungi, with high melanin content and therefore highly resistant to space radiation. The visual inspection and analysis of the images taken before and after the short and the long term experiments have shown that all biocontainers were returned to Earth without damages. Microscope images of the lids of the culture plates revealed that the spores of all species were actually not detached from the surface of the wafers and did not contaminate the lids. From the adhesion point of view all types of wafers can be used in space experiments, with a special comment on the viability in the particular case of iron wafers when used for spores that belong to B. halophila (halophilic strain). This is encouraging in performing experiments with fungi without risking contamination. The spore viability was lower in the experiment for long time to ISS conditions than that of the short experiment. From the observations, it is suggested that the environment of the enclosed biocontainer, as well as the species'specific behaviour have an important effect, reducing the viability in time. Even the spores were not detached from the surface of the wafers, it was observed that spores used in the

  4. Fungal Spores Viability on the International Space Station

    Science.gov (United States)

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

    2016-04-01

    In this study we investigated the security of a spaceflight experiment from two points of view: spreading of dried fungal spores placed on the different wafers and their viability during short and long term missions on the International Space Station (ISS). Microscopic characteristics of spores from dried spores samples were investigated, as well as the morphology of the colonies obtained from spores that survived during mission. The selected fungal species were: Aspergillus niger, Cladosporium herbarum, Ulocladium chartarum, and Basipetospora halophila. They have been chosen mainly based on their involvement in the biodeterioration of different substrate in the ISS as well as their presence as possible contaminants of the ISS. From biological point of view, three of the selected species are black fungi, with high melanin content and therefore highly resistant to space radiation. The visual inspection and analysis of the images taken before and after the short and the long term experiments have shown that all biocontainers were returned to Earth without damages. Microscope images of the lids of the culture plates revealed that the spores of all species were actually not detached from the surface of the wafers and did not contaminate the lids. From the adhesion point of view all types of wafers can be used in space experiments, with a special comment on the viability in the particular case of iron wafers when used for spores that belong to B. halophila (halophilic strain). This is encouraging in performing experiments with fungi without risking contamination. The spore viability was lower in the experiment for long time to ISS conditions than that of the short experiment. From the observations, it is suggested that the environment of the enclosed biocontainer, as well as the species'specific behaviour have an important effect, reducing the viability in time. Even the spores were not detached from the surface of the wafers, it was observed that spores used in the

  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.; Beilen, J.van; Caspers, M.; O'Brien, A.; Koster, C.de; Oomes, S.; Smelt, J.; Kort, R.; Beek, A.ter

    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. Dna stability and survival of bacillus subtilis spores in extreme dryness

    Science.gov (United States)

    Dose, Klaus; Gill, Markus

    1995-06-01

    The inactivation of Bacillus subtilis spores during long-term exposure (up to several months) to extreme dryness (especially vacuum) is strain-dependent, through only to a small degree. During a first phase (lasting about four days) monolayers of spores lose about 20% of their viability, regardless of the strain studied. During this phase loss in viability can be equally attributed both to damages of hydrophobic structures (membranes and proteins) and DNA. During a second phase lasting for the remaining time of experimental observation (weeks, months and years) the loss in viability is slowed. A viability of 55% to 75% (depending on the strain) is attained after a total exposure of 36 days. The loss in viability during the second phase can be correlated with the occurrence of DNA double strand breaks. Also covalent DNA-protein cross-links are formed by vacuum exposure. If the protein moiety of these cross-links is degraded by proteinase K-treatment in vitro additional DNA double strand breaks result. The data are also discussed with respect to survival on Mars and in near Earth orbits.

  7. Characteristics Comparison of Acid-soluble and Pepsin-soluble Collagens from bond of Lateolabrax j aponicus C%鲈鱼骨酸溶性和酶溶性胶原的性质比较

    Institute of Scientific and Technical Information of China (English)

    马国红; 张延华; 宋理平

    2015-01-01

    以鲈鱼骨为原料提取得到酸溶性胶原(ASC)和酶溶性胶原(PSC),对ASC和PSC的性质进行比较。粘度测定结果表明,ASC的变性温度25~30℃,PSC的变性温度为30~35℃;电泳结果表明,ASC和 PSC都属于Ⅰ型胶原且纯度比较高;氨基酸分析结果显示,羟脯氨酸和脯氨酸含量ASC均低于PSC ,氨基酸检测结果与粘度检测结果相一致。%Acid-soluble and pepsin-soluble collagens (ASC and PSC) were extracted from the bond of L ate‐olabrax j aponicus C and partially characterized .The compositions and certain properties of Acid-soluble and pepsin-soluble collagens (ASC and PSC) of the bond of Lateolabrax japonicus C .were strdied .The denat‐uration temperature (Td) of collagens were researched by viscosity of collagen solution .The Td of ASC and PSC from bone were 25~30 ℃ and 30~35 ℃ ,respectively .Electrophoretic patterns of collagens from bone were high purity Collagen typeⅠ,hydroxyproline and proline were lower the ASC than the PSC .which was in accordance with the results of composition analysis and Td .

  8. Inactivation of chemical and heat-resistant spores of Bacillus and Geobacillus by nitrogen cold atmospheric plasma evokes distinct changes in morphology and integrity of spores.

    Science.gov (United States)

    van Bokhorst-van de Veen, Hermien; Xie, Houyu; Esveld, Erik; Abee, Tjakko; Mastwijk, Hennie; Nierop Groot, Masja

    2015-02-01

    Bacterial spores are resistant to severe conditions and form a challenge to eradicate from food or food packaging material. Cold atmospheric plasma (CAP) treatment is receiving more attention as potential sterilization method at relatively mild conditions but the exact mechanism of inactivation is still not fully understood. In this study, the biocidal effect by nitrogen CAP was determined for chemical (hypochlorite and hydrogen peroxide), physical (UV) and heat-resistant spores. The three different sporeformers used are Bacillus cereus a food-borne pathogen, and Bacillus atrophaeus and Geobacillus stearothermophilus that are used as biological indicators for validation of chemical sterilization and thermal processes, respectively. The different spores showed variation in their degree of inactivation by applied heat, hypochlorite, hydrogen peroxide, and UV treatments, whereas similar inactivation results were obtained with the different spores treated with nitrogen CAP. G. stearothermophilus spores displayed high resistance to heat, hypochlorite, hydrogen peroxide, while for UV treatment B. atrophaeus spores are most tolerant. Scanning electron microscopy analysis revealed distinct morphological changes for nitrogen CAP-treated B. cereus spores including etching effects and the appearance of rough spore surfaces, whereas morphology of spores treated with heat or disinfectants showed no such changes. Moreover, microscopy analysis revealed CAP-exposed B. cereus spores to turn phase grey conceivably because of water influx indicating damage of the spores, a phenomenon that was not observed for non-treated spores. In addition, data are supplied that exclude UV radiation as determinant of antimicrobial activity of nitrogen CAP. Overall, this study shows that nitrogen CAP treatment has a biocidal effect on selected Bacillus and Geobacillus spores associated with alterations in spore surface morphology and loss of spore integrity. PMID:25481059

  9. 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

  10. Effect of Nanofibers on Spore Penetration and Lunar Dust Filtration

    Directory of Open Access Journals (Sweden)

    Phil Gibson, Ph.D.

    2008-06-01

    Full Text Available The results of two separate studies on biological spore penetration and simulated lunar dust filtration illustrate the use of nanofibers in some nonstandard filtration applications (nanofibers are generally defined as having diameters of less than a micron. In the first study, a variety of microporous liners containing microfibers and nanofibers were combined with cotton-based fabrics in order to filter aerosolized spores. The aerosol penetration resistance of the nanofiber-lined fabrics was measured, and some analysis was conducted of where the particles are captured within the fabric layers. Testing was conducted with aerosolized living spores, in order to evaluate the efficacy of various fabric treatments on spore viability within the fabric layers after exposure. Reported are the results of studies on fabrics with and without a removable electrospun nanofiber liner, and the fate of the spores within the fabric layers. In the second study, non-instrumented filtration testing using simulated lunar dust determined the comparative filtration efficiency of various nonwoven filtration media. Nanofiber witness media, combined with scanning electron microscope images, showed that an electrospun nonwoven filter layer effectively filtered out all the large and fine particles of the simulated lunar dust.

  11. 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.

  12. Environmental Persistence of Bacillus anthracis and Bacillus subtilis Spores

    Science.gov (United States)

    Wood, Joseph P.; Meyer, Kathryn M.; Kelly, Thomas J.; Choi, Young W.; Rogers, James V.; Riggs, Karen B.; Willenberg, Zachary J.

    2015-01-01

    There is a lack of data for how the viability of biological agents may degrade over time in different environments. In this study, experiments were conducted to determine the persistence of Bacillus anthracis and Bacillus subtilis spores on outdoor materials with and without exposure to simulated sunlight, using ultraviolet (UV)-A/B radiation. Spores were inoculated onto glass, wood, concrete, and topsoil and recovered after periods of 2, 14, 28, and 56 days. Recovery and inactivation kinetics for the two species were assessed for each surface material and UV exposure condition. Results suggest that with exposure to UV, decay of spore viability for both Bacillus species occurs in two phases, with an initial rapid decay, followed by a slower inactivation period. The exception was with topsoil, in which there was minimal loss of spore viability in soil over 56 days, with or without UV exposure. The greatest loss in viable spore recovery occurred on glass with UV exposure, with nearly a four log10 reduction after just two days. In most cases, B. subtilis had a slower rate of decay than B. anthracis, although less B. subtilis was recovered initially. PMID:26372011

  13. A cationic lipid-formulated plasmid DNA vaccine confers sustained antibody-mediated protection against aerosolized anthrax spores

    OpenAIRE

    Hermanson, G; Whitlow, V.; Parker,S; Tonsky, K.; Rusalov, D.; Ferrari, M.; Lalor, P; Komai, M.; Mere, R.; Bell, M.; Brenneman, K; Mateczun, A.; Evans, T.; Kaslow, D.; Galloway, D

    2004-01-01

    DNA vaccines provide an attractive technology platform against bioterrorism agents due to their safety record in humans and ease of construction, testing, and manufacture. We have designed monovalent and bivalent anthrax plasmid DNA (pDNA) vaccines encoding genetically detoxified protective antigen (PA) and lethal factor (LF) proteins and tested their immunogenicity and ability to protect rabbits from an aerosolized inhalation spore challenge. Immune responses after two or three injections of...

  14. Antibacterial action of gramicidin S and tyrocidines in relation to active transport, in vitro transcription, and spore outgrowth.

    OpenAIRE

    Danders, W; Marahiel, M A; Krause, M.; Kosui, N; Kato, T.; Izumiya, N; Kleinkauf, H

    1982-01-01

    The cyclopeptide antibiotic gramicidin S or tyrocidine in concentrations of 2 to 4 mumol/mg of membrane protein inhibited the active transport of [3H]alanine and [3H]uridine in membrane vesicles isolated from Bacillus brevis and Bacillus subtilis. We used one analog of gramicidin S and two of tyrocidine A to study the relationship between peptide structure and antibacterial action as seen in inhibiting active transport and in vitro transcription and in delaying spore outgrowth. The data showe...

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

    Science.gov (United States)

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

    2016-07-01

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

  16. BzpF is a CREB-like transcription factor that regulates spore maturation and stability in Dictyostelium

    OpenAIRE

    Huang, Eryong; Hughes, Timothy R; Katoh, Mariko; Shaulsky, Gad; Zupan, Blaž; Talukder, Shaheynoor; Curk, Tomaž

    2015-01-01

    The cAMP response element-binding protein (CREB) is a highly conserved transcription factor that integrates signaling through the cAMP-dependent protein kinase A (PKA) in many eukaryotes. PKA plays a critical role in Dictyostelium development but no CREB homologue has been identified in this system. Here we show that Dictyostelium utilizes a CREB-like protein, BzpF, to integrate PKA signaling during late development. bzpF– mutants produce compromised spores, which are extremely unstable and g...

  17. 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.

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

    Science.gov (United States)

    Zhu, Chunmao; Kawamura, Kimitaka; Fukuda, Yasuro; Mochida, Michihiro; Iwamoto, Yoko

    2016-06-01

    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 levels in daytime than nighttime, indicating substantial photochemical SOA formation. Using tracer-based methods, we estimated that fungal spores account for 45 % of organic carbon (OC) in nighttime and 22 % in daytime, whereas BVOC oxidation products account for 15 and 19 %, respectively. To our knowledge, we present for the first time highly time-resolved results that fungal spores overwhelmed BVOC oxidation products in contributing to OA especially in nighttime. This study emphasizes the importance of both PBAPs and SOAs in forming forest organic aerosols.

  19. Determination of fungal spore release from wet building materials

    DEFF Research Database (Denmark)

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

    2003-01-01

    of typical indoor fungi has been measured under controlled conditions. The fungi were cultivated for a period of 4-6 weeks on sterilized wet wallpapered gypsum boards at a relative humidity (RH) of approximately 97%. A specially designed small chamber (P-FLEC) was placed on the gypsum board. The release...... and Trichoderma harzianum ), the number of spores produced on the gypsum board and subsequently released was quantified. Also the relationship between air velocities from 0.3 to 3 m/s over the surface and spore release has been measured. The method was found to give very reproducible results for each fungal...

  20. The ecology of anthrax spores: tough but not invincible.

    OpenAIRE

    Dragon, D C; Rennie, R P

    1995-01-01

    Bacillus anthracis is the causative agent of anthrax, a serious and often fatal disease of wild and domestic animals. Central to the persistence of anthrax in an area is the ability of B. anthracis to form long-lasting, highly resistant spores. Understanding the ecology of anthrax spores is essential if one hopes to control epidemics. Studies on the ecology of anthrax have found a correlation between the disease and specific soil factors, such as alkaline pH, high moisture, and high organic c...

  1. Decontamination of Anthrax spores in critical infrastructure and critical assets.

    Energy Technology Data Exchange (ETDEWEB)

    Boucher, Raymond M.; Crown, Kevin K.; Tucker, Mark David; Hankins, Matthew Granholm

    2010-05-01

    Decontamination of anthrax spores in critical infrastructure (e.g., subway systems, major airports) and critical assets (e.g., the interior of aircraft) can be challenging because effective decontaminants can damage materials. Current decontamination methods require the use of highly toxic and/or highly corrosive chemical solutions because bacterial spores are very difficult to kill. Bacterial spores such as Bacillus anthracis, the infectious agent of anthrax, are one of the most resistant forms of life and are several orders of magnitude more difficult to kill than their associated vegetative cells. Remediation of facilities and other spaces (e.g., subways, airports, and the interior of aircraft) contaminated with anthrax spores currently requires highly toxic and corrosive chemicals such as chlorine dioxide gas, vapor- phase hydrogen peroxide, or high-strength bleach, typically requiring complex deployment methods. We have developed a non-toxic, non-corrosive decontamination method to kill highly resistant bacterial spores in critical infrastructure and critical assets. A chemical solution that triggers the germination process in bacterial spores and causes those spores to rapidly and completely change to much less-resistant vegetative cells that can be easily killed. Vegetative cells are then exposed to mild chemicals (e.g., low concentrations of hydrogen peroxide, quaternary ammonium compounds, alcohols, aldehydes, etc.) or natural elements (e.g., heat, humidity, ultraviolet light, etc.) for complete and rapid kill. Our process employs a novel germination solution consisting of low-cost, non-toxic and non-corrosive chemicals. We are testing both direct surface application and aerosol delivery of the solutions. A key Homeland Security need is to develop the capability to rapidly recover from an attack utilizing biological warfare agents. This project will provide the capability to rapidly and safely decontaminate critical facilities and assets to return them to

  2. Proton dynamics in bacterial spores, a neutron scattering investigation

    Directory of Open Access Journals (Sweden)

    Noue Alexandre Colas de la

    2015-01-01

    Full Text Available Results from first neutron scattering experiments on bacterial spores are reported. The elastic intensities and mean square displacements have a non-linear behaviour as function of temperature, which is in agreement with a model presenting more pronounced variations at around 330 K (57 ∘C and 400 K (127 ∘C. Based on the available literature on thermal properties of bacterial spores, mainly referring to differential scanning calorimetry, they are suggested to be associated to main endothermic transitions induced by coat and/or core bacterial response to heat treatment.

  3. Flow-cytometric Analysis of Bacillus anthracis Spores

    Directory of Open Access Journals (Sweden)

    D. V. Kamboj

    2006-11-01

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

  4. Characteristics of spore germination and protonemal development in Hypnum pacleseens

    Institute of Scientific and Technical Information of China (English)

    HUANG Shiliang; LI Min; ZHAO Jiancheng; ZHANG Yuanming; WANG Zhenjie

    2006-01-01

    The spore germination,protonemal development,and gametophyte differentiation of Hypnum pacleseens were observed in cultivation.Photomicrographs showed that spore germination of Hypnum pacleseens occured within the exospore.Its protonema is massive with filamentous chloronema formed inside.The terminal part of the chloronema differentiated into filamentous caulonema and its rhizoid was derived from the apical cell of the filamentous chloronema.The initial cell of gametophyte differentiated from chloronema and caulonema.Sporeling type of Hypnum pacleseens is developmentally similar to Glyphmitrium-type.

  5. Systematic Assessment of Nonproteolytic Clostridium botulinum Spores for Heat Resistance

    Science.gov (United States)

    Stringer, Sandra C.; Barker, Gary C.; Peck, Michael W.

    2016-01-01

    ABSTRACT Heat treatment is an important controlling factor that, in combination with other hurdles (e.g., pH, aw), is used to reduce numbers and prevent the growth of and associated neurotoxin formation by nonproteolytic C. botulinum in chilled foods. It is generally agreed that a heating process that reduces the spore concentration by a factor of 106 is an acceptable barrier in relation to this hazard. The purposes of the present study were to review the available data relating to heat resistance properties of nonproteolytic C. botulinum spores and to obtain an appropriate representation of parameter values suitable for use in quantitative microbial risk assessment. In total, 753 D values and 436 z values were extracted from the literature and reveal significant differences in spore heat resistance properties, particularly those corresponding to recovery in the presence or absence of lysozyme. A total of 503 D and 338 z values collected for heating temperatures at or below 83°C were used to obtain a probability distribution representing variability in spore heat resistance for strains recovered in media that did not contain lysozyme. IMPORTANCE In total, 753 D values and 436 z values extracted from literature sources reveal significant differences in spore heat resistance properties. On the basis of collected data, two z values have been identified, z = 7°C and z = 9°C, for spores recovered without and with lysozyme, respectively. The findings support the use of heat treatment at 90°C for 10 min to reduce the spore concentration by a factor of 106, providing that lysozyme is not present during recovery. This study indicates that greater heat treatment is required for food products containing lysozyme, and this might require consideration of alternative recommendation/guidance. In addition, the data set has been used to test hypotheses regarding the dependence of spore heat resistance on the toxin type and strain, on the heating technique used, and on the

  6. [The flotation characteristics of Bacillus cells and spores].

    Science.gov (United States)

    Stabnikova, E V; Gregirchak, N N; Taranenko, T O

    1991-01-01

    Variations in hydrophobicity of the surface of bacillary cells and their capacity to flotation in the process of batch cultivation have been studied. It is shown that hydrophobicity of the cell surface increases in the course of batch cultivation of Bacillus thuringiensis, B. licheniformis and B. megaterium. Hydrophobicity of spores of the mentioned cultures is considerably higher than that of the vegetative cells. The increase of hydrophobicity of bacillary cells positively correlated with their capacity to flotation. That is why the use of flotation for the age fractionation of bacillary cells is possible: spores are concentrated in the foam while vegetative cells remain in the culture liquid. PMID:1779906

  7. 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...

  8. A novel, inducible, citral lyase purified from spores of Penicillium digitatum

    NARCIS (Netherlands)

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

    2002-01-01

    A novel lyase, combining hydratase and aldolase activity, that converts citral into methylheptenone and acetaldehyde, was purified from spores of Penicillium digitatum. Remarkably, citral lyase activity was induced 118-fold by incubating nongerminating spores with the substrate, citral. This cofacto

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

    OpenAIRE

    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 s...

  10. Characterization of fungal spores in ambient particulate matter: A study from the Himalayan region

    Science.gov (United States)

    Kumar, Ajay; Attri, Arun K.

    2016-10-01

    Fungal spores as a constituent of ambient particulate matter (PM) is of concern; they not only display the physical traits of a particle, but are also potential allergens and health risk. An investigation over fourteen month was undertaken at a rural site located in the Western Himalayan region, to evaluate the PM associated fungal spores' concentration and diversity. The season-wise change in the fungal spores concentration in the Coarse Particulate Matter (CPM) fraction (aerodynamic diameter > 10 μm) varied from 500 to 3899 spores m-3. Their average concentration over 14 months was 1517 spores m-3. Significant diversity of fungal spores in the CPM samples was observed; 27 individual genera of fungal spores were identified, of which many were known allergens. Presence of Ascomycota and Basidiomycota fungal spores was dominant in the samples; ∼20% of the spores were un-characterized. The season-wise variability in fungal spores showed a statistically significant high correlation with CPM load. Maximum number concentration of the spores in CPM was recorded in the summer, while minimum in the winter. The high diversity of spores occurred during monsoon and post monsoon months. The meteorological factors played an important role in the fungal spores' distribution profile. The temporal profile of the spores showed significant correlation with the ambient temperature (T), relative humidity (RH), wind speed (WS) and planetary boundary layer (PBL) height. Strong correlation of WS with fungal spores and CPM, and wind back trajectories suggest that re-suspension and wind assisted transport of PM contributes to ambient CPM associated fungal spores.

  11. Insect feeding on spores of a bracket fungus, Elfvingia applanata (Pers.) Karst. (Ganodermataceae, Aphyllophorales)

    OpenAIRE

    Tuno, Nobuko

    1999-01-01

    Insects visiting sporocarps of Elfvingia applanata, a wood-rotting bracket fungus, were examined in Kyoto, central Japan. Mycodrosophila flies (Drosophilidae: Diptera) were predominant and visited the spore-producing sporocarps exclusively. They were observed feeding on the spores, and a number of spores seemed to be alive even after having passed through insects' digestive tracts. In addition, the insects attached a number of spores on their body surfaces. In a rearing experiment with insect...

  12. Multinucleate spores contribute to evolutionary longevity of asexual glomeromycota.

    Science.gov (United States)

    Jany, Jean-Luc; Pawlowska, Teresa E

    2010-04-01

    Arbuscular mycorrhizal fungi (Glomeromycota) are the dominant symbionts of land plants and one of the oldest multicellular lineages that exist without evidence of sexual reproduction. The mechanisms that protect these organisms from extinction due to accumulation of deleterious mutations in the absence of sexual recombination are unclear. Glomeromycota reproduce by spores containing hundreds of nuclei, which represents a departure from the typical eukaryotic developmental pattern, where a multicellular organism is re-created from a uninucleate propagule. To understand whether the multinucleate spore makeup may have contributed to the evolutionary success of Glomeromycota, we examined the dynamics of spore nuclei in Glomus etunicatum using live three-dimensional imaging and mathematical models. We show that the spores are populated by an influx of a stream of nuclei from the surrounding mycelium rather than by divisions of a single founder nucleus. We present evidence that mechanisms of selection are likely to operate at the level of individual nuclei. On the basis of mathematical analyses of the effects that these nuclear dynamics have on the population mutation load, we postulate that the developmental patterns of sporogenesis have adaptive significance for moderating the accumulation of deleterious mutations and may have contributed to the evolutionary longevity of Glomeromycota. PMID:20170364

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

    International Nuclear Information System (INIS)

    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)

  14. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores.

    Science.gov (United States)

    Edmonds, Jason; Lindquist, H D Alan; Sabol, Jonathan; Martinez, Kenneth; Shadomy, Sean; Cymet, Tyler; Emanuel, Peter

    2016-01-01

    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. PMID:27123934

  15. Changes in spore chemistry and appearance with increasing maturity

    NARCIS (Netherlands)

    W.T. Fraser; J.S. Watson; M.A. Sephton; B.H. Lomax; G. Harrington; W.D. Gosling; S. Self

    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 heat

  16. Phospholipase C delta regulates germination of Dictyostelium spores

    NARCIS (Netherlands)

    Van Dijken, P.; Van Haastert, PJM

    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 i

  17. 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.

  18. 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.

  19. 9 CFR 113.66 - Anthrax Spore Vaccine-Nonencapsulated.

    Science.gov (United States)

    2010-01-01

    ... for 24 hours to 28 hours. Each plate having uniformly distributed colonies shall be counted and an..., DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD... test animals (30 vaccinates and 12 controls). (2) An arithmetic mean spore count of vaccine...

  20. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores

    Science.gov (United States)

    Edmonds, Jason; Lindquist, H. D. Alan; Sabol, Jonathan; Martinez, Kenneth; Shadomy, Sean; Cymet, Tyler; Emanuel, Peter

    2016-01-01

    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. PMID:27123934

  1. Ascoaphaera osmophila sp.nov. An Australian Spore Cyst

    DEFF Research Database (Denmark)

    Skou, Jens-Peder; King, , J.

    1984-01-01

    Ascosphaera osmophila sp. nov. is described. Septa occur often close together and remain intact when the mycelium disintegrates. A fairly good production of mature spore cysts occurs only on media containing 10% sugar or more. A. osmophila lives in association with the mason bee, Chalicodoma...

  2. 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 na

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

    NARCIS (Netherlands)

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

    2015-01-01

    Spoilage of heat treated foods can be caused by the presence of surviving spore-formers. It is virtually impossible to prevent contamination at the primary production level as spores are ubiquitous present in the environment and can contaminate raw products. As a result spore inactivation treatments

  4. Effect of temperature on green spore longevity for the ferns Equisetum ramosissimum and Osmunda regalis

    Science.gov (United States)

    Some fern species produce chlorophyllic or green spores. Green spores lose viability quickly compared to nongreen spores, and so need specialized treatment for long term conservation in germplasm banks. Dry storage at different temperatures (25 ºC, 4 ºC, -25 ºC, -80 ºC and -196 ºC) was studied in ...

  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. The Ice Nucleation Ability of Selected Atmospherically Abundant Fungal Spores

    Science.gov (United States)

    Iannone, R.; Chernoff, D. I.; Bertram, A. K.

    2010-12-01

    Ice clouds are widely recognized for their roles in the earth’s radiation budget and climate systems. However, their formation mechanisms are poorly understood thus constituting an uncertainty in the evaluation of the global radiation budget. An important mechanism of ice cloud formation is heterogeneous nucleation on aerosol particles. The surface properties of these particles, called ice nuclei (IN), directly affect the temperature at which ice nucleation occurs. There is a growing emphasis on the study of bioaerosols (e.g., bacteria, fungi, pollen) as IN since they are ubiquitous in the atmosphere. The focus of the current study is to determine the ice nucleation properties of spores obtained from a variety of fungi. Aerosolized spores were impacted onto a hydrophobic glass substrate and immersed in ultrapure water. A technique involving an optical light microscope coupled to a flow cell was used to precisely control temperature and humidity within the cell. A digital camera captured high-resolution video of the particles undergoing ice nucleation, allowing for the analyses of freezing events and particle sizes. The first experimental results using spores obtained from the fungal genera Cladosporium and Penicillium reveal an average temperature increase of ~1-5 K in the ice nucleation temperature compared to homogeneous nucleation (i.e., freezing of pure liquid water). Furthermore, there appears to be a relationship between the amount of spores present per droplet and the freezing temperature of water. These results are presented and discussed, and the potential contribution of these data to further the understanding of heterogeneous nucleation in the atmosphere is provided. Box plot summarizing freezing data for homogeneous nucleation experiments (leftmost box) and binned data from heterogeneous nucleation experiments involving spores of Cladosporium. Freezing data are distributed into 200 µm2 bins that represent the total area of all observable inclusions

  7. 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

  8. 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

  9. Intact Cell/Spore Mass Spectrometry of Fusarium Macro Conidia for Fast Isolate and Species Differentiation

    Science.gov (United States)

    Dong, Hongjuan; Marchetti-Deschmann, Martina; Winkler, Wolfgang; Lohninger, Hans; Allmaier, Guenter

    The focus of this paper is the development of an approach called intact cell mass spectrometry (ICMS) or intact spore mass spectrometry (ISMS) based on the technique matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the rapid differentiation and identification of Fusarium species. Several parameters, which are known to affect the quality of IC mass spectra, have been investigated in detail by varying the MALDI matrix as well as the solvent system, in which the matrix has been dissolved, the solvent system for sample purification and the type of sample/MALDI matrix deposition technique. In the end characteristic as well as highly reproducible IC or IS mass spectra or peptide/protein fingerprints of three Fusarium species (F. cerealis, F. graminearum and F. poae) including 16 Fusarium isolates derived from different hosts and geographical locations have been obtained. Unscaled hierarchical cluster analysis based on ICMS data of eight selected Fusarium isolates of two species F. graminearum and F. poae revealed significant difference among the peptide/protein pattern of them. The results of the applied cluster analysis proved that, ICMS is a powerful approach for the rapid differentiation of Fusarium species. In addition, an on-target tryptic digestion was applied to Fusarium macro conidia spores to identify proteins using MALDI post source decay (PSD) fragment ion analysis. Two kinds of trypsin, namely bead-immobilized - to favor cleavage of surface-associated proteins - and non-immobilized trypsin were applied and compared. The results showed that the latter is more suitable for generating sequence tags by PSD fragment ion analysis.

  10. Investigation of ice-assisted sonication on the microstructure and chemical quality of Ganoderma lucidum spores.

    Science.gov (United States)

    Zhao, Ding; Chang, Ming-Wei; Li, Jing-Song; Suen, William; Huang, Jie

    2014-11-01

    Ganoderma lucidum spores (GLS) are well known for disease treatment and vitality enhancement, and have been shown to contain a variety of bioactive components, such as polysaccharides and triterpenes. However, the resilient bilayer sporoderm structure of GLS restricts the release of bioactive components and limits its complete pharmacological effects. The current study was aimed to improve the quality of GLS by means of a customized sonication technique, particularly, the effect of sonication processing parameters on GLS-breaking efficiencies was investigated. Significant morphological changes, such as cracked, fractured, and disintegrated GLS were observed using scanning electron microscopy (SEM) after sonication treatment. The performance for breaking GLS sporoderm was obtained at ultrasonic power density of 23.7 W/cm(2) , duty cycle 100%, and 90-min processing time. Through the combination of sonication in an ice bath, sporoderm breaking efficiency can be further increased from 45% to almost 75%. FTIR analysis revealed an increase in bioactive components of polysaccharide, protein, and fatty acid from the sonication processed GLS when compared to ground spores available commercially. The current results indicated that the ice bath combined sonication method is more effective in delivering GLS ingredients and could be an economic technique for the production of high-quality broken sporoderm GLS.

  11. Effect of Carbon and Nitrogen Availability on Metabolism of Amino Acids in Germinating Spores of Arbuscular Mycorrhizal Fungi

    Institute of Scientific and Technical Information of China (English)

    JIN Hai-Ru; JIANG Dong-Hua; ZHANG Ping-Hua

    2011-01-01

    The effects of carbon (C) and nitrogen (N) sources on N utilization and biosynthesis of amino acids were examined in the germinating spores of the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith after exposure to various N substrates,CO2,glucose,and/or root exudates.The N uptake and de novo biosynthesis of amino acids were analyzed using stable isotopic labeling with mass spectrometric detection.High-performance liquid chromatography-based analysis was used to measure amino acid levels.In the absence of exogenous N sources and in the presence of 25 mL L-1 CO2,the germinating AM fungal spores utilized internal N storage as well as C skeletons derived from the degradation of storage lipids to biosynthesize the free amino acids,in which serine and glycine were produced predominantly.The concentrations of internal amino acids increased gradually as the germination time increased from 0 to 1 or 2 weeks.However,asparagine and glutamine declined to the low levels; both degraded to provide the biosynthesis of other amino acids with C and N donors.The availability of exogenous inorganic N (ammonium and nitrate) and organic N (urea,arginine,and glutamine) to the AM fungal spores using only CO2 for germination generated more than 5 times more internal free amino acids than those in the absence of exogenous N.A supply of exogenous nitrate to the AM fungal spores with only CO2 gave rise to more than 10 times more asparagine than that without exogenous N.In contrast,the extra supply of exogenous glucose to the AM fungal spores generated a significant enhancement in the uptake of exogenous N sources,with more than 3 times more free amino acids being produced than those supplied with only exogenous CO2.Meanwhile,arginine was the most abundant free amino acid produced and it was incorporated into the proteins of AM fungal spores to serve as an N storage compound.

  12. Arabitol and mannitol as tracers for the quantification of airborne fungal spores

    Science.gov (United States)

    Bauer, Heidi; Claeys, Magda; Vermeylen, Reinhilde; Schueller, Elisabeth; Weinke, Gert; Berger, Anna; Puxbaum, Hans

    Fungal spores constitute a sizeable fraction of coarse organic carbon (OC) in the atmospheric aerosol. In order to avoid tedious spore count methods, tracers for quantifying the spore-OC in atmospheric aerosol are sought. Arabitol and mannitol have been proposed as such tracers, since no other emission sources for these compounds have been reported. By parallel investigations of spore counts and tracer determinations from PM 10 filter samples we could derive quantitative relationships between the amounts of tracer compounds and the numbers of spores in the atmosphere for different sites in the area of Vienna. We obtained over all average relationships of 1.2 pg arabitol spore -1, with a range of 0.8-1.8, and 1.7 pg mannitol spore -1, with a range of 1.2-2.4, with a clear site dependence. Thus, using these conversion factors from spore counts to spore-OC and spore-mass, along with analytical data for arabitol or mannitol in filter samples, the contribution of fungal spores to the OC and to the mass balance of atmospheric aerosol particles can be estimated.

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

    Directory of Open Access Journals (Sweden)

    Jacobus C. de Roode

    2013-08-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2006-11-14

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Lawrence Livermore National Laboratory

    2006-12-11

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

  16. Breaking and Characteristics of Ganoderma Lucidum Spores by High Speed Entrifugal Shearing Pulverizer

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The spores of Ganoderma lucidum were ground and broken to ultrafine particles by high speed centrifugal shearing(HSCS) pulverizer. The characteristics of Ganoderma lucidum spores were analyzed by scanning electron microscope (SEM), Fourier transform infrared spectrophotometry (FTIR). Ultraviolet-visible pectrophotometer was used to determine the extraction ratio of aqueous solubility polysaccharide between the raw and broken spores. The immunological function on the mice before and after the breaking of spores was investigated. The experimental results show that after being ground, the sporoderm-broken ratio reachs 100%,the original active ingredients of ganoderma lucidum spores do not change, and the extraction ratio of aqueous solubility polysaccharide is greatly increased by 40.08%. The broken spores show much higher immunological activity comparing with original spores of Ganoderma lucidum.

  17. BzpF is a CREB-like transcription factor that regulates spore maturation and stability in Dictyostelium.

    Science.gov (United States)

    Huang, Eryong; Talukder, Shaheynoor; Hughes, Timothy R; Curk, Tomaz; Zupan, Blaz; Shaulsky, Gad; Katoh-Kurasawa, Mariko

    2011-10-01

    The cAMP response element-binding protein (CREB) is a highly conserved transcription factor that integrates signaling through the cAMP-dependent protein kinase A (PKA) in many eukaryotes. PKA plays a critical role in Dictyostelium development but no CREB homologue has been identified in this system. Here we show that Dictyostelium utilizes a CREB-like protein, BzpF, to integrate PKA signaling during late development. bzpF(-) mutants produce compromised spores, which are extremely unstable and germination defective. Previously, we have found that BzpF binds the canonical CRE motif in vitro. In this paper, we determined the DNA binding specificity of BzpF using protein binding microarray (PBM) and showed that the motif with the highest specificity is a CRE-like sequence. BzpF is necessary to activate the transcription of at least 15 PKA-regulated, late-developmental target genes whose promoters contain BzpF binding motifs. BzpF is sufficient to activate two of these genes. The comparison of RNA sequencing data between wild type and bzpF(-) mutant revealed that the mutant fails to express 205 genes, many of which encode cellulose-binding and sugar-binding proteins. We propose that BzpF is a CREB-like transcription factor that regulates spore maturation and stability in a PKA-related manner. PMID:21810415

  18. Yeast Interacting Proteins Database: YDL239C, YHR184W [Yeast Interacting Proteins Database

    Lifescience Database Archive (English)

    Full Text Available it as prey (1) YHR184W SSP1 Protein involved in the control of meiotic nuclear division and coordination... the control of meiotic nuclear division and coordination of meiosis with spore formation; transcription is

  19. Fed-batch production of gluconic acid by terpene-treated Aspergillus niger spores.

    Science.gov (United States)

    Ramachandran, Sumitra; Fontanille, Pierre; Pandey, Ashok; Larroche, Christian

    2008-12-01

    Aspergillus niger spores were used as catalyst in the bioconversion of glucose to gluconic acid. Spores produced by solid-state fermentation were treated with 15 different terpenes including monoterpenes and monoterpenoids to permeabilize and inhibit spore germination. It was found that spore membrane permeability is significantly increased by treatment with terpenoids when compared to monoterpenes. Best results were obtained with citral and isonovalal. Studies were carried out to optimize spores concentration (10(7)-10(10) spores/mL), terpene concentrations in the bioconversion medium and time of exposure (1-18 h) needed for permeabilization of spores. Fed-batch production of gluconate was done in a bioreactor with the best conditions [10(9) spores/mL of freeze-thawed spores treated with citral (3% v/v) for 5 h] followed by sequential additions of glucose powder and pH-regulated with a solution containing 2 mol/L of either NaOH or KOH. Bioconversion performance of the spore enzyme was compared with the commercial glucose oxidase at 50, 60, and 70 degrees C. Results showed that the spore enzyme was comparatively stable at 60 degrees C. It was also found that the spores could be reutilized for more than 14 cycles with almost similar reaction rate. Similar biocatalytic activity was rendered by spores even after its storage of 1 year at -20 degrees C. This study provided an experimental evidence of the significant catalytic role played by A. niger spore in bioconversion of glucose to gluconic acid with high yield and stability, giving protection to glucose oxidase.

  20. Nanoscale structural and mechanical analysis of Bacillus anthracis spores inactivated with rapid dry heating.

    Science.gov (United States)

    Xing, Yun; Li, Alex; Felker, Daniel L; Burggraf, Larry W

    2014-03-01

    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.

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

    Science.gov (United States)

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

    2016-05-01

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

  2. Self-inhibition of spore germination via reactive oxygen in the fungus Cladosporium cucumerinum, causal agent of cucurbit scab

    Science.gov (United States)

    Cladosporium cucumerinum spore germination in vitro depended on spore suspension density. Different fungal isolates displayed maximum germination at different spore concentrations. For one isolate, maximum spore density was observed at both 18 and 25 °C, although germination percentage increased sli...

  3. Cytokine Response to Infection with Bacillus anthracis Spores

    OpenAIRE

    Pickering, Alison K.; Osorio, Manuel; Lee, Gloria M.; Grippe, Vanessa K.; Bray, Mechelle; Merkel, Tod J.

    2004-01-01

    Bacillus anthracis, the etiological agent of anthrax, is a gram-positive, spore-forming bacterium. The inhalational form of anthrax is the most severe and is associated with rapid progression of the disease and the outcome is frequently fatal. Transfer from the respiratory epithelium to regional lymph nodes appears to be an essential early step in the establishment of infection. This transfer is believed to occur by means of carriage within alveolar macrophages following phagocytosis. Therefo...

  4. Modelling the heat stress and the recovery of bacterial spores.

    Science.gov (United States)

    Mafart, P; Leguérinel, I

    1997-07-22

    After heat treatment, the temperature incubation and the medium composition, (pH and sodium chloride content) influence the capacity of injured spores to repair heat damage. The concept of heat resistance D- (decimal reduction time) and z-values (temperature increase which results in a ten fold reduction of the D value) is not sufficient and the ratio of spore recovery after incubation should be considered in calculations used in thermal processing of food. This paper aims to derive a model describing the recovery of injured spores as a function of both the heat treatment intensity and the environmental conditions. According to data from numerous investigators, when spores are incubated in unfavorable conditions, the ratio of cell recovery and the apparent D-value are reduced. Moreover the ratio of the apparent D-value and the estimated in optimal incubation D-value is constant and independent of the heat treatment conditions. Beyond these observations it is shown that the ratio of cell recovery with respect to the heat treatment F-value (exposure time, in minutes, at 121.1 degrees C which results in the same destruction ratio that the considered heat treatment does) is linear and can be quantified by using two factors independent of the heat treatment: the gamma-factor reflects the degree of precariousness due to the heat stress while the epsilon-factor reflects more intrinsically the incubation conditions without previous heat treatment. The gamma-factor varies as a function of the incubation temperature according to an Arrhenius law.

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

    Directory of Open Access Journals (Sweden)

    Cho Moo

    2011-05-01

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

  6. Distribution of sterols in the fungi. I - Fungal spores

    Science.gov (United States)

    Weete, J. D.; Laseter, J. L.

    1974-01-01

    Mass spectrometry was used to examine freely extractable sterols from spores of several species of fungi. Ergosterol was the most common sterol produced by any individual species, but it was completely absent from two species belonging to apparently distantly related groups of fungi: the aquatic Phycomycetes and the rust fungi. This fact could have taxonomic or phylogenetic implications. The use of glass capillary columns in the resolution of the sterols is shown to eliminate some of the difficulty inherent in this process.

  7. Setting risk-informed environmental standards for Bacillus anthracis spores.

    Science.gov (United States)

    Hong, Tao; Gurian, Patrick L; Ward, Nicholas F Dudley

    2010-10-01

    In many cases, human health risk from biological agents is associated with aerosol exposures. Because air concentrations decline rapidly after a release, it may be necessary to use concentrations found in other environmental media to infer future or past aerosol exposures. This article presents an approach for linking environmental concentrations of Bacillus. anthracis (B. anthracis) spores on walls, floors, ventilation system filters, and in human nasal passages with human health risk from exposure to B. anthracis spores. This approach is then used to calculate example values of risk-informed concentration standards for both retrospective risk mitigation (e.g., prophylactic antibiotics) and prospective risk mitigation (e.g., environmental clean up and reoccupancy). A large number of assumptions are required to calculate these values, and the resulting values have large uncertainties associated with them. The values calculated here suggest that documenting compliance with risks in the range of 10(-4) to 10(-6) would be challenging for small diameter (respirable) spore particles. For less stringent risk targets and for releases of larger diameter particles (which are less respirable and hence less hazardous), environmental sampling would be more promising.

  8. Mutagenesis of Bacillus subtilis spores exposed to simulated space environment

    Science.gov (United States)

    Munakata, N.; Natsume, T.; Takahashi, K.; Hieda, K.; Panitz, C.; Horneck, G.

    Bacterial spores can endure in a variety of extreme earthly environments. However, some conditions encountered during the space flight could be detrimental to DNA in the spore, delimiting the possibility of transpermia. We investigate the genetic consequences of the exposure to space environments in a series of preflight simulation project of EXPOSE. Using Bacillus subtilis spores of repair-proficient HA101 and repair-deficient TKJ6312 strains, the mutations conferring resistance to rifampicin were detected, isolated and sequenced. Most of the mutations were located in a N-terminal region of the rpoB gene encoding RNA polymerase beta-subunit. Among several potentially mutagenic factors, high vacuum, UV radiation, heat, and accelerated heavy ions induced mutations with varying efficiencies. A majority of mutations induced by vacuum exposure carried a tandem double-base change (CA to TT) at a unique sequence context of TCAGC. Results indicate that the vacuum and high temperature may act synergistically for the induction of mutations.

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

    Directory of Open Access Journals (Sweden)

    Michael B Francis

    Full Text Available Infections caused by Clostridium difficile have increased steadily over the past several years. While studies on C. difficile virulence and physiology have been hindered, in the past, by lack of genetic approaches and suitable animal models, newly developed technologies and animal models allow these processes to be studied in detail. One such advance is the generation of a mouse-model of C. difficile infection. The development of this system is a major step forward in analyzing the genetic requirements for colonization and infection. While important, it is equally as important in understanding what differences exist between mice and humans. One of these differences is the natural bile acid composition. Bile acid-mediated spore germination is an important step in C. difficile colonization. Mice produce several different bile acids that are not found in humans. These muricholic acids have the potential to impact C. difficile spore germination. Here we find that the three muricholic acids (α-muricholic acid, β-muricholic acid and ω-muricholic acid inhibit C. difficile spore germination and can impact the growth of vegetative cells. These results highlight an important difference between humans and mice and may have an impact on C. difficile virulence in the mouse-model of C. difficile infection.

  10. Quantum dot incorporated Bacillus spore as nanosensor for viral infection.

    Science.gov (United States)

    Zhang, Xinya; Zhou, Qian; Shen, Zhongfeng; Li, Zheng; Fei, Ruihua; Ji, Eoon Hye; Hu, Shen; Hu, Yonggang

    2015-12-15

    In this paper, we report a high-throughput biological method to prepare spore-based monodisperse microparticles (SMMs) and then form the nanocomposites of CdTe quantum dot (QD)-loaded SMMs by utilizing the endogenous functional groups from Bacillus spores. The SMMs and QD-incorporated spore microspheres (QDSMs) were characterized by using transmission electron microscopy, high-resolution transmission electron microscopy, fluorescence microscopy, fluorescence and UV-visible absorption spectroscopy, zeta potential analysis, Fourier-transform infrared spectroscopy, potentiometric titrations, X-ray photo-electron spectroscopy. The thermodynamics of QD/SMM interaction and antigen/QDSM interaction was also investigated by isothermal titration microcalorimetry (ITC). Fluorescent QDSMs coded either with a single luminescence color or with multiple colors of controlled emission intensity ratios were obtained. Green QDSMs were used as a model system to detect porcine parvovirus antibody in swine sera via flow cytometry, and the results demonstrated a great potential of QDSMs in high-throughput immunoassays. Due to the advantages such as simplicity, low cost, high throughput and eco-friendliness, our developed platform may find wide applications in disease detection, food safety evaluation and environmental assessment.

  11. Spore production of Penicillium roqueforti by solid state fermentation: Stoichiometry, growth and sporulation behavior.

    Science.gov (United States)

    Desfarges, C; Larroche, C; Gros, J B

    1987-06-01

    The development of Penicillium roqueforti on buckwheat seeds proceeds roughly into four steps, involving a lag phase and three growth phases. First, it appears as a spore germination and external colonization of the grains by the mycelium. Then, mainly external sporulation and internal colonization of the seeds occur and finally internal sporulation takes place. The Stoichiometry of the growth and the sporulation is established. Kinetic experiments performed in a fixed bed reactor show that the growth of the microorganism (biomass production) may be estimated by the protein content of the medium. This growth occurs with a very low mu(max) value close to 0.030 h(-1). The chitin content of the medium is an indicator of the sporulation, just as the metabolic liquor (mainly water) produced during the course of a cultivation. The values of the observed respiratory quotient are close to those predicted by stoichiometry.

  12. The protein composition of reconstituted 30S ribosomal subunits: the effects of single protein omission.

    Science.gov (United States)

    Buck, M A; Olah, T V; Perrault, A R; Cooperman, B S

    1991-06-01

    Using reverse phase HPLC, we have been able to quantify the protein compositions of reconstituted 30S ribosomal subunits, formed either with the full complement of 30S proteins in the reconstitution mix or with a single protein omitted. We denote particles formed in the latter case as SPORE (single protein omission reconstitution) particles. An important goal in 30S reconstitution studies is the formation of reconstituted subunits having uniform protein composition, preferably corresponding to one copy of each protein per reconstituted particle. Here we describe procedures involving variation of the protein:rRNA ratio that approach this goal. In SPORE particles the omission of one protein often results in the partial loss in uptake of other proteins. We also describe procedures to increase the uptake of such proteins into SPORE particles, thus enhancing the utility of the SPORE approach in defining the role of specific proteins in 30S structure and function. The losses of proteins other than the omitted protein provide a measure of protein:protein interaction within the 30S subunit. Most of these losses are predictable on the basis of other such measures. However, we do find evidence for several long-range protein:protein interactions (S6:S3, S6:S12, S10:S16, and S6:S4) that have not been described previously.

  13. Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi

    OpenAIRE

    Jolly, Carrie E.; Leonard, Cory A.; J. Russell Hayman

    2010-01-01

    Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02_0100; “C1”) was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and...

  14. Genetic Diversity and Association Characters of Bacteria Isolated from Arbuscular Mycorrhizal Fungal Spore Walls.

    Science.gov (United States)

    Selvakumar, Gopal; Krishnamoorthy, Ramasamy; Kim, Kiyoon; Sa, Tong-Min

    2016-01-01

    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. PMID:27479250

  15. 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.

  16. Enhancement of intrinsic antitumor activity in spore-endotoxin mixtures of Bacillus thuringiensis by exposure to ultraviolet radiation

    International Nuclear Information System (INIS)

    Irradiation of spore-endotoxin mixtures from Bacillus thuringiensis cultures at 254 nm (60 μW cm-2) enhances their intrinsic antitumor potency as well as that of either component. The extent of enhancement depends on the length of exposure (optimum: 35 min) and may thus be due to photochemical changes of the endotoxin protein or/and to photoproduction of additional compounds with antitumor activity. Antitumor effects, expressed as survival rates of C57BL/6 mice inoculated with Lewis' mouse lung carcinoma and subjected to treatments 24 h later, depended on the number of doses of preparations administered (mixture, separated components). (author)

  17. Enhancement of intrinsic antitumor activity in spore-endotoxin mixtures of Bacillus thuringiensis by exposure to ultraviolet radiation

    Energy Technology Data Exchange (ETDEWEB)

    Zamola, B.; Karminski-Zamola, G.; Fuks, Z.; Kubovic, M. (Zagreb Univ. (Yugoslavia)); Wrishcer, M. (Institut Rudjer Boskovic, Zagreb (Yugoslavia))

    1985-03-01

    Irradiation of spore-endotoxin mixtures from Bacillus thuringiensis cultures at 254 nm (60 ..mu..W cm/sup -2/) enhances their intrinsic antitumor potency as well as that of either component. The extent of enhancement depends on the length of exposure (optimum: 35 min) and may thus be due to photochemical changes of the endotoxin protein or/and to photoproduction of additional compounds with antitumor activity. Antitumor effects, expressed as survival rates of C57BL/6 mice inoculated with Lewis' mouse lung carcinoma and subjected to treatments 24 h later, depended on the number of doses of preparations administered (mixture, separated components).

  18. Gene discovery in EST sequences from the wheat leaf rust fungus Puccinia triticina sexual spores, asexual spores and haustoria, compared to other rust and corn smut fungi

    Directory of Open Access Journals (Sweden)

    Wynhoven Brian

    2011-03-01

    . tritici (Pst, and poplar leaf rust Melampsora species, and the corn smut fungus, Ustilago maydis (Um. While extensive homologies were found, many genes appeared novel and species-specific; over 40% of genes did not match any known sequence in existing databases. Focusing on spore stages, direct comparison to Um identified potential functional homologs, possibly allowing heterologous functional analysis in that model fungus. Many potentially secreted protein genes were identified by similarity searches against genes and proteins of Pgt and Melampsora spp., revealing apparent orthologs. Conclusions The current set of Pt unigenes contributes to gene discovery in this major cereal pathogen and will be invaluable for gene model verification in the genome sequence.

  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. 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.

  1. 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

  2. Comparative Study of Pressure-Induced Germination of Bacillus subtilis Spores at Low and High Pressures

    OpenAIRE

    Wuytack, Elke Y.; Boven, Steven; Michiels, Chris W.

    1998-01-01

    We have studied pressure-induced germination of Bacillus subtilis spores at moderate (100 MPa) and high (500 to 600 MPa) pressures. Although we found comparable germination efficiencies under both conditions by using heat sensitivity as a criterion for germination, the sensitivity of pressure-germinated spores to some other agents was found to depend on the pressure used. Spores germinated at 100 MPa were more sensitive to pressure (>200 MPa), UV light, and hydrogen peroxide than were those g...

  3. PCR Assay To Detect Bacillus anthracis Spores in Heat-Treated Specimens

    OpenAIRE

    Fasanella, A.; Losito, S.; Adone, R.; Ciuchini, F.; Trotta, T.; Altamura, S. A.; D. Chiocco; Ippolito, G

    2003-01-01

    Recent interest in anthrax is due to its potential use in bioterrorism and as a biowarfare agent against civilian populations. The development of rapid and sensitive techniques to detect anthrax spores in suspicious specimens is the most important aim for public health. With a view to preventing exposure of laboratory workers to viable Bacillus anthracis spores, this study evaluated the suitability of PCR assays for detecting anthrax spores previously inactivated at 121°C for 45 min. The resu...

  4. Measurements of the pH within dormant and germinated bacterial spores.

    OpenAIRE

    Setlow, B; Setlow, P

    1980-01-01

    The pH within the core or central region of dormant spores of Bacillus cereus and B. megaterium is 6.3-6.4 irrespective of the external pH. However, the spore's internal pH rises to 7.3-7.5 upon germination. The low internal pH of the dormant spore may be a contributing factor to its metabolic dormancy.

  5. Preferential utilization of exogenously supplied leucine for protein synthesis in estradiol-induced and uninduced cockerel liver explants.

    Science.gov (United States)

    Gehrke, L; Ilan, J

    1983-01-01

    A cockerel liver explant system has been used to study protein synthesis and ribosome transit times. After a 2-hr preincubation of explant tissue in the presence of a large concentration of nonradioactive leucine, a small quantity of [3H]leucine was added and the kinetics of uptake of [3H]leucine into the intracellular acid-soluble leucine pool was compared to the incorporation of [3H]leucine into protein. Incorporation of [3H]leucine into protein reaches a linear rate almost immediately after addition of label, whereas the acid-soluble pool does not reach constant specific activity until much later. The length of time needed to reach a linear rate of incorporation of [3H]leucine into protein is approximately equal to the length of time needed to equilibrate nascent polypeptide chains with labeled precursor--that is, one average ribosome transit time. Therefore, it seems that the immediate precursor pool for protein synthesis reaches constant specific activity almost instantly after addition of [3H]leucine. The results indicate that at least part of the supply of leucine for protein synthesis is derived directly from the exogenous incubation medium and not from the intracellular acid-soluble amino acid pool. Images PMID:6574484

  6. Rapid inactivation of Penicillium digitatum spores using high-density nonequilibrium atmospheric pressure plasma

    Science.gov (United States)

    Iseki, Sachiko; Ohta, Takayuki; Aomatsu, Akiyoshi; Ito, Masafumi; Kano, Hiroyuki; Higashijima, Yasuhiro; Hori, Masaru

    2010-04-01

    A promising, environmentally safe method for inactivating fungal spores of Penicillium digitatum, a difficult-to-inactivate food spoilage microorganism, was developed using a high-density nonequilibrium atmospheric pressure plasma (NEAPP). The NEAPP employing Ar gas had a high electron density on the order of 1015 cm-3. The spores were successfully and rapidly inactivated using the NEAPP, with a decimal reduction time in spores (D value) of 1.7 min. The contributions of ozone and UV radiation on the inactivation of the spores were evaluated and concluded to be not dominant, which was fundamentally different from the conventional sterilizations.

  7. Fungal spore content of the atmosphere of the Cave of Nerja (southern Spain): diversity and origin.

    Science.gov (United States)

    Docampo, Silvia; Trigo, M Mar; Recio, Marta; Melgar, Marta; García-Sánchez, José; Cabezudo, Baltasar

    2011-01-15

    Fungal spores are of great interest in aerobiology and allergy due to their high incidence in both outdoor and indoor environments and their widely recognized ability to cause respiratory diseases and other pathologies. In this work, we study the spore content of the atmosphere of the Cave of Nerja, a karstic cavity and an important tourist attraction situated on the eastern coast of Malaga (southern Spain), which receives more than half a million visitors every year. This study was carried out over an uninterrupted period of 4 years (2002-2005) with the aid of two Hirst-type volumetric pollen traps (Lanzoni VPPS 2000) situated in different halls of the cave. In the atmosphere of the Cave of Nerja, 72 different spore types were detected during the studied period and daily mean concentrations of up to 282,195 spores/m(3) were reached. Thirty-five of the spore types detected are included within Ascomycota and Basidiomycota (19 and 16 types, respectively). Of the remaining spore types, 32 were categorized within the group of so-called imperfect fungi, while Oomycota and Myxomycota were represented by 2 and 3 spore types, respectively. Aspergillus/Penicillium was the most abundant spore type with a yearly mean percentage that represented 50% of the total, followed by Cladosporium. Finally, the origin of the fungal spores found inside the cave is discussed on the basis of the indoor/outdoor concentrations and the seasonal behaviour observed. PMID:21138779

  8. Effects of meteorological factors on the levels of Alternaria spores on a potato crop

    Science.gov (United States)

    Escuredo, Olga; Seijo, Maria Carmen; Fernández-González, Maria; Iglesias, Isabel

    2011-03-01

    Alternaria solani Soraeur produces early blight in Solanum tuberosum L., leading to significant agricultural losses. The current study was carried out on the extensive potato crop situated in north-western of Spain during 2007, 2008 and 2009. In this area potato crops are the most important source of income. In this work we used a Hirst-type volumetric spore-trap for the aerobiological monitoring of Alternaria spores. The highest spore concentrations were recorded during the 2009 cycle (10,555 spores), and the lowest concentrations were recorded during the 2008 cycle (5,471 spores). Over the 3 years of study, the highest concentrations were registered during the last stage of the crop. The aim of the study was to observe the influence of meteorological factors on the concentration of Alternaria spores, which can lead to serious infection and early blight. Prediction of the stages during which a crop is particularly vulnerable to infection allows for adjustment of the application of fungicide and is of environmental and agricultural importance. For this reason, we tested three models (P-Days, DD and IWP) to predict the first treatment and decrease the negative effect that these spores have on potato crops. The parameter that showed the most significant correlation with spore concentrations was minimum temperature. We used ARIMA (autoregressive integrated model of running mean) time-series models to determine the forecast. We considered weather data as predictor variables and the concentration of spores on the previous day as the fixed variable.

  9. 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.

  10. Control of Bacillus licheniformis spores isolated from dairy materials in yogurt production.

    Science.gov (United States)

    Tanaka, Takashi; Ito, Akiko; Kamikado, Hideaki

    2012-01-01

    To determine the effects of sporulation temperature and period on Bacillus licheniformis spore heat resistance, B. licheniformis strain No.25 spores were sporulated at 30, 37, 42, or 50°C for 11 d and at 50°C for 1.7, 4, 7, or 11 d. The heat resistance of B. licheniformis strain No.25 spores at 110°C increased with an increase in the sporulation temperature. Spores sporulated at 50°C were 1.4-fold more heat resistant than those sporulated at 30°C. Furthermore, the heat resistance of B. licheniformis strain No.25 spores at 110°C increased with an increase in the sporulation period. Spores sporulated for 11 d were 5.3-fold more heat resistant than those sporulated for 1.7 d. The heat resistance of B. licheniformis strain No.25 spores at 110°C increased with increases in the sporulation temperature and sporulation period. The results presented in this study can be applied to the pasteurization process to control B. licheniformis spores. Pasteurization at 110°C for about 60sec. is effective in controlling B. licheniformis spores isolated from dairy materials in yogurt production.

  11. Dry heat exposures of surface exposed and embedded Bacillus spores

    Science.gov (United States)

    Schubert, Wayne

    Dry heat microbial reduction (DHMR) is the primary technique used to reduce the microbial load of spacecraft and component parts. Often, manufacturing procedures require heating flight hardware to high temperatures for purposes other than planetary protection DHMR. The existing specifications, however, do not allow for additional planetary protection bioburden reduction credit if the hardware is exposed without controlled relative humidity. The intent of this study was to provide adequate data on the DHMR technique to support modification of four aspects of current requirements; expansion of acceptable time and temperature combinations used for spacecraft dry heat microbial reduction processes above 125° C, determining the effect that humidity has on spore lethality as a function of temperature, understanding the lethality for spores with exceptionally high thermal resistance and to investigate the extended exposure requirement for materials that might contain embedded microorganisms. Spores from two bacterial species were tested, B. atrophaeus ATCC 9372 and B. sp. ATCC 29669, under three conditions encompassing 5 temperature points. Embedded experiments utilized a silicone rubber polymer that is commonly used on robotic spacecraft, and surface exposed experiments were performed under both ambient and vacuum-controlled humidity conditions. The results obtained support the use of DHMR protocols that extend the maximum temperature range from 125° C to 170° C, with either controlled or ambient humidity. If implemented, this will give projects bioburden reduction credit for shorter treatments at extended temperatures, and allow spacecraft to be processed in more readily available and less expensive facilities that do not have humidity control, with significant cost and schedule benefits. The study also demonstrated that the required heating time for materials presumed to have embedded bioburden is conservative.

  12. Efficacy of oxonia active against selected spore formers.

    Science.gov (United States)

    Blakistone, B; Chuyate, R; Kautter, D; Charbonneau, J; Suit, K

    1999-03-01

    Alternatives to hydrogen peroxide are being sought for use in aseptic packaging systems because this sterilant is efficacious at temperatures higher than some of the newer packaging materials can tolerate. Earlier in this century, peracetic acid was known to be bactericidal, sporicidal, and virucidal but was not widely used because of handling, toxicity, and stability problems. Sanitizer suppliers have capitalized on the efficacy of hydrogen peroxide, acetic acid, and peracetic acid stabilized with a sequestering agent. Formulations have been improved and marketed as Oxonia Active, and its use as an alternative sterilant to hydrogen peroxide merits evaluation. Oxonia was assessed at a concentration of 2% and a temperature of 40 degrees C against a number of spore-forming organisms, including foodborne pathogens. Spores tested in aqueous suspension showed an order of sensitivity (least to greatest) to Oxonia as follows: Bacillus cereus > B. subtilis A > B. stearothermophilus > B. subtilis var. globigii > B. coagulans > Clostridium sporogenes (PA3679) > C. butyricum > C. botulinum type B (nonproteolytic) > C. botulinum type B (proteolytic) = C. botulinum type A = C. botulinum type E. B. subtilis A and B. stearothermophilus spores tested in the dry state were less sensitive to Oxonia than when tested in aqueous suspension. B. cereus, a foodborne pathogen, proved to be markedly less sensitive to Oxonia under the described test conditions. The decreased sensitivity to Oxonia by the foodborne pathogen B. cereus raises concern about the efficacy of the sterilant for aseptic packaging of low-acid foods. Further work will be needed to determine if this decreased sensitivity is an inherent property of the organism that affords unusual protection against Oxonia or if the challenge parameters selected were at the minimum conditions for efficacy. PMID:10090246

  13. Atividade da invertase ácida solúvel e da insolúvel em tubérculos de batata recondicionados após o armazenamento sob diferentes temperaturas Acid soluble invertase and insoluble activity in potato tuber subsequent reconditioning after storage in different temperatures

    Directory of Open Access Journals (Sweden)

    Ladislau Soares Ferreira

    2007-12-01

    Full Text Available Objetivou-se no trabalho determinar a atividade das enzimas invertase ácida solúvel e invertase insolúvel em tubérculos de batata de genótipos Atlantic, Pérola, Asterix e C-1786-6-94, armazenados em diferentes temperaturas com posterior recondicionamento. Determinou-se a atividade das enzimas invertase ácida solúvel e insolúvel aos 0, 30 e 60 dias de armazenamento. Aos 30 dias de armazenamento a atividade das duas enzimas foi estimulada pela temperatura de 4ºC. O recondicionamento de 4ºC para 20ºC diminuiu a atividade das enzimas em todos os cultivares. O armazenamento a 12ºC bem como o recondicionamento de 12ºC para 20ºC pouco influenciou na atividade da invertase ácida solúvel. A atividade da invertase insolúvel sob 12ºC foi aumentada aos 30 e 60 dias de armazenamento, no clone C1786-6-94 e na cultivar Atlantic. O armazenamento a 20ºC não alterou a atividade das enzimas invertase ácida solúvel e insolúvel para as cultivares em estudo.The aim of this work was to quantify the activity of the acid soluble invertase enzymes and insoluble invertase in tubers of potato genotypes: Atlantic, Pérola, Asterix and C-1786-6-94 stored in different temperatures with recondition. One determined the activity of the acid soluble enzymes and insoluble invertase at 0 30 and 60 days of storage. In 30 days of storage the activity of the two enzymes was stimulated by the temperature at 4ºC. The recondition from 4º C to 20º C reduced the activity of the insoluble and acid soluble invertase in all cultivars, however in different way among them. The storage at 12ºC and the reconditioned from 12º C to 20º C had little influence on the activity of the acid soluble invertase. The activity of the insoluble invertase under 12º C was increased at 30 and 60 days of recondition in C-1786-6-94 clone and Atlantic cultivar. The storage at 20ºC did not alter the activity of the acid soluble invertase enzyme and insoluble invertase for the

  14. Spore production of Beauveria bassiana from agro-industrial residues

    OpenAIRE

    Herta Stutz Dalla Santa; Osmar Roberto Dalla Santa; Débora Brand; Luciana Porto de Souza Vandenberghe; Carlos Ricardo Soccol

    2005-01-01

    The purpose of this work was to produce Beauveria bassiana by Solid-State Fermentation using agro-industrial residues and optimizing the cultivation conditions. Refused potatoes, coffee husks and sugar-cane bagasse were tested. The blend of refused potatoes and sugar-cane bagasse (60-40%) with particle size in the range of 0.8-2 mm was used in the fermentation experiments. In Erlenmeyer flasks the best spore production was achieved with the following conditions: incubation temperature 26º C; ...

  15. Spore production of Beauveria bassiana from agro-industrial residues

    Directory of Open Access Journals (Sweden)

    Herta Stutz Dalla Santa

    2005-06-01

    Full Text Available The purpose of this work was to produce Beauveria bassiana by Solid-State Fermentation using agro-industrial residues and optimizing the cultivation conditions. Refused potatoes, coffee husks and sugar-cane bagasse were tested. The blend of refused potatoes and sugar-cane bagasse (60-40% with particle size in the range of 0.8-2 mm was used in the fermentation experiments. In Erlenmeyer flasks the best spore production was achieved with the following conditions: incubation temperature 26º C; initial pH 6.0; inoculum concentration 10(7 spores.g-1.dw and initial moisture 75%. In the column type reactor using forced aeration under the optimized conditions, the maximum production (1.07x10(10spores.g-1.dw was obtained at the 10th day of fermentation. The respirometric analyses of the fermentation showed a strong correlation between fungal growth and spore production.O objetivo deste trabalho foi produzir Beauveria bassiana por fermentação no estado sólido em resíduos agro-industriais e otimizar as condições de cultivo. Batata-refugo, polpa de café e bagaço de cana de açúcar foram testados. A mistura de batata-refugo e de bagaço de cana de açúcar (60:40%, com granulometria de 2 a 0,8 mm foi escolhida como melhor substrato/suporte. Em frascos de Erlenmeyer a produção de esporos foi maior com as seguintes condições: pH 6,0; temperatura de incubação de 26º C; taxa de inóculo de 10(7 esporos.g-1 de matéria seca; e umidade inicial de 75%. Em bioreator do tipo coluna com aeração forçada, as condições otimizadas possibilitaram uma produção máxima de esporos no 10º dia de fermentação, obtendo-se 1,07x10(10 esporos.g-1 de matéria seca. A análise respirométrica desta fermentação permitiu correlacionar o desenvolvimento do fungo com a produção de esporos.

  16. An Optical Biosensor for Bacillus Cereus Spore Detection

    Science.gov (United States)

    Li, Chengquan; Tom, Harry W. K.

    2005-03-01

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

  17. Ice Nucleation of Fungal Spores from the Classes Agaricomycetes, Ustilaginomycetes, and Eurotiomycetes, and the effect on the Atmospheric Transport of these Spores

    Energy Technology Data Exchange (ETDEWEB)

    Haga, D. I.; Burrows, Susannah M.; Iannone, R.; Wheeler, M. J.; Mason, R.; Chen, J.; Polishchuk, E. A.; Poschl, U.; Bertram, Allan K.

    2014-08-26

    Ice nucleation on fungal spores may affect the frequency and properties of ice and mixed-phase clouds. We studied the ice nucleation properties of 12 different species of fungal spores chosen from three classes: Agaricomycetes, Ustilagomycetes, and Eurotiomycetes. Agaricomycetes include many types of mushroom species and are cosmopolitan all over the globe. Ustilagomycetes are agricultural pathogens and have caused widespread damage to crops. Eurotiomycetes are found on all types of decaying material and include important human allergens. We focused on these classes since they are thought to be abundant in the atmosphere and because there is very little information on the ice nucleation ability of these classes of spores in the literature. All of the fungal spores investigated were found to cause freezing of water droplets at temperatures warmer than homogeneous freezing. The cumulative number of ice nuclei per spore was 0.001 at temperatures between -19 °C and -29 °C, 0.01 between -25.5 °C and -31 °C, and 0.1 between -26 °C and -36 °C. On average, the order of ice nucleating ability for these spores is Ustilagomycetes > Agaricomycetes ≅ Eurotiomycetes. We show that at temperatures below -20 °C, all of the fungal spores studied here are less efficient ice nuclei compared to Asian mineral dust on a per surface area basis. We used our new freezing results together with data in the literature to compare the freezing temperatures of spores from the phyla Basidiomycota and Ascomycota, which together make up 98 % of known fungal species found on Earth. The data show that within both phyla (Ascomycota and Basidiomycota) there is a wide range of freezing properties, and also that the variation within a phylum is greater than the variation between the average freezing properties of the phyla. Using a global chemistry-climate transport model, we investigated whether ice nucleation on the studied spores, followed by precipitation, can influence the atmospheric

  18. Comparison of the antitumor activity on spore polysaccharides (G/-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum

    Institute of Scientific and Technical Information of China (English)

    Peng-yunWANG; Zhi-binLIN

    2004-01-01

    AIM: To compare the antitumor activity of spore polysaccharides (GI-SP) and broken spore polysaccharides (GI-BSP) from spores of Ganoderma lucidum (Leyss ex fr) Karst. METHODS: BALB/c mice were implanted with Sarcoma 180 and administered intragastrically with Gl-SP or G/-BPS (50, 100, 200 mg/kg)respectively for 14 d. At the end of experiment, the tumor were removed and weighted. At the same time, spleens of tumorbearing mice were prepared to observe the effect of Gl-SP and

  19. Localization of the Germination Protein GerD to the Inner Membrane in Bacillus subtilis Spores▿

    OpenAIRE

    Pelczar, Patricia L.; Setlow, Peter

    2008-01-01

    GerD of Bacillus subtilis is a protein essential for normal spore germination with either l-alanine or a mixture of l-asparagine, d-glucose, d-fructose, and potassium ions. GerD's amino acid sequence suggests that it may be a lipoprotein, indicating a likely location in a membrane. Location in the spore's outer membrane seems unlikely, since removal of this membrane does not result in a gerD spore germination phenotype, suggesting that GerD is likely in the spore's inner membrane. In order to...

  20. 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

  1. 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.

  2. Activate to eradicate: inhibition of Clostridium difficile spore outgrowth by the synergistic effects of osmotic activation and nisin.

    Directory of Open Access Journals (Sweden)

    Michelle M Nerandzic

    Full Text Available BACKGROUND: Germination is the irreversible loss of spore-specific properties prior to outgrowth. Because germinating spores become more susceptible to killing by stressors, induction of germination has been proposed as a spore control strategy. However, this strategy is limited by superdormant spores that remain unaffected by germinants. Harsh chemicals and heat activation are effective for stimulating germination of superdormant spores but are impractical for use in a hospital setting, where Clostridium difficile spores present a challenge. Here, we tested whether osmotic activation solutes will provide a mild alternative for stimulation of superdormant C. difficile spores in the presence of germinants as previously demonstrated in several species of Bacillus. In addition, we tested the hypothesis that the limitations of superdormancy can be circumvented with a combined approach using nisin, a FDA-approved safe bacteriocin, to inhibit outgrowth of germinated spores and osmotic activation solutes to enhance outgrowth inhibition by stimulating superdormant spores. PRINCIPAL FINDINGS: Exposure to germination solution triggered ~1 log(10 colony forming units (CFU of spores to germinate, and heat activation increased the spores that germinated to >2.5 log(10CFU. Germinating spores, in contrast to dormant spores, became susceptible to inhibition by nisin. The presence of osmotic activation solutes did not stimulate germination of superdormant C. difficile spores exposed to germination solution. But, in the absence of germination solution, osmotic activation solutes enhanced nisin inhibition of superdormant spores to >3.5 log(10CFU. The synergistic effects of osmotic activation solutes and nisin were associated with loss of membrane integrity. CONCLUSIONS: These findings suggest that the synergistic effects of osmotic activation and nisin bypass the limitations of germination as a spore control strategy, and might be a novel method to safely and

  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. Draft Genome Sequences of Seven Thermophilic Spore-Forming Bacteria Isolated from Foods That Produce Highly Heat-Resistant Spores, Comprising Geobacillus spp., Caldibacillus debilis, and Anoxybacillus flavithermus

    NARCIS (Netherlands)

    Berendsen, Erwin M; Wells-Bennik, Marjon H J; Krawczyk, Antonina O; de Jong, Anne; van Heel, Auke; Holsappel, Siger; Eijlander, Robyn T; Kuipers, Oscar P

    2016-01-01

    Here, we report the draft genomes of five strains of Geobacillus spp., one Caldibacillus debilis strain, and one draft genome of Anoxybacillus flavithermus, all thermophilic spore-forming Gram-positive bacteria.

  6. Draft Genome Sequences of Seven Thermophilic Spore-Forming Bacteria Isolated from Foods That Produce Highly Heat-Resistant Spores, Comprising Geobacillus spp., Caldibacillus debilis, and Anoxybacillus flavithermus

    Science.gov (United States)

    Berendsen, Erwin M.; Wells-Bennik, Marjon H. J.; Krawczyk, Antonina O.; de Jong, Anne; van Heel, Auke; Holsappel, Siger; Eijlander, Robyn T.

    2016-01-01

    Here, we report the draft genomes of five strains of Geobacillus spp., one Caldibacillus debilis strain, and one draft genome of Anoxybacillus flavithermus, all thermophilic spore-forming Gram-positive bacteria. PMID:27151781

  7. Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi

    Directory of Open Access Journals (Sweden)

    Carrie E. Jolly

    2010-01-01

    Full Text Available Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02_0100; “C1” was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and specific antiserum was generated. Two-dimensional Western blotting analysis showed that the purified antibodies were monospecific. Immunoelectron microscopy of developing and mature E. cuniculi spores revealed that the protein localized to internal structures and not to the spore surface. In spore adherence inhibition assays, the anti-C1 antibodies did not inhibit spore adherence to host cell surfaces, whereas antibodies to a known surface adhesin (EnP1 did so. In future studies, the antibodies to the ‘C1’ Hsp70 will be used to delineate spore surface protein expression.

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

    DEFF Research Database (Denmark)

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

    to the evolution of Bacillus cereus group bacteria (e.g. B. cereus, B. anthracis, B. thuringiensis) as a pathogen. It has been hypothesized that the spore stage protects against digestion by predating protists. Indeed, B. thuringiensis spores have been shown to be readily ingested by ciliated protists but failed...

  9. Gas discharge plasmas are effective in inactivating Bacillus and Clostridium spores.

    Science.gov (United States)

    Tseng, Shawn; Abramzon, Nina; Jackson, James O; Lin, Wei-Jen

    2012-03-01

    Bacterial spores are the most resistant form of life and have been a major threat to public health and food safety. Nonthermal atmospheric gas discharge plasma is a novel sterilization method that leaves no chemical residue. In our study, a helium radio-frequency cold plasma jet was used to examine its sporicidal effect on selected strains of Bacillus and Clostridium. The species tested included Bacillus subtilis, Bacillus stearothermophilus, Clostridium sporogenes, Clostridium perfringens, Clostridium difficile, and Clostridium botulinum type A and type E. The plasmas were effective in inactivating selected Bacillus and Clostridia spores with D values (decimal reduction time) ranging from 2 to 8 min. Among all spores tested, C. botulinum type A and C. sporogenes were significantly more resistant to plasma inactivation than other species. Observations by phase contrast microscopy showed that B. subtilis spores were severely damaged by plasmas and the majority of the treated spores were unable to initiate the germination process. There was no detectable fragmentation of the DNA when the spores were treated for up to 20 min. The release of dipicolinic acid was observed almost immediately after the plasma treatment, indicating the spore envelope damage could occur quickly resulting in dipicolinic acid release and the reduction of spore resistance.

  10. Detection of Fungal Spores Using a Generic Surface Plasmon Resonance Immunoassay

    DEFF Research Database (Denmark)

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

    2007-01-01

    . Spiked Pst samples were further examined in a background of a related spore and it was found that Pst quantification was possible in this mixture. This study represent the first use of SPR technology for fungal spore detection as well as the first report of a successful biosensor-based detection strategy...

  11. Release of elicitors from rice blast spores under the action of reactive oxygen species

    Science.gov (United States)

    The effects of reactive oxygen species (ROS) on secretion of hypothesized elicitors from spores of rice blast causal fungus Magnaporthe grisea were studied. For spore exposure to exogenous ROS, they were germinated for 5 h in 50 µM H2O2 followed by addition of catalase E.C. 1.11.1.6 (to decompose pe...

  12. The characterisation of Bacillus spores occurring in the manufacturing of (low acid) canned products

    NARCIS (Netherlands)

    Oomes, S.J.C.M.; Zuijlen, A.C.M. van; Hehenkamp, J.O.; Witsenboer, H.; Vossen, J.M.B.M. van der; Brul, S.

    2007-01-01

    Spore-forming bacteria can be a problem in the food industry, especially in the canning industry. Spores present in ingredients or present in the processing environment severely challenge the preservation process since their thermal resistance may be very high. We therefore asked the question which

  13. 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.

    hydrostatic pressure, low temperature and low nutrients. Effects of these on germination of spores from a few deep-sea Aspergillus isolates are examined. Spores from most of the fungi germinated under elevated hydrostatic pressure at 30 degrees C. The ambient...

  14. 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.

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

    DEFF Research Database (Denmark)

    Stockmarr, Anders; Andreasen, Viggo; Østergård, Hanne

    2007-01-01

    A new modeling framework for particle dispersal is explored in the context of the particles being fungal spores dispersed within a field. The model gives rise to both exponentially decreasing and polynomially decreasing two-dimensional densities of deposited fungal spores. We reformulate the mode...

  16. Inactivation of Spores of Bacillus anthracis Sterne, Bacillus cereus, and Bacillus thuringiensis subsp. israelensis by Chlorination

    OpenAIRE

    Rice, E W; Adcock, N. J.; Sivaganesan, M; Rose, L. J.

    2005-01-01

    Three species of Bacillus were evaluated as potential surrogates for Bacillus anthracis for determining the sporicidal activity of chlorination as commonly used in drinking water treatment. Spores of Bacillus thuringiensis subsp. israelensis were found to be an appropriate surrogate for spores of B. anthracis for use in chlorine inactivation studies.

  17. 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.

  18. 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...

  19. Minimizing the level of butyric acid bacteria spores in farm tank milk

    NARCIS (Netherlands)

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

    2007-01-01

    A year-long survey of 24 dairy farms was conducted to determine the effects of farm management on the concentrations of butyric acid bacteria (BAB) spores in farm tank milk (FTM). The results were used to validate a control strategy derived from model simulations. The BAB spore concentrations were m

  20. Gas discharge plasmas are effective in inactivating Bacillus and Clostridium spores.

    Science.gov (United States)

    Tseng, Shawn; Abramzon, Nina; Jackson, James O; Lin, Wei-Jen

    2012-03-01

    Bacterial spores are the most resistant form of life and have been a major threat to public health and food safety. Nonthermal atmospheric gas discharge plasma is a novel sterilization method that leaves no chemical residue. In our study, a helium radio-frequency cold plasma jet was used to examine its sporicidal effect on selected strains of Bacillus and Clostridium. The species tested included Bacillus subtilis, Bacillus stearothermophilus, Clostridium sporogenes, Clostridium perfringens, Clostridium difficile, and Clostridium botulinum type A and type E. The plasmas were effective in inactivating selected Bacillus and Clostridia spores with D values (decimal reduction time) ranging from 2 to 8 min. Among all spores tested, C. botulinum type A and C. sporogenes were significantly more resistant to plasma inactivation than other species. Observations by phase contrast microscopy showed that B. subtilis spores were severely damaged by plasmas and the majority of the treated spores were unable to initiate the germination process. There was no detectable fragmentation of the DNA when the spores were treated for up to 20 min. The release of dipicolinic acid was observed almost immediately after the plasma treatment, indicating the spore envelope damage could occur quickly resulting in dipicolinic acid release and the reduction of spore resistance. PMID:22075631

  1. 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…

  2. Detecting Clostridium difficile spores from inanimate surfaces of the hospital environment: which method is best?

    Science.gov (United States)

    Claro, Tânia; Daniels, Stephen; Humphreys, Hilary

    2014-09-01

    The recovery of Clostridium difficile spores from hospital surfaces was assessed using rayon swabs, flocked swabs, and contact plates. The contact plate method was less laborious, achieved higher recovery percentages, and detected spores at lower inocula than swabs. Rayon swabs were the least efficient method. However, further studies are required in health care settings.

  3. 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...

  4. DESTRUCTION OF ASPERGILLUS VERSICOLOR, PENICILLIUM CRYSOGENUM, STACHYBOTRYS CHARTARUM, AND CLADOSPORIUM CLADOSPORIDES SPORES USING CHEMICAL OXIDATION TREATMENT PROCESS

    Science.gov (United States)

    The survival of aqueous suspensions of Penicillium chrysogenum, Stachybotrys chartarum, Aspergillus versicolor, and Cladosporium cladosporioides spores was evaluated using various combinations of hydrogen peroxide and iron (II) as catalyst. Spores were suspended in water and trea...

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

    Energy Technology Data Exchange (ETDEWEB)

    Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Tsouris, Costas [ORNL

    2011-01-01

    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. 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.

  6. The Adsorption Properties of Bacillus atrophaeus Spore on Functionalized Carbon Nanotubes

    Directory of Open Access Journals (Sweden)

    P. Cortes

    2010-01-01

    Full Text Available An equilibrium study of Bacillus atrophaeus (B.a spores on functionalized Single-Wall Carbon Nanotubes (SWCNTs has been performed in order to characterize the adsorption properties of the spores/nanotubes complex. The carbon nanotubes here investigated were subjected to a two-step purification and functionalization treatment in order to introduce chemical groups on their basal planes. The inclusion of carboxyl functional groups on the nanotubes was corroborated by Raman and infrared spectroscopy. These carboxyl groups appear to enhance the nanotube-B.a. interaction by reacting with the proteinaceous pili appendages present on the spore surface. The adsorption data demonstrate that bacillus spores diffuse faster on functionalized carbon nanotubes than on as-received and purified nanomaterials. Transmission Electron Microscopy also shows that the chemically treated nanotubes resulted in a swollen nano-network which seems to further enhance the bacillus adsorption due to a more extensive spore-nanotube contact area.

  7. 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

  8. 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. PMID:23658025

  9. Cold-air atmospheric pressure plasma against Clostridium difficile spores: a potential alternative for the decontamination of hospital inanimate surfaces.

    Science.gov (United States)

    Claro, Tânia; Cahill, Orla J; O'Connor, Niall; Daniels, Stephen; Humphreys, Hilary

    2015-06-01

    Clostridium difficile spores survive for months on environmental surfaces and are highly resistant to decontamination. We evaluated the effect of cold-air plasma against C. difficile spores. The single-jet had no effect while the multi-jet achieved 2-3 log10 reductions in spore counts and may augment traditional decontamination.

  10. The use of bacterial spore formers as probiotics.

    Science.gov (United States)

    Hong, Huynh A; Duc, Le Hong; Cutting, Simon M

    2005-09-01

    The field of probiosis has emerged as a new science with applications in farming and aqaculture as alternatives to antibiotics as well as prophylactics in humans. Probiotics are being developed commercially for both human use, primarily as novel foods or dietary supplements, and in animal feeds for the prevention of gastrointestinal infections, with extensive use in the poultry and aquaculture industries. The impending ban of antibiotics in animal feed, the current concern over the spread of antibiotic resistance genes, the failure to identify new antibiotics and the inherent problems with developing new vaccines make a compelling case for developing alternative prophylactics. Among the large number of probiotic products in use today are bacterial spore formers, mostly of the genus Bacillus. Used primarily in their spore form, these products have been shown to prevent gastrointestinal disorders and the diversity of species used and their applications are astonishing. Understanding the nature of this probiotic effect is complicated, not only because of the complexities of understanding the microbial interactions that occur within the gastrointestinal tract (GIT), but also because Bacillus species are considered allochthonous microorganisms. This review summarizes the commercial applications of Bacillus probiotics. A case will be made that many Bacillus species should not be considered allochthonous microorganisms but, instead, ones that have a bimodal life cycle of growth and sporulation in the environment as well as within the GIT. Specific mechanisms for how Bacillus species can inhibit gastrointestinal infections will be covered, including immunomodulation and the synthesis of antimicrobials. Finally, the safety and licensing issues that affect the use of Bacillus species for commercial development will be summarized, together with evidence showing the growing need to evaluate the safety of individual Bacillus strains as well as species on a case by case by basis

  11. 分光光度法测定生物质水解液中糠醛、酸溶木质素、还原糖的含量%FURFURAL, ACID-SOLUBLE LIGNIN AND REDUCING SUGAR CONTENT IN BIOMASS HYDROLYSATE BY SPECTROPHOTOMETRIC METHOD

    Institute of Scientific and Technical Information of China (English)

    孙绍晖; 宋勇; 孙培勤; 陈俊武

    2013-01-01

    使用紫外可见分光光度法测定糠醛、酸溶木质素、还原糖浓度,通过选择合适的波长、空白和测定顺序,消除各组分之间的相互干扰.优化的测定方法为:首先用双波长法测定糠醛和酸溶木质素浓度,再在530nm处测定酸水解液与DNS的吸光值扣除糠醛与DNS的吸光值即为还原糖与DNS的吸光值.可简便快速准确测定生物质水解液中糠醛、酸溶木质素、还原糖浓度.%The concentrations of furfural, acid-soluble lignin and reducing sugar were detected using UV-visible spectrophotometry. The interaction between the components was eliminated by selecting the appropriate wavelength, the blank and detection order. The optimum detection method was as follows: first, detecting the concentration of furfural and acid-soluble lignin using dual-wavelength method; then, detecting the absorbance of acid hydrolysate and DNS at 530 nm, the exact absorbance of sugar and DNS was obtained by deducting absorbance of furfural and DNS. The concentration of the furfural, acid-soluble lignin, reducing sugar in woody biomass hydrolyzed solution can be accurately determined by this method.

  12. Mechanistic studies of the spore photoproduct lyase via a single cysteine mutation.

    Science.gov (United States)

    Yang, Linlin; Lin, Gengjie; Nelson, Renae S; Jian, Yajun; Telser, Joshua; Li, Lei

    2012-09-11

    5-Thyminyl-5,6-dihydrothymine (also called spore photoproduct or SP) is the exclusive DNA photodamage product in bacterial endospores. It is repaired by a radical SAM (S-adenosylmethionine) enzyme, the spore photoproduct lyase (SPL), at the bacterial early germination phase. Our previous studies proved that SPL utilizes the 5'-dA• generated by the SAM cleavage reaction to abstract the H(6proR) atom to initiate the SP repair process. The resulting thymine allylic radical was suggested to take an H atom from an unknown protein source, most likely cysteine 141. Here we show that C141 can be readily alkylated in the native SPL by an iodoacetamide treatment, suggesting that it is accessible to the TpT radical. SP repair by the SPL C141A mutant yields TpTSO(2)(-) and TpT simultaneously from the very beginning of the reaction; no lag phase is observed for TpTSO(2)(-) formation. Should any other protein residue serve as the H donor, its presence would result in TpT being the major product at least for the first enzyme turnover. These observations provide strong evidence to support C141 as the direct H atom donor. Moreover, because of the lack of this intrinsic H donor, the C141A mutant produces TpT via an unprecedented thymine cation radical reduction (proton-coupled electron transfer) process, contrasting to the H atom transfer mechanism in the wild-type (WT) SPL reaction. The C141A mutant repairs SP at a rate that is ~3-fold slower than that of the WT enzyme. Formation of TpTSO(2)(-) and TpT exhibits a V(max) deuterium kinetic isotope effect (KIE) of 1.7 ± 0.2, which is smaller than the (D)V(max) KIE of 2.8 ± 0.3 determined for the WT SPL reaction. These findings suggest that removing the intrinsic H atom donor disturbs the rate-limiting process during enzyme catalysis. As expected, the prereduced C141A mutant supports only ~0.4 turnover, which is in sharp contrast to the >5 turnovers exhibited by the WT SPL reaction, suggesting that the enzyme catalytic cycle (SAM

  13. Spore test parameters matter: Mesophilic and thermophilic spore counts detected in raw milk and dairy powders differ significantly by test method.

    Science.gov (United States)

    Kent, D J; Chauhan, K; Boor, K J; Wiedmann, M; Martin, N H

    2016-07-01

    United States dairy industry exports have steadily risen in importance over the last 10yr, with dairy powders playing a particularly critical role. Currently, approximately half of US-produced nonfat dry milk and skim milk powder is exported. Reaching new and expanding existing export markets relies in part on the control of endospore-forming bacteria in dairy powders. This study reports baseline mesophilic and thermophilic spore counts and spore populations from 55 raw material samples (primarily raw milk) and 33 dairy powder samples from dairy powder processors across the United States. Samples were evaluated using various spore testing methodologies and included initial heat treatments of (1) 80°C for 12 min; (2) 100°C for 30 min; and (3) 106°C for 30 min. Results indicate that significant differences in both the level and population of spores were found for both raw milk and dairy powders with the various testing methods. Additionally, on average, spore counts were not found to increase significantly from the beginning to the end of dairy powder processing, most likely related to the absence of biofilm formation by processing plant-associated sporeformers (e.g., Anoxybacillus sp.) in the facilities sampled. Finally, in agreement with other studies, Bacillus licheniformis was found to be the most prevalent sporeformer in both raw materials and dairy powders, highlighting the importance of this organism in developing strategies for control and reduction of spore counts in dairy powders. Overall, this study emphasizes the need for standardization of spore enumeration methodologies in the dairy powder industry.

  14. Spore test parameters matter: Mesophilic and thermophilic spore counts detected in raw milk and dairy powders differ significantly by test method.

    Science.gov (United States)

    Kent, D J; Chauhan, K; Boor, K J; Wiedmann, M; Martin, N H

    2016-07-01

    United States dairy industry exports have steadily risen in importance over the last 10yr, with dairy powders playing a particularly critical role. Currently, approximately half of US-produced nonfat dry milk and skim milk powder is exported. Reaching new and expanding existing export markets relies in part on the control of endospore-forming bacteria in dairy powders. This study reports baseline mesophilic and thermophilic spore counts and spore populations from 55 raw material samples (primarily raw milk) and 33 dairy powder samples from dairy powder processors across the United States. Samples were evaluated using various spore testing methodologies and included initial heat treatments of (1) 80°C for 12 min; (2) 100°C for 30 min; and (3) 106°C for 30 min. Results indicate that significant differences in both the level and population of spores were found for both raw milk and dairy powders with the various testing methods. Additionally, on average, spore counts were not found to increase significantly from the beginning to the end of dairy powder processing, most likely related to the absence of biofilm formation by processing plant-associated sporeformers (e.g., Anoxybacillus sp.) in the facilities sampled. Finally, in agreement with other studies, Bacillus licheniformis was found to be the most prevalent sporeformer in both raw materials and dairy powders, highlighting the importance of this organism in developing strategies for control and reduction of spore counts in dairy powders. Overall, this study emphasizes the need for standardization of spore enumeration methodologies in the dairy powder industry. PMID:27085396

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

    Science.gov (United States)

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

    2016-05-01

    After severe wildfires, pine recovery depends on ectomycorrhizal (ECM) fungal spores surviving and serving as partners for regenerating forest trees. We took advantage of a large, severe natural forest fire that burned our long-term study plots to test the response of ECM fungi to fire. We sampled the ECM spore bank using pine seedling bioassays and high-throughput sequencing before and after the California Rim Fire. We found that ECM spore bank fungi survived the fire and dominated the colonization of in situ and bioassay seedlings, but there were specific fire adapted fungi such as Rhizopogon olivaceotinctus that increased in abundance after the fire. The frequency of ECM fungal species colonizing pre-fire bioassay seedlings, post-fire bioassay seedlings and in situ seedlings were strongly positively correlated. However, fire reduced the ECM spore bank richness by eliminating some of the rare species, and the density of the spore bank was reduced as evidenced by a larger number of soil samples that yielded uncolonized seedlings. Our results show that although there is a reduction in ECM inoculum, the ECM spore bank community largely remains intact, even after a high-intensity fire. We used advanced techniques for data quality control with Illumina and found consistent results among varying methods. Furthermore, simple greenhouse bioassays can be used to determine which fungi will colonize after fires. Similar to plant seed banks, a specific suite of ruderal, spore bank fungi take advantage of open niche space after fires. PMID:26473720

  16. Detecting invisible bacillus spores on surfaces using a portable surface-enhanced Raman analyzer

    Science.gov (United States)

    Farquharson, Stuart; Inscore, Frank; Sperry, Jay F.

    2006-10-01

    Since the distribution of anthrax causing spores through the U.S. Postal System in the autumn of 2001, numerous methods have been developed to detect spores with the goal of minimizing casualties. During and following an attack it is also important to detect spores on surfaces, to assess extent of an attack, to quantify risk of infection by contact, as well as to evaluate post-attack clean-up. To perform useful measurements, analyzers and/or methods must be capable of detecting as few as 10 spores/cm2, in under 5-minutes, with little or no sample preparation or false-positive responses, using a portable device. In an effort to develop such a device, we have been investigating the ability of surfaceenhanced Raman spectroscopy (SERS) to detect dipicolinic acid (DPA) as a chemical signature of bacilli spores. In 2003 we employed SERS to measure DPA extracted from a 10,000 spores per μL sample using hot dodecylamine. Although the entire measurement was performed in 2 minutes, the need to heat the dodecylamine limits field portability of the method. Here we describe the use of a room temperature digesting agent in combination with SERS to detect 220 spores collected from a surface in a 1 μL sample within 3 minutes.

  17. Effect of ethanol perturbation on viscosity and permeability of an inner membrane in Bacillus subtilis spores.

    Science.gov (United States)

    Loison, Pauline; Gervais, Patrick; Perrier-Cornet, Jean-Marie; Kuimova, Marina K

    2016-09-01

    In this work, we investigated how a combination of ethanol and high temperature (70°C), affect the properties of the inner membrane of Bacillus subtilis spores. We observed membrane permeabilization for ethanol concentrations ≥50%, as indicated by the staining of the spores' DNA by the cell impermeable dye Propidium Iodide. The loss of membrane integrity was also confirmed by a decrease in the peak corresponding to dipicolinic acid using infrared spectroscopy. Finally, the spore refractivity (as measured by phase contrast microscopy) was decreased after the ethanol-heat treatment, suggesting a partial rehydration of the protoplast. Previously we have used fluorescent lifetime imaging microscopy (FLIM) combined with the fluorescent molecular rotor Bodipy-C12 to study the microscopic viscosity in the inner membrane of B. subtilis spores, and showed that at normal conditions it is characterized by a very high viscosity. Here we demonstrate that the ethanol/high temperature treatment led to a decrease of the viscosity of the inner membrane, from 1000cP to 860cP for wild type spores at 50% of ethanol. Altogether, our present work confirms the deleterious effect of ethanol on the structure of B. subtilis spores, as well as demonstrates the ability of FLIM - Bodipy-C12 to measure changes in the microviscosity of the spores upon perturbation. PMID:27267704

  18. 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.

  19. Inactivation of Alicyclobacillus acidoterrestris spores in apple and orange juice concentrates by gamma irradiation.

    Science.gov (United States)

    Lee, Su-Yeon; Park, Sang-Hyun; Kang, Dong-Hyun

    2014-02-01

    The objective of this study was to evaluate the effect of different concentrations of reconstituted apple and orange juice on reduction of Alicyclobacillus acidoterrestris spores by gamma irradiation. Spores of A. acidoterrestris were inoculated into three concentrations of apple (18, 36, and 72 °Brix) and orange (11, 33, and 66 °Brix) juice and subjected to five radiation doses (1, 3, 5, 7, and 10 kGy). No significant reductions (P > 0.05) in spores were observed after the 1-kGy treatment for all apple and orange concentrations. Spores in 18, 36, and 72 °Brix apple juice concentrates subjected to 10 kGy were reduced to 4.34, 3.9, and 3.84 log CFU/ml, respectively. Similar results were observed for orange juice. When 10 kGy was applied to 11 °Brix orange juice, populations of spores were reduced by 5 log CFU/ml. The reduction of spores in 33 and 66 °Brix orange juice concentrates exposed to 10-kGy gamma irradiation was 4.54 and 3.85 log CFU/ml, respectively. Juice concentration did not affect (P > 0.05) the number of surviving A. acidoterrestris spores from the same kGy treatment. Gamma irradiation treatment did not change the pH or water activity of the juice (P > 0.05).

  20. 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.

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

    Science.gov (United States)

    Poppinga, Simon; Haushahn, Tobias; Warnke, Markus; Masselter, Tom; Speck, Thomas

    2015-01-01

    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. PMID:26444002

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

    Science.gov (United States)

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

    2013-03-01

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

  3. New insights in the bacterial spore resistance to extreme terrestrial and extraterrestrial factors

    Science.gov (United States)

    Moeller, Ralf; Horneck, Gerda; Reitz, Guenther

    Based on their unique resistance to various space parameters, Bacillus endospores are one of the model systems used for astrobiological studies. The extremely high resistance of bacterial endospores to environmental stress factors has intrigued researchers since long time and many characteristic spore features, especially those involved in the protection of spore DNA, have already been uncovered. The disclosure of the complete genomic sequence of Bacillus subtilis 168, one of the often used astrobiological model system, and the rapid development of tran-scriptional microarray techniques have opened new opportunities of gaining further insights in the enigma of spore resistance. Spores of B. subtilis were exposed to various extreme ter-restrial and extraterrestrial stressors to reach a better understanding of the DNA protection and repair strategies, which them to cope with the induced DNA damage. Following physical stress factors of environmental importance -either on Earth or in space -were selected for this thesis: (i) mono-and polychromatic UV radiation, (ii) ionizing radiation, (iii) exposure to ultrahigh vacuum; and (iv) high shock pressures simulating meteorite impacts. To reach a most comprehensive understanding of spore resistance to those harsh terrestrial or simulated extraterrestrial conditions, a standardized experimental protocol of the preparation and ana-lyzing methods was established including the determination of the following spore responses: (i) survival, (ii) induced mutations, (iii) DNA damage, (iv) role of different repair pathways by use of a set of repair deficient mutants, and (v) transcriptional responses during spore germi-nation by use of genome-wide transcriptome analyses and confirmation by RT-PCR. From this comprehensive set of data on spore resistance to a variety of environmental stress parameters a model of a "built-in" transcriptional program of bacterial spores in response to DNA damaging treatments to ensure DNA restoration

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

    Science.gov (United States)

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

    2016-01-01

    Elimination of Bacillus anthracis spores from the environment is a difficult and costly process due in part to the toxicity of current sporicidal agents. For this reason we investigated the ability of the spore germinants L-alanine (100 mM) and inosine (5 mM) to reduce the concentration of peracetic acid (PAA) required to inactivate B. anthracis spores. While L-alanine significantly enhanced (p = 0.0085) the bactericidal activity of 500 ppm PAA the same was not true for inosine suggesting some form of negative interaction. In contrast the germinant combination proved most effective at 100 ppm PAA (p = 0.0009). To determine if we could achieve similar results in soil we treated soil collected from the burial site of an anthrax infected animal which had been supplemented with spores of the Sterne strain of B. anthracis to increase the level of contamination to 10(4) spores/g. Treatment with germinants followed 1 h later by 5000 ppm PAA eliminated all of the spores. In contrast direct treatment of the animal burial site using this approach delivered using a back pack sprayer had no detectable effect on the level of B. anthracis contamination or on total culturable bacterial numbers over the course of the experiment. It did trigger a significant, but temporary, reduction (p < 0.0001) in the total spore count suggesting that germination had been triggered under real world conditions. In conclusion, we have shown that the application of germinants increase the sensitivity of bacterial spores to PAA. While the results of the single field trial were inconclusive, the study highlighted the potential of this approach and the challenges faced when attempting to perform real world studies on B. anthracis spores contaminated sites. PMID:26858699

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

    Science.gov (United States)

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

    2016-01-01

    Elimination of Bacillus anthracis spores from the environment is a difficult and costly process due in part to the toxicity of current sporicidal agents. For this reason we investigated the ability of the spore germinants L-alanine (100 mM) and inosine (5 mM) to reduce the concentration of peracetic acid (PAA) required to inactivate B. anthracis spores. While L-alanine significantly enhanced (p = 0.0085) the bactericidal activity of 500 ppm PAA the same was not true for inosine suggesting some form of negative interaction. In contrast the germinant combination proved most effective at 100 ppm PAA (p = 0.0009). To determine if we could achieve similar results in soil we treated soil collected from the burial site of an anthrax infected animal which had been supplemented with spores of the Sterne strain of B. anthracis to increase the level of contamination to 104 spores/g. Treatment with germinants followed 1 h later by 5000 ppm PAA eliminated all of the spores. In contrast direct treatment of the animal burial site using this approach delivered using a back pack sprayer had no detectable effect on the level of B. anthracis contamination or on total culturable bacterial numbers over the course of the experiment. It did trigger a significant, but temporary, reduction (p < 0.0001) in the total spore count suggesting that germination had been triggered under real world conditions. In conclusion, we have shown that the application of germinants increase the sensitivity of bacterial spores to PAA. While the results of the single field trial were inconclusive, the study highlighted the potential of this approach and the challenges faced when attempting to perform real world studies on B. anthracis spores contaminated sites. PMID:26858699

  6. At-line determination of spore inoculum quality in Penicillium chrysogenum bioprocesses.

    Science.gov (United States)

    Ehgartner, Daniela; Herwig, Christoph; Neutsch, Lukas

    2016-06-01

    Spore inoculum quality in filamentous bioprocesses is a critical parameter influencing pellet morphology and, consequently, process performance. It is essential to determine the concentration of viable spores before inoculation, to implement quality control and decrease batch-to-batch variability. The ability to assess the spore physiologic status with close-to-real time resolution would offer interesting perspectives enhanced process analytical technology (PAT) and quality by design (QbD) strategies. Up to now, the parameters contributing to spore inoculum quality are not clearly defined. The state-of-the-art method to investigate this variable is colony-forming unit (CFU) determination, which assesses the number of growing spores. This procedure is tedious, associated with significant inherent bias, and not applicable in real time.Here, a novel method is presented, based on the combination of viability staining (propidium iodide and fluorescein diacetate) and large-particle flow cytometry. It is compatible with the complex medium background often observed in filamentous bioprocesses and allows for a classification of the spores into different subpopulations. Next to viable spores with intact growth potential, dormant or inactive as well as physiologically compromised cells are accurately determined. Hence, a more holistic few on spore inoculum quality and early-phase biomass composition is provided, offering enhanced information content.In an industrially relevant model bioprocess, good correlation to CFU counts was found. Morphological parameters (e.g. spore swelling) that are not accessible via standard monitoring tools were followed over the initial process phase with close temporal resolution. PMID:26820651

  7. Fungal spores as potential ice nuclei in fog/cloud water and snow

    Science.gov (United States)

    Bauer, Heidi; Goncalves, Fabio L. T.; Schueller, Elisabeth; Puxbaum, Hans

    2010-05-01

    INTRODUCTION: In discussions about climate change and precipitation frequency biological ice nucleation has become an issue. While bacterial ice nucleation (IN) is already well characterized and even utilized in industrial processes such as the production of artificial snow or to improve freezing processes in food industry, less is known about the IN potential of fungal spores which are also ubiquitous in the atmosphere. A recent study performed at a mountain top in the Rocky Mountains suggests that fungal spores and/or pollen might play a role in increased IN abundance during periods of cloud cover (Bowers et al. 2009). In the present work concentrations of fungal spores in fog/cloud water and snow were determined. EXPERIMENTAL: Fog samples were taken with an active fog sampler in 2008 in a traffic dominated area and in a national park in São Paulo, Brazil. The number concentrations of fungal spores were determined by microscopic by direct enumeration by epifluorescence microscopy after staining with SYBR Gold nucleic acid gel stain (Bauer et al. 2008). RESULTS: In the fog water collected in the polluted area at a junction of two highly frequented highways around 22,000 fungal spores mL-1 were counted. Fog in the national park contained 35,000 spores mL-1. These results were compared with cloud water and snow samples from Mt. Rax, situated at the eastern rim of the Austrian Alps. Clouds contained on average 5,900 fungal spores mL-1 cloud water (1,300 - 11,000) or 2,200 spores m-3 (304 - 5,000). In freshly fallen snow spore concentrations were lower than in cloud water, around 1,000 fungal spores mL-1 were counted (Bauer et al. 2002). In both sets of samples representatives of the ice nucleating genus Fusarium could be observed. REFERENCES: Bauer, H., Kasper-Giebl, A., Löflund, M., Giebl, H., Hitzenberger, R., Zibuschka, F., Puxbaum, H. (2002). The contribution of bacteria and fungal spores to the organic carbon content of cloud water, precipitation and aerosols

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

    Directory of Open Access Journals (Sweden)

    Bazzle LJ

    2014-12-01

    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

  9. Mechanisms of sorbate inhibition of Bacillus cereus T and Clostridium botulinum 62A spore germination.

    OpenAIRE

    Smoot, L A; Pierson, M. D.

    1981-01-01

    The mechanism by which potassium sorbate inhibits Bacillus cereus T and Clostridium botulinum 62A spore germination was investigated. Spores of B. cereus T were germinated at 35 degrees C in 0.08 M sodium-potassium phosphate buffers (pH 5.7 and 6.7) containing various germinants (L-alanine, L-alpha-NH2-n-butyric acid, and inosine) and potassium sorbate. Spores of C. botulinum 62A were germinated in the same buffers but with 10 mM L-lactic acid, 20 mM sodium bicarbonate, L-alanine or L-cystein...

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

    Science.gov (United States)

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

    2016-01-01

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

  11. 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).

  12. Gender Influenced Spore Dimorphism in Nosema bombycis Nageli Causing Pebrine Disease in Mulberry Silkworm, Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Satadal CHAKRABARTY

    2013-04-01

    Full Text Available Nosema bombycis is a pathogen causing pebrine disease of the mulberry silkworm, Bombyx mori. The disease spreads mainly through transovarian transmission of environmental spore and secondarily through contaminated food, rearing appliances, etc. by primary spores. Ultra-structure studies using Scanning and Transmission Electron Microscopy of the two spores revealed differences in the primary spore which contained a Short Polar Tube (ST with a thin wall (< 200 nm, and the environmental spore which contained a Long Polar Tube (LT with a thick wall (> 200 nm. It is observed that the yield of spore with LT is highest in female moths, whereas, it is spores with ST are highest in male moths. Besides ultra-structures, the development pattern of the two types of spores is also different. It is an interesting finding in the present study that, spores of N. bombycis produced two types of spores and multiplied in different gender under the influence of the host’s reproductive role and physiology for transmission of disease. The detailed study on ultra-structure of disporous N. bombycis in both the sexes of B. mori along with their development in the life cycle stages of silkworms with special reference to the inoculum concentration of spore is discussed.  

  13. Nematicidal spore-forming Bacilli share similar virulence factors and mechanisms.

    Science.gov (United States)

    Zheng, Ziqiang; Zheng, Jinshui; Zhang, Zhengming; Peng, Donghai; Sun, Ming

    2016-01-01

    In the soil environment, Bacilli can affect nematode development, fecundity and survival. However, although many Bacillus species can kill nematodes, the virulence mechanisms Bacilli utilize remain unknown. In this study, we collected 120 strains comprising 30 species across the Bacillaceae and Paenibacillaceae families of the Bacillales order and measured their nematicidal activities in vitro. Comparison of these strains' nematicidal capacities revealed that nine species, including Bacillus thuringiensis, B. cereus, B. subtilis, B. pumilus, B. firmus, B. toyonensis, Lysinibacillus sphaericus, Brevibacillus laterosporus and B. brevis, were highly nematicidal, the first of which showed the highest activity. Genome sequencing and analysis identified many potential virulence factors, which grouped into five types. At least four possible mechanisms were deduced on the basis of the combination of these factors and the bacterial nematicidal activity, including a pore-forming mechanism of crystal proteins, an inhibition-like mechanism of thuringiensin and a degradation mechanism of proteases and/or chitinases. Our results demonstrate that 120 spore-forming Bacilli across different families share virulence factors that may contribute to their nematicidal capacity. PMID:27539267

  14. Nematicidal spore-forming Bacilli share similar virulence factors and mechanisms

    Science.gov (United States)

    Zheng, Ziqiang; Zheng, Jinshui; Zhang, Zhengming; Peng, Donghai; Sun, Ming

    2016-01-01

    In the soil environment, Bacilli can affect nematode development, fecundity and survival. However, although many Bacillus species can kill nematodes, the virulence mechanisms Bacilli utilize remain unknown. In this study, we collected 120 strains comprising 30 species across the Bacillaceae and Paenibacillaceae families of the Bacillales order and measured their nematicidal activities in vitro. Comparison of these strains’ nematicidal capacities revealed that nine species, including Bacillus thuringiensis, B. cereus, B. subtilis, B. pumilus, B. firmus, B. toyonensis, Lysinibacillus sphaericus, Brevibacillus laterosporus and B. brevis, were highly nematicidal, the first of which showed the highest activity. Genome sequencing and analysis identified many potential virulence factors, which grouped into five types. At least four possible mechanisms were deduced on the basis of the combination of these factors and the bacterial nematicidal activity, including a pore-forming mechanism of crystal proteins, an inhibition-like mechanism of thuringiensin and a degradation mechanism of proteases and/or chitinases. Our results demonstrate that 120 spore-forming Bacilli across different families share virulence factors that may contribute to their nematicidal capacity. PMID:27539267

  15. 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

  16. Investigation of Sterilization Mechanism for Geobacillus stearothermophilus Spores with Plasma-Excited Neutral Gas

    Science.gov (United States)

    Matsui, Kei; Ikenaga, Noriaki; Sakudo, Noriyuki

    2015-09-01

    We investigate the mechanism of the sterilization with plasma-excited neutral gas that uniformly sterilizes both the space and inner wall of the reactor chamber at atmospheric pressure. Only reactive neutral species such as plasma-excited gas molecules and radicals are separated from the plasma and sent to the reactor chamber for chemical sterilization. The plasma source gas uses humidified mixture of nitrogen and oxygen. Geobacillus stearothermophilus spores and tyrosine which is amino acid are treated by the plasma-excited neutral gas. Shape change of the treated spore is observed by SEM, and chemical modification of the treated tyrosine is analyzed by HPLC. As a result, the surface of the treated spore shows depression. Hydroxylation and nitration of tyrosine are shown after the treatment. For these reasons, we believe that the sterilization with plasma-excited neutral gas results from the deformation of spore structure due to the chemical modification of amino acid.

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

    International Nuclear Information System (INIS)

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

  18. Investigation of the effect of VUV radiation on the viability of microfungi spores

    Science.gov (United States)

    Zvereva, G.; Kirtsideli, I.; Benken, K.; Saifitdinova, A.; Galkina, S.; Parfenov, V.

    2015-12-01

    Irradiation of various types of microfungi spores by vacuum ultraviolet radiation (VUV) (λ = 172 nm, 2 mW/cm2 ) was carried out in this work. It was found, that the VUV radiation leads to inactivation of spores in the dose range 10-240 mJ/cm2 , depending on the fungal species. Shadowing effect of overlapping layers of spores was observed. Protective property of melanin at λ = 172 nm has been proven experimentally. Presence of melanin in the cell structure led to an increase of the inactivation dose of VUV radiation in more than one order. Fluorescence microscopy have revealed differences in the structure of the membrane of control and irradiated spore. VUV irradiation of DNA samples (200 ng) at λ=172 nm showed almost complete absence of double-stranded DNA parts at doses of more than 240 mJ/cm2 .

  19. Radiosensitization of Bacillus cereus spores in minced meat treated with cinnamaldehyde

    International Nuclear Information System (INIS)

    Minced meat beef inoculated with Bacillus cereus spores was treated with four essential oil constituents. The active compounds were sprayed separately onto the meat in order to determine the concentration needed to reduce by 1 log the population of B. cereus spores. Cinnamaldehyde was the best antimicrobial compound selected. It was mixed with ascorbic acid and/or sodium pyrophosphate decahydrate and tested for its efficiency to increase the relative radiation sensitivity (RRS) of B. cereus spores in minced meat packed under air. Results demonstrated that the radiation treatment in presence of the cinnamaldehyde and sodium phosphate decahydrate increased the RRS of B. cereus spores by two fold. The study revealed also that the irradiation of raw beef meat pre-treated with cinnamaldehyde produced an inhibition of the growth of B. cereus count during refrigerated storage. This technology seems to be compatible with industrial meat processing.

  20. Radiosensitization of Bacillus cereus spores in minced meat treated with cinnamaldehyde

    Science.gov (United States)

    Ayari, S.; Dussault, D.; Jerbi, T.; Hamdi, M.; Lacroix, M.

    2012-08-01

    Minced meat beef inoculated with Bacillus cereus spores was treated with four essential oil constituents. The active compounds were sprayed separately onto the meat in order to determine the concentration needed to reduce by 1 log the population of B. cereus spores. Cinnamaldehyde was the best antimicrobial compound selected. It was mixed with ascorbic acid and/or sodium pyrophosphate decahydrate and tested for its efficiency to increase the relative radiation sensitivity (RRS) of B. cereus spores in minced meat packed under air. Results demonstrated that the radiation treatment in presence of the cinnamaldehyde and sodium phosphate decahydrate increased the RRS of B. cereus spores by two fold. The study revealed also that the irradiation of raw beef meat pre-treated with cinnamaldehyde produced an inhibition of the growth of B. cereus count during refrigerated storage. This technology seems to be compatible with industrial meat processing.

  1. 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...... place. However, the segregation patterns for cycloheximide resistance and several auxotrophic markers were random, suggesting that normal meiosis indeed occurs. To explain the lack of second-division segregation of mating types, we concluded that crossing-over does not occur between the mating......-type locus and the centromere, and that meiosis I is tied to spore formation, which explains why the number of spores is limited to two. The latter assumption was also supported by fluorescence microscopy. The second meiotic division takes place inside the spores and is followed by the resorption of two...

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

    Directory of Open Access Journals (Sweden)

    J. P. Roux

    1992-12-01

    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.

  3. Assessing the activity of microbicides against bacterial spores: knowledge and pitfalls.

    Science.gov (United States)

    Leggett, M J; Setlow, P; Sattar, S A; Maillard, J-Y

    2016-05-01

    Bacterial endospores (spores) have a higher intrinsic resistance to microbicides as compared to other microbial forms, most likely due to their impermeable outer layers and low water content. Though structural differences between the spores of various bacterial species may account for observed variations in their resistance to microbicides, flaws in methods for testing the sporicidal activity of microbicides often exaggerate the differences. This has major implications when considering the selection of one or more surrogates to assess microbicides against clinically relevant spore-formers such as Clostridium difficile. The mounting significance of Cl. difficile as a pathogen is leading to a corresponding increase in the number of commercially available microbicidal formulations claiming activity against its spores without proper differentiation between the product's sporistatic and sporicidal actions. In this review we critically assess the situation and the implications of product claims on the field use of microbicidal products. PMID:26784857

  4. Comparative analysis of the immunologic response induced by the Sterne 34F2 live spore Bacillus anthracis vaccine in a ruminant model.

    Science.gov (United States)

    Ndumnego, Okechukwu C; Köhler, Susanne M; Crafford, Jannie; van Heerden, Henriette; Beyer, Wolfgang

    2016-10-01

    The Sterne 34F2 live spore vaccine (SLSV) developed in 1937 is the most widely used veterinary vaccine against anthrax. However, literature on the immunogenicity of this vaccine in a target ruminant host is scarce. In this study, we evaluated the humoral response to the Bacillus anthracis protective antigen (rPA), a recombinant bacillus collagen-like protein of anthracis (rBclA), formaldehyde inactivated spores (FIS) prepared from strain 34F2 and a vegetative antigen formulation prepared from a capsule and toxin deficient strain (CDC 1014) in Boer goats. The toxin neutralizing ability of induced antibodies was evaluated using an in vitro toxin neutralization assay. The protection afforded by the vaccine was also assessed in vaccinates. Anti-rPA, anti-FIS and lethal toxin neutralizing titres were superior after booster vaccinations, compared to single vaccinations. Qualitative analysis of humoral responses to rPA, rBclA and FIS antigens revealed a preponderance of anti-FIS IgG titres following either single or double vaccinations with the SLSV. Antibodies against FIS and rPA both increased by 350 and 300-fold following revaccinations respectively. There was no response to rBclA following vaccinations with the SLSV. Toxin neutralizing titres increased by 80-fold after single vaccination and 700-fold following a double vaccination. Lethal challenge studies in naïve goats indicated a minimum infective dose of 36 B. anthracis spores. Single and double vaccination with the SLSV protected 4/5 and 3/3 of goats challenged with>800 spores respectively. An early booster vaccination following the first immunization is suggested in order to achieve a robust immunity. Results from this study indicate that this crucial second vaccination can be administered as early as 3 months after the initial vaccination. PMID:27496738

  5. Efficacy of a Vaccine Based on Protective Antigen and Killed Spores against Experimental Inhalational Anthrax▿ ‡

    OpenAIRE

    Gauthier, Yves P.; Tournier, Jean-Nicolas; Paucod, Jean-Charles; Corre, Jean-Philippe; Mock, Michèle; Goossens, Pierre L.; Vidal, Dominique R.

    2008-01-01

    Protective antigen (PA)-based anthrax vaccines acting on toxins are less effective than live attenuated vaccines, suggesting that additional antigens may contribute to protective immunity. Several reports indicate that capsule or spore-associated antigens may enhance the protection afforded by PA. Addition of formaldehyde-inactivated spores (FIS) to PA (PA-FIS) elicits total protection against cutaneous anthrax. Nevertheless, vaccines that are effective against cutaneous anthrax may not be so...

  6. Draft Genome Sequences of Four Thermophilic Spore Formers Isolated from a Dairy-Processing Environment

    Science.gov (United States)

    Caspers, Martien P. M.; Boekhorst, Jos; de Jong, Anne; Kort, Remco; Nierop Groot, Masja

    2016-01-01

    Spores of thermophilic spore-forming bacteria are a common cause of contamination in dairy products. Here, we report draft genome sequences of four thermophilic strains from a milk-processing plant or standard milk, namely, a Geobacillus thermoglucosidans isolate (TNO-09.023), Geobacillus stearothermophilus TNO-09.027, and two Anoxybacillus flavithermus isolates (TNO-09.014 and TNO-09.016). PMID:27516503

  7. Draft Genome Sequences of Four Thermophilic Spore Formers Isolated from a Dairy-Processing Environment.

    Science.gov (United States)

    Caspers, Martien P M; Boekhorst, Jos; de Jong, Anne; Kort, Remco; Nierop Groot, Masja; Abee, Tjakko

    2016-01-01

    Spores of thermophilic spore-forming bacteria are a common cause of contamination in dairy products. Here, we report draft genome sequences of four thermophilic strains from a milk-processing plant or standard milk, namely, a Geobacillus thermoglucosidans isolate (TNO-09.023), Geobacillus stearothermophilus TNO-09.027, and two Anoxybacillus flavithermus isolates (TNO-09.014 and TNO-09.016). PMID:27516503

  8. Germination properties as marker events characterizing later stages of Bacillus subtilis spore formation.

    OpenAIRE

    Dion, P; Mandelstam, J

    1980-01-01

    At various stages during spore formation sporangia were shocked by cold treatment or with toluene, and the germination requirements of the prespores were examined. Up to 5 h after induction of sporulation (t5) germination was spontaneous; i.e., it occurred without any added germinants. After t5, during stages V and VI, the capacity for spontaneous germination diminished progressively, and the spores acquired a need for externally added germinants. At t6 this need was satisfied by either L-ala...

  9. Inactivation of Bacillus anthracis Spores during Laboratory-Scale Composting of Feedlot Cattle Manure.

    Science.gov (United States)

    Xu, Shanwei; Harvey, Amanda; Barbieri, Ruth; Reuter, Tim; Stanford, Kim; Amoako, Kingsley K; Selinger, Leonard B; McAllister, Tim A

    2016-01-01

    Anthrax outbreaks in livestock have social, economic and health implications, altering farmer's livelihoods, impacting trade and posing a zoonotic risk. Our study investigated the survival of Bacillus thuringiensis and B. anthracis spores sporulated at 15, 20, or 37°C, over 33 days of composting. Spores (∼7.5 log10 CFU g(-1)) were mixed with manure and composted in laboratory scale composters. After 15 days, the compost was mixed and returned to the composter for a second cycle. Temperatures peaked at 71°C on day 2 and remained ≥55°C for an average of 7 days in the first cycle, but did not exceed 55°C in the second. For B. thuringiensis, spores generated at 15 and 21°C exhibited reduced (P < 0.05) viability of 2.7 and 2.6 log10 CFU g(-1) respectively, as compared to a 0.6 log10 CFU g(-1) reduction for those generated at 37°C. For B. anthracis, sporulation temperature did not impact spore survival as there was a 2.5, 2.2, and 2.8 log10 CFU g(-1) reduction after composting for spores generated at 15, 21, and 37°C, respectively. For both species, spore viability declined more rapidly (P < 0.05) in the first as compared to the second composting cycle. Our findings suggest that the duration of thermophilic exposure (≥55°C) is the main factor influencing survival of B. anthracis spores in compost. As sporulation temperature did not influence survival of B. anthracis, composting may lower the viability of spores associated with carcasses infected with B. anthracis over a range of sporulation temperatures. PMID:27303388

  10. [Concentration of allergic fungi spores in the air of flats in Lódź].

    Science.gov (United States)

    Krawczyk, P; Kowalski, M L; Ochecka-Szymańska, A

    1999-01-01

    The real contribution of moulds to the pathogenesis of allergic diseases remains unknown, although positive skin prick tests and/or specific serum IgE to moki allergens can be detected in 1-5% of atopic patients. A significant problem in assesment of exposure to mould allergens, resulting with difficulty in standarization of methods. The aim of this work was to assess the concentration of spores of 8 mould species in flats inhabited by peoples who Bont show any symptoms of allergy. The Open Petri Dish (OPD) method involving sedimentation of participles contained in the column of air over the dish was used to assess the number of spores in 1 m3 of indoor atmospheres. All colonies were counted, but only 8 mould species implicated in inhaled allergy were identified, ie.: Alternaria tenuis, Cladosporium herbarum, Helminthosporum halodes, Pullularia pullulans, Penicillium notatam, Rhizopus nigricans, Mucor mucedo, Aspergillus fumigatus. The tests were carried out in 10 flats located in various quarters of the cify of Lodź during three consecutive days of September 1995 between 5:00 pm and 6:04 pm. In analyzing the percentage of spores of each of the eight mould species tested we determined that, independent of fiat and test day, C. herbarum predominated. It is good agreement with the observations of other authors who report that among large quantities of fungi that are detected in late summer, usually C. herbarum spores dominate. This is the season when the incidence of the Cladosporium spores in the atmospheric air increases. Spores of H. halodes were detected least frequently. Our study demonstrated the presence of substantial amounts of mould spores in indoor air of houses in Lódź. The spores belong to species with documented allergenicity, suggesting that they may play a role in development of allergic sensitization in susceptible subjects.

  11. Inactivation of Bacillus anthracis Spores during Laboratory-Scale Composting of Feedlot Cattle Manure

    Science.gov (United States)

    Xu, Shanwei; Harvey, Amanda; Barbieri, Ruth; Reuter, Tim; Stanford, Kim; Amoako, Kingsley K.; Selinger, Leonard B.; McAllister, Tim A.

    2016-01-01

    Anthrax outbreaks in livestock have social, economic and health implications, altering farmer’s livelihoods, impacting trade and posing a zoonotic risk. Our study investigated the survival of Bacillus thuringiensis and B. anthracis spores sporulated at 15, 20, or 37°C, over 33 days of composting. Spores (∼7.5 log10 CFU g-1) were mixed with manure and composted in laboratory scale composters. After 15 days, the compost was mixed and returned to the composter for a second cycle. Temperatures peaked at 71°C on day 2 and remained ≥55°C for an average of 7 days in the first cycle, but did not exceed 55°C in the second. For B. thuringiensis, spores generated at 15 and 21°C exhibited reduced (P < 0.05) viability of 2.7 and 2.6 log10 CFU g-1 respectively, as compared to a 0.6 log10 CFU g-1 reduction for those generated at 37°C. For B. anthracis, sporulation temperature did not impact spore survival as there was a 2.5, 2.2, and 2.8 log10 CFU g-1 reduction after composting for spores generated at 15, 21, and 37°C, respectively. For both species, spore viability declined more rapidly (P < 0.05) in the first as compared to the second composting cycle. Our findings suggest that the duration of thermophilic exposure (≥55°C) is the main factor influencing survival of B. anthracis spores in compost. As sporulation temperature did not influence survival of B. anthracis, composting may lower the viability of spores associated with carcasses infected with B. anthracis over a range of sporulation temperatures. PMID:27303388

  12. Increased levels of ambient fungal spores in Taiwan are associated with dust events from China

    Science.gov (United States)

    Wu, Pei-Chih; Tsai, Jui-Chen; Li, Fang-Chun; Lung, Shih-Chun; Su, Huey-Jen

    2004-09-01

    Fungi are ubiquitous in nature and their spores are often dispersed into the atmosphere through turbulent airstreams. As yellow sandstorm blown from deserts in China had affected the ambient air quality with increasing levels of ambient particulates, often including significant amounts of biologically active particles has therefore become imperative for concerns of their health implications. Our study was aimed to examine the effects of yellow sandstorm events on the fungal composition and concentrations in ambient air. Atmospheric fungal spores were continuously collected using Burkard Volumetric Spore Trap. Samples collected between December 2000 and April 2001 were selected for priority analysis from days when the yellow sandstorms were reported to affect Taiwan according to the Central Weather Bureau in Taiwan. The composition of dominant spores such as Basidiospore, Penicillium/Aspergillus, Nigrospora, Arthrinium, Curvularia, Rusts, Stemphylium, Cercospora, Pithomyces, and unidentified fungi were significantly higher than those of background days. The increase of Basidiospore, Penicillium/Aspergillus, Nigrospora, and those unidentified fungi seems to be significantly associated with the increase of ambient particulate levels with regression coefficients ranging from 0.887 to 31.98. Our study has identified increasing ambient concentrations during sandstorm episodes are observed for some major fungi, Basidiospore, Penicillium, Aspergillus, and those unidentified fungi and the trends of the increase seems to associate with ambient particulate levels. Further efforts to clarify the relationship between those high fungal spore exposures and clinical adverse health effects are suggested in the future. In addition, effects of climatic factors and other particulate levels on the variation of ambient fungal spore levels are also desired in further study. Additional monitoring of ambient fungal spores in the first line of west coastline is hoped to assist in

  13. 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).

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

    Directory of Open Access Journals (Sweden)

    Momein H. El-Katatny

    2012-06-01

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

  15. Prevalence of culturable airborne spores of selected allergenic and pathogenic fungi in outdoor air

    Science.gov (United States)

    O'Gorman, Céline M.; Fuller, Hubert T.

    2008-06-01

    Temporal and spatial variations in airborne spore concentrations of selected allergenic and pathogenic fungi were examined in Dublin, Ireland, in 2005. Air samples were taken at four outdoor locations in the city every 2 weeks, coupled with measurements of meteorological conditions. Total culturable airborne fungal spore concentrations in Dublin ranged from 30-6800 colony forming units per cubic metre of air (CFU m-3) over the 12-month period. Cladosporium, Penicillium, Aspergillus and Alternaria spores were constantly present in the Dublin atmosphere, representing >20% of the total culturable spore count. Concentrations of Cladosporium increased significantly in summer and reached allergenic threshold levels, peaking at over 3200 CFU m-3 in August. Penicillium spore concentrations never reached allergenic threshold levels, with average concentrations of Cryptococcus neoformans nor Stachybotrys chartarum were detected, but airborne basidiospores of Schizophyllum commune were evidenced by the dikaryotization of monokaryon tester strains following exposure to the air. The relationships between airborne fungal spore concentrations and meteorological factors were analysed by redundancy analysis and revealed positive correlations between temperature and Cladosporium and relative humidity and Penicillium and Aspergillus.

  16. Lung dendritic cells rapidly mediate anthrax spore entry through the pulmonary route.

    Science.gov (United States)

    Cleret, Aurélie; Quesnel-Hellmann, Anne; Vallon-Eberhard, Alexandra; Verrier, Bernard; Jung, Steffen; Vidal, Dominique; Mathieu, Jacques; Tournier, Jean-Nicolas

    2007-06-15

    Inhalational anthrax is a life-threatening infectious disease of considerable concern, especially because anthrax is an emerging bioterrorism agent. The exact mechanisms leading to a severe clinical form through the inhalational route are still unclear, particularly how immobile spores are captured in the alveoli and transported to the lymph nodes in the early steps of infection. We investigated the roles of alveolar macrophages and lung dendritic cells (LDC) in spore migration. We demonstrate that alveolar macrophages are the first cells to phagocytose alveolar spores, and do so within 10 min. However, interstitial LDCs capture spores present in the alveoli within 30 min without crossing the epithelial barrier suggesting a specific mechanism for rapid alveolus sampling by transepithelial extension. We show that interstitial LDCs constitute the cell population that transports spores into the thoracic lymph nodes from within 30 min to 72 h after intranasal infection. Our results demonstrate that LDCs are central to spore transport immediately after infection. The rapid kinetics of pathogen transport may contribute to the clinical features of inhalational anthrax. PMID:17548636

  17. Measurement and analysis on optical characteristics of Aspergillus oryzae spores in infrared band

    Science.gov (United States)

    Li, Le; Hu, Yihua; Gu, Youlin; Chen, Wei; Xu, Shilong; Zhao, Xinying

    2015-10-01

    Spore is an important part of bioaerosols. The optical characteristics of spore is a crucial parameter for study on bioaerosols. The reflection within the waveband of 2.5 to15μm were measured by squash method. Based on the measured data, Complex refractive index of Aspergillus oryzae spores within the waveband of 3 to 5μm and 8 to 14 μm were calculated by using Krames-Kronig (K-K) relationship. Then,the mass extinction coefficient of Aspergillus oryzae spores within the waveband of 3 to 5μm and 8 to 14μm were obtained by utilizing Mie scattering theory, and the results were analyzed and discussed. The average mass extinction coefficient of Aspergillus oryzae spores is 0.51 m2/g in the range of 3 to 5μm and 0.48m2/g in the range of 8 to 14μm. Compared with common inorganic compounds, Aspergillus oryzae spores possesses a good extinction performance in infrared band.

  18. The aluminium and iodine pentoxide reaction for the destruction of spore forming bacteria.

    Science.gov (United States)

    Clark, Billy R; Pantoya, Michelle L

    2010-10-21

    The threat of biological weapons is a major concern in the present day and has led to studying methods to neutralize spore forming bacteria. A new technique involves the use of a thermite reaction that exhibits biocidal properties to limit bacterial growth. The objective was to examine the influence on bacteria growth upon spore exposure to thermite reactions with and without biocidal properties. Three thermites are considered: two that have biocidal properties (aluminium (Al) combined with iodine pentoxide (I(2)O(5)) and Al combined with silver oxide (Ag(2)O)); and, one that produces a highly exothermic reaction but has no biocidal properties (Al combined with iron oxide (Fe(2)O(3))). Results show that Al + I(2)O(5) is extremely effective at neutralizing spores after only one hour of exposure. The temperature generated by the reaction was not determined to be an influential factor affecting spore growth kinetics. Further analysis of the thermite reactions revealed that the Al + I(2)O(5) reaction produces iodine gas that effectively interacts with the spores and neutralizes bacteria growth, while the Al + Ag(2)O reaction temperature does not vaporize silver. In the condensed phase silver does not interact with the spores enough to neutralize bacteria growth. This study gives evidence that a thermite can be used as a stable transportation and delivery system for biocidal gas. PMID:20730185

  19. Spore sensitivity to sunlight and freezing can restrict dispersal in wood-decay fungi.

    Science.gov (United States)

    Norros, Veera; Karhu, Elina; Nordén, Jenni; Vähätalo, Anssi V; Ovaskainen, Otso

    2015-08-01

    Assessment of the costs and benefits of dispersal is central to understanding species' life-history strategies as well as explaining and predicting spatial population dynamics in the changing world. While mortality during active movement has received much attention, few have studied the costs of passive movement such as the airborne transport of fungal spores. Here, we examine the potential of extreme environmental conditions to cause dispersal mortality in wood-decay fungi. These fungi play a key role as decomposers and habitat creators in forest ecosystems and the populations of many species have declined due to habitat loss and fragmentation. We measured the effect of simulated solar radiation (including ultraviolet A and B) and freezing at -25°C on the spore germinability of 17 species. Both treatments but especially sunlight markedly reduced spore germinability in most species, and species with thin-walled spores were particularly light sensitive. Extrapolating the species' laboratory responses to natural irradiance conditions, we predict that sunlight is a relevant source of dispersal mortality at least at larger spatial scales. In addition, we found a positive effect of spore size on spore germinability, suggesting a trade-off between dispersal distance and establishment. We conclude that freezing and particularly sunlight can be important sources of dispersal mortality in wood-decay fungi which can make it difficult for some species to colonize isolated habitat patches and habitat edges. PMID:26380666

  20. Assessment of Gamma Radiation Resistance of Spores Isolated from the Spacecraft Assembly Facility During MSL Assembly

    Science.gov (United States)

    Chopra, Arsh; Ramirez, Gustavo A.; Vaishampayan, Parag A.; Venkateswaran, Kasthuri J.

    2011-01-01

    Spore forming bacteria, a common inhabitant of spacecraft assembly facilities, are known to tolerate extreme environmental conditions such as radiation, desiccation, and high temperatures. Since the Viking era (early 1970's), spores have been utilized to assess the degree and level of microbiological contamination on spacecraft and their associated spacecraft assembly facilities. There is a growing concern that desiccation and extreme radiation resistant spore forming microorganisms associated with spacecraft surfaces can withstand space environmental conditions and subsequently proliferate on another solar body. Such forward contamination would certainly jeopardize future life detection or sample return technologies. It is important to recognize that different classes of organisms are critical while calculating the probability of contamination, and methods must be devised to estimate their abundances. Microorganisms can be categorized based on radiation sensitivity as Type A, B, C, and D. Type C represents spores resistant to radiation (10% or greater survival above 0.8 Mrad gamma radiation). To address these questions we have purified 96 spore formers, isolated during planetary protection efforts of Mars Science Laboratory assembly for gamma radiation resistance. The spores purified and stored will be used to generate data that can be used further to model and predict the probability of forward contamination.

  1. The Adsorption Properties of Bacillus atrophaeus Spores on Single-Wall Carbon Nanotubes

    Directory of Open Access Journals (Sweden)

    P. Cortes

    2009-01-01

    Full Text Available An adsorption equilibrium and a kinetic study of Bacillus atrophaeus on Single-Wall Carbon Nanotubes (SWCNTs were here performed to provide the basis for developing biosensor devices for detecting threatening micro-organisms in water supply systems. B. atrophaeus spores and carbon nanotubes were subjected to a batch adsorption process to document their equilibria and kinetics. Here, commercial nanotubes were either studied as received or were acid-purified before adsorption experiments. The Bacillus spores appear to show higher affinity towards the purified nanotubes than to the as-received nanomaterial. The effective diffusivity of the spores onto the purified nanotubes was found to be approximately 30 percent higher than onto the as-received nanotubes. It seems that the removal of amorphous carbon from the as-received nanotubes through a purification process yielded an intimate nantoubes-spore interaction as revealed by transmission electron microscopy. Freundlich model successfully correlated the adsorption equilibrium data for the nanotubes-spore interaction. Transmission electron micrographs showed extensive contact between the Bacillus and the purified nanotubes, but the association appeared less intimate between the spores and the as-received nanotubes.

  2. Resistance of Bacillus subtilis spores to 12C ion beams, stimulation of high-energy charged particles in space

    Science.gov (United States)

    Zhang, Li; Dang, Bingrong; Li, Junxiong; Chen, Jinsong; Liu, Mei; Liu, Zhiheng; Zhang, Lixin

    To monitor the response of live microbes in space radiation environment with high-energy charged particles, we carry out ground stimulation radiation experiments. Spores of Bacillus (CGMCC 1.1849) species are one of the model systems used for astro- and radiobiological studies. (12) C ion beams served as stimulated space radiation from 5gry, 10gry, 20gry, 40gry, to 80gry at a rate of 15gry/min Death rates are measured and mutant strains are isolated. Five representative strains are analyzed for their corresponding gene sequences, protein sequences and gene expression index of DNA repair system gene recA and recO. The statistic results showed the strains resistance to (12) C ion beams radiation is partially due to the increase of gene expression index of recA and recO. In conclusion, our research provide a surrogate system to monitor the live microbial response in resistant to space radiation environment.

  3. Monitoring and assessment of airborne Cladosporium Link and Alternaria Nées spores in Sivrihisar (Eskisehir), Turkey.

    Science.gov (United States)

    Erkara, Ismuhan Potoglu; Ilhan, Semra; Oner, Setenay

    2009-01-01

    The spores of Cladosporium spp. and Alternaria spp., commonly described as the most allergenic spores, were collected by means of Durham gravimetric sampler from the Sivrihisar (Eskisehir) atmosphere throughout 2005 to 2006. The weekly variations in spores/cm(2) of Cladosporium and Alternaria were recorded. During this period, a total of 6,198 spores belonging to Cladosporium spp. and Alternaria spp. were recorded. Of these spores, 2,969 were identified in 2005 and 3,229 in 2006. While 69.55% of the total spores were those of Cladosporium spp., 30.45% were Alternaria spp. Relationships between airborne fungal spore presence and weather conditions were examined statistically. A Shapiro-Wilk test revealed that the airborne spores of Cladosporium spp. and Alternaria spp. had a normal distribution. Following this, Chi-square test, t test and Pearson correlation analysis were performed. The effects of temperature and relative humidity on the spore numbers of Cladosporium spp. and Alternaria spp. were significant according to the month in which they were collected (p < 0.01). The spore concentrations of each species reached to their highest levels in June 2006.

  4. Candidate genes that may be responsible for the unusual resistances exhibited by Bacillus pumilus SAFR-032 spores.

    Directory of Open Access Journals (Sweden)

    Madhan R Tirumalai

    Full Text Available The spores of several Bacillus species, including Bacillus pumilus SAFR-032 and B. safensis FO-36b, which were isolated from the spacecraft assembly facility at NASA's Jet Propulsion Laboratory, are unusually resistant to UV radiation and hydrogen peroxide. In order to identify candidate genes that might be associated with these resistances, the whole genome of B. pumilus SAFR-032, and the draft genome of B. safensis FO-36b were compared in detail with the very closely related type strain B. pumilus ATCC7061(T. 170 genes are considered characteristic of SAFR-032, because they are absent from both FO-36b and ATCC7061(T. Forty of these SAFR-032 characteristic genes are entirely unique open reading frames. In addition, four genes are unique to the genomes of the resistant SAFR-032 and FO-36b. Fifty three genes involved in spore coat formation, regulation and germination, DNA repair, and peroxide resistance, are missing from all three genomes. The vast majority of these are cleanly deleted from their usual genomic context without any obvious replacement. Several DNA repair and peroxide resistance genes earlier reported to be unique to SAFR-032 are in fact shared with ATCC7061(T and no longer considered to be promising candidates for association with the elevated resistances. Instead, several SAFR-032 characteristic genes were identified, which along with one or more of the unique SAFR-032 genes may be responsible for the elevated resistances. These new candidates include five genes associated with DNA repair, namely, BPUM_0608 a helicase, BPUM_0652 an ATP binding protein, BPUM_0653 an endonuclease, BPUM_0656 a DNA cytosine-5- methyltransferase, and BPUM_3674 a DNA helicase. Three of these candidate genes are in immediate proximity of two conserved hypothetical proteins, BPUM_0654 and BPUM_0655 that are also absent from both FO-36b and ATCC7061(T. This cluster of five genes is considered to be an especially promising target for future experimental

  5. Large-spored Alternaria pathogens in section Porri disentangled.

    Science.gov (United States)

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

    2014-09-01

    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

  6. Novel Species of Non-Spore-Forming Bacteria

    Science.gov (United States)

    Briegel, Ariane; Osman, Shariff; Moissl, Christine; Hosoya,Naofumi; Venkateswaran, Kasthuri; Satomi, Masataka; Mayilraj, Shanmugam

    2008-01-01

    While cataloging cultivatable microbes from the airborne biological diversity of the atmosphere of the Regenerative Enclosed life-support Module Simulator (REMS) system at Marshall Space Flight Center, two strains that belong to one novel bacterial species were isolated. Based on 16S rRNA gene sequencing and the unique morphology and the taxonomic characteristics of these strains, it is shown that they belong to the family Intrasporangiaceae, related to the genus Tetrasphaera, with phylogenetic distances from any validly described species of the genus Tetrasphaera ranging from 96.71 to 97.76 percent. The fatty acid profile supported the affiliation of these novel strains to the genus Tetrasphaera except for the presence of higher concentrations of octadecenoic acid (C18:0) and cis-9-octadecenoic acid (C18:1), which discriminates these strains from other valid species. In addition, DNA-DNA hybridization studies indicate that these strains belong to a novel species that could be readily distinguished from its nearest neighbor, Tetrasphaera japonica AMC 5116T, with less than 20 percent DNA relatedness. Physiological and biochemical tests show few phenotypic dissimilarities, but genotypic analysis allowed the differentiation of these gelatin-liquefying strains from previously reported strains. The name Tetrasphaera remsis sp. Nov. is proposed with the type strain 3-M5-R-4(sup T) (=ATCC BAA-1496(sup T)=CIP 109413(sup T). The cells are Gram-positive, nonmotile, cocci, in tetrad arrangement and clusters. Spore formation is not observed. No species of Tetrashpaera has ever been isolated from airborne samples. Previous discoveries have come from soil and activated sludge samples. As other species of this genus have demonstrated enhanced biological phosphorus removal activity, further tests are required to determine if this newly discovered species would have bioremediation applications.

  7. HPLC法同时测定食品中的乙酰磺胺酸钾、苯甲酸、山梨酸、糖精钠%Simultaneous determination of acesulfame patassium, benzoic acid, sorbic acid, soluble saccharin in foods by HPLC

    Institute of Scientific and Technical Information of China (English)

    胡丽华

    2013-01-01

    目的:建立同时检测食品中乙酰磺胺酸钾、苯甲酸、山梨酸、糖精钠的HPLC方法.方法:按GB/T23459-2009《食品中苯甲酸、山梨酸、糖精钠的测定-高效液相色谱法》的提取方法,调整流动相比例使四种物质得到良好分离,并测定其在不同食品中的加标回收率,做方法学论证.结果:乙酰磺胺酸钾、苯甲酸、山梨酸、糖精钠在2.0μg/ml ~ 40.0 μg/ml范围内线性良好(r>0.999),检出限<0.086 mg/kg,加标回收率在87.4% ~ 106.2%之间,重复测定的RSD<1.3% (n =6).结论:该方法简便、快捷、准确,适合批量样品检测.%Objective:Simultaneous determination of acesulfame patassium,benzoic acid,sorbic acid,soluble saccharin in foods by HPLC was established.Methods:According to GB/T 23459-2009 “Determination of benzoic acid,sorbic acid,soluble saccharin in foods-High performance liquid chromatography method”,the four substances were well separated by adjusting the mobile phase proportion and determined the recoveries in different foods for method science demonstration.Results:Acesulfame potassium,benzoic acid,sorbic acid,soluble saccharin showed good linearity(r >0.999) in the range of 2.0 μg/ml ~40.0 μg/ml,the limit of the detection was less than 0.086 mg/kg,the recovery rates were 87.4% ~ 106.2%,the relative standard deviation was less than 1.3% (n =6).Conclusion:The method is simple,fast,accurate,and suitable to test bulk samples.

  8. Hematotoxicity and genotoxicity evaluations in Swiss mice intraperitoneally exposed to Bacillus thuringiensis (var kurstaki) spore crystals genetically modified to express individually Cry1Aa, Cry1Ab, Cry1Ac, or Cry2Aa.

    Science.gov (United States)

    Mezzomo, Bélin Poletto; Miranda-Vilela, Ana Luisa; Barbosa, Lilian Carla Pereira; Albernaz, Vanessa Lima; Grisolia, Cesar Koppe

    2016-08-01

    Bacillus thuringiensis (Bt) has been widely used in foliar sprays as part of integrated pest management strategies against insect pests of agricultural crops. Since the advent of genetically modified plants expressing Bt δ-endotoxins, the bioavailability of Cry proteins has increased, and therefore for biosafety reasons their adverse effects should be studied, mainly for nontarget organisms. We evaluated, in Swiss mice, the hematotoxicity and genotoxicity of the genetically modified strains of Bt spore crystals Cry1Aa, 1Ab, 1Ac, or 2Aa at 27 mg/kg, and Cry1Aa, 1Ab and 2Aa also at 136 and 270 mg/kg, administered with a single intraperitoneal injection 24 h before euthanasia. Controls received filtered water or cyclophosphamide. Blood samples collected by cardiac puncture were used to perform hemogram, and bone marrow was extracted for the micronucleus test. Bt spore crystals presented toxicity for lymphocytes when in higher doses, which varied according to the type of spore crystal studied, besides promoting cytotoxic and genotoxic effects for the erythroid lineage of bone marrow, mainly at highest doses. Although the profile of such adverse side effects can be related to their high level of exposure, which is not commonly found in the environment, results indicated that these Bt spore crystals were not harmless to mice. This suggests that a more specific approach should be taken to increase knowledge about their toxicological properties and to establish the toxicological risks to nontarget organisms. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 970-978, 2016.

  9. Four quantitative trait loci associated with low Nosema ceranae (Microsporidia) spore load in the honeybee Apis mellifera

    DEFF Research Database (Denmark)

    Huang, Qiang; Kryger, Per; Le Conte, Yves;

    2014-01-01

    Nosema ceranae has been recently introduced into the honeybee Apis mellifera as a novel microsporidian gut parasite. To locate the genetic region involved in N. ceranae infection tolerance, we fed N. ceranae spores to haploid drones of a F1 hybrid queen produced from a cross between a queen...... of a Nosema-resistant bred strain and drones of susceptible colonies. The spore loads of the infected F1 drones were used as the phenotype to identify quantitative trait loci (QTLs) associated with N. ceranae spore load. One hundred forty-eight infected drones were individually genotyped with microsatellite...... markers at an average marker distance of 20 cM along the genome. Four QTLs were significantly associated with low spore load, explaining 20.4 % of total spore load variance. Moreover, a candidate gene Aubergine (Aub) within the major QTL region was significantly overexpressed in drones with low spore...

  10. 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 (spices had high levels of aerobic spores (> 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 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.

  11. 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 in...

  12. 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 in...

  13. Plant spore walls as a record of long-term changes in ultraviolet-B radiation

    Science.gov (United States)

    Lomax, Barry H.; Fraser, Wesley T.; Sephton, Mark A.; Callaghan, Terry V.; Self, Stephen; Harfoot, Michael; Pyle, John A.; Wellman, Charles H.; Beerling, David J.

    2008-09-01

    Stratospheric ozone screens the Earth's surface from harmful ultraviolet-B radiation. Concentrations of stratospheric ozone are governed by a variety of natural and anthropogenic factors, including solar cycles, volcanic aerosols, ozone-depleting substances and climate change. However, assessing this variability before instrumental records has proved difficult owing to the lack of a well-constrained proxy. Here, we use microspectroscopy to analyse the chemical composition of herbarium samples of clubmoss (Lycophyta) spores originating from high- and low-latitude localities, where they were exposed to different ultraviolet-B histories. We show that the concentration of two ultraviolet-B-absorbing compounds in the walls of high-northern- and southern-latitude spores is strongly regulated by historical variations in ultraviolet-B radiation. Conversely, we find little change in the concentration of these compounds in spores originating from tropical Ecuador, where ultraviolet levels have remained relatively stable. Using spores from Greenland, we reconstruct past (1907-1993) changes in ozone concentration and ultraviolet-B flux; we reveal strong similarities between spore-wall reconstructions, and independent instrumental records and model results. Our findings suggest that ultraviolet-B-absorbing compounds in plant spore walls have the potential to act as a proxy for past changes in terrestrial ultraviolet-B radiation and stratospheric ozone. The chemical signature of plant spore walls in herbaria, and possibly also in sedimentary and ice-core archives, may therefore prove valuable for reconstructing past variations in stratospheric ozone and their connections with changes in solar radiation and climate.

  14. 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.

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

    International Nuclear Information System (INIS)

    Highlights: • A fungal biofilter/photoreactor was evaluated in terms of hexane and spore removal. • Biofilter supported elimination capacities of ≈35 g m−3 h−1 and CO2 yields of ≈75%. • The photocatalytic process slightly boosted the hexane abatement performance. • Biofilter emitted fungal spores at concentrations of 2.4 × 103–9.0 × 104 CFU m−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−3 h−1. Steady n-hexane elimination capacities of 30–40 g m−3 h−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 × 103 to 9.0 × 104 CFU m−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 biotechnologies

  16. Direct Probing of the Surface Ultrastructure and Molecular Interactions of Dormant and Germinating Spores of Phanerochaete chrysosporium

    OpenAIRE

    Dufrêne, Yves; Boonaert, C J; Gerin, Patrick A.; Asther, M.; Rouxhet, Paul

    1999-01-01

    Atomic force microscopy (AFM) has been used to probe, under physiological conditions, the surface ultrastructure and molecular interactions of spores of the filamentous fungus Phanerochaete chrysosporium. High-resolution images revealed that the surface of dormant spores was uniformly covered with rodlets having a periodicity of 10 +/- 1 nm, which is in agreement with earlier freeze-etching measurements. In contrast, germinating spores had a very smooth surface partially covered with rough gr...

  17. 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.

  18. 远源地下水入侵阿拉善高原沙漠:酸溶物 Sr 同位素证据%Invasion of Remote Groundwater into Deserts on the Alxa Plateau:Acid Soluble Sr Isotopic Evidence

    Institute of Scientific and Technical Information of China (English)

    潘耀东; 饶文波; 王啸

    2014-01-01

    选取北方西-东向气候剖面上物质组成相似的4个沙漠(塔克拉玛干、巴丹吉林、腾格里和毛乌素),测定其地表沙酸溶物87 Sr/86 Sr 比值(质量之比)及 Ca2+和 Sr2+质量比.结果表明,Ca2+和 Sr2+质量比相关性极好,在降雨量低的塔克拉玛干沙漠中最高,向东逐渐降低,在降雨量多的毛乌素沙漠中最低.降雨量是控制阳离子浓度变化的主导因素.酸溶物87 Sr/86 Sr 比值在4个沙漠中有不同于阳离子变化的区域特征.酸溶物87 Sr/86 Sr 比值在塔克拉玛干沙漠中低于0.7100,在巴丹吉林和腾格里沙漠中大于0.7112,而在毛乌素沙漠中介于0.7100~0.7112之间.塔克拉玛干和毛乌素沙漠地表沙酸溶物87 Sr/86 Sr 比值仅受气候变化的制约.巴丹吉林和腾格里沙漠除了受气候因素影响外还受其它因素的制约,远源地下水对巴丹吉林沙漠的影响为47%,对腾格里沙漠为44%,接近气候因素的影响.%87 Sr/ 86 Sr ratios and Ca2 + ,Sr2 + concentrations of acid soluble fractions of surface sands are measured for four deserts(Taklimakan,Badain Jaran,Tengger and Mu Us)with similar mineral and geochemical compositions in the W-E direction over North China. Ca2 + and Sr2 + concentrations have an excellent correlation,are the highest in the Taklimakan Desert with the low rainfall,decreasing eastwards,and the lowest in the Mu Us sandy land with the high rainfall. The rainfall is the main factor controlling cationic concentration. Acid soluble 87 Sr/ 86 Sr ratios have regional characteristics different from cationic change in the four deserts. Acid soluble 87 Sr/ 86 Sr ratios are low-er than 0. 71 for the Taklimakan desert,more than 0. 711 2 for the Badain Jaran and Tengger deserts,and between 0. 71 and 0. 711 2 in the Mu Us sandy land. Acid soluble 87 Sr/ 86 Sr ratios of surface sands in the Taklimakan desert and the Mu Us sandy land were only affected by climate factor. The Badain

  19. [Sporogenesis and spores of Equisetum bogotense (Equisetaceae) from mountain areas of Colombia].

    Science.gov (United States)

    Rincón-Baron, Edgar Javier; Torres, Gerardo Andrés; Rolleri, Cristina Hilda

    2013-09-01

    Studies on some reproductive traits in Equisetum species are scarce and valuable to understand species distribution. Therefore, a detailed study of the sporogenesis process and spore development in E. bogotense is presented, with an analysis of the main events during meiosis, maturation of spores, spore wall ultrastructure, orbicules and elaters. Specimens were collected from 500 to 4500 m in Cauca, Colombia. Strobili at different maturation stages were fixed, dehydrated, embedded in resin, and ultra-microtome obtained sections were stained with Toluidine blue. Observations were made with optical microscopy with differential interference contrast illumination technique (DIC), transmission and scanning electron microscopy (TEM and SEM). Ultrathin sections (70-80 microm) for TEM observations were stained with uranyl acetate and lead citrate; while samples for SEM observations, were fixed, dehydrated in 2.2-dimethoxypropane and dried at critical point as in standard methods. Strobili have numerous mature sporangiophores, each one with a peltate structure, the scutellum, bearing five-six sessile sporangia attached to the axis of strobilus by the manubrium. Immature sporocytes (spore mother cells) are tightly packed within the young sporangia. The sporocytes quickly undergo meiosis, by passing the stage of archesporium and give origin to tetrads of spores. The tapetum loses histological integrity during early stages of sporogenesis, intrudes as a plasmodial mass into the cavity of the sporangium, partially surrounding premeiotic sporocytes, and then, tetrads and adult spores. The tapetum disintegrates towards the end of the sporogenesis, leaving spores free within the sporangial cavity. Spores present several cytological changes that allow them to achieve greater size and increase the number of plastids, before reaching the adult stage. Sporoderm includes three layers external to the cytoplasmic membrane of the spore cell, and they are pseudoendospore, exospore and

  20. Inactivation Process of Penicillium digitatum Spores Treated with Non-equilibrium Atmospheric Pressure Plasma

    Science.gov (United States)

    Hashizume, Hiroshi; Ohta, Takayuki; Mori, Takumi; Iseki, Sachiko; Hori, Masaru; Ito, Masafumi

    2013-05-01

    To investigate the inactivation process of Penicillium digitatum spores treated with a non-equilibrium atmospheric pressure plasma, the spores were observed using a fluorescent microscope and compared with those treated with ultraviolet (UV) light or moist heat. The treated spores were stained with two fluorescent dyes, 1,1'-dioctadecyl-3,3,Y,3'-tetramethylindocarbocyanine perchlorate (DiI) and diphenyl-1-pyrenylphosphine (DPPP). The intracellular organelles as well as cell membranes in the spores treated with the plasma were stained with DiI without a major morphological change of the membranes, while the organelles were never stained in the spores treated with UV light or moist heat. Moreover, DPPP staining revealed that organelles were oxidized by plasma treatment unlike UV light or moist heat treatments. These results suggest that only plasma treatment induces a minor structural change or functional inhibition of cell membranes, which leads to the oxidation of the intracellular organelles without a major deformation of the membranes through the penetration of reactive oxygen species generated by the plasma into the cell.

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

    International Nuclear Information System (INIS)

    The biosorption of radionuclide 241Am from solution was studied by a. niger spore and hyphae, and the effects of the operational conditions on the treatment were investigated. The results showed the treatment by A. niger spore and hyphae were very efficient. An average of 96% of the total 241Am was removed from 241Am solutions of 5.6-111 MBq/L (C0), with adsorption capacities (W) of 7.2-142.4 MBq/g biomass, 5.2-106.5 MBq/g, respectively. The biosorption equilibrium was achieved within 1 h and the optimum pH value ranged 3-0.1 mol/L HNO3 and 3-2 for spore and hyphae of A. niger, respectively. No significant effects on 241Am biosorption were observed at 15 degree C-45 degree C, or challenged with containing Au3+ or Ag+, even 2000 times above 241Am amount. the index relationship between concentrations and adsorption capacities of 241Am indicated that the 241Am biosorption by A. niger spore and hyphae obey to Freundlich adsorption equation. The adsorption behavior of A. niger spore and hyphae were basically coincident

  2. Meteorological and agricultural effects on airborne Alternaria and Cladosporium spores and clinical aspects in Valladolid (Spain).

    Science.gov (United States)

    Reyes, Estefanía Sánchez; de la Cruz, David Rodríguez; Merino, Ma Eugenia Sanchís; Sánchez, José Sánchez

    2009-01-01

    The aeropalynological monitoring was carried out from 1 February 2005-31 January 2007. The total number of spores collected during the main spore season (MSS) in 2005 was 4,500 for Alternaria and 93,744 in the case of Cladosporium, whereas in 2006 values were increased (8,385 for Alternaria and 150,144 for Cladosporium), reaching the maximum concentrations on 18 July and 17 June 2006 with 344 and 5,503 spores, respectively. The influence of the main meteorological parameters on spore concentrations was studied, resulting in a positive correlation with temperature. Rainfall, relative humidity and frequency of calms obtained negative correlations in the case of Alternaria, and positive for Cladosporium, the total daily hours of sunshine having an inverse influence on them. The intra-diurnal pattern was very similar for both genera, with a greater representation towards the central hours of the day and at night. Finally, some clinical aspects for the Alternaria spore type were analyzed, with a low percentage of sensitized patients though (9.5%). Only one patient showed positive skin test reaction to Cladosporium.

  3. 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

  4. 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.

  5. Fertile structures with in situ spores of a dipterid fern from the Triassic in southern China.

    Science.gov (United States)

    Wang, Yongdong; Li, Liqin; Guignard, Gaëtan; Dilcher, David L; Xie, Xiaoping; Tian, Ning; Zhou, Ning; Wang, Yan

    2015-05-01

    Clathropteris was a typical dipterid fern with well documented fossil record and was widely dispersed during the Mesozoic; however, our knowledge of fertile structures including in situ spores for this genus is still very limited. Here we report well-preserved compression specimens of Clathropteris obovata Oishi from the Late Triassic of Guangyuan, Sichuan Province, China. The specimens show round to oval and exindusiate sori, vertical to oblique annuli in sporangia, and in situ trilete spores with verrucate and baculate sculptures, which are comparable to dispersed spore genera of Converrucosisporites and Conbaculatisporites. Comparisons of relevant fossil taxa suggest that specimens of C. obovata from Triassic of China provide for the first time in Asia the detailed fertile structures with in situ spore characters of dipterid fossil Clathropteris. Unlike living Dipteris, Mesozoic fossils of Dipteridaceae show a high diversity and a range of complex morphology of in situ spores, thus are significant for the evolutionary links between Dipteridaceae and other related fern clade, including Gleicheniaceae and Matoniaceae of the Gleicheniales. PMID:25773305

  6. Detection of Anthrax Simulants with Microcalorimetric Spectroscopy: Bacillus subtilis and Bacillus cereus Spores

    Science.gov (United States)

    Arakawa, Edward T.; Lavrik, Nickolay V.; Datskos, Panos G.

    2003-04-01

    Recent advances in the development of ultrasensitive micromechanical thermal detectors have led to the advent of novel subfemtojoule microcalorimetric spectroscopy (CalSpec). On the basis of principles of photothermal IR spectroscopy combined with efficient thermomechanical transduction, CalSpec provides acquisition of vibrational spectra of microscopic samples and absorbates. We use CalSpec as a method of identifying nanogram quantities of biological micro-organisms. Our studies focus on Bacillus subtilis and Bacillus cereus spores as simulants for Bacillus anthracis spores. Using CalSpec, we measured IR spectra of B. subtilis and B. cereus spores present on surfaces in nanogram quantities (approximately 100 -1000 spores). The spectra acquired in the wavelength range of 690 -4000 cm-1 (2.5 -14.5 μm) contain information-rich vibrational signatures that reflect the different ratios of biochemical makeup of the micro-organisms. The distinctive features in the spectra obtained for the two types of micro-organism can be used to distinguish between the spores of the Bacillus family. As compared with conventional IR and Fourier-transform IR microscopic spectroscopy techniques, the advantages of the present technique include significantly improved sensitivity (at least a full order of magnitude), absence of expensive IR detectors, and excellent potential for miniaturization.

  7. Increased resistance to detachment of adherent microspheres and Bacillus spores subjected to a drying step.

    Science.gov (United States)

    Faille, Christine; Bihi, Ilyesse; Ronse, Annette; Ronse, Gilles; Baudoin, Michael; Zoueshtiagh, Farzam

    2016-07-01

    In various environments, including that of food processing, adherent bacteria are often subjected to drying conditions. These conditions have been shown to result in changes in the ability of biofilms to cross-contaminate food in contact with them. In this study, we investigated the consequences of a drying step on the further ability of adherent bacterial spores to resist detachment. An initial series of experiment was set up with latex microspheres as a model. A microsphere suspension was deposited on a glass slide and incubated at 25, 35 and 50°C for times ranging from 1h to 48h. By subjecting the dried slides to increasing water flow rates, we showed that both time and temperature affected the ease of microsphere detachment. Similar observations were made for three Bacillus spores despite differences in their surface properties, especially regarding their surface physicochemistry. The differences in ease of adherent spore detachment could not be clearly linked to the minor changes in spore morphology, observed after drying in various environmental conditions. In order to explain the increased interaction between spheres or spores and glass slides, the authors made several assumptions regarding the possible underlying mechanisms: the shape of the liquid bridge between the sphere and the substratum, which is greatly influenced by the hydrophilic/hydrophobic characters of both surfaces; the accumulation of soil at the liquid/air interface; the presence of trapped nano-bubbles around and/or under the sphere. PMID:27022869

  8. 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. PMID:27052695

  9. The decision to germinate is regulated by divergent molecular networks in spores and seeds.

    Science.gov (United States)

    Vesty, Eleanor F; Saidi, Younousse; Moody, Laura A; Holloway, Daniel; Whitbread, Amy; Needs, Sarah; Choudhary, Anushree; Burns, Bethany; McLeod, Daniel; Bradshaw, Susan J; Bae, Hansol; King, Brian Christopher; Bassel, George W; Simonsen, Henrik Toft; Coates, Juliet C

    2016-08-01

    Dispersal is a key step in land plant life cycles, usually via formation of spores or seeds. Regulation of spore- or seed-germination allows control over the timing of transition from one generation to the next, enabling plant dispersal. A combination of environmental and genetic factors determines when seed germination occurs. Endogenous hormones mediate this decision in response to the environment. Less is known about how spore germination is controlled in earlier-evolving nonseed plants. Here, we present an in-depth analysis of the environmental and hormonal regulation of spore germination in the model bryophyte Physcomitrella patens (Aphanoregma patens). Our data suggest that the environmental signals regulating germination are conserved, but also that downstream hormone integration pathways mediating these responses in seeds were acquired after the evolution of the bryophyte lineage. Moreover, the role of abscisic acid and diterpenes (gibberellins) in germination assumed much greater importance as land plant evolution progressed. We conclude that the endogenous hormone signalling networks mediating germination in response to the environment may have evolved independently in spores and seeds. This paves the way for future research about how the mechanisms of plant dispersal on land evolved. PMID:27257104

  10. Determination of thermobacteriological parameters and size of Bacillus stearothermophilus ATCC 7953 spores

    Directory of Open Access Journals (Sweden)

    Marcos Fraiha

    2010-12-01

    Full Text Available In order to determine thermobacteriological parameters for B. stearothermophilus spores, they were diluted in a saline solution medium and in ground corn-soybean mix, distributed in TDT tube, and submitted to heat for a specific period of time. The D value (time to reduce 1 log cycle of microbial count under a certain temperature and z value (variation of temperature to cause 10-fold change in D value were estimated. To estimate their dimensions, the spores were visualized by using a scanning electron microscope. D121.1 ºC and z values for these spores, as determined in the saline solution, were 8.8 minutes and 12.8 ºC, respectively. D121,1 ºC and z values determined in the corn-soy mix were 14.2 minutes and 23.7 ºC, respectively. The micrographs indicated that the spores have homogeneous shape and size, with length and diameter of 2 and 1 µm, respectively. These results confirm that the spore is highly thermal-resistant, and it is a good biological indicator to evaluate the extrusion process as a feed sterilizer.

  11. Effect of Bacillus subtilis spore (GalliPro® nutrients equivalency value on broiler chicken performance

    Directory of Open Access Journals (Sweden)

    Mojtaba Zaghari

    2015-02-01

    Full Text Available The experiment was conducted to evaluate the nutrients equivalency value of Bacillus subtilis spore (GalliPro® for broiler chickens and its potential for decreasing feed nutrients concentration and cost. A total of 720 day old Ross 308 broiler chicks was allocated in 6 treatments (2 sexes×3 diets with 6 replication for 7 weeks. Dietary treatments: main treatment (MT was routine broiler diet added 0.2 g/kg GalliPro® (Bacillus subtilis 4×109 CFU/g DSM 17299 and using nutrients equivalency of GalliPro® for feed formulation; negative control (NC was the same as main treatment without GalliPro® (subtracted the nutrients equivalent value of GalliPro®; positive control (PC was the same as MT diet in nutrients content but without GalliPro®. Effect of dietary treatments on body weight (BW was not significant. However, the average BW of male and female chicks receiving negative control diet was 2.0% (68 g lower than PC and MT groups (P>0.05. Dietary treatments had no significant effect on average daily feed intake. Feed conversion ratio of chicks receiving PC and MT diets were 2.7% better than NC chicks (P<0.01. Male chicks were superior to female in all measured traits (P<0.01. Effect of treatments on carcass characteristics was not significant. There was no interaction between factors on measured parameters. Performance of chicks receiving diet with GalliPro® compared with PC showed that GalliPro® liberated 0.4 crude protein from MT diet and consequently decreased the broiler feeding cost.

  12. Antibody responses to a spore carbohydrate antigen as a marker of nonfatal inhalation anthrax in rhesus macaques.

    Science.gov (United States)

    Saile, Elke; Boons, Geert-Jan; Buskas, Therese; Carlson, Russell W; Kannenberg, Elmar L; Barr, John R; Boyer, Anne E; Gallegos-Candela, Maribel; Quinn, Conrad P

    2011-05-01

    The Bacillus anthracis exosporium protein BclA contains an O-linked antigenic tetrasaccharide whose terminal sugar is known as anthrose (J. M. Daubenspeck et al., J. Biol. Chem. 279:30945-30953, 2004). We hypothesized that serologic responses to anthrose may have diagnostic value in confirming exposure to aerosolized B. anthracis. We evaluated the serologic responses to a synthetic anthrose-containing trisaccharide (ATS) in a group of five rhesus macaques that survived inhalation anthrax following exposure to B. anthracis Ames spores. Two of five animals (RM2 and RM3) were treated with ciprofloxacin starting at 48 hours postexposure and two (RM4 and RM5) at 72 h postexposure; one animal (RM1) was untreated. Infection was confirmed by blood culture and detection of anthrax toxin lethal factor (LF) in plasma. Anti-ATS IgG responses were determined at 14, 21, 28, and 35 days postexposure, with preexposure serum as a control. All animals, irrespective of ciprofloxacin treatment, mounted a specific, measurable anti-ATS IgG response. The earliest detectable responses were on days 14 (RM1, RM2, and RM5), 21 (RM4), and 28 (RM3). Specificity of the anti-ATS responses was demonstrated by competitive-inhibition enzyme immunoassay (CIEIA), in which a 2-fold (wt/wt) excess of carbohydrate in a bovine serum albumin (BSA) conjugate of the oligosaccharide (ATS-BSA) effected >94% inhibition, whereas a structural analog lacking the 3-hydroxy-3-methyl-butyryl moiety at the C-4" of the anthrosyl residue had no inhibition activity. These data suggest that anti-ATS antibody responses may be used to identify aerosol exposure to B. anthracis spores. The anti-ATS antibody responses were detectable during administration of ciprofloxacin. PMID:21389148

  13. Proteomics Reveals that Proteins Expressed During the Early Stage of Bacillus anthracis Infection Are Potential Targets for the Development of Vaccines and Drugs

    Institute of Scientific and Technical Information of China (English)

    Chun-Ming Huang; Craig A. Elmets; De-chu C. Tang; Fuming Li; Nabiha Yusuf

    2004-01-01

    In this review, we advance a new concept in developing vaccines and/or drugs to target specific proteins expressed during the early stage of Bacillus anthracis (an thrax) infection and address existing challenges to this concept. Three proteins (immune inhibitor A, GPR-like spore protease, and alanine racemase) initially identified by proteomics in our laboratory were found to have differential expres sions during anthrax spore germination and early outgrowth. Other studies of different bacillus strains indicate that these three proteins are involved in either germination or cytotoxicity of spores, suggesting that they may serve as potential targets for the design of anti-anthrax vaccines and drugs.

  14. The mechanism of DNA ejection in the Bacillus anthracis spore-binding phage 8a revealed by cryo-electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Fu, Xiaofeng [Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, Houston, TX 77030 (United States); Walter, Michael H. [Department of Biology, University of Northern Iowa, Cedar Falls, IA 50614 (United States); Paredes, Angel [Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, Houston, TX 77030 (United States); Morais, Marc C., E-mail: mcmorais@utmb.edu [Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX 77555 (United States); Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555 (United States); Liu, Jun, E-mail: Jun.Liu.1@uth.tmc.edu [Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, Houston, TX 77030 (United States)

    2011-12-20

    The structure of the Bacillus anthracis spore-binding phage 8a was determined by cryo-electron tomography. The phage capsid forms a T = 16 icosahedron attached to a contractile tail via a head-tail connector protein. The tail consists of a six-start helical sheath surrounding a central tail tube, and a structurally novel baseplate at the distal end of the tail that recognizes and attaches to host cells. The parameters of the icosahedral capsid lattice and the helical tail sheath suggest protein folds for the capsid and tail-sheath proteins, respectively, and indicate evolutionary relationships to other dsDNA viruses. Analysis of 2518 intact phage particles show four distinct conformations that likely correspond to four sequential states of the DNA ejection process during infection. Comparison of the four observed conformations suggests a mechanism for DNA ejection, including the molecular basis underlying coordination of tail sheath contraction and genome release from the capsid.

  15. Quantitative clarification of inactivation mechanism of Penicillium digitatum spores treated with neutral oxygen radicals

    Science.gov (United States)

    Hashizume, Hiroshi; Ohta, Takayuki; Takeda, Keigo; Ishikawa, Kenji; Hori, Masaru; Ito, Masafumi

    2015-01-01

    We have quantitatively investigated the oxidative inactivation process of Penicillium digitatum spores including intracellular nanostructural changes through neutral oxygen species with a flux-defined atmospheric-pressure oxygen radical source, using fluorescent confocal-laser microscopy and transmission electron microscopy (TEM). The results suggest that neutral oxygen species, particularly ground-state atomic oxygen [O(3Pj)], which is an effective species for inactivating P. digitatum spores, inhibit the function of the cell membrane of spores without causing major superficial morphological changes at a low O(3Pj) dose of ˜2.1 × 1019 cm-2 under an O(3Pj) flux of 2.3 × 1017 cm-2 s-1, following the oxidation of intracellular organelles up to an O(3Pj) dose of ˜1.0 × 1020 cm-2. Finally, intracellular nanostructures are degraded by excess oxygen radicals over an O(3Pj) dose of ˜1.0 × 1020 cm-2.

  16. 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.

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

    International Nuclear Information System (INIS)

    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)

  18. Strategies For 2-Heptanone Biosynthesis From Octanoic Acid By Penicillium Roqueforti Spores

    Directory of Open Access Journals (Sweden)

    M Arpah

    2003-08-01

    Full Text Available This paper reviews two strategies of 2-heptanone (blue cheese aroma compound biosynthesis from octanoic acid by Penicillium roqueforti spores. First, the production and preparation of fungal spores are discussed a long with effect of spores treatment on their biocatalytic activity. Following this the first strategy of 2-heptanone production i.e batch production of 2-heptanone by submerged bioconversion process is discussed. Furthermore 2-heptanone loss by air stream stripping due to its hight volatility is evaluated and then the second strategy, that is continuous biosynthesis in aerated stirred reactor, is addressed. An option for controlling pH of biosynthesis medium is also discussed. Finally, in the concluding paragraphs, utility of the strategies are presented.

  19. Seasonal prevalence of air-borne pollen and spores in Kuala Lumpur, Malaysia.

    Science.gov (United States)

    Ho, T M; Tan, B H; Ismail, S; Bujang, M K

    1995-06-01

    Aerosampling using Rotorod samplers was conducted in the Institute for Medical Research, Kuala Lumpur, Malaysia, from December 1991 to November 1993. Samples were collected twice a week between 10.00 hours to 12.00 hours. Rods were stained and examined microscopically. A total of 8 and 20 types of pollens and mold spores were collected, respectively. More mold spores were collected than pollens. Grass pollen constituted more than 40 percent of total pollen counts. Gramineae pollen counts peaked in March and September. The most abundant mold spore was Cladosporium followed by Rust, Nigrospora, Curvularia and Smut. Cladosporium counts peaked in February and August. Rust counts peaked in June and December whereas counts for Nigrospora peaked in February and October. Highest counts of Smut were recorded in March and October. Curvularia counts peaked in January, June and September.

  20. 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.

  1. Acute phase proteins in experimentally induced pregnancy toxemia in goats.

    Science.gov (United States)

    González, Félix H D; Hernández, Fuensanta; Madrid, Josefa; Martínez-Subiela, Silvia; Tvarijonaviciute, Asta; Cerón, José J; Tecles, Fernando

    2011-01-01

    The present work aimed to study the behavior of acute phase proteins (haptoglobin, serum amyloid A, acid soluble glycoprotein, fibrinogen, and albumin) in fasting-induced pregnancy toxemia in goats and their relationship with classical indicators of this disorder such as beta-hydroxybutyrate and nonesterified fatty acids in the blood and decreased urine pH and ketonuria. Twelve adult Murciano-Granadina goats at the final stage of gestation were used in this experiment. Pregnancy toxemia was induced in 6 goats by fasting for 72 hr. The other 6 animals were used as control group. Ketonuria was present in 4 out of 5 fasting animals at 24 hr and in all fasting animals at 48 hr of fasting. Serum nonesterified fatty acids were significantly increased at 24, 48, and 72 hr of fasting. Beta-hydroxybutyrate and haptoglobin achieved significantly increased concentrations at 48 hr and 72 hr, respectively, remaining increased during the entire study. Serum amyloid A, acid soluble glycoprotein, fibrinogen, and albumin were not affected by fasting. In conclusion, acute phase proteins (including haptoglobin) seemed not to have an advantage over traditional markers in diagnosis of fasting-induced pregnancy toxemia in goats. PMID:21217028

  2. Kinetics of Mn(II) oxidation by spores of the marine Bacillus sp. SG-1

    Science.gov (United States)

    Toyoda, Kazuhiro; Tebo, Bradley M.

    2016-09-01

    The kinetics of Mn(II) oxidation by spores of the marine Bacillus sp. SG-1 was measured under controlled conditions of the initial Mn(II) concentration, spore concentration, chemical speciation, pH, O2, and temperature. Mn(II) oxidation experiments were performed with spore concentrations ranging from 0.7 to 11 × 109 spores/L, a pH range from 5.8 to 8.1, temperatures between 4 and 58 °C, a range of dissolved oxygen from 2 to 270 μM, and initial Mn(II) concentrations from 1 to 200 μM. The Mn(II) oxidation rates were directly proportional to the spore concentrations over these ranges of concentration. The Mn(II) oxidation rate increased with increasing initial Mn(II) concentration to a critical concentration, as described by the Michaelis-Menten model (Km = ca. 3 μM). Whereas with starting Mn(II) concentrations above the critical concentration, the rate was almost constant in low ionic solution (I = 0.05, 0.08). At high ionic solution (I = 0.53, 0.68), the rate was inversely correlated with Mn(II) concentration. Increase in the Mn(II) oxidation rate with the dissolved oxygen concentration followed the Michaelis-Menten model (Km = 12-19 μM DO) in both a HEPES-buffered commercial drinking (soft) water and in artificial and natural seawater. Overall, our results suggest that the mass transport limitations of Mn(II) ions due to secondary Mn oxide products accumulating on the spores cause a significant decrease of the oxidation rate at higher initial Mn(II) concentration on a spore basis, as well as in more concentrated ionic solutions. The optimum pH for Mn(II) oxidation was approximately 7.0 in low ionic solutions (I = 0.08). The high rates at the alkaline side (pH > 7.5) may suggest a contribution by heterogeneous reactions on manganese bio-oxides. The effect of temperature on the Mn(II) oxidation rate was studied in three solutions (500 mM NaCl, ASW, NSW solutions). Thermal denaturation occurred at 58 °C and spore germination was evident at 40 °C in all three

  3. Effect of artificial UV irradiation on spore content of stall air and fattening pig breeding

    International Nuclear Information System (INIS)

    The influence of a continuous UV irradiation (emitter NN 33/89 original Hanau) during the fattening periods primarily in the bactericide region of 253.7 nm of various intensities on the spore content of air, on the state of health and on the fattening breeding of pigs was tested in two fattening procedures. The high spore number per m3 air of over 700 000 upon occupying the stall in the first fattening procedure was reduced by 90.5% to about 70 000 after 1 week of UV irradiation, and in the second procedure, from 111 500 to 16 000, i.e. a reduction of 85.5%. The spore content of the stall air then exhibited large deviations reducing and increasing. The same deviations were recorded for dust content. There was no absolute correlation between dust and spore content of the air until the 11th week after starting UV irradiation in either test. The spore content sank in the reference stalls also without UV irradiation, by 29.9% in the first fattening procedure 1 week after occupying the stall and even by 75% in the second procedure. The spore content of the air in the reference stalls also then exhibited deviations sinking and rising as in the test stalls with UV irradiation. Here too, there was no correlation between dust and spore content of the air. The spore content in the air was 2 to 7 times higher in the reference stalls than in the test stalls. One may conclude from the tests that the promoting irradiation strength is between 15 and 20 μW/cm2 and that short-term stool production in danish stalling, 60 μW/cm2 are not harmful. Air disinfection with UV irradiation, can only be part of the total hygiene measures taken in veterinary medicine and may only be considered as an important link in the chain of the health promoting and increased efficient hygiene measures in the intensification of aggriculturally useful animals. (orig./AJ)

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

    OpenAIRE

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

    2015-01-01

    Little has been done to study microbial enhanced oil recovery (MEOR) in chalk reservoirs. The present study focused on core flooding experiments to see microbial plugging and its effect on oil recovery. A pressure tapped core holder with pressure ports at 1.2 cm, 3.8 cm, and 6.3 cm from the inlet 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 ...

  5. Viability of Clostridium sporogenes spores after CaO hygienization of meat waste

    OpenAIRE

    Justyna Bauza-Kaszewska; Zbigniew Paluszak; Krzysztof Skowron

    2014-01-01

    The occurrence of the pathogenic species [i]C. perfringens[/i] and [i]C. botulinum spores[/i] in animal by-products poses a potential epidemiological hazard. Strong entero- and neurotoxins produced by these bacteria adversely affect human health. To inactivate pathogens present in animal by-products, waste must be subjected to various methods of sanitization. The aim of the presented study was to estimate the effect of different doses of CaO on the viability of spores [i] Clostridium sporogen...

  6. Protoplast water content of bacterial spores determined by buoyant density sedimentation.

    OpenAIRE

    Lindsay, J A; Beaman, T C; Gerhardt, P

    1985-01-01

    Protoplast wet densities (1.315 to 1.400 g/ml), determined by buoyant density sedimentation in Metrizamide gradients, were correlated inversely with the protoplast water contents (26.4 to 55.0 g of water/100 g of wet protoplast) of nine diverse types of pure lysozyme-sensitive dormant bacterial spores. The correlation equation provided a precise method for obtaining the protoplast water contents of other spore types with small impure samples and indicated that the average protoplast dry densi...

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

    OpenAIRE

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

    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 × 10(5) L mol(-1). The addition of calcium dipicolinate, largely found in bacterial spores, changes the color of the aqueous solution of [Eu(Bn)(+)] from orange t...

  8. A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores.

    Science.gov (United States)

    Nerandzic, Michelle M; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J

    2016-01-01

    Background.  Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5-2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods.  We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results.  Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200-2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions.  Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin. PMID:26885539

  9. A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores

    Science.gov (United States)

    Nerandzic, Michelle M.; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J.

    2016-01-01

    Background. Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5–2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods. We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results. Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200–2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions. Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin. PMID:26885539

  10. Die another day: Fate of heat-treated Geobacillus stearothermophilus ATCC 12980 spores during storage under growth-preventing conditions.

    Science.gov (United States)

    Mtimet, Narjes; Trunet, Clément; Mathot, Anne-Gabrielle; Venaille, Laurent; Leguérinel, Ivan; Coroller, Louis; Couvert, Olivier

    2016-06-01

    Geobacillus stearothermophilus spores are recognized as one of the most wet-heat resistant among aerobic spore-forming bacteria and are responsible for 35% of canned food spoilage after incubation at 55 °C. The purpose of this study was to investigate and model the fate of heat-treated survivor spores of G. stearothermophilus ATCC 12980 in growth-preventing environment. G. stearothermophilus spores were heat-treated at four different conditions to reach one or two decimal reductions. Heat-treated spores were stored in nutrient broth at different temperatures and pH under growth-preventing conditions. Spore survival during storage was evaluated by count plating over a period of months. Results reveal that G. stearothermophilus spores surviving heat treatment lose their viability during storage under growth-preventing conditions. Two different subpopulations were observed during non-thermal inactivation. They differed according to the level of their resistance to storage stress, and the proportion of each subpopulation can be modulated by heat treatment conditions. Finally, tolerance to storage stress under growth-preventing conditions increases at refrigerated temperature and neutral pH regardless of heat treatment conditions. Such results suggest that spore inactivation due to heat treatment could be completed by storage under growth-preventing conditions. PMID:26919821

  11. Demonstrated Efficacy of a Pilot Heterologous Whole-Spore Vaccine against Microsporidial Gill Disease in Rainbow Trout

    OpenAIRE

    Harkness, J. E.; Guselle, N. J.; Speare, D J

    2013-01-01

    Intraperitoneal vaccines using whole viable spores of the microsporidian Glugea anomala or Glugea hertwigi reduced the numbers of branchial xenomas by 80% and 91%, respectively, after a standard experimental infection of juvenile rainbow trout with the microsporidian Loma salmonae. Similar significant results were obtained when killed-spore preparations were used.

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

    NARCIS (Netherlands)

    Voort, van der M.; Abee, T.

    2013-01-01

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

  13. Effect of ultraviolet radiation on spore viability and mosquitocidal activity of an indigenous ISPC-8 Bacillus sphaericus Neide strain.

    Science.gov (United States)

    Hadapad, A B; Vijayalakshmi, N; Hire, R S; Dongre, T K

    2008-08-01

    Effects of UV-A, UV-B and their combination on spore viability and larvicidal activity of an indigenous isolate of Bacillus sphaericus Neide, ISPC-8 were studied under laboratory conditions. The UV sensitivity of ISPC-8 was compared with standard strain 1593 and larvicidal activity was tested against third instar larvae of mosquito, Culex quinquefasciatus Say. No significant adverse effects on viability as well as larvicidal activity of both strains were observed when spores were exposed to UV-A for 6h. However, exposure to UV-B for a few minutes adversely affected the spore viability as well as larvicidal activity and this adverse effect was more pronounced on spore viability. In both strains about 50% larvicidal activity was retained after exposure of the spores to UV-B for 8h. However, spore viability at this exposure of time was drastically reduced to 2.5% in ISPC-8 and 0.3% in 1593. The spore viability and larvicidal activity patterns were found to be similar to UV-B treatment when spores were exposed to a combination of UV-A and UV-B. Our study hence, shows the adverse effect of UV radiation on ISPC-8 and 1593 indicating the need to incorporate eco-friendly UV protectants in formulations so that the efficacy of biopesticides based on these entomopathogens can be prolonged under field conditions, especially in tropical countries.

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

    Science.gov (United States)

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

    2016-01-01

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

  15. Novel handwashes are superior to soap and water in removal of Clostridium difficile spores from the hands.

    Science.gov (United States)

    Isaacson, Dylan; Haller, Barbara; Leslie, Hannah; Roemer, Marguerite; Winston, Lisa

    2015-05-01

    We examined the efficacy of 5 experimental handwash formulations in removing nontoxigenic Clostridium difficile spores from the hands of health care workers. Handwashing with sand resulted in an additional 0.5-log reduction in spore recovery compared with the current standard of soap and water.

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

    International Nuclear Information System (INIS)

    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 108, 4 x 103 and 5 x 101 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)

  17. Relationships between airborne fungal spore concentration of Cladosporium and the summer climate at two sites in Britain

    Science.gov (United States)

    Hollins, P. D.; Kettlewell, P. S.; Atkinson, M. D.; Stephenson, D. B.; Corden, J. M.; Millington, W. M.; Mullins, J.

    Cladosporium conidia have been shown to be important aeroallergens in many regions throughout the world, but annual spore concentrations vary considerably between years. Understanding these annual fluctuations may be of value in the clinical management of allergies. This study investigates the number of days in summer when spore concentration exceeds the allergenic threshold in relation to regional temperature and precipitation at two sites in England and Wales over 27 years. Results indicate that number of days in summer when the Cladosporium spores are above the allergenic concentration is positively correlated with regional temperature and negatively correlated with precipitation for both sites over the study period. Further analysis used a winter North Atlantic Oscillation index to explore the potential for long-range forecasting of the aeroallergen. For both spore measurement sites, a positive correlation exists between the winter North Atlantic Oscillation index and the number of days in summer above the allergenic threshold for Cladosporium spore concentration.

  18. Dynamics of cap opening in Agaricus bisporus and changes in spore numbers vs. cap opening and radiation treatment

    Energy Technology Data Exchange (ETDEWEB)

    Kovacs, E.; Voeroes, Zs.; Farkas, J. (Koezponti Elelmiszeripari Kutato Intezet, Budapest (Hungary))

    1981-01-01

    The spore count in Agaricus bisporus treated with radiation doses of 1.5 and 2.5 kGy was studied as a function of cap opening and radiation treatment (at 14-16 deg C and RH 80-90%). It was established that spore formation starts in the mushroom while the cap is still closed and the opening of the pileus starts when the spore number begins to increase. In treated mushrooms not only the cap remains closed but in lack of the development of gills the development of spores is also inhibited. Maturation retardation becomes apparent in the inhibition of the increase in number and of the darkening of the spores.

  19. Comparison of four different methods for extraction of Stachybotrys chartarum spore DNA and verification by real-time PCR.

    Science.gov (United States)

    Black, J A; Foarde, K K

    2007-07-01

    A comparison of four different methods for the extraction of spore DNA from Stachybotrys chartarum was conducted. Spore DNA was extracted and purified using either one of three different commercial kits or water. All preparations utilized bead milling. Genomic DNA extracted from 10(1) to 10(7) spores was assessed by both agarose gel electrophoresis and real-time quantitative polymerase chain reaction (qPCR) performed against multi-copy (rRNA) and single-(tubulin) gene targets. The spore isolation technique we employed was verified to be pure by light microscopy. Although all preparatory methods led to successful detection by qPCR, S. chartarum spore DNA prepared using the Qiagen Plant kit was notably better over the extraction range.

  20. Inhibition of fungal spore adhesion by zosteric Acid as the basis for a novel, nontoxic crop protection technology.

    Science.gov (United States)

    Stanley, Michele S; Callow, Maureen E; Perry, Ruth; Alberte, Randall S; Smith, Robert; Callow, James A

    2002-04-01

    ABSTRACT To explore the potential for nontoxic crop protection technologies based on the inhibition of fungal spore adhesion, we have tested the effect of synthetic zosteric acid (p-(sulfo-oxy) cinnamic acid), a naturally occurring phenolic acid in eelgrass (Zostera marina L.) plants, on spore adhesion and infection in two pathosystems: rice blast caused by Magnaporthe grisea and bean anthracnose caused by Colletotrichum lindemuthianum. We have shown that zosteric acid inhibits spore adhesion to model and host leaf surfaces and that any attached spores fail to develop appressoria, and consequently do not infect leaf cells. Low concentrations of zosteric acid that are effective in inhibiting adhesion are not toxic to either fungus or to the host. The inhibition of spore adhesion in the rice blast pathogen is fully reversible. On plants, zosteric acid reduced (rice) or delayed (bean) lesion development. These results suggest that there is potential for novel and environmentally benign crop protection technologies based on manipulating adhesion.

  1. Yeast Interacting Proteins Database: YDL239C, YLR423C [Yeast Interacting Proteins Database

    Lifescience Database Archive (English)

    Full Text Available of a Don1p-containing structure at the leading edge of the prospore membrane via interaction with spindle p...cription Protein required for spore wall formation, thought to mediate assembly of a Don1p-containing structure at the leading

  2. Yeast Interacting Proteins Database: YDR176W, YDL239C [Yeast Interacting Proteins Database

    Lifescience Database Archive (English)

    Full Text Available a Don1p-containing structure at the leading edge of the prospore membrane via interaction with spindle pole...ining structure at the leading edge of the prospore membrane via interaction with spindle pole body componen...DY3 Prey description Protein required for spore wall formation, thought to mediate assembly of a Don1p-conta

  3. Cell growth stimulating effect of Ganoderma lucidum spores and their potential application for Chinese hamster ovary K1 cell cultivation.

    Science.gov (United States)

    Li, Ding; Zhong, Qi; Liu, Tingting; Wang, Jufang

    2016-06-01

    In this work, water-soluble extracts of Ganoderma lucidum spores (Gls), a Chinese medicinal herb that possesses cell growth stimulating function, were found to be an effective growth factor for Chinese hamster ovary (CHO) cell cultivation. The Gls extract was prepared and supplemented to CHO K1 cell culture media with various serum levels. Our results obtained from both the static culture and the spinner-flask suspension culture showed that use of small-amount Gls extract effectively promoted cell growth and suppressed cell apoptosis induced by serum deprivation with normal cell cycle maintained in a low-serum medium. The low-serum medium containing 1 % (v/v) fetal bovine serum (FBS) and 0.01 % (w/v) Gls extract showed a comparable performance on both cell growth and fusion protein productivity with the conventional CHO culture medium containing 10 % (v/v) FBS and a commercial serum-free medium. This is the first study of the potential of Gls extracts for use as an alternative cell growth factor and nutrient for CHO cells. The findings have presented a new approach to economic cultivation of CHO cells for therapeutic protein production.

  4. Cell growth stimulating effect of Ganoderma lucidum spores and their potential application for Chinese hamster ovary K1 cell cultivation.

    Science.gov (United States)

    Li, Ding; Zhong, Qi; Liu, Tingting; Wang, Jufang

    2016-06-01

    In this work, water-soluble extracts of Ganoderma lucidum spores (Gls), a Chinese medicinal herb that possesses cell growth stimulating function, were found to be an effective growth factor for Chinese hamster ovary (CHO) cell cultivation. The Gls extract was prepared and supplemented to CHO K1 cell culture media with various serum levels. Our results obtained from both the static culture and the spinner-flask suspension culture showed that use of small-amount Gls extract effectively promoted cell growth and suppressed cell apoptosis induced by serum deprivation with normal cell cycle maintained in a low-serum medium. The low-serum medium containing 1 % (v/v) fetal bovine serum (FBS) and 0.01 % (w/v) Gls extract showed a comparable performance on both cell growth and fusion protein productivity with the conventional CHO culture medium containing 10 % (v/v) FBS and a commercial serum-free medium. This is the first study of the potential of Gls extracts for use as an alternative cell growth factor and nutrient for CHO cells. The findings have presented a new approach to economic cultivation of CHO cells for therapeutic protein production. PMID:26921102

  5. Bacillus anthracis spores germinate extracellularly at air–liquid interface in an in vitro lung model under serum‐free conditions

    OpenAIRE

    Powell, J D; Hutchison, J.R.; Hess, B.M.; Straub, T.M.

    2015-01-01

    Abstract Aims To better understand the parameters that govern spore dissemination after lung exposure using in vitro cell systems. Methods and Results We evaluated the kinetics of uptake, germination and proliferation of Bacillus anthracis Sterne spores in association with human primary lung epithelial cells, Calu‐3 and A549 cell lines. We also analysed the influence of various cell culture medium formulations related to spore germination. Conclusions We found negligible spore uptake by epith...

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

    Directory of Open Access Journals (Sweden)

    Letícia Christina Pires Gonçalves

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

  7. 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 × 10(5) L mol(-1). The addition of calcium dipicolinate, largely found in bacterial spores, changes the color of the aqueous solution of [Eu(Bn)(+)] from orange to magenta. The limit of detection (LOD) of calcium dipicolinate is around 2.0 × 10(-6) mol L(-1) and the LOD determined for both spores, B. cereus and B. anthracis, is (1.1 ± 0.3)× 10(6) spores mL(-1). This simple, green, fast and low cost colorimetric assay was selective for calcium dipicolinate when compared to several analogous compounds. The importance of this work relies on the potential use of betalains, raw natural pigments, as colorimetric sensors for biological applications. PMID:24019934

  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. Genome Sequences of Three Spore-Forming Bacteria Isolated from the Feces of Organically Raised Chickens

    Science.gov (United States)

    Kennedy, Victoria; Van Laar, Tricia A.; Aleru, Omoshola; Thomas, Michael; Ganci, Michelle

    2016-01-01

    Antibiotic feed supplements have been implicated in the rise of multidrug-resistant bacteria. An alternative to antibiotics is probiotics. Here, we report the genome sequences of two Bacillus and one Solibacillus species, all spore-forming, Gram-positive bacteria, isolated from the feces organically raised chicken feces, with potential to serve as probiotics. PMID:27587809

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

    NARCIS (Netherlands)

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

    2010-01-01

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

  11. Leveraging a high resolution microfluidic assay reveals insights into pathogenic fungal spore germination.

    Science.gov (United States)

    Barkal, Layla J; Walsh, Naomi M; Botts, Michael R; Beebe, David J; Hull, Christina M

    2016-05-16

    Germination of spores into actively growing cells is a process essential for survival and pathogenesis of many microbes. Molecular mechanisms governing germination, however, are poorly understood in part because few tools exist for evaluating and interrogating the process. Here, we introduce an assay that leverages developments in microfluidic technology and image processing to quantitatively measure germination with unprecedented resolution, assessing both individual cells and the population as a whole. Using spores from Cryptococcus neoformans, a leading cause of fatal fungal disease in humans, we developed a platform to evaluate spores as they undergo morphological changes during differentiation into vegetatively growing yeast. The assay uses pipet-accessible microdevices that can be arrayed for efficient testing of diverse microenvironmental variables, including temperature and nutrients. We discovered that temperature influences germination rate, a carbon source alone is sufficient to induce germination, and the addition of a nitrogen source sustains it. Using this information, we optimized the assay for use with fungal growth inhibitors to pinpoint stages of germination inhibition. Unexpectedly, the clinical antifungal drugs amphotericin B and fluconazole did not significantly alter the process or timing of the transition from spore to yeast, indicating that vegetative growth and germination are distinct processes in C. neoformans. Finally, we used the high temporal resolution of the assay to determine the precise defect in a slow-germination mutant. Combining advances in microfluidics with a robust fungal molecular genetic system allowed us to identify and alter key temporal, morphological, and molecular events that occur during fungal germination. PMID:27026574

  12. Comparative analysis of Bacillus weihenstephanensis KBAB4 spores obtained at different temperatures

    NARCIS (Netherlands)

    Garcia, D.; Voort, van der M.; Abee, T.

    2010-01-01

    The impact of Bacillus weihenstephanensis KBAB4 sporulation temperature history was assessed on spore heat resistance, germination and outgrowth capacity at a temperature range from 7 to 30 °C. Sporulation rate and efficiency decreased at low temperature, as cells sporulated at 12, 20 and 30 °C with

  13. 40 CFR 180.1011 - Viable spores of the microorganism Bacillus thuringiensis Berliner; exemption from the...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Viable spores of the microorganism Bacillus thuringiensis Berliner; exemption from the requirement of a tolerance. 180.1011 Section 180.1011 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL...

  14. The spatiotemporal distributions and determinants of ambient fungal spores in the Greater Taipei area

    International Nuclear Information System (INIS)

    Airborne fungal spores, a type of bioaerosols, are significant air pollutants. We conducted a study to determine the spatiotemporal distributions of ambient fungi in the Greater Taipei area and develop land use regression (LUR) models for total and major fungal taxa. Four seasonal sampling campaigns were conducted over a year at 44 representative sites. Multiple regressions were performed to construct the LUR models. Ascospores were the most prevalent category, followed by Aspergillus/Penicillium, basidiospores, and Cladosporium. The highest fungal concentrations were found in spring. According to the LUR models, higher concentrations of Aspergillus/Penicillium and basidiospores were respectively present in residential/commercial areas and in areas with shorter road lengths. Various meteorological factors, particulates with aerodynamic diameters of ≤10 μm, and elevation also had significant relationships with fungal concentrations. The LUR models developed in this study can be used to assess spatiotemporal fungal distribution in the Greater Taipei area. - Highlights: • Spatiotemporal distributions of ambient fungal spores were evaluated. • Land-use regression models for ambient fungal spores were developed. • The major predictors were meteorological factors and particulates. • Commercial and residential areas are predictor of Aspergillus/Penicillium levels. • Road length is predictor of basidiospores levels. - The first land-use regression models of ambient fungal spores were developed which can be used to assess spatiotemporal fungal distributions in the Greater Taipei area

  15. 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 f

  16. Genome Sequences of Three Spore-Forming Bacteria Isolated from the Feces of Organically Raised Chickens.

    Science.gov (United States)

    Kennedy, Victoria; Van Laar, Tricia A; Aleru, Omoshola; Thomas, Michael; Ganci, Michelle; Rawat, Mamta

    2016-01-01

    Antibiotic feed supplements have been implicated in the rise of multidrug-resistant bacteria. An alternative to antibiotics is probiotics. Here, we report the genome sequences of two Bacillus and one Solibacillus species, all spore-forming, Gram-positive bacteria, isolated from the feces organically raised chicken feces, with potential to serve as probiotics. PMID:27587809

  17. The effect of turf cutting on plant and arbuscular mycorrhizal spore recolonisation: Implications for heathland restoration

    NARCIS (Netherlands)

    Vergeer, P.; Berg, van den L.J.L.; Baar, J.; Ouborg, N.J.; Roelofs, J.G.M.

    2006-01-01

    In two natural heathland vegetations, we analysed the effect of turf cutting on spore numbers of arbuscular mycorrhizal fungi (AMF). Next to this, we performed a controlled factorial experiment to examine the role of AMF for germination and establishment of Arnica montana in both turf cut and non-tu

  18. 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.

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

    NARCIS (Netherlands)

    M. Farenhorst; B.G.J. Knols

    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 s

  20. 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 type

  1. The role of aquaporins in pH-Dependent germination of Rhizopus delemar spores

    Science.gov (United States)

    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 u...

  2. The decision to germinate is regulated by divergent molecular networks in spores and seeds

    DEFF Research Database (Denmark)

    Vesty, Eleanor F.; Saidi, Younousse; Moody, Laura A.;

    2016-01-01

    , the role of abscisic acid and diterpenes (gibberellins) in germination assumed much greater importance as land plant evolution progressed. We conclude that the endogenous hormone signalling networks mediating germination in response to the environment may have evolved independently in spores and seeds...

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

    Science.gov (United States)

    Owens, Trevor

    2012-01-01

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

  4. Development and Use of Fluorescent Antibody and qPCR Protocols for the Electrostatic Spore Trap

    Science.gov (United States)

    Fluorescent antibody (FA) and qPCR protocols were evaluated for the newly developed aerobiological sampler (Ionic Spore Trap), which depends upon electrostatic deposition of particulates onto a 25 mm aluminum disk (stub). This device was originally designed for assessment of captured particulates by...

  5. Modelling the effect of sub(lethal) heat treatment of Bacillus subtilis spores on germination rate and outgrowth to exponentially growing vegetative cells

    NARCIS (Netherlands)

    Smelt, J.P.P.M.; Bos, A.P.; Kort, R.; Brul, S.

    2008-01-01

    Spores of Bacillus subtilis were subjected to relatively mild heat treatments in distilled water and properties of these spores were studied. These spores had lost all or part of their dipicolinic acid (DPA) depending on the severity of the heat treatment. Even after relatively mild heat treatments

  6. 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

  7. Rapid Detection of Viable Bacillus anthracis Spores in Environmental Samples by Using Engineered Reporter Phages.

    Science.gov (United States)

    Sharp, Natasha J; Molineux, Ian J; Page, Martin A; Schofield, David A

    2016-04-01

    Bacillus anthracis, the causative agent of anthrax, was utilized as a bioterrorism agent in 2001 when spores were distributed via the U.S. postal system. In responding to this event, the Federal Bureau of Investigation used traditional bacterial culture viability assays to ascertain the extent of contamination of the postal facilities within 24 to 48 h of environmental sample acquisition. Here, we describe a low-complexity, second-generation reporter phage assay for the rapid detection of viableB. anthracis spores in environmental samples. The assay uses an engineered B. anthracis reporter phage (Wβ::luxAB-2) which transduces bioluminescence to infected cells. To facilitate low-level environmental detection and maximize the signal response, expression of luxABin an earlier version of the reporter phage (Wβ::luxAB-1) was optimized. These alterations prolonged signal kinetics, increased light output, and improved assay sensitivity. Using Wβ::luxAB-2, detection of B. anthracis spores was 1 CFU in 8 h from pure cultures and as low as 10 CFU/g in sterile soil but increased to 10(5)CFU/g in unprocessed soil due to an unstable signal and the presence of competing bacteria. Inclusion of semiselective medium, mediated by a phage-expressed antibiotic resistance gene, maintained signal stability and enabled the detection of 10(4)CFU/g in 6 h. The assay does not require spore extraction and relies on the phage infecting germinating cells directly in the soil sample. This reporter phage displays promise for the rapid detection of low levels of spores on clean surfaces and also in grossly contaminated environmental samples from complex matrices such as soils. PMID:26873316

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

    Directory of Open Access Journals (Sweden)

    Kelly A Fimlaid

    2015-10-01

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

  9. Chemokine-Releasing Microparticles Improve Bacterial Clearance and Survival of Anthrax Spore-Challenged Mice

    Science.gov (United States)

    Espina, Virginia; Liotta, Lance A.; Popov, Serguei G.

    2016-01-01

    In this study the hydrogel microparticles (MPs) were used to enhance migration of neutrophils in order to improve outcome of anthrax infection in a mouse model. Two MP formulations were tested. In the first one the polyacrylamide gel MPs were chemically coupled with Cibacron Blue (CB) affinity bait. In the second one the bait molecules within the MPs were additionally loaded with neutrophil-attracting chemokines (CKs), human CXCL8 and mouse CCL3. A non-covalent interaction of the bait with the CKs provided their gradual release after administration of the MPs to the host. Mice were challenged into footpads with Bacillus anthracis Sterne spores and given a dose of MPs a few hours before and/or after the spores. Pre-treatment with a single dose of CK-releasing MPs without any additional intervention was able to induce influx of neutrophils to the site of spore inoculation and regional lymph nodes correlating with reduced bacterial burden and decreased inflammatory response in footpads. On average, in two independent experiments, up to 53% of mice survived over 13 days. All control spore-challenged but MP-untreated mice died. The CB-coupled particles were also found to improve survival likely due to the capacity to stimulate release of endogenous CKs, but were less potent at decreasing the inflammatory host response than the CK-releasing MPs. The CK post-treatment did not improve survival compared to the untreated mice which died within 4 to 6 days with a strong inflammation of footpads, indicating quick dissemination of spores though the lymphatics after challenge. This is the first report on the enhanced innate host resistance to anthrax in response to CKs delivered and/or endogenously induced by the MPs. PMID:27632537

  10. Assessment of Clostridium perfringens spore response to high hydrostatic pressure and heat with nisin.

    Science.gov (United States)

    Gao, Yulong; Qiu, Weifen; Wu, Ding; Fu, Qiang

    2011-08-01

    The elimination of spores from low-acid foods presents food-processing and food-safety challenges to high-pressure processing (HPP) developers as bacterial spores are extremely resistant to pressure. Therefore, the effects of pressure (400-800 MPa), temperature (35-95 °C), and nisin (0-496 IU/mL) on the inactivation of Clostridium perfringens AS 64701 spores at various pressure-holding times (7.5-17.5 min) were explored. A second-order polynomal equation for HPP- and nisin-induced inactivation of C. perfringens spores was constructed with response surface methodology. Experiment results showed that the experimental values were shown to be significantly in agreement with the predicted values because the adjusted determination coefficient (R (Adj)²) was 0.9708 and the level of significance was P process parameters (obtained by solving the quadratic polynomal equation) for a six-log cycle reduction of C. perfringens AS 64701 spores were pressure of 654 Mpa, temperature of 74 °C, pressure-holding time of 13.6 min, and nisin concentration of 328 IU/mL. The validation of the model equation for predicting the optimum response values was verified effectively by ten test points that were not used in the establishment of the model. Compared with conventional HPP techniques, the main process advantages of HPP-nisin combination sterilization in the UHT milk are, lower pressure, temperature, natural preservative (nisin), and in a shorter treatment time. The synergistic inactivation of bacteria by HPP-nisin combination is a promising and natural method to increase the efficiency and safety of high-pressure pasteurization.

  11. Viability of Clostridium sporogenes spores after CaO hygienization of meat waste

    Directory of Open Access Journals (Sweden)

    Justyna Bauza-Kaszewska

    2014-09-01

    Full Text Available The occurrence of the pathogenic species [i]C. perfringens[/i] and [i]C. botulinum spores[/i] in animal by-products poses a potential epidemiological hazard. Strong entero- and neurotoxins produced by these bacteria adversely affect human health. To inactivate pathogens present in animal by-products, waste must be subjected to various methods of sanitization. The aim of the presented study was to estimate the effect of different doses of CaO on the viability of spores [i] Clostridium sporogenes[/i] in meat wastes category 3. During the research, two doses of burnt lime were added to the poultry mince meat and meat mixed with swine blood contaminated with [i]Clostridium sporogenes[/i] spore suspension. Half of the samples collected for microbiological analyses were buffered to achieve the pH level ~7, the other were examined without pH neutralization. To estimate the spore number, 10-fold dilution series in peptone water was prepared and heat-treated at 80 °C for 10 min. After cooling-down, one milliliter of each dilution was pour-plated onto DRCM medium solidified with agar. Statistical analysis were performed using the Statistica software. Application of 70% CaO caused complete inactivation of [i]Clostridium spores[/i] in meat wastes after 48 hours. The highest temperature achieved during the experiment was 67 °C. Rapid alkalization of the biomass resulted in increasing pH to values exceeding 12. The effect of liming was not dependent on the meat wastes composition nor CaO dose. The experiment proved the efficiency of liming as a method of animal by-products sanitization. Application of the obtained results may help reduce the epidemiological risk and ensure safety to people handling meat wastes at each stage of their processing and utilization.

  12. Rapid Detection of Viable Bacillus anthracis Spores in Environmental Samples by Using Engineered Reporter Phages

    Science.gov (United States)

    Sharp, Natasha J.; Molineux, Ian J.; Page, Martin A.

    2016-01-01

    Bacillus anthracis, the causative agent of anthrax, was utilized as a bioterrorism agent in 2001 when spores were distributed via the U.S. postal system. In responding to this event, the Federal Bureau of Investigation used traditional bacterial culture viability assays to ascertain the extent of contamination of the postal facilities within 24 to 48 h of environmental sample acquisition. Here, we describe a low-complexity, second-generation reporter phage assay for the rapid detection of viable B. anthracis spores in environmental samples. The assay uses an engineered B. anthracis reporter phage (Wβ::luxAB-2) which transduces bioluminescence to infected cells. To facilitate low-level environmental detection and maximize the signal response, expression of luxAB in an earlier version of the reporter phage (Wβ::luxAB-1) was optimized. These alterations prolonged signal kinetics, increased light output, and improved assay sensitivity. Using Wβ::luxAB-2, detection of B. anthracis spores was 1 CFU in 8 h from pure cultures and as low as 10 CFU/g in sterile soil but increased to 105 CFU/g in unprocessed soil due to an unstable signal and the presence of competing bacteria. Inclusion of semiselective medium, mediated by a phage-expressed antibiotic resistance gene, maintained signal stability and enabled the detection of 104 CFU/g in 6 h. The assay does not require spore extraction and relies on the phage infecting germinating cells directly in the soil sample. This reporter phage displays promise for the rapid detection of low levels of spores on clean surfaces and also in grossly contaminated environmental samples from complex matrices such as soils. PMID:26873316

  13. Prevalence, intensity and complications of Microsporidium spores amongst HIV-positive hospital patients in Ilorin, Nigeria

    Directory of Open Access Journals (Sweden)

    Amase Nyamngee

    2013-03-01

    Full Text Available Background: Microsporidiasis, which is of great concern for immunocompromised patients, is poorly studied in developing countries.Objectives: A study was carried out amongst HIV-positive hospital patients and HIV-negative hospital controls in Ilorin, Nigeria, between January 2009 and July 2010 to determine the prevalence and intensity of Microsporidium spores and the complications associated with their presence.Method: Stool samples from 750 HIV-positive patients and 375 HIV-negative patients were studied using the Chromotrope-2R staining technique. Determination of CD4+ count was performed on the Partec Cyflow SL-3 CD4/8 instrument. Intensity of spores was determined by counting the total number of the spores in a 10 μl stained smear of stool. Images were captured with Phenix Microimage Analysis Software and data obtained were analysed using the Statistical Package for the Social Sciences.Results: The prevalence of Microsporidium isolates amongst the HIV-positive hospital patients was significantly higher (42.4% than amongst the HIV-negative controls (19.2%(p < 0.05. The intensity of microsporidial spores amongst HIV-positive hospital patients was also significantly higher than amongst the controls (p < 0.05. However, the difference in the intensity of spores amongst HIV-positive patients who were on antiretroviral therapy(n = 411 and those who were not (n = 339 was not significant (p = 0.236. Microsporidiasis in HIV infection infection was common amongst patients with with low CD4+ counts, diarrhoea, body rashes and cough.Conclusion: Both the prevalence and intensity of Microsporidiasis are high amongst HIV-positive hospital patients; campaigns to promote awareness, prevention and control are required. Laboratory testing for microsporidia in HIV patients should be performed routinely so as to identify the organism for prompt medical attention.

  14. Adhesion and cellsurface properties of wild species of spore formers against enteric pathogens

    Institute of Scientific and Technical Information of China (English)

    Anu P Sebastian; Keerthi T. R

    2013-01-01

    Objective:To investigate the adhesion potential and cell surface properties against enteric pathogensSalmonella typhi,Salmonella para typhiA andVibrio cholera.Methods:Adhesion potentials of spore and vegetative phase were studied separately for the isolates.Hydrophobic nature was measured on the basis of affinity towards the xylene.Autoaggregation and coaggregation were studied on the basis of clumping of cells.In vitro adhesion studies were done on mucouswhich were prepared from infant child faeces.Biofilm production of superior adhesive isolate was confirmed bySEM analysis.Results:Spore and vegetative phases of isolates possessed a different rate of adhesion potentials on intestinal mucous, which indicated that cell surface properties were involved in adhesion process.Spores showed a higher hydrophobicity than their vegetative cells which remainedless or non hydrophobic.Vegetative phases showed capabilities for autoaggregation and coaggregation.Spores were found to be more adhesive on intestinal mucous than vegetative phase.Among enteric pathogensVibrio cholera registered higher adhesion potentials with supporting cell surface properties.Among the five sporeforming isolates, isolateBM-3 possess superior adhesion than enteric pathogens and also exhibited biofilm formation which enhances colonization potential.Conclusions:Spore and vegetative cell phases shows differences in adhesion potentials.Cell surface properties and adhesion studies reveals that isolateBM-3 can be selected as superior isolate which is capable for biofilm production.In short, isolateBM-3 possesses an enhanced adhesion potential than enteric pathogens towards intestinal mucous which is a desirable probiotic character.

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

    International Nuclear Information System (INIS)

    The polyols arabitol and mannitol, recently proposed as source tracers for fungal spores, were used in this study to estimate fungal contributions to atmospheric aerosol. Airborne particulate matter (PM2.5 and PM10) was collected at Jianfengling Mountain, a tropical rainforest on Hainan Island situated off the south China coast, during spring and analyzed for arabitol and mannitol by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The average concentrations of arabitol and mannitol exhibited high values with averages of 7.0 and 16.0 ng m-3 respectively in PM2.5 and 44.0 and 71.0 ng m-3 in PM10. The two tracers correlated well with each other, especially in the coarse mode aerosol (PM2.5-10), indicating they were mainly associated with coarse aerosol particles and had common sources. Arabitol and mannitol in PM10 showed significant positive correlations with relative humidity, as well as positive correlations with average temperature, suggesting a wet emissions mechanism of biogenic aerosol in the form of fungal spores. We made estimations of the contribution of fungal spores to ambient PM mass and to organic carbon, based on the observed ambient concentrations of these two tracers. The relative contributions of fungal spores to the PM10 mass were estimated to range from 1.6 to 18.2%, with a rather high mean value of 7.9%, and the contribution of fungal spores to organic carbon in PM10 ranged from 4.64 to 26.1%, with a mean value of 12.1%, implying that biological processes are important sources of atmospheric aerosol.

  16. Physiological role of germicidins in spore germination and hyphal elongation in Streptomyces coelicolor A3(2).

    Science.gov (United States)

    Aoki, Yuu; Matsumoto, Daisuke; Kawaide, Hiroshi; Natsume, Masahiro

    2011-09-01

    Four germicidin homologs were isolated from a liquid culture of Streptomyces coelicolor A3(2). These were identified as germicidins A, B and C, and surugapyrone A (germicidin D). Absolute stereochemistry of the chiral center in germicidins A and C is determined to be S. All germicidins inhibited germination of S. coelicolor A3(2) spores above 1 μg ml(-1). S. coelicolor A3(2) spores collected from a single petri dish (9 cm i.d.) contained 5.4 μg of germicidin A (-2.7 × 10(-14) g per spore), which accounts for 2.3% of the spore extract, and contents of germicidins B, C and D were 0.2-0.8 μg. The activity of the spore extract corresponded well with the sum of the activity of each germicidin, which was estimated from the content and dose-response curve, which indicates that germicidins functions as self-germination inhibitors in S. coelicolor A3(2). Inhibitory action of germicidin A on spore germination was reversible and germicidin A inhibited not only spore germination but also hyphal elongation.

  17. Greater enhancement of Bacillus subtilis spore yields in submerged cultures by optimization of medium composition through statistical experimental designs.

    Science.gov (United States)

    Chen, Zhen-Min; Li, Qing; Liu, Hua-Mei; Yu, Na; Xie, Tian-Jian; Yang, Ming-Yuan; Shen, Ping; Chen, Xiang-Dong

    2010-02-01

    Bacillus subtilis spore preparations are promising probiotics and biocontrol agents, which can be used in plants, animals, and humans. The aim of this work was to optimize the nutritional conditions using a statistical approach for the production of B. subtilis (WHK-Z12) spores. Our preliminary experiments show that corn starch, corn flour, and wheat bran were the best carbon sources. Using Plackett-Burman design, corn steep liquor, soybean flour, and yeast extract were found to be the best nitrogen source ingredients for enhancing spore production and were studied for further optimization using central composite design. The key medium components in our optimization medium were 16.18 g/l of corn steep liquor, 17.53 g/l of soybean flour, and 8.14 g/l of yeast extract. The improved medium produced spores as high as 1.52 +/- 0.06 x 10(10) spores/ml under flask cultivation conditions, and 1.56 +/- 0.07 x 10(10) spores/ml could be achieved in a 30-l fermenter after 40 h of cultivation. To the best of our knowledge, these results compared favorably to the documented spore yields produced by B. subtilis strains. PMID:19697022

  18. Survivability of bare, individual Bacillus subtilis spores to high-velocity surface impact: Implications for microbial transfer through space

    Science.gov (United States)

    Barney, Brandon L.; Pratt, Sara N.; Austin, Daniel E.

    2016-06-01

    Laboratory experiments show that endospores of Bacillus subtilis survive impact against a solid surface at velocities as high as 299 ±28 m/s. During impact, spores experience and survive accelerations of at least 1010 m/s2. The spores were introduced into a vacuum chamber using an electrospray source and accelerated to a narrow velocity distribution by entrainment in a differentially pumped gas flow. Different velocity ranges were studied by modifying the gas flow parameters. The spores were electrically charged, allowing direct measurement of the velocity of each spore as it passed through an image charge detector prior to surface impact. Spores impacted a glass surface and were collected for subsequent analysis by culturing. Most spores survived impact at all measured velocities. These experiments differ fundamentally from other studies that show either shock or impact survivability of bacteria embedded within or on the surface of a projectile. Bacteria in the present experiments undergo a single interaction with a solid surface at the full impact velocity, in the absence of any other effects such as cushioning due to microbe agglomerations, deceleration due to air or vapor, or transfer of impact shock through solid or liquid media. During these full-velocity impact events, the spores experience extremely high decelerations. This study is the first reported instance of accelerations of this magnitude experienced during a bacteria impact event. These results are discussed in the context of potential transfer of viable microbes in space and other scenarios involving surface impacts at high velocities.

  19. Survival and Germinability of Bacillus subtilis Spores Exposed to Simulated Mars Solar Radiation: Implications for Life Detection and Planetary Protection

    Science.gov (United States)

    Tauscher, Courtney; Schuerger, Andrew C.; Nicholson, Wayne L.

    2006-08-01

    Bacterial spores have been considered as microbial life that could survive interplanetary transport by natural impact processes or human spaceflight activity. Deposition of terrestrial microbes or their biosignature molecules onto the surface of Mars could negatively impact life detection experiments and planetary protection measures. Simulated Mars solar radiation, particularly the ultraviolet component, has been shown to reduce spore viability, but its effect on spore germination and resulting production of biosignature molecules has not been explored. We examined the survival and germinability of Bacillus subtilis spores exposed to simulated martian conditions that include solar radiation. Spores of B. subtilis that contain luciferase resulting from expression of an sspB-luxAB gene fusion were deposited on aluminum coupons to simulate deposition on spacecraft surfaces and exposed to simulated Mars atmosphere and solar radiation. The equivalent of 42 min of simulated Mars solar radiation exposure reduced spore viability by nearly 3 logs, while germination-induced bioluminescence, a measure of germination metabolism, was reduced by less than 1 log. The data indicate that spores can retain the potential to initiate germination-associated metabolic processes and produce biological signature molecules after being rendered nonviable by exposure to Mars solar radiation.

  20. Co-occurrence of airborne allergenic pollen grains and fungal spores in Rzeszów, Poland (2000-2002

    Directory of Open Access Journals (Sweden)

    Idalia Kasprzyk

    2012-12-01

    Full Text Available The co-occurrence of airborne allergenic pollen grains and fungal spores was estimated in Rzeszów in the years 2000-2002. The volumetric method was used in this aerobiological study. Six taxa of pollen grains and five types of fungal spores characterized by strong allergenicity and/or high concentrations in the air were analyzed. The time series of pollen grains and fungal spores were compared using PCA analysis. The periods of the greatest concentrations of tree pollen did not coincide with similar periods for herbaceous plants and fungal spores. From February to mid-March, Alnus pollen dominated in the air. The second period was characterized by Betula pollen. It occurred in April. Herbaceous pollen and fungal spores occurred in the air simultaneously (from mid May to the end of August, creating a risky situation for sensitized people. The periods of the highest concentrations of Epicoccum and Ganoderma fungal spores did not coincide with the same period for the examined plant taxa. In Rzeszów the probability of becoming exposed to very high concentrations of allergenic pollen and fungal spores at the same time was high, especially in July, when the highest concentrations of Poaceae, Alternaria and Cladosporium were noted. The hypersensitivity to only one plant or fungal allergen is rarely encountered. Under the present scenarios of global warming, pollen seasons of many taxa will be longer and sufferers will have year-long symptoms.

  1. Adhesion of B. subtilis spores and vegetative cells onto stainless steel--DLVO theories and AFM spectroscopy.

    Science.gov (United States)

    Harimawan, Ardiyan; Zhong, Shaoping; Lim, Chwee-Teck; Ting, Yen-Peng

    2013-09-01

    Interactions between the bacterium Bacillus subtilis (either as vegetative cells or as spores) and stainless steel 316 (SS-316) surfaces were quantified using the classical Derjaguin-Landau-Verwey-Overbeek (DLVO) theory and extended DLVO (xDLVO) approach in conjunction with live force spectroscopy using an Atomic Force Microscope (AFM). The xDLVO approach accounts for acid-base (polar) interactions that are not considered in the classical DLVO theory. AFM results revealed that spores manifested stronger attraction interactions to stainless steel compared to their vegetative cells counterparts due to lower energy barrier as predicted by both the theoretical approaches as well as the higher hydrophobicity on the spore surfaces. Both DLVO and xDLVO theories predict that vegetative cells manifest weaker attachment on the surfaces compared to spores. Results of AFM force measurement corroborate these findings; spores recorded significantly higher adhesion force (2.92±0.4 nN) compared to vegetative cells (0.65±0.2 nN). The adhesion of spores presents greater challenges in biofilm control owing to its stronger attachment and persistence when the spores are formed under adverse environmental conditions. PMID:23777862

  2. Observations on the Inactivation Efficacy of a MALDI-TOF MS Chemical Extraction Method on Bacillus anthracis Vegetative Cells and Spores.

    Directory of Open Access Journals (Sweden)

    Simon A Weller

    Full Text Available A chemical (ethanol; formic acid; acetonitrile protein extraction method for the preparation of bacterial samples for matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS identification was evaluated for its ability to inactivate bacterial species. Initial viability tests (with and without double filtration of the extract through 0.2 μM filters, indicated that the method could inactivate Escherichia coli MRE 162 and Klebsiella pneumoniae ATCC 35657, with or without filtration, but that filtration was required to exclude viable, avirulent, Bacillus anthracis UM23CL2 from extracts. Multiple, high stringency, viability experiments were then carried out on entire filtered extracts prepared from virulent B. anthracis Vollum vegetative cells and spores ranging in concentration from 10(6-10(8 cfu per extract. B. anthracis was recovered in 3/18 vegetative cell extracts and 10/18 spore extracts. From vegetative cell extracts B. anthracis was only recovered from extracts that had undergone prolonged Luria (L-broth (7 day and L-agar plate (a further 7 days incubations. We hypothesise that the recovery of B. anthracis in vegetative cell extracts is due to the escape of individual sub-lethally injured cells. We discuss our results in view of working practises in clinical laboratories and in the context of recent inadvertent releases of viable B. anthracis.

  3. Observations on the Inactivation Efficacy of a MALDI-TOF MS Chemical Extraction Method on Bacillus anthracis Vegetative Cells and Spores.

    Science.gov (United States)

    Weller, Simon A; Stokes, Margaret G M; Lukaszewski, Roman A

    2015-01-01

    A chemical (ethanol; formic acid; acetonitrile) protein extraction method for the preparation of bacterial samples for matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) identification was evaluated for its ability to inactivate bacterial species. Initial viability tests (with and without double filtration of the extract through 0.2 μM filters), indicated that the method could inactivate Escherichia coli MRE 162 and Klebsiella pneumoniae ATCC 35657, with or without filtration, but that filtration was required to exclude viable, avirulent, Bacillus anthracis UM23CL2 from extracts. Multiple, high stringency, viability experiments were then carried out on entire filtered extracts prepared from virulent B. anthracis Vollum vegetative cells and spores ranging in concentration from 10(6)-10(8) cfu per extract. B. anthracis was recovered in 3/18 vegetative cell extracts and 10/18 spore extracts. From vegetative cell extracts B. anthracis was only recovered from extracts that had undergone prolonged Luria (L)-broth (7 day) and L-agar plate (a further 7 days) incubations. We hypothesise that the recovery of B. anthracis in vegetative cell extracts is due to the escape of individual sub-lethally injured cells. We discuss our results in view of working practises in clinical laboratories and in the context of recent inadvertent releases of viable B. anthracis. PMID:26633884

  4. Sporoderm-Broken Spores of Ganoderma lucidum Inhibit the Growth of Lung Cancer: Involvement of the Akt/mTOR Signaling Pathway.

    Science.gov (United States)

    Chen, Yali; Lv, Jing; Li, Kun; Xu, Jing; Li, Mingyan; Zhang, Wen; Pang, Xiufeng

    2016-10-01

    The sporoderm-broken spores of Ganoderma lucidum (SBGS) and their extracts exhibited a wide range of biological activities. In the present study, we prepare ethanol/ethanol extract (E/E-SBGS) and ethanol/aqueous extract (E/A-SBGS) from SBGS and examine their antitumor activities against human lung cancer. Our results showed that E/E-SBGS, not E/A-SBGS, inhibited the survival and migration of lung cancer cells in a dose-dependent manner. E/E-SBGS arrested cell cycle at G2/M phase and triggered apoptosis by decreasing the expression and activity of cell cycle regulators, cyclin B1 and cdc2, as well as anti-apoptotic proteins, Bcl-2 and Bcl-xl. Consequently, colony formation of lung cancer cells was markedly blocked by E/E-SBGS at subtoxic concentrations. Oral administration of both E/E-SBGS and SBGS significantly suppressed tumor volume and tumor weight without gross toxicity in mice. Mechanism study showed that E/E-SBGS dose-dependently suppressed the activation of Akt, the mammalian target of rapamycin (mTOR) and their downstream molecules S6 kinase and 4E-BP1 in treated tumor cells. Taken together, these results indicate that the ethanol extract of sporoderm-broken spores of G. lucidum suppresses the growth of human lung cancer, at least in part, through inhibition of the Akt/mTOR signaling pathway, suggesting its potential role in cancer treatments.

  5. Ganoderma lucidum spore powder modulates Bcl-2 and Bax expression in the hippocampus and cerebral cortex, and improves learning and memory in pentylenetetrazole-kindled rats

    Institute of Scientific and Technical Information of China (English)

    Shuang Zhao; Shengchang Zhang; Shuqiu Wang

    2011-01-01

    We studied the effects of Ganoderma lucidum spore powder on Bax and Bcl-2 expression and neuronal apoptosis in pentylenetetrazole-kindled epileptic rats. Sixty adult rats were randomly divided into a control group, an epileptic group (kindled) and three medication groups ( 150, 300,450 mg/kg given to kindled rats). Bax and Bcl-2 immunohistochemistry and TUNEL labeling show ed that the number of Bax- and TUNEL-positive cells in the hippocampus and cerebral cortex decreased significantly in the high-dose medication group, while the number of Bcl-2immunoreactive cells increased. The Morris water maze test showed that high-dose treatment significantly shortened escape latency and increased spatial probe trial performance. Our findings indicate that a high dose of Ganoderma lucidum spore powder upregulates the expressionof antiapoptotic Bcl-2 protein in the hippocampus and cerebral cortex, inhibits proapoptotic Bax expression, and decreases seizure-induced neuronal apoptosis. Further,Ganoderma lucidum appears to protect against epilepsy-related learning and memory impairments.

  6. Significant contributions of fungal spores to the organic carbon and to the aerosol mass balance of the urban atmospheric aerosol

    Science.gov (United States)

    Bauer, Heidi; Schueller, Elisabeth; Weinke, Gert; Berger, Anna; Hitzenberger, Regina; Marr, Iain L.; Puxbaum, Hans

    Fungal spores are ubiquitous components of atmospheric aerosols and are therefore also contributors to the organic carbon (OC) component and to the mass of PM 10 (PM—particulate matter) aerosols. In this study we use spore counts and an experimentally derived factor of 13 pg C and of 33 pg fresh weight per spore for assessing quantitatively the contribution to OC and PM 10. The concentrations of airborne fungal spores were determined at a suburban (Schafberg) and a traffic-dominated urban site (Rinnböckstrasse) in Vienna, Austria, during spring and summer. Fungal spores OC ranged from 22 to 677 ng m -3 with a summer mean value of around 350 ng m -3 at the suburban site and 300 ng m -3 at the urban traffic site. At the suburban site fungal spores contributed on average 6% in spring and 14% in summer to aerosol OC mass concentration. At the traffic-dominated site fungal spores accounted for 2% of OC in spring and for 8% in summer. The fungal contribution to PM 10 was also notable and amounted to 3% and 7% at the suburban and to 1% and 4% at the urban site in spring and summer, respectively. Impactor measurements of OC at the suburban site showed that in summer fungal spores were predominant contributors to the coarse aerosol OC, and accounted on average for 60% of the OC in the PM 2-10 fraction. Fungal spores thus can be regarded as main components to PM 10, total OC and, most importantly, coarse OC even in urban areas.

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

    Directory of Open Access Journals (Sweden)

    Brandes Ammann Andrea

    2011-07-01

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

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

    Science.gov (United States)

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

  9. Real-time in situ electron spin resonance measurements on fungal spores of Penicillium digitatum during exposure of oxygen plasmas

    Science.gov (United States)

    Ishikawa, Kenji; Mizuno, Hiroko; Tanaka, Hiromasa; Tamiya, Kazuhiro; Hashizume, Hiroshi; Ohta, Takayuki; Ito, Masafumi; Iseki, Sachiko; Takeda, Keigo; Kondo, Hiroki; Sekine, Makoto; Hori, Masaru

    2012-07-01

    We report the kinetic analysis of free radicals on fungal spores of Penicillium digitatum interacted with atomic oxygen generated plasma electric discharge using real time in situ electron spin resonance (ESR) measurements. We have obtained information that the ESR signal from the spores was observed and preliminarily assignable to semiquinone radical with a g-value of around 2.004 and a line width of approximately 5G. The decay of the signal is possibly linked to the inactivation of the fungal spore. The real-time in situ ESR has proven to be a useful method to elucidate plasma-induced surface reactions on biological specimens.

  10. Real-time In Situ Electron Spin Resonance Measurements on Fungal Spores of Penicillium digitatum during Exposure of Oxygen Plasmas

    CERN Document Server

    Ishikawa, Kenji; Tanaka, Hiromasa; Tamiya, Kazuhiro; Hashizume, Hiroshi; Ohta, Takayuki; Ito, Masafumi; Iseki, Sachiko; Takeda, Keigo; Kondo, Hiroki; Sekine, Makoto; Hori, Masaru

    2012-01-01

    We report the kinetic analysis of free radicals on fungal spores of Penicillium digitatum interacted with atomic oxygen generated plasma electric discharge using real time in situ electron spin resonance (ESR) measurements. We have obtained information that the ESR signal from the spores was observed and preliminarily assignable to semiquinone radical with a g-value of around 2.004 and a line width of approximately 5G. The decay of the signal is possibly linked to the inactivation of the fungal spore. The real-time in situ ESR has proven to be a useful method to elucidate plasma-induced surface reactions on biological specimens.

  11. Production of spores of Trichoderma harzianum on sugar cane molasses and bagasse pith in solid state fermentation for biocontrol

    OpenAIRE

    Jose A. Rodríguez-León; Domenech, F.; M. León; Méndez, T.; D. E. Rodríguez; Ashok Pandey

    1999-01-01

    Solid state fermentation was carried out for the production of spores from Trichoderma harzianum No 53 using sugar cane bagasse pith as solid matrix and sugar cane molasses as carbon and energy source. Different nitrogen sources such as urea, (NH4)2SO4 , NH4H2PO4 and (NH4)2HPO4 were added in the media to test their effect on spores production. Among these, urea was found most useful that resulted high no of spores (1x10(9)/gDM). The influence of temperature and initial moisture of the substra...

  12. Studies on the physiology of Funaria hygrometrica spore germination. Part IV. The effect of blue and red light on the germination of spores

    Directory of Open Access Journals (Sweden)

    J. Krupa

    2015-05-01

    Full Text Available The effect of radiation with wave-lengths 654 nm and 425 nm on the germination of Funaria hydgrometrica spores has been studied. The wave-length bands used for the radiation treatment were employed for various durations and in various combinations. The effects of red and blue light were different in the individual phases of germination when after an inductive radiation treatment light in the 425 nm wave-length was supplied. Differences in the effect of red and blue light are observable in the "post-effects" land in the germination morphology.

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

    Science.gov (United States)

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

    2013-01-15

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

  14. 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

  15. Bacillus subtilis spore survival and expression of germination-induced bioluminescence after prolonged incubation under simulated Mars atmospheric pressure and composition: implications for planetary protection and lithopanspermia

    Science.gov (United States)

    Nicholson, Wayne L.; Schuerger, Andrew C.

    2005-01-01

    Bacterial endospores in the genus Bacillus are considered good models for studying interplanetary transfer of microbes by natural or human processes. Although spore survival during transfer itself has been the subject of considerable study, the fate of spores in extraterrestrial environments has received less attention. In this report we subjected spores of a strain of Bacillus subtilis, containing luciferase resulting from expression of an sspB-luxAB gene fusion, to simulated martian atmospheric pressure (7-18 mbar) and composition (100% CO(2)) for up to 19 days in a Mars simulation chamber. We report here that survival was similar between spores exposed to Earth conditions and spores exposed up to 19 days to simulated martian conditions. However, germination-induced bioluminescence was lower in spores exposed to simulated martian atmosphere, which suggests sublethal impairment of some endogenous spore germination processes.

  16. A Recombination Directionality Factor Controls the Cell Type-Specific Activation of σK and the Fidelity of Spore Development in Clostridium difficile

    Science.gov (United States)

    Serrano, Mónica; Kint, Nicolas; Pereira, Fátima C.; Saujet, Laure; Boudry, Pierre; Dupuy, Bruno; Martin-Verstraete, Isabelle

    2016-01-01

    The strict anaerobe Clostridium difficile is the most common cause of nosocomial diarrhea, and the oxygen-resistant spores that it forms have a central role in the infectious cycle. The late stages of sporulation require the mother cell regulatory protein σK. In Bacillus subtilis, the onset of σK activity requires both excision of a prophage-like element (skinBs) inserted in the sigK gene and proteolytical removal of an inhibitory pro-sequence. Importantly, the rearrangement is restricted to the mother cell because the skinBs recombinase is produced specifically in this cell. In C. difficile, σK lacks a pro-sequence but a skinCd element is present. The product of the skinCd gene CD1231 shares similarity with large serine recombinases. We show that CD1231 is necessary for sporulation and skinCd excision. However, contrary to B. subtilis, expression of CD1231 is observed in vegetative cells and in both sporangial compartments. Nevertheless, we show that skinCd excision is under the control of mother cell regulatory proteins σE and SpoIIID. We then demonstrate that σE and SpoIIID control the expression of the skinCd gene CD1234, and that this gene is required for sporulation and skinCd excision. CD1231 and CD1234 appear to interact and both proteins are required for skinCd excision while only CD1231 is necessary for skinCd integration. Thus, CD1234 is a recombination directionality factor that delays and restricts skinCd excision to the terminal mother cell. Finally, while the skinCd element is not essential for sporulation, deletion of skinCd results in premature activity of σK and in spores with altered surface layers. Thus, skinCd excision is a key element controlling the onset of σK activity and the fidelity of spore development. PMID:27631621

  17. Multifactorial Resistance of Bacillus subtilis Spores to High-Energy Proton Radiation: Role of Spore Structural Components and the Homologous Recombination and Non-Homologous End Joining DNA Repair Pathways

    OpenAIRE

    Moeller, Ralf; Reitz, Günther; Li, Zuofeng; Klein, Stuart; Nicholson, Wayne L.

    2012-01-01

    The space environment contains high-energy charged particles (e.g., protons, neutrons, electrons, α-particles, heavy ions) emitted by the Sun and galactic sources or trapped in the radiation belts. Protons constitute the majority (87%) of high-energy charged particles. Spores of Bacillus species are one of the model systems used for astro- and radiobiological studies. In this study, spores of different Bacillus subtilis strains were used to study the effects of high energetic proton irradiati...

  18. Variable Lymphocyte Receptor Recognition of the Immunodominant Glycoprotein of Bacillus anthracis Spores

    Energy Technology Data Exchange (ETDEWEB)

    Kirchdoerfer, Robert N.; Herrin, Brantley R.; Han, Byung Woo; Turnbough, Jr., Charles L.; Cooper, Max D.; Wilson, Ian A. (SNU); (Scripps); (Emory); (UAB); (Emory Vaccine)

    2012-07-25

    Variable lymphocyte receptors (VLRs) are the adaptive immune receptors of jawless fish, which evolved adaptive immunity independent of other vertebrates. In lieu of the immunoglobulin fold-based T and B cell receptors, lymphocyte-like cells of jawless fish express VLRs (VLRA, VLRB, or VLRC) composed of leucine-rich repeats and are similar to toll-like receptors (TLRs) in structure, but antibodies (VLRB) and T cell receptors (VLRA and VLRC) in function. Here, we present the structural and biochemical characterization of VLR4, a VLRB, in complex with BclA, the immunodominant glycoprotein of Bacillus anthracis spores. Using a combination of crystallography, mutagenesis, and binding studies, we delineate the mode of antigen recognition and binding between VLR4 and BclA, examine commonalities in VLRB recognition of antigens, and demonstrate the potential of VLR4 as a diagnostic tool for the identification of B. anthracis spores.

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

    DEFF Research Database (Denmark)

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

    2016-01-01

    resulted in the following conclusions. In order to obtain sufficient local concentrations of surfactant, substantial amounts of substrate should be supplied; however, massive growth of bacteria increases the risk for clogging at the well inlet areas, causing injectivity loss. In such areas, starvation may....... One of the ways of bacterial survival and propagation under harsh reservoir conditions is formation of spores. A model has been developed that accounts for bacterial growth, substrate consumption, surfactant production, attachment/filtering out, sporulation, and reactivation. Application of spore......-forming bacteria is an advantageous novelty of the present approach. The mathematical setup is a set of 1D transport equations involving reactions and attachment. Characteristic sigmoidal curves are used to describe sporulation and reactivation in response to substrate concentrations. The role of surfactant...

  20. Identification of spores in the polycentric anaerobic gut fungi which enhance their ability to survive.

    Science.gov (United States)

    Brookman; Ozkose; Rogers; Trinci; Theodorou

    2000-03-01

    Two new isolates of the gut fungi were obtained from the rumen digesta and faeces of a cow. These isolates, designated Anaeromyces following rDNA typing, displayed a polycentric growth habit but differed from all other gut fungi in that they were able to survive in the laboratory for considerable periods without the need for sub-culture. Light microscopy of preparations from old liquid-grown cultures revealed the presence of DNA-containing spores with two or four chambers. A comparative evaluation of the growth produced when fresh media were inoculated with a sample originating from young or old cultures revealed that active growth was delayed with the inoculum from the older culture. We propose that the chambered spores observed in these cultures provide an alternative path in the life cycle of these fungi and may function as a resting stage within the anaerobic environment of the herbivore gut. PMID:10719208

  1. 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

    in higher oil production from the heterogeneous chalk rock. In all cases, an incubation period ('shut-in') after the bacterial and/or nutrient injection was needed to give sufficient time for the bacteria to grow inside the core and to produce more oil. Our findings show potential application of bacteria......Little has been done to study microbial enhanced oil recovery (MEOR) in chalk reservoirs. The present study focuses on core flooding experiments designed to see microbial plugging and its effect on oil recovery. A pressure tapped core holder was used for this purpose. A spore forming bacteria...... 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...

  2. Meriderma species (Myxomycetes from the Polish Carpathians: a taxonomic revision using SEM-visualized spore ornamentation

    Directory of Open Access Journals (Sweden)

    Paulina Janik

    2016-04-01

    Full Text Available Meriderma represents a recently described genus of nivicolous myxomycetes with high morphological variability. Due to many complications in its taxonomy and species recognition in the past, the group was considered a morphologically variable complex. Recent clarifications and recognition of morphological boundaries into species and morphotypes has fostered a classification revision of specimens found in the Carpathians. Material used in this study was systematically collected in the Polish part of the Carpathians from 2004 to 2009. As a result of micro- and macroscopic observations of 54 collections, we recorded nine taxa of Meriderma. Seven of these (all but M. carestiae and M. cribrarioides are the first records for Poland and for the Carpathians overall. Our observations based on analysis of spore ornamentation by SEM are in accordance with recently proposed classification and confirm segregation of taxa based on spore ornamentation pattern.

  3. Comparison of methods for the enumeration of Clostridium perfringens spores in water.

    Science.gov (United States)

    Junqueira, Valéria Christina Amstalden; Neto, Romeu Cantúsio; da Silva, Neusely; Terra, Juliana Hirata

    2012-01-01

    Four methods for enumerating Clostridium perfringens spores in water were evaluated: (1) the IMM (Iron Milk Medium) method (MPN); (2) the LS (Lactose Sulfite Broth) method (MPN); (3) the m-CP (membrane filtration Clostridium perfringens Agar) method (membrane filtration); and (4) the TSC (Tryptose Sulfite Cycloserine Agar) method (membrane filtration). The performance of these methods was compared with that of the DRCM (Differential Reinforced Clostridium Medium) method (MPN) as adopted by CETESB (Brazil's Environmental Sanitation Technology Company) for the analysis of C. perfringens spores in water. Statistical analysis was performed according to ISO 17994:2004 (Water Quality - Criteria for Establishing Equivalence between Microbiological Methods). The LS, m-CP, and TSC methods were considered not equivalent to the DRCM method, as they gave significantly lower results. The IMM showed inconclusive results and, according to ISO 17994:2004, analysis of a greater number of samples is needed to draw definitive conclusions comparing IMM and DRCM.

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

    Directory of Open Access Journals (Sweden)

    Celenk Molva

    2015-12-01

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

  5. Fate of Enteromorpha flexuosa (Wulfen) J. Agardh and its spores in darkness: Implications for ballast water management

    Digital Repository Service at National Institute of Oceanography (India)

    Kolwalkar, J.P.; Sawant, S.S.; Dhargalkar, V.K.

    Effects of darkness on the filaments and spores of a marine alga, Enteromorpha flexuosa (Wulfen) J. Agardh was evaluated through incubation experiments. Sporulation in the algal filaments was observed even after dark exposure of 108 days...

  6. Hematotoxicity and genotoxicity evaluations in Swiss mice intraperitoneally exposed to Bacillus thuringiensis (var kurstaki) spore crystals genetically modified to express individually Cry1Aa, Cry1Ab, Cry1Ac, or Cry2Aa.

    Science.gov (United States)

    Mezzomo, Bélin Poletto; Miranda-Vilela, Ana Luisa; Barbosa, Lilian Carla Pereira; Albernaz, Vanessa Lima; Grisolia, Cesar Koppe

    2016-08-01

    Bacillus thuringiensis (Bt) has been widely used in foliar sprays as part of integrated pest management strategies against insect pests of agricultural crops. Since the advent of genetically modified plants expressing Bt δ-endotoxins, the bioavailability of Cry proteins has increased, and therefore for biosafety reasons their adverse effects should be studied, mainly for nontarget organisms. We evaluated, in Swiss mice, the hematotoxicity and genotoxicity of the genetically modified strains of Bt spore crystals Cry1Aa, 1Ab, 1Ac, or 2Aa at 27 mg/kg, and Cry1Aa, 1Ab and 2Aa also at 136 and 270 mg/kg, administered with a single intraperitoneal injection 24 h before euthanasia. Controls received filtered water or cyclophosphamide. Blood samples collected by cardiac puncture were used to perform hemogram, and bone marrow was extracted for the micronucleus test. Bt spore crystals presented toxicity for lymphocytes when in higher doses, which varied according to the type of spore crystal studied, besides promoting cytotoxic and genotoxic effects for the erythroid lineage of bone marrow, mainly at highest doses. Although the profile of such adverse side effects can be related to their high level of exposure, which is not commonly found in the environment, results indicated that these Bt spore crystals were not harmless to mice. This suggests that a more specific approach should be taken to increase knowledge about their toxicological properties and to establish the toxicological risks to nontarget organisms. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 970-978, 2016. PMID:25899034

  7. Anthrax vaccine design: strategies to achieve comprehensive protection against spore, bacillus, and toxin

    OpenAIRE

    Roehrl, Michael H.; Wang, Jun-Xia

    2005-01-01

    The successful use of Bacillus anthracis as a lethal biological weapon has prompted renewed research interest in the development of more effective vaccines against anthrax. The disease consists of three critical components: spore, bacillus, and toxin, elimination of any of which confers at least partial protection against anthrax. Current remedies rely on postexposure antibiotics to eliminate bacilli and pre- and postexposure vaccination to target primarily toxins. Vaccines effective against ...

  8. Morphological and material properties of polyethyleneterephthalate (PET) fibres with spores incorporated

    OpenAIRE

    Ciera, Lucy Wanjiru; Beladjal, Lynda; Almeras, Xavier; Gheysens, Tom; Van Landuyt, Lieve; Mertens, Johan; Nierstrasz, Vincent; Van Langenhove, Lieva

    2014-01-01

    Owing to the current demand for textiles with new functionalities and improved properties, there has been a continuous effort to modify Polyethylene terephthalate (PET) materials. In our previous study, we demonstrated that Bacillus amyloliquefaciens spores can be incorporated into PET fibres during extrusion. However, the extent to which they can be incorporated without fundamentally changing the properties of the fibres is unknown. In this work, scanning electron microscopy (SEM), transmiss...

  9. Application of gaseous disinfectants ozone and chlorine dioxide for inactivation of Bacillus subtilis spores

    OpenAIRE

    Aydogan, Ahmet

    2006-01-01

    A terrorist attack involving chemical and/or biological warfare agents is a growing possibility. Since anthrax is considered as an immediate public-health threat that can be created by a warfare agent, it is imperative to investigate the potential remediation technologies effective against this threat. In this study, the effectiveness of two gaseous disinfectants, ozone and chlorine dioxide, to inactivate B.subtilis spores - as surrogate to B.anthracis that can cause the infectious anthrax di...

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

    OpenAIRE

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

    2010-01-01

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

  11. Incorporation of Specific Fatty Acid Precursors During Spore Germination and Outgrowth in Bacillus thuringiensis

    OpenAIRE

    Nickerson, Kenneth W.; Bulla, Lee A

    1980-01-01

    The selective incorporation of precursors specific for individual fatty acids in germinating and outgrowing spores of Bacillus thuringiensis is described. The specific precursors utilized were [14C]butyrate, -isobutyrate, -valerate, and -isovalerate, which were incorporated into even-numbered normal-chain isomers, even-numbered iso-isomers, odd-numbered normal-chain acids, and odd-numbered isohomologs, respectively. This preferential incorporation by B. thuringiensis allows the terminal carbo...

  12. Structural Characterization and Chemical Modification of a Glucan from Spores of Ganoderma lucidum

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A linear α-D-(1→3)-glucan, named PSG, was obtained from the spores of Ganoderma lucidum. The synthesis of positively and negatively charged polyelectrolytes from PSG was developed. Amine groups and carboxymethyl groups were introduced through nucleophilic substitution with 3-chloropropylamine or chloroacetic acid, respectively. Reaction conditions were varied to obtain insight into the influence of variables on the degree of substitution.

  13. Induced Sporicidal Activity of Chlorhexidine against Clostridium difficile Spores under Altered Physical and Chemical Conditions

    OpenAIRE

    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 Bacil...

  14. A genomic update on clostridial phylogeny: Gram-negative spore-formers and other misplaced clostridia

    OpenAIRE

    Yutin, Natalya; Michael Y. Galperin

    2013-01-01

    The class Clostridia in the phylum Firmicutes (formerly low-G+C Gram-positive bacteria) includes diverse bacteria of medical, environmental, and biotechnological importance. The Selenomonas-Megasphaera-Sporomusa branch, which unifies members of the Firmicutes with Gram-negative-type cell envelopes, was recently moved from Clostridia to a separate class Negativicutes. However, draft genome sequences of the spore-forming members of the Negativicutes revealed typically clostridial sets of sporul...

  15. Decontamination of Bacillus subtilis var.niger spores on selected surfaces by chlorine dioxide gas

    Institute of Scientific and Technical Information of China (English)

    Yan-ju LI; Neng ZHU; Hai-quan JIA; Jin-hui WU; Ying YI; Jian-cheng QI

    2012-01-01

    Objective:Chlorine dioxide (CD) gas has been used as a fumigant in the disinfection of biosafety laboratories.In this study,some experiments were conducted to assess the inactivation of spores inoculated on six materials [stainless steel (SS),painted steel (PS),polyvinyl chlorid (PVC),polyurethane (PU),glass (GS),and cotton cloth (CC)] by CD gas.The main aims of the study were to determine the sporicidal efficacy of CD gas and the effect of prehumidification before decontamination on sporicidal efficacy.Methods:Material coupons (1.2 cm diameter of SS,PS,and PU; 1.0 cm×1.0 cm for PVC,GS,and CC) were contaminated with 10 μl of Bacillus subtilis var.niger(ATCC 9372) spore suspension in mixed organic burden and then dried in a biosafety cabinet for 12 h.The spores were recovered by soaking the coupons in 5 ml of extraction liquid for 1 h and then vortexing the liquid for 1 min.Results:The log reductions in spore numbers on inoculated test materials exposed to CD gas [0.080% (volume ratio,v/v) for 3 h]were in the range of from 1.80 to 6.64.Statistically significant differences were found in decontamination efficacies on test material coupons of SS,PS,PU,and CC between with and without a 1-h prehumidification treatment.With the extraction method,there were no statistically significant differences in the recovery ratios between the porous and non-porous materials.Conclusions:The results reported from this study could provide information for developing decontamination technology based on CD gas for targeting surface microbial contamination.

  16. 1-octanol, a self-inhibitor of spore germination in Penicillium camemberti

    OpenAIRE

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

    2016-01-01

    Penicillium camemberti is a technologically relevant fungus used to manufacture mould-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 ...

  17. Quantitative Evaluation of the Impact of Bather Density on Levels of Human-Virulent Microsporidian Spores in Recreational Water▿

    OpenAIRE

    Graczyk, Thaddeus K.; Sunderland, Deirdre; Tamang, Leena; Shields, Timothy M.; LUCY Frances E.; Breysse, Patrick N.

    2007-01-01

    Microsporidial gastroenteritis, a serious disease of immunocompromised people, can have a waterborne etiology. During summer months, samples of recreational bathing waters were tested weekly for human-virulent microsporidian spores and water quality parameters in association with high and low bather numbers during weekends and weekdays, respectively. Enterocytozoon bieneusi spores were detected in 59% of weekend (n = 27) and 30% of weekday (n = 33) samples, and Encephalitozoon intestinalis sp...

  18. Testing Nucleoside Analogues as Inhibitors of Bacillus anthracis Spore Germination In Vitro and in Macrophage Cell Culture ▿

    OpenAIRE

    Alvarez, Zadkiel; Lee, Kyungae; Abel-Santos, Ernesto

    2010-01-01

    Bacillus anthracis, the etiological agent of anthrax, has a dormant stage in its life cycle known as the endospore. When conditions become favorable, spores germinate and transform into vegetative bacteria. In inhalational anthrax, the most fatal manifestation of the disease, spores enter the organism through the respiratory tract and germinate in phagosomes of alveolar macrophages. Germinated cells can then produce toxins and establish infection. Thus, germination is a crucial step for the i...

  19. Bacillus anthracis spore interactions with mammalian cells: Relationship between germination state and the outcome of in vitro

    OpenAIRE

    Stojkovic Bojana; Prouty Angela M; Tamilselvam Batcha; Gut Ian M; Czeschin Stephanie; van der Donk Wilfred A; Blanke Steven R

    2011-01-01

    Abstract Background During inhalational anthrax, internalization of Bacillus anthracis spores by host cells within the lung is believed to be a key step for initiating the transition from the localized to disseminated stages of infection. Despite compelling in vivo evidence that spores remain dormant within the bronchioalveolar spaces of the lungs, and germinate only after uptake into host cells, most in vitro studies of infection have been conducted under conditions that promote rapid germin...

  20. Bacillus anthracis spore interactions with mammalian cells: Relationship between germination state and the outcome of in vitro

    Directory of Open Access Journals (Sweden)

    Stojkovic Bojana

    2011-02-01

    Full Text Available Abstract Background During inhalational anthrax, internalization of Bacillus anthracis spores by host cells within the lung is believed to be a key step for initiating the transition from the localized to disseminated stages of infection. Despite compelling in vivo evidence that spores remain dormant within the bronchioalveolar spaces of the lungs, and germinate only after uptake into host cells, most in vitro studies of infection have been conducted under conditions that promote rapid germination of spores within the culture medium. Results Using an in vitro model of infection, we evaluated the influence of the germination state of B. anthracis spores, as controlled by defined culture conditions, on the outcome of infection. Spores prepared from B. anthracis Sterne 7702 germinated in a variety of common cell culture media supplemented with fetal bovine serum (FBS while, in the absence of FBS, germination was strictly dependent on medium composition. RAW264.7 macrophage-like cells internalized spores to the same extent in either germinating or non-germinating media. However, significantly more viable, intracellular B. anthracis were recovered from cells infected under non-germinating conditions compared to germinating conditions. At the same time, RAW264.7 cells demonstrated a significant loss in viability when infected under non-germinating conditions. Conclusions These results suggest that the outcome of host cell infection is sensitive to the germination state of spores at the time of uptake. Moreover, this study demonstrates the efficacy of studying B. anthracis spore infection of host cells within a defined, non-germinating, in vitro environment.

  1. ON THE USE OF FROTH FLOTATION IN THE RECOVERY OF Bacillus sphaericus SPORES

    Directory of Open Access Journals (Sweden)

    E.M. RIOS

    1997-06-01

    Full Text Available Abstract - The recovery of Bacillus sphaericus strain 2362 spores from fermented medium by batch flotation was tested under different conditions. Flotation kinetic studies were performed at 800 rpm and 3 l air/min. The pH values were adjusted at the following set of values: 5.0, 7.0 and 9.0. The results showed that the spore removal rate is influenced by the pH value. At pH equal to 5.0 we observe an adverse effect on the spore concentrate obtention. In this situation the maximum value of the concentration factor was 1.4 when the recuperation percentage was 99%. At pH equal to 7.0 the concentration factor reached the highest value, 7.0, but the recuperation percentage stayed around 96%. Field experiments with the floated material demonstrated that its larvicide activity was sufficient to keep a Culex quinquefasciatus larvae population under control at in a breeding site, during 3 months with 2 applications

  2. Nisin is an effective inhibitor of Clostridium difficile vegetative cells and spore germination.

    Science.gov (United States)

    Le Lay, Christophe; Dridi, Larbi; Bergeron, Michel G; Ouellette, Marc; Fliss, Ismaı L

    2016-02-01

    Clostridium difficile is the most frequently identified enteric pathogen in patients with nosocomial antibiotic-associated diarrhoea and pseudomembranous colitis. Several clinically isolated C. difficile strains are resistant to antibiotics other than metronidazole and vancomycin. Recently, bacteriocins of lactic acid bacteria have been proposed as an alternative or complementary treatment. The aim of this study was to investigate the inhibitory effect of nisin, a bacteriocin produced by several strains of Lactococcus lactis, against clinical isolates of C. difficile. Nisin Z obtained from culture of L. lactis subsp. lactis biovar. diacetylactis was tested along with commercial nisin A. The effect of nisin A on C. difficile spores was also examined. Nisin A and Z both inhibited the growth of all C. difficile isolates, and MICs were estimated at 6.2 μg ml(-1) for nisin Z and 0.8 μg ml(-1) for nisin A. In addition, C. difficile spores were also susceptible to nisin A (25.6 μg ml(-1)), which reduced spore viability by 40-50%. These results suggested that nisin and hence nisin-producing Lactococcus strains could be used to treat C. difficile-associated diarrhoea.

  3. Study on the ice nucleation activity of fungal spores (Ascomycota and Basidiomycota)

    Science.gov (United States)

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

    2012-04-01

    Biogenic ice nucleation (IN) in the atmosphere is a topic of growing interest, as, according to IPCC, the impact of IN on global climate is crucial to perform reliable climate model calculations. About 20 years ago IN activity of a few lichen and Fusarium species [1,2] was reported, while all other investigated fungi were IN-negative. However, as the fungal kingdom is vast, many abundant species, especially the Basidiomycota (most mushrooms), were not tested before. Furthermore, the focus of the past studies was on the IN activity of the mycelium as a cryoprotective mechanism, and not on the airborne spores. We carried out oil immersion measurements [3] with spores from 17 different fungal species of ecological, economical or sanitary importance. Most of these species have not been investigated before, like exponents of Aspergillus, Trichoderma and Agaricales (most mushrooms). Apart from F. avenaceum, spores of all measured species showed moderate or no IN activity, supporting the hypothesis that significant IN activity is a rather exclusive property of only a few species within the fungal kingdom. [1] Kieft TL and Ruscetti T: J. Bacteriol. 172, 3519-3523, 1990. [2] Pouleur S et al.: Appl. Environ. Microbiol., 58, 2960-2964, 1992. [3] Marcolli C et al.: Atmos. Chem. Phys. 7, 5081-5091, 2007.

  4. A ketoreductase gene from Streptomyces mycarofaciens 1748 DNA involved in biosynthesis of a spore pigment

    Institute of Scientific and Technical Information of China (English)

    夏焕章; 王以光

    1997-01-01

    An efficient plasmid transformation system for S. mycarofaciens 1748 has been established. In order to determine the function of MKR gene in S. mycarofaciens 1748, the gene disruption experiment was carried out. For this purpose the plasmid pKC1139 was used. A recombinant strain with white spore appeared, in contrast to the grey-colour spore of S. mycarofaciens 1748. This suggested that homologous recombination between plasmid-borne MKR gene sequence and the chromosome of S. mycarofaciens 1748 had occurred. A Southern hybridization experiment using α- P-labelled MKR gene as probe indicated that the desired integration event had occurred in the re-combinant. The result of gene disruption showed that the alteration of this gene in the chromosome of S. mycarofa-ciens 1748 made sporulating colonies remain white instead of taking on the typical grey colour of sporulating wild type colonies, suggesting that MKR gene is involved in the biosynthesis of a spore pigment. The recombinant strain was in-cubated wit

  5. Inactivation, mutation induction and repair in Bacillus subtilis spores irradiated with heavy ions

    Science.gov (United States)

    Horneck, G.; Bücker, H.

    Studies on the response of bacterial spores to accelerated heavy ions (HZE particles) help in understanding problems of space radiobiology and exobiology. Layers of spores of Bacillus subtilis strains, differing in repair capabilities, were irradiated with accelerated boron, carbon and neon ions of linear energy transfer (LET) values up to 14000 MeV cm2/g. Inactivation as measured by loss of colony forming ability and induction of mutations as measured by reversion to histidine prototrophy and resistance to 150 μg/ml sodium azide were tested, as well as the influence of repair processes on these effects. For inactivation, the cross-sectional values σ plotted as a function of LET follow a saturation curve. The plateau, which is reached around a LET of 2000 MeV cm2/g, occurs at 2.5 × 10-9 cm2, a value in good agreement with the dimensions of the spore protoplast. Lethal damage produced at LET values < 2000 MeV cm2/g is reparable. Recombination repair is more effective than excision repair. At higher LET values, lethal damage could not be reconstituted by the repair mechanisms studied. In addition, at these high LET values, the frequency of induced mutations was drastically decreased. The data support the assumption of at least two qualitatively different types of lesion, depending on the LET of the affecting heavy ion.

  6. Femtosecond Spectroscopy of Calcium Dipicolinate-A Major Component of Bacterial Spores.

    Science.gov (United States)

    Mundt, Ramona; Torres Ziegenbein, Christian; Fröbel, Sascha; Weingart, Oliver; Gilch, Peter

    2016-09-01

    Bacterial spores are rich in calcium dipicolinate (CaDPA). The role of this compound in the high UV resistance of spore DNA and their unique DNA photochemistry is not yet clarified. Here, the photophysical properties of CaDPA dissolved in water are studied by means of steady-state and time-resolved spectroscopy as well as quantum chemistry. Upon 255 nm excitation, a fluorescence emission with a yield of 1.7 × 10(-5) is detected. This low yield is in line with a measured fluorescence lifetime of 110 fs. Transient absorption experiments point to further transitions with time constants of 92 ps and 6.8 μs. The microsecond time constant is assigned to the decay of a triplet state. The yield of this state is close to unity. With the aid of quantum chemistry (TD-DFT, DFT-MRCI), the following transitions are identified. The primarily excited (1)ππ* state depletes within 110 fs. The depletion results in the population of an energetically close lying (1)nπ* state. An El-Sayed allowed intersystem crossing process with a time constant of 92 ps ensues. Implications of these findings on the interaction between photoexcited CaDPA and spore DNA are discussed. PMID:27571143

  7. Inhibition of Ageratina adenophora on Spore Germination and Gametophyte Development of Macrothelypteris torresiana

    Institute of Scientific and Technical Information of China (English)

    Kai-Mei Zhang; Lei Shi; Chuang-Dao Jiang; Zhen-Yu Li

    2008-01-01

    Allalopathy of Ageratina adenophora plays an important role in its invasion. However, we have little knowledge of its allelpathic effects on ferns. In Petri dish bioassays, the inhibitory potential of aqueous leachates from roots, stems and leaves of A. adenophora was studied on the spore germination and gametophyte development of Macrothelypteris torresiana. All leachates Inhibited the spore germination and growth of the first rhizoid of M. torresiana and inhibitory effects Increased with increasing leachate concentrations. Root leachates proved most inhibitory. Gametophyte rhizoids of M. torresiana treated with stem and leaf leachates ofA. adenophora were erect, which was similar to those of the control. However, gametophyte rhizoids of M. torresiana treated with root leachates of A. adenophora were erect, but also curving or swollen. Moreover, curving and swollen rhizoids Increased with Increasing concentrations. As time went by, rhizoids treated with root leachates were not so curved and the swelling almost disappeared. Possible causes are discussed in the present study. The increasing concentrations of leaf leachates also delayed the stages of gametophyte development. With the treatment of root leachates, the delay was more obvious. Thus A. adenophora inhibited the spore germination and gametophyte development of M. torresiana and the root leachates were most inhibitory.

  8. Differential Inactivation of Fungal Spores in Water and on Seeds by Ozone and Arc Discharge Plasma

    Science.gov (United States)

    Kang, Min Ho; Pengkit, Anchalee; Choi, Kihong; Jeon, Seong Sil; Choi, Hyo Won; Shin, Dong Bum; Choi, Eun Ha; Uhm, Han Sup; Park, Gyungsoon

    2015-01-01

    Seed sterilization is essential for preventing seed borne fungal diseases. Sterilization tools based on physical technologies have recently received much attention. However, available information is very limited in terms of efficiency, safety, and mode of action. In this study, we have examined antifungal activity of ozone and arc discharge plasma, potential tools for seed sterilization. In our results, ozone and arc discharge plasma have shown differential antifungal effects, depending on the environment associated with fungal spores (freely submerged in water or infected seeds). Ozone inactivates Fusarium fujikuroi (fungus causing rice bakanae disease) spores submerged in water more efficiently than arc discharge plasma. However, fungal spores associated with or infecting rice seeds are more effectively deactivated by arc discharge plasma. ROS generated in water by ozone may function as a powerful fungicidal factor. On the other hand, shockwave generated from arc discharge plasma may have greatly contributed to antifungal effects on fungus associated with rice seeds. In support of this notion, addition of ultrasonic wave in ozone generating water has greatly increased the efficiency of seed disinfection. PMID:26406468

  9. Differential Inactivation of Fungal Spores in Water and on Seeds by Ozone and Arc Discharge Plasma.

    Directory of Open Access Journals (Sweden)

    Min Ho Kang

    Full Text Available Seed sterilization is essential for preventing seed borne fungal diseases. Sterilization tools based on physical technologies have recently received much attention. However, available information is very limited in terms of efficiency, safety, and mode of action. In this study, we have examined antifungal activity of ozone and arc discharge plasma, potential tools for seed sterilization. In our results, ozone and arc discharge plasma have shown differential antifungal effects, depending on the environment associated with fungal spores (freely submerged in water or infected seeds. Ozone inactivates Fusarium fujikuroi (fungus causing rice bakanae disease spores submerged in water more efficiently than arc discharge plasma. However, fungal spores associated with or infecting rice seeds are more effectively deactivated by arc discharge plasma. ROS generated in water by ozone may function as a powerful fungicidal factor. On the other hand, shockwave generated from arc discharge plasma may have greatly contributed to antifungal effects on fungus associated with rice seeds. In support of this notion, addition of ultrasonic wave in ozone generating water has greatly increased the efficiency of seed disinfection.

  10. Application of nisin and pediocin against resistance and germination of Bacillus spores in sous vide products.

    Science.gov (United States)

    Cabo, M L; Torres, B; Herrera, J J R; Bernárdez, M; Pastoriza, L

    2009-03-01

    Sous vide and other mild preservation techniques are increasingly demanded by consumers. However, spores often will survive in minimally processed foods, causing both spoilage and safety problems. The main objective of the present work was to solve an industrial spoilage problem associated with two sous vide products: mushrooms and shellfish salad. Bacillus subtilis and Bacillus licheniformis predominated as the most heat-resistant organisms isolated from mushrooms and shellfish salad, respectively. The combined effects of nisin and pediocin against resistance and germination of both Bacillus species were described by empirical equations. Whereas nisin was more effective for decreasing thermal resistance of B. subtilis spores, pediocin was more effective against B. licheniformis. However, a significant positive interaction between both biopeptides for decreasing the proportion of vegetative cells resulting from thermoresistant spores was demonstrated in later experiments, thus indicating the increased efficacy of applying high concentrations of both bacteriocins. This efficacy was further demonstrated in additional challenge studies carried out at 15 degrees C in the two sous vide products: mushrooms and shellfish salad. Whereas no vegetative cells were detected after 90 days in the presence of bacteriocins, almost 100% of the population in nontreated samples of mushrooms and shellfish salad was in the vegetative state after 17 and 43 days of storage at 15 degrees C, respectively. PMID:19343939

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

    Science.gov (United States)

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

    2015-03-01

    Ecologists have long acknowledged the importance of seed banks; yet, despite the fact that many plants rely on mycorrhizal fungi for survival and growth, the structure of ectomycorrhizal (ECM) fungal spore banks remains poorly understood. The primary goal of this study was to assess the geographic structure in pine-associated ECM fungal spore banks across the North American continent. Soils were collected from 19 plots in forests across North America. Fresh soils were pyrosequenced for fungal internal transcribed spacer (ITS) amplicons. Adjacent soil cores were dried and bioassayed with pine seedlings, and colonized roots were pyrosequenced to detect resistant propagules of ECM fungi. The results showed that ECM spore banks correlated strongly with biogeographic location, but not with the identity of congeneric plant hosts. Minimal community overlap was found between resident ECM fungi vs those in spore banks, and spore bank assemblages were relatively simple and dominated by Rhizopogon, Wilcoxina, Cenococcum, Thelephora, Tuber, Laccaria and Suillus. Similar to plant seed banks, ECM fungal spore banks are, in general, depauperate, and represent a small and rare subset of the mature forest soil fungal community. Yet, they may be extremely important in fungal colonization after large-scale disturbances such as clear cuts and forest fires. PMID:25557275

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

    Directory of Open Access Journals (Sweden)

    Elise A Lamont

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

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

    Science.gov (United States)

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

    2009-06-01

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

  14. Study of increasing spore content of Beauveria bassiana%提高白僵菌孢子含量的研究

    Institute of Scientific and Technical Information of China (English)

    刘杰; 陈荣; 孙惠杰

    2001-01-01

    Adding some protein of Antheraea pernyi pupae as the replacement of powder of bean cake and meize flour into Beauveria bassiana raw material and culturing solid in three degrees in iron plate can significantly increase the spores content of Beauveria bassiana,reduce the cost and raise the effect.%在生产白僵菌的原料中,加入一定量的柞蚕蛹蛋白代替豆饼粉或玉米粉,采用铁盘三级固体培养,可显著提高白僵菌的含孢量、缩短生产周期、降低成本、提高对害虫的杀伤速率和效果。

  15. Enhanced Immune Response to DNA Vaccine Encoding Bacillus anthracis PA-D4 Protects Mice against Anthrax Spore Challenge.

    Directory of Open Access Journals (Sweden)

    Na Young Kim

    Full Text Available Anthrax has long been considered the most probable bioweapon-induced disease. The protective antigen (PA of Bacillus anthracis plays a crucial role in the pathogenesis of anthrax. In the current study, we evaluated the efficiency of a genetic vaccination with the fourth domain (D4 of PA, which is responsible for initial binding of the anthrax toxin to the cellular receptor. The eukaryotic expression vector was designed with the immunoglobulin M (IgM signal sequence encoding for PA-D4, which contains codon-optimized genes. The expression and secretion of recombinant protein was confirmed in vitro in 293T cells transfected with plasmid and detected by western blotting, confocal microscopy, and enzyme-linked immunosorbent assay (ELISA. The results revealed that PA-D4 protein can be efficiently expressed and secreted at high levels into the culture medium. When plasmid DNA was given intramuscularly to mice, a significant PA-D4-specific antibody response was induced. Importantly, high titers of antibodies were maintained for nearly 1 year. Furthermore, incorporation of the SV40 enhancer in the plasmid DNA resulted in approximately a 15-fold increase in serum antibody levels in comparison with the plasmid without enhancer. The antibodies produced were predominantly the immunoglobulin G2 (IgG2 type, indicating the predominance of the Th1 response. In addition, splenocytes collected from immunized mice produced PA-D4-specific interferon gamma (IFN-γ. The biodistribution study showed that plasmid DNA was detected in most organs and it rapidly cleared from the injection site. Finally, DNA vaccination with electroporation induced a significant increase in immunogenicity and successfully protected the mice against anthrax spore challenge. Our approach to enhancing the immune response contributes to the development of DNA vaccines against anthrax and other biothreats.

  16. Enhanced Immune Response to DNA Vaccine Encoding Bacillus anthracis PA-D4 Protects Mice against Anthrax Spore Challenge.

    Science.gov (United States)

    Kim, Na Young; Chang, Dong Suk; Kim, Yeonsu; Kim, Chang Hwan; Hur, Gyeung Haeng; Yang, Jai Myung; Shin, Sungho

    2015-01-01

    Anthrax has long been considered the most probable bioweapon-induced disease. The protective antigen (PA) of Bacillus anthracis plays a crucial role in the pathogenesis of anthrax. In the current study, we evaluated the efficiency of a genetic vaccination with the fourth domain (D4) of PA, which is responsible for initial binding of the anthrax toxin to the cellular receptor. The eukaryotic expression vector was designed with the immunoglobulin M (IgM) signal sequence encoding for PA-D4, which contains codon-optimized genes. The expression and secretion of recombinant protein was confirmed in vitro in 293T cells transfected with plasmid and detected by western blotting, confocal microscopy, and enzyme-linked immunosorbent assay (ELISA). The results revealed that PA-D4 protein can be efficiently expressed and secreted at high levels into the culture medium. When plasmid DNA was given intramuscularly to mice, a significant PA-D4-specific antibody response was induced. Importantly, high titers of antibodies were maintained for nearly 1 year. Furthermore, incorporation of the SV40 enhancer in the plasmid DNA resulted in approximately a 15-fold increase in serum antibody levels in comparison with the plasmid without enhancer. The antibodies produced were predominantly the immunoglobulin G2 (IgG2) type, indicating the predominance of the Th1 response. In addition, splenocytes collected from immunized mice produced PA-D4-specific interferon gamma (IFN-γ). The biodistribution study showed that plasmid DNA was detected in most organs and it rapidly cleared from the injection site. Finally, DNA vaccination with electroporation induced a significant increase in immunogenicity and successfully protected the mice against anthrax spore challenge. Our approach to enhancing the immune response contributes to the development of DNA vaccines against anthrax and other biothreats. PMID:26430894

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

    Directory of Open Access Journals (Sweden)

    Alicja K Warda

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Bagge, Elisabeth

    2009-07-01

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

  19. N-Lauroylation during the Expression of Recombinant N-Myristoylated Proteins: Implications and Solutions.

    Science.gov (United States)

    Flamm, Andrea Gabriele; Le Roux, Anabel-Lise; Mateos, Borja; Díaz-Lobo, Mireia; Storch, Barbara; Breuker, Kathrin; Konrat, Robert; Pons, Miquel; Coudevylle, Nicolas

    2016-01-01

    Incorporation of myristic acid onto the N terminus of a protein is a crucial modification that promotes membrane binding and correct localization of important components of signaling pathways. Recombinant expression of N-myristoylated proteins in Escherichia coli can be achieved by co-expressing yeast N-myristoyltransferase and supplementing the growth medium with myristic acid. However, undesired incorporation of the 12-carbon fatty acid lauric acid can also occur (leading to heterogeneous samples), especially when the available carbon sources are scarce, as it is the case in minimal medium for the expression of isotopically enriched samples. By applying this method to the brain acid soluble protein 1 and the 1-185 N-terminal region of c-Src, we show the significant, and protein-specific, differences in the membrane binding properties of lauroylated and myristoylated forms. We also present a robust strategy for obtaining lauryl-free samples of myristoylated proteins in both rich and minimal media.

  20. Protein Accumulation Changes Associated with Germination of the Uromyces appendiculatus Uredospore

    Science.gov (United States)

    Uromyces appendiculatus is transmitted when asexual uredospores are released from an infected dry bean leaf and spread by wind to other beans where spores germinate and cause rust disease. We have used Multidimensional Protein Identification Technology (MudPIT) to survey proteins in germinating spo...