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Sample records for acid-soluble spore proteins

  1. Small acid soluble proteins for rapid spore identification.

    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. Small, acid-soluble proteins bound to DNA protect Bacillus subtilis spores from being killed by freeze-drying.

    Fairhead, H; Setlow, B; Waites, W M; Setlow, P

    1994-01-01

    Wild-type spores of Bacillus subtilis were resistant to eight cycles of freeze-drying, whereas about 90% of spores lacking the two major DNA-binding proteins (small, acid-soluble proteins alpha and beta) were killed by three to four cycles of freeze-dryings, with significant mutagenesis and DNA damage accompanying the killing. This role for alpha/beta-type small, acid-soluble proteins in spore resistance to freeze-drying may be important in spore survival in the environment.

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

    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...

  4. Restoration of Brain Acid Soluble Protein 1 Inhibits Proliferation and Migration of Thyroid Cancer Cells

    Guo, Run-Sheng; Yu, Yue; Chen, Jun; Chen, Yue-Yu; Shen, Na; Qiu, Ming

    2016-01-01

    Background: Brain acid soluble protein 1 (BASP1) is identified as a novel potential tumor suppressor in several cancers. However, its role in thyroid cancer has not been investigated yet. In the present study, the antitumor activities of BASP1 against the growth and migration of thyroid cancer cells were evaluated. Methods: BASP1 expression in thyroid cancer tissues and normal tissues were examined by immunohistochemical staining and the association between its expression and prognosis was analyzed. pcDNA-BASP1 carrying full length of BASP1 cDNA was constructed to restore the expression of BASP1 in thyroid cancer cell lines (BHT-101 and KMH-2). The cell proliferation in vitro and in vivo was evaluated by WST-1 assay and xenograft tumor models, respectively. Cell cycle distribution after transfection was analyzed using flow cytometry. Cell apoptosis after transfection was examined by annexin V/propidium iodide assay. The migration was examined using transwell assay. Results: BASP1 expression was abundant in normal tissues while it is significantly decreased in cancer tissues (P = 0.000). pcDNA-BASP1 restored the expression of BASP1 and significantly inhibited the growth of BHT-101 and KMH-2 cells as well as xenograft tumors in nude mice (P = 0.000). pcDNA-BASP1 induced G1 arrest and apoptosis in BHT-101 and KMH-2 cells. In addition, pcDNA-BASP1 significantly inhibited the cell migration. Conclusions: Downregulation of BASP1 expression may play a role in the tumorigenesis of thyroid cancer. Restoration of BASP1 expression exerted extensive antitumor activities against growth and migration of thyroid cancer cells, which suggested that BASP1 gene might act as a potential therapeutic agent for the treatment of thyroid cancer. PMID:27270539

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

    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.

  6. Spores

    A spore is a cell that certain fungi, plants (moss, ferns), and bacteria produce. Spores are involved in reproduction. Certain bacteria make spores as a way to defend themselves. These spores have thick walls. They can resist high temperatures, ...

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

    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

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

    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.

  9. Detection of spore coat protein of Bacillus subtilis by immunological method

    The spore coat protein of Bacillus subtilis was separated, and the qualitative assay for the spore coat protein was made by use of the immunological technique. The immunological method was found to be useful for judging the maturation of spore coat in the course of sporulation. The spore coat protein antigen appeared at t2 stage of sporulation. The addition of rifampicin at the earlier stages of sporulation inhibited the increase in content of the spore coat antigen. (auth.)

  10. Biomarkers of Aspergillus spores: Strain typing and protein identification

    Šulc, Miroslav; Pešlová, Kateřina; Žabka, Martin; Hajdúch, M.; Havlíček, Vladimír

    2009-01-01

    Roč. 280, 1-3 (2009), s. 162-168. ISSN 1387-3806 R&D Projects: GA MŠk LC07017; GA ČR GP203/05/P575 Institutional research plan: CEZ:AV0Z50200510 Keywords : aspergillus * spore * protein Subject RIV: EE - Microbiology, Virology Impact factor: 2.117, year: 2009

  11. Relationship between the germination and spore wall proteins in Nosema bombycis

    Tan, Xiao-Hui; Pan, Guo-Qing; WU Zheng-Li; Li, Yan-hong; ZHANG Rui-Zhi; XU Jin-Shan; ZHOU Ze-Yang

    2008-01-01

    To study the composition of spore wall proteins (SWPs) in Nosema bombycis and the relevance with spore germination,we firstly developed a method called GDGC, which is spores germination in vitro activated by 0.1 mol/L K2CO3, combined with Density Gradient Centrifugation to purify the germinated spore coats. Using this method, we obtained the spore coats, and then comparatively analyzed the composition of SWPs among the supernatant after spore germination, the purified spore coats and normal s...

  12. Effect of protein kinase inhibitors on protein phosphorylation and germination of aerial spores from Streptomyces coelicolor.

    Palecková, P; Kontrová, F; Kofronová, O; Bobek, J; Benada, O; Mikulík, K

    2007-01-01

    In vitro phosphorylation reaction using extracts prepared from cells in the exponential phase of growth and aerial spores of Streptomyces coelicolor displayed the presence of multiply phosphorylated proteins. Effect of protein kinase inhibitors (PKIs) (geldanamycin, wortmannin, apigenin, genistein, roscovitine, methyl 2,5-dihydroxycinnamate, rapamycin, staurosporine) was determined on protein phosphorylation and on germination of spores. The in vitro experiments showed differences in phosphoprotein pattern due to the presence of PKIs. Cultivation of aerial spores with PKIs led to a significant delay in germ tube emergence and filament formation. However, none of the tested PKIs completely blocked the germination process. These results indicate that protein kinases of spores form complex networks sharing common modulating site that plays an important role in proper timing of early developmental events. PMID:17702458

  13. Protein Composition of Infectious Spores Reveals Novel Sexual Development and Germination Factors in Cryptococcus.

    Huang, Mingwei; Hebert, Alexander S; Coon, Joshua J; Hull, Christina M

    2015-08-01

    Spores are an essential cell type required for long-term survival across diverse organisms in the tree of life and are a hallmark of fungal reproduction, persistence, and dispersal. Among human fungal pathogens, spores are presumed infectious particles, but relatively little is known about this robust cell type. Here we used the meningitis-causing fungus Cryptococcus neoformans to determine the roles of spore-resident proteins in spore biology. Using highly sensitive nanoscale liquid chromatography/mass spectrometry, we compared the proteomes of spores and vegetative cells (yeast) and identified eighteen proteins specifically enriched in spores. The genes encoding these proteins were deleted, and the resulting strains were evaluated for discernable phenotypes. We hypothesized that spore-enriched proteins would be preferentially involved in spore-specific processes such as dormancy, stress resistance, and germination. Surprisingly, however, the majority of the mutants harbored defects in sexual development, the process by which spores are formed. One mutant in the cohort was defective in the spore-specific process of germination, showing a delay specifically in the initiation of vegetative growth. Thus, by using this in-depth proteomics approach as a screening tool for cell type-specific proteins and combining it with molecular genetics, we successfully identified the first germination factor in C. neoformans. We also identified numerous proteins with previously unknown functions in both sexual development and spore composition. Our findings provide the first insights into the basic protein components of infectious spores and reveal unexpected molecular connections between infectious particle production and spore composition in a pathogenic eukaryote. PMID:26313153

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

    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...

  15. 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

    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.

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

    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.

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

    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.

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

    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...

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

    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.

  20. A new spore differentiation factor (SDF) secreted by Dictyostelium cells is phosphorylated by the cAMP dependent protein kinase.

    Anjard, C; van Bemmelen, M; Véron, M; Reymond, C D

    1997-10-01

    Upon starvation, Dictyostelium discoideum unicellular amoebae form a multicellular organism leading to the development of a fruiting body containing spores. Single cells of sporogenous mutants, unlike wild type cells, are able to differentiate into spores under specific conditions. We show in this report that overexpression of the catalytic subunit of the cAMP dependent protein kinase (PKA), not only renders the cells sporogenous, but is also accompanied by the production/release of a diffusible spore differentiation factor (SDF). SDF is a small, thermostable phospho-polypeptide. In vitro dephosphorylation reduces SDF spore differentiation capacity, which can be regained in vitro by PKA phosphorylation. These results indicate that SDF is a PKA substrate and might be activated in vivo by this protein kinase. Since spore differentiation requires PKA catalytic subunit activation, we conclude that the response of prespore cells to SDF involves an intracellular pathway dependent on PKA. PMID:9373946

  1. Effect of the osmotic conditions during sporulation on the subsequent resistance of bacterial spores.

    Nguyen Thi Minh, Hue; Guyot, Stéphane; Perrier-Cornet, Jean-Marie; Gervais, Patrick

    2008-08-01

    The causes of Bacillus spore resistance remain unclear. Many structures including a highly compact envelope, low hydration of the protoplast, high concentrations of Ca-chelated dipicolinic acid, and the presence of small acid-soluble spore proteins seem to contribute to resistance. To evaluate the role of internal protoplast composition and hydration, spores of Bacillus subtilis were produced at different osmotic pressures corresponding to water activities of 0.993 (standard), 0.970, and 0.950, using the two depressors (glycerol or NaCl). Sporulation of Bacillus subtilis was slower and reduced in quantity when the water activity was low, taking 4, 10, and 17 days for 0.993, 0.970, and 0.950 water activity, respectively. The spores produced at lower water activity were smaller and could germinate on agar medium at lower water activity than on standard spores. They were also more sensitive to heat (97 degrees C for 5-60 min) than the standard spores but their resistance to high hydrostatic pressure (350 MPa at 40 degrees C for 20 min to 4 h) was not altered. Our results showed that the water activity of the sporulation medium significantly affects spore properties including size, germination capacity, and resistance to heat but has no role in bacterial spore resistance to high hydrostatic pressure. PMID:18506440

  2. Two proteins of the Dictyostelium spore coat bind to cellulose in vitro.

    Zhang, Y; Brown, R D; West, C M

    1998-07-28

    The spore coat of Dictyostelium contains nine different proteins and cellulose. Interactions between protein and cellulose were investigated using an in vitro binding assay. Proteins extracted from coats with urea and 2-mercaptoethanol could, after removal of urea by gel filtration, efficiently bind to particles of cellulose (Avicel), but not Sephadex or Sepharose. Two proteins, SP85 and SP35, were enriched in the reconstitution, and they retained their cellulose binding activities after purification by ion exchange chromatography under denaturing conditions to suppress protein--protein interactions. Neither protein exhibited cellulase activity, though under certain conditions SP85 copurified with a cellulase activity which appeared after germination. Amino acid sequencing indicated that SP85 and SP35 are encoded by the previously described pspB and psvA genes. This was confirmed for SP85 by showing that natural M(r) polymorphisms correlated with changes in the number of tetrapeptide-encoding sequence repeats in pspB. Using PCR to reconstruct missing elements from the recombinogenic middle region of pspB, SP85 was shown to consist of three sequence domains separated by two groups of the tetrapeptide repeats. Expression of partial pspB cDNAs in Escherichia coli showed that cellulose-binding activity resided in the Cys-rich COOH-terminal domain of SP85. This cellulose-binding activity can explain SP85's ultrastructural colocalization with cellulose in vivo. Amino acid composition and antibody binding data showed that SP35 is derived from the Cys-rich N-terminal region of the previously described psvA protein. SP85 and SP35 may link other proteins to cellulose during coat assembly and germination. PMID:9692967

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

    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...

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

    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...

  5. 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.

    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

  6. Gene activity during germination of spores of the fern, Onoclea sensibilis: RNA and protein synthesis and the role of stored mRNA

    Raghavan, V.

    1991-01-01

    Pattern of 3H-uridine incorporation into RNA of spores of Onoclea sensibilis imbibed in complete darkness (non-germinating conditions) and induced to germinate in red light was followed by oligo-dT cellulose chromatography, gel electrophoresis coupled with fluorography and autoradiography. In dark-imbibed spores, RNA synthesis was initiated about 24 h after sowing, with most of the label accumulating in the high mol. wt. poly(A) -RNA fraction. There was no incorporation of the label into poly(A) +RNA until 48 h after sowing. In contrast, photo-induced spores began to synthesize all fractions of RNA within 12 h after sowing and by 24 h, incorporation of 3H-uridine into RNA of irradiated spores was nearly 70-fold higher than that into dark-imbibed spores. Protein synthesis, as monitored by 3H-arginine incorporation into the acid-insoluble fraction and by autoradiography, was initiated in spores within 1-2 h after sowing under both conditions. Autoradiographic experiments also showed that onset of protein synthesis in the cytoplasm of the germinating spore is independent of the transport of newly synthesized nuclear RNA. One-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis of 35S-methionine-labelled proteins revealed a good correspondence between proteins synthesized in a cell-free translation system directed by poly(A) +RNA of dormant spores and those synthesized in vivo by dark-imbibed and photo-induced spores. These results indicate that stored mRNAs of O. sensibilis spores are functionally competent and provide templates for the synthesis of proteins during dark-imbibition and germination.

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

    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

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

    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.))

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

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

    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

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

    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 ...

  11. Purification and partial characterization of a novel calcium-binding protein from Bacillus cereus T spores and inhibition of germination by calmodulin antagonists

    A novel calcium-binding protein has been purified from the dormant spores of Bacillus cereus T. B. cereus T spores were extensively washed, broken, and heated at 90 degree C for 2 min. Using calcium-dependent hydrophobic interaction chromatography plus DEAE-cellulose and hydroxylapatite columns, a single protein was obtained which possessed calcium-binding capacity and some characteristics of calmodulin. This heat-stable protein was retained by hydrophobic matrices or a calmodulin antagonist in a calcium-dependent manner. The crude spore extract displaced bovine brain calmodulin from its antibody in a radioimmunoassay and the immunoreactive specific activity of the partially purified fraction which eluted from phenyl-Sepharose was ca. 200-fold greater than the crude spore extract. Purity of this protein was verified by sodium dodecyl sulfate-polyarcylamide gel electrophoresis and reversed-phase HPLC. Calcium-binding ability was verified with a competitive calcium binding assay using Chelex-100 resin and 45Ca autoradiography. SDS-PAGE and amino acid composition indicated the molecular weight of the protein was 24-kDa. The effects of two calmodulin antagonists, trifluoperazine (TFP) and N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W-7) on L-alanine-induced germination of Bacillus cereus T spores were examined by measuring commitment to germination, loss of heat resistance, release of calcium, decrease in optical density at 660 nm and phase-contrast microscopy

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

    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

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

    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.

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

    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.

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

    Thomas James Schwartz; Martin Lawoko

    2010-01-01

    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% efficie...

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

    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...

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

    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

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

    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...

  19. Distinction of broken cellular wall Ganoderma lucidum spores and G. lucidum spores using FTIR microspectroscopy

    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.

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

    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.)

  1. Bacillus subtilis Spore Inner Membrane Proteome.

    Zheng, Linli; Abhyankar, Wishwas; Ouwerling, Natasja; Dekker, Henk L; van Veen, Henk; van der Wel, Nicole N; Roseboom, Winfried; de Koning, Leo J; Brul, Stanley; de Koster, Chris G

    2016-02-01

    The endospore is the dormant form of Bacillus subtilis and many other Firmicutes. By sporulation, these spore formers can survive very harsh physical and chemical conditions. Yet, they need to go through germination to return to their growing form. The spore inner membrane (IM) has been shown to play an essential role in triggering the initiation of germination. In this study, we isolated the IM of bacterial spores, in parallel with the isolation of the membrane of vegetative cells. With the use of GeLC-MS/MS, over 900 proteins were identified from the B. subtilis spore IM preparations. By bioinformatics-based membrane protein predictions, ca. one-third could be predicted to be membrane-localized. A large number of unique proteins as well as proteins common to the two membrane proteomes were identified. In addition to previously known IM proteins, a number of IM proteins were newly identified, at least some of which are likely to provide new insights into IM physiology, unveiling proteins putatively involved in spore germination machinery and hence putative germination inhibition targets. PMID:26731423

  2. Localization of the Cortex Lytic Enzyme CwlJ in Spores of Bacillus subtilis

    Bagyan, Irina; Setlow, Peter

    2002-01-01

    The enzyme CwlJ is involved in the depolymerization of cortex peptidoglycan during germination of spores of Bacillus subtilis. CwlJ with a C-terminal His tag was functional and was extracted from spores by procedures that remove spore coat proteins. However, this CwlJ was not extracted from disrupted spores by dilute buffer, high salt concentrations, Triton X-100, Ca2+-dipicolinic acid, dithiothreitol, or peptidoglycan digestion, disappeared during spore germination, and was not present in co...

  3. Rapid onsite assessment of spore viability.

    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.

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

    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 ...

  5. A study of Ganoderma lucidum spores by FTIR microspectroscopy

    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.

  6. Influence of neutral salts on the hydrothermal stability of acid-soluble collagen.

    Brown, E M; Farrell, H M; Wildermuth, R J

    2000-02-01

    The thermal stability of acid-soluble collagens was studied by circular dichroism (CD) spectroscopy. Adult bovine dermal collagen (BDC), rat-tail tendon collagen (RTC), and calf skin collagen (CSC) were compared. Despite some variability in amino acid composition and apparent molecular weight, the CD spectra for helical and unordered collagen structures were essentially the same for all the sources. The melting of these collagens occurs as a two-stage process characterized by a pretransition (Tp) followed by complete denaturation (Td). The characteristic temperatures vary with the source of the collagen; for mature collagens (BDC, RTC) Tp = 30 degrees C and Td = 36 degrees C, and for CSC Tp = 34 degrees C and Td = 40 degrees C. Neutral salts, NaCl or KCl, at low concentrations (0.02-0.2 M) appear to bind to the collagens and shift the thermal transitions of these collagens to lower temperatures. PMID:10945432

  7. Interlaboratory evaluation of cellulosic acid-soluble internal air sampling capsules for multi-element analysis.

    Andrews, Ronnee N; Feng, H Amy; Ashley, Kevin

    2016-01-01

    An interlaboratory study was carried out to evaluate the use of acid-soluble cellulosic air sampling capsules for their suitability in the measurement of trace elements in workplace atmospheric samples. These capsules are used as inserts to perform closed-face cassette sample collection for occupational exposure monitoring. The interlaboratory study was performed in accordance with NIOSH guidelines that describe statistical procedures for evaluating measurement accuracy of air monitoring methods. The performance evaluation materials used consisted of cellulose acetate capsules melded to mixed-cellulose ester filters that were dosed with multiple elements from commercial standard aqueous solutions. The cellulosic capsules were spiked with the following 33 elements of interest in workplace air monitoring: Ag, Al, As, Ba, Be, Ca, Cd, Co, Cr, Cu, Fe, In, K, La, Li, Mg, Mn, Mo, Ni, P, Pb, Sb, Se, Sn, Sr, Te, Ti, Tl, V, W, Y, Zn, Zr. The elemental loading levels were certified by an accredited provider of certified reference materials. Triplicates of media blanks and multielement-spiked capsules at three different elemental loadings were sent to each participating laboratory; the elemental loading levels were not revealed to the laboratories. The volunteer participating laboratories were asked to prepare the samples by acid dissolution and to analyze aliquots of extracted samples by inductively coupled plasma atomic emission spectrometry in accordance with NIOSH methods. It was requested that the study participants report their analytical results in units of μg of each target element per internal capsule sample. For the majority of the elements investigated (30 out of 33), the study accuracy estimates obtained satisfied the NIOSH accuracy criterion (A < 25%). This investigation demonstrates the utility of acid-soluble internal sampling capsules for multielement analysis by atomic spectrometry. PMID:26308974

  8. 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

    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

  9. Fifth international fungus spore conference

    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.

  10. Effect of irradiation of bacterial spores on their thermoresistance

    Spores of the species: Bac. subtilis, Bac. cerus, Cl. perfringens and Cl. botulinum were studied. The spores were irradiated in PBS (physiological buffer solution) and in bouillon (about 1.0% of protein) with X-rays at doses: Bac. subtilis and Bac. coreus - 5, 10, 50 and 100 K.radiation, Cl. perfringens - 50, 100 and 200 K-radiation, Cl. botulinum - 100, 200 and 300 K-radiation. Directly after irradiation the suspension was heated at temperatures causing death of a part of spores. The effects of the experiments were determined by quantitative bacteriological cultures. The results obtained indicate that irradiation of bacterial spores with relatively small doses of X-radiation decreases with increased irradiation doses. Synergetic action of irradiation and heating of spores was found, which also increased with increased doses. The greatest cOanges in thermoresistance occurred in spores of Bac. subtilis, and the smallest in Cl. botulinum. The experiments carried out with Cl. perfringens and Cl. botulinum show that thermoresistance of spores decreases more intensively on their irradiation in a non-proteininc medium (PBS). The presence of protein in the medium (bouillon) increases also radioresistance and thermoresistance of spores. (author)

  11. Anthrax Spores under a microscope

    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.

  12. Spore: Spawning Evolutionary Misconceptions?

    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.

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

    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.

  14. 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

    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.

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

    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.

  16. Vacuum-induced Mutations In Bacillus Subtilis Spores

    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.

  17. Ultrastructure and properties of Paecilomyces lilacinus spores

    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)

  18. Ultrastructure and properties of Paecilomyces lilacinus spores.

    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

  19. Hydrazine vapor inactivates Bacillus spores

    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.

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

    Raghavan, V.

    1992-01-01

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

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

    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)

  2. Spore and the sociocultural moment

    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.

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

    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...

  4. Spore Coat Architecture of Clostridium novyi-NT spores

    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.

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

    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

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

    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

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

    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

  8. Handling technique of spore-forming bacteria in radiation sterilization. 1. Preparation of spores

    This paper deals with a handling technique of spore-forming bacteria in radiation sterilization. An explanation is given under three sections: (1) life cycle of spore-forming bacteria, medium to form bacterial spores, and colony and purification methods of bacterial spores; (2) methods for measuring the number of bacterial spores and resistance against gamma radiation (D values); and (3) a test method for identifying spore-forming bacteria and a simple identification method. (N.K.)

  9. Stem rust spores elicit rapid RPG1 phosphorylation

    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...

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

    In the present study we investigated the dynamical properties of entire bacterial spores by neutron scattering, in order to compare the dynamics observed on the picosecond time-scale to results extracted from other physical measurements. The main objective was to provide a better understanding of the uncommon resistance properties of bacterial spores due to the peculiarities of their core. Elastic incoherent neutron scattering (EINS) measurements as a function of temperature were performed on the thermal (λ = 2.23 Angstroms) high-energy resolution backscattering spectrometer IN13 (Institut Laue-Langevin, Grenoble, France). From elastic incoherent measurements, it is possible to extract atomic mean square displacements (MSD), which represent the sample's flexibility at a given temperature, and the effective force constant that is a measure of protein resilience. We used a temperature range that progressively led to the complete inactivation of the spore population. This procedure allowed us to follow the dynamics of the spore components along with major structural changes, notably originating from the core deep rearrangement. Results show that 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 and 400 K. 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

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

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

  12. Triple fixation of Bacillus subtilis dormant spores.

    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.

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

    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...

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

    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.

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

    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.

  16. Natamycin and the germinating spore

    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

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

    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 ...

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

    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

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

    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.

  20. Ultraviolet-Resistant Bacterial Spores

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

    2007-01-01

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

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

    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

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

    Full Text Available 9C ADY3 Protein required for spore wall formation, thought to mediate assembly of...DY3 Prey description Protein required for spore wall formation, thought to mediate assembly of a Don1p-conta

  3. Yeast Interacting Proteins Database: YDL239C, YLR072W [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly... Bait ORF YDL239C Bait gene name ADY3 Bait description Protein required for spore wall formation, thought

  4. Yeast Interacting Proteins Database: YDL239C, YPL070W [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...cription Protein required for spore wall formation, thought to mediate assembly of a Don1p-containing struct

  5. Yeast Interacting Proteins Database: YDL239C, YML042W [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...iption Protein required for spore wall formation, thought to mediate assembly of a Don1p-containing structur

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

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...C Bait ORF YDL239C Bait gene name ADY3 Bait description Protein required for spore wall formation, thought

  7. Yeast Interacting Proteins Database: YDL239C, YAL028W [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...39C Bait ORF YDL239C Bait gene name ADY3 Bait description Protein required for spore wall formation, thought

  8. Yeast Interacting Proteins Database: YDL239C, YDR148C [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...239C Bait gene name ADY3 Bait description Protein required for spore wall formation, thought to mediate asse

  9. Yeast Interacting Proteins Database: YDL239C, YDR273W [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...ption Protein required for spore wall formation, thought to mediate assembly of a Don1p-containing structure

  10. Yeast Interacting Proteins Database: YDL239C, YOR324C [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...it gene name ADY3 Bait description Protein required for spore wall formation, thought to mediate assembly of

  11. Yeast Interacting Proteins Database: YDL239C, YPL255W [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...ait ORF YDL239C Bait gene name ADY3 Bait description Protein required for spore wall formation, thought to m

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

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...cription Protein required for spore wall formation, thought to mediate assembly of a Don1p-containing struct

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

    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.

  14. FoSTUA, Encoding a Basic Helix-Loop-Helix Protein, Differentially Regulates Development of Three Kinds of Asexual Spores, Macroconidia, Microconidia, and Chlamydospores, in the Fungal Plant Pathogen Fusarium oxysporum

    Ohara, Toshiaki; Tsuge, Takashi

    2004-01-01

    The soil-borne fungus Fusarium oxysporum causes vascular wilt of a wide variety of plant species. F. oxysporum produces three kinds of asexual spores, macroconidia, microconidia, and chlamydospores. Falcate macroconidia are formed generally from terminal phialides on conidiophores and rarely from intercalary phialides on hyphae. Ellipsoidal microconidia are formed from intercalary phialides on hyphae. Globose chlamydospores with thick walls are developed by the modification of hyphal and coni...

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

    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

  16. Identification and Characterization of Glycoproteins on the Spore Surface of Clostridium difficile

    Strong, Philippa C. R.; Fulton, Kelly M.; Aubry, Annie; Foote, Simon; Twine, Susan M; Logan, Susan M.

    2014-01-01

    In this study, we identify a major spore surface protein, BclA, and provide evidence that this protein is glycosylated. Following extraction of the spore surface, solubilized proteins were separated by one-dimensional PAGE and stained with glycostain to reveal a reactive high-molecular-mass region of approximately 600 kDa. Tandem mass spectrometry analysis of in-gel digests showed this band to contain peptides corresponding to a putative exosporangial glycoprotein (BclA3) and identified a num...

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

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

  18. Ptaquiloside in Bracken Spores from Britain

    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...

  19. Ptaquiloside in bracken spores from Britain

    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...

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

    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.

  1. Transfer of Bacillus cereus spores from packaging paper into food.

    Ekman, Jaakko; Tsitko, Irina; Weber, Assi; Nielsen-LeRoux, Christina; Lereclus, Didier; Salkinoja-Salonen, Mirja

    2009-11-01

    Food packaging papers are not sterile, as the manufacturing is an open process, and the raw materials contain bacteria. We modeled the potential transfer of the Bacillus cereus spores from packaging paper to food by using a green fluorescent protein-expressing construct of Bacillus thuringiensis Bt 407Cry(-) [pHT315Omega(papha3-gfp)], abbreviated BT-1. Paper (260 g m(-2)) containing BT-1 was manufactured with equipment that allowed fiber formation similar to that of full-scale manufactured paper. BT-1 adhered to pulp during papermaking and survived similar to an authentic B. cereus. Rice and chocolate were exposed to the BT-1-containing paper for 10 or 30 days at 40 or 20 degrees C at relative air humidity of 10 to 60%. The majority of the spores remained immobilized inside the fiber web; only 0.001 to 0.03% transferred to the foods. This amount is low compared with the process hygiene criteria and densities commonly found in food, and it does not endanger food safety. To measure this, we introduced BT-1 spores into the paper in densities of 100 to 1,000 times higher than the amounts of the B. cereus group bacteria found in commercial paper. Of BT-1 spores, 0.03 to 0.1% transferred from the paper to fresh agar surface within 5 min of contact, which is more than to food during 10 to 30 days of exposure. The findings indicate that transfer from paper to dry food is restricted to those microbes that are exposed on the paper surface and readily detectable with a contact agar method. PMID:19903384

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

    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

  3. Survival of Bacillus pumilus spores for a prolonged period of time in real space conditions.

    Vaishampayan, Parag A; Rabbow, Elke; Horneck, Gerda; Venkateswaran, Kasthuri J

    2012-05-01

    To prevent forward contamination and maintain the scientific integrity of future life-detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated surfaces, spores of Bacillus pumilus SAFR-032 exhibited unusually high resistance to decontamination techniques such as UV radiation and peroxide treatment. Subsequently, B. pumilus SAFR-032 was flown to the International Space Station (ISS) and exposed to a variety of space conditions via the European Technology Exposure Facility (EuTEF). After 18 months of exposure in the EXPOSE facility of the European Space Agency (ESA) on EuTEF under dark space conditions, SAFR-032 spores showed 10-40% survivability, whereas a survival rate of 85-100% was observed when these spores were kept aboard the ISS under dark simulated martian atmospheric conditions. In contrast, when UV (>110 nm) was applied on SAFR-032 spores for the same time period and under the same conditions used in EXPOSE, a ∼7-log reduction in viability was observed. A parallel experiment was conducted on Earth with identical samples under simulated space conditions. Spores exposed to ground simulations showed less of a reduction in viability when compared with the "real space" exposed spores (∼3-log reduction in viability for "UV-Mars," and ∼4-log reduction in viability for "UV-Space"). A comparative proteomics analysis indicated that proteins conferring resistant traits (superoxide dismutase) were present in higher concentration in space-exposed spores when compared to controls. Also, the first-generation cells and spores derived from space-exposed samples exhibited elevated UVC resistance when compared with their ground control counterparts. The data generated are important for calculating the probability and mechanisms of microbial survival in space conditions and assessing microbial contaminants

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

    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.

  5. 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

    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

  6. Time course gene expression profiling of yeast spore germination reveals a network of transcription factors orchestrating the global response

    Geijer Cecilia

    2012-10-01

    Full Text Available Abstract Background Spore germination of the yeast Saccharomyces cerevisiae is a multi-step developmental path on which dormant spores re-enter the mitotic cell cycle and resume vegetative growth. Upon addition of a fermentable carbon source and nutrients, the outer layers of the protective spore wall are locally degraded, the tightly packed spore gains volume and an elongated shape, and eventually the germinating spore re-enters the cell cycle. The regulatory pathways driving this process are still largely unknown. Here we characterize the global gene expression profiles of germinating spores and identify potential transcriptional regulators of this process with the aim to increase our understanding of the mechanisms that control the transition from cellular dormancy to proliferation. Results Employing detailed gene expression time course data we have analysed the reprogramming of dormant spores during the transition to proliferation stimulated by a rich growth medium or pure glucose. Exit from dormancy results in rapid and global changes consisting of different sequential gene expression subprograms. The regulated genes reflect the transition towards glucose metabolism, the resumption of growth and the release of stress, similar to cells exiting a stationary growth phase. High resolution time course analysis during the onset of germination allowed us to identify a transient up-regulation of genes involved in protein folding and transport. We also identified a network of transcription factors that may be regulating the global response. While the expression outputs following stimulation by rich glucose medium or by glucose alone are qualitatively similar, the response to rich medium is stronger. Moreover, spores sense and react to amino acid starvation within the first 30 min after germination initiation, and this response can be linked to specific transcription factors. Conclusions Resumption of growth in germinating spores is characterized by

  7. Sensitive, Rapid Detection of Bacterial Spores

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

    2009-01-01

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

  8. Dothistroma septosporum: spore production and weather conditions

    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.

  9. Virulence Plasmids of Spore-Forming Bacteria.

    Adams, Vicki; Li, Jihong; Wisniewski, Jessica A; Uzal, Francisco A; Moore, Robert J; McClane, Bruce A; Rood, Julian I

    2014-12-01

    Plasmid-encoded virulence factors are important in the pathogenesis of diseases caused by spore-forming bacteria. Unlike many other bacteria, the most common virulence factors encoded by plasmids in Clostridium and Bacillus species are protein toxins. Clostridium perfringens causes several histotoxic and enterotoxin diseases in both humans and animals and produces a broad range of toxins, including many pore-forming toxins such as C. perfringens enterotoxin, epsilon-toxin, beta-toxin, and NetB. Genetic studies have led to the determination of the role of these toxins in disease pathogenesis. The genes for these toxins are generally carried on large conjugative plasmids that have common core replication, maintenance, and conjugation regions. There is considerable functional information available about the unique tcp conjugation locus carried by these plasmids, but less is known about plasmid maintenance. The latter is intriguing because many C. perfringens isolates stably maintain up to four different, but closely related, toxin plasmids. Toxin genes may also be plasmid-encoded in the neurotoxic clostridia. The tetanus toxin gene is located on a plasmid in Clostridium tetani, but the botulinum toxin genes may be chromosomal, plasmid-determined, or located on bacteriophages in Clostridium botulinum. In Bacillus anthracis it is well established that virulence is plasmid determined, with anthrax toxin genes located on pXO1 and capsule genes on a separate plasmid, pXO2. Orthologs of these plasmids are also found in other members of the Bacillus cereus group such as B. cereus and Bacillus thuringiensis. In B. thuringiensis these plasmids may carry genes encoding one or more insecticidal toxins. PMID:26104459

  10. Inactivation of Bacterial Spore, Endotoxin, Lipid A, Normal Prion and Abnormal Prion by Exposures to Several Sorts of Gases Plasma.

    Shintani, Hideharu

    2016-01-01

    This review discusses the application of several sorts of non-equilibrium gas plasma discharges for sterilization and disinfection treatments against spores or bioburden on/in the healthcare products or biological indicators. The basic properties of electrical discharges are briefly reviewed and thereafter the paper discusses the interactions of gas plasma with several sorts of biological systems such as bacteria, bacterial spores, endotoxins, lipid A and normal and abnormal prion proteins. PMID:27009504

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

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

  12. High-Resolution Spore Coat Architecture and Assembly of Bacillus Spores

    Malkin, A J; Elhadj, S; Plomp, M

    2011-03-14

    Elucidating the molecular architecture of bacterial and cellular surfaces and its structural dynamics is essential to understanding mechanisms of pathogenesis, immune response, physicochemical interactions, environmental resistance, and provide the means for identifying spore formulation and processing attributes. I will discuss the application of in vitro atomic force microscopy (AFM) for studies of high-resolution coat architecture and assembly of several Bacillus spore species. We have demonstrated that bacterial spore coat structures are phylogenetically and growth medium determined. We have proposed that strikingly different species-dependent coat structures of bacterial spore species are a consequence of sporulation media-dependent nucleation and crystallization mechanisms that regulate the assembly of the outer spore coat. Spore coat layers were found to exhibit screw dislocations and two-dimensional nuclei typically observed on inorganic and macromolecular crystals. This presents the first case of non-mineral crystal growth patterns being revealed for a biological organism, which provides an unexpected example of nature exploiting fundamental materials science mechanisms for the morphogenetic control of biological ultrastructures. We have discovered and validated, distinctive formulation-specific high-resolution structural spore coat and dimensional signatures of B. anthracis spores (Sterne strain) grown in different formulation condition. We further demonstrated that measurement of the dimensional characteristics of B. anthracis spores provides formulation classification and sample matching with high sensitivity and specificity. I will present data on the development of an AFM-based immunolabeling technique for the proteomic mapping of macromolecular structures on the B. anthracis surfaces. These studies demonstrate that AFM can probe microbial surface architecture, environmental dynamics and the life cycle of bacterial and cellular systems at near

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

    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...

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

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

    2015-01-01

    Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger maximizes the diameter of the vesicle and the spore chain length. The length and width of the conidiophore stalk and vesicle were reduced nearly twofold in a ΔveA strain. The latter implies a fourfo...

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

    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...

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

    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

  17. Fifth international fungus spore conference. [Abstracts]: Final technical report

    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.

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

    Plomp, M; Malkin, A J

    2008-06-02

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

  19. Requirements for in vitro germination of Paenibacillus larvae spores.

    Alvarado, Israel; Phui, Andy; Elekonich, Michelle M; Abel-Santos, Ernesto

    2013-03-01

    Paenibacillus larvae is the causative agent of American foulbrood (AFB), a disease affecting honey bee larvae. First- and second-instar larvae become infected when they ingest food contaminated with P. larvae spores. The spores then germinate into vegetative cells that proliferate in the midgut of the honey bee. Although AFB affects honey bees only in the larval stage, P. larvae spores can be distributed throughout the hive. Because spore germination is critical for AFB establishment, we analyzed the requirements for P. larvae spore germination in vitro. We found that P. larvae spores germinated only in response to l-tyrosine plus uric acid under physiologic pH and temperature conditions. This suggests that the simultaneous presence of these signals is necessary for spore germination in vivo. Furthermore, the germination profiles of environmentally derived spores were identical to those of spores from a biochemically typed strain. Because l-tyrosine and uric acid are the only required germinants in vitro, we screened amino acid and purine analogs for their ability to act as antagonists of P. larvae spore germination. Indole and phenol, the side chains of tyrosine and tryptophan, strongly inhibited P. larvae spore germination. Methylation of the N-1 (but not the C-3) position of indole eliminated its ability to inhibit germination. Identification of the activators and inhibitors of P. larvae spore germination provides a basis for developing new tools to control AFB. PMID:23264573

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

    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.

  1. 9 CFR 113.66 - Anthrax Spore Vaccine-Nonencapsulated.

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Anthrax Spore Vaccine-Nonencapsulated... REQUIREMENTS Live Bacterial Vaccines § 113.66 Anthrax Spore Vaccine—Nonencapsulated. Anthrax Spore Vaccine... in 9 CFR 113.64 and the requirements in this paragraph. Any serial or subserial found...

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

    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

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

    Zimmerman, Kolea; Levitis, Daniel; Pringle, Anne

    2014-01-01

    various ascospore characteristics. Mixed effects models of these data show that the female parent accounts for the majority of variation in perithecial production, number of spores produced, and spore germination. Surprisingly, both sexes equally influence the percentage of spores that are pigmented. In...

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

    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 bacterial spores due to insufficient handwashing during daily patient care. PMID:27236515

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

    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.

  6. Determination of fungal spore release from wet building materials

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

    2003-01-01

    release of fungal spores was induced by well-defined jets of air impacting from rotating nozzles. The spores and other particles released from the surface were transported by the air flowing from the chamber through a top outlet to a particle counter and sizer. For two of the fungi (Penicillium......The release and transport of fungal spores from water-damaged building materials is a key factor for understanding the exposure to particles of fungal origin as a possible cause of adverse health effects associated to growth of fungi indoors. In this study, the release of spores from nine species...... each fungal isolate, whereas the spore release is very different for different fungi under identical conditions. Also, the relationship between air velocity and spore release depends on the fungus. For some fungi a significant number of particles smaller than the spore size were released. The method...

  7. Mushroom spore dispersal by convectively-driven winds

    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.

  8. 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

    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

  9. Spore development and nuclear inheritance in arbuscular mycorrhizal fungi

    Hijri Mohamed

    2011-02-01

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

  10. Carboniferous and Devonian Polysporia and its spores

    Bek, Jiří; Dašková, Jiřina; Shyamala, Ch.; Drábková, J.

    Prague : Institute of Geology, Academy of Science, 2006 - (Bek, J.; Brocke, R.; Dašková, J.; Fatka, O.). s. 12-13 ISBN 80-903511-3-1. [Palaeozoic Palynology in Space and Time : CIMP General meeting 2006. 02.09.2006-06.09.2006, Prague] Institutional research plan: CEZ:AV0Z30130516 Keywords : Polysporia * spores * palaeobotany Subject RIV: DB - Geology ; Mineralogy

  11. Airborne Spore Analysis of Karabük Atmosphere

    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.

  12. Activation and killing of Dictyostelium discoideum spores with urea.

    Cotter, D A; O'Connell, R W

    1976-12-01

    The optimal conditions for activation of Dictyostellium discoideum spores are an 8 M urea treatment for 30 min. The lag between activation and swelling is 45 min. Lower concentrations of urea do not activate entire spore populations. Incubating spores in 8 M urea for 60 min or treatment with 10 M urea for 30 min results in a lengthening of the post-activation lag and a decrease in the final percentage of germination. Urea-activated spores can be deactivated by azide, cyanide, osmotic pressure, and low-temperature incubation. Activated spores do not germinate if incubated in 1 M urea for 24 h but will complete germination upon resuspension in urea-free buffer. Shocking spores at 45 degrees C in 8 M urea or incubating spores in 4-8 M urea for 10 h at 23.5 degrees C causes inactivation. When suspended in urea-free buffer, a larger percentage of these dead spores release spheroplasts through a longitudinal split in the spore case. Sequential enzyme treatment of spheroplasts with cellulase and pronase causes them to release lysable protoplasts. The data of these experiments suggest that shedding of the outer and middle wall layers during physiological spore swelling may be a physical process rather than an enzymatic one. PMID:1034498

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

    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

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

    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

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

    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...

  16. Clustering of spore-specific genes in Aspergillus nidulans.

    Orr, W C; Timberlake, W E

    1982-01-01

    We have investigated the chromosomal organization of genes that are expressed specifically in the asexual spores (conidia) of the Ascomycete fungus Aspergillus nidulans, using two experimental approaches. In the first, 30 different recombinant clones, containing long nuclear DNA inserts and at least one spore-specific gene, were selected randomly. The total number of spore-specific genes present in each clone was then determined by RNA blot analysis. In the second approach, several chromosoma...

  17. Heat Resistance and Population Stability of Lyophilized Bacillus subtilis Spores

    Odlaug, Theron E.; Caputo, Ross A.; Graham, Gary S.

    1981-01-01

    Bacillus subtilis 5230 spores were lyophilized in 0.067 M phosphate buffer and stored at 2 to 8°C for 9 to 27 months. The lyophilized spores were reconstituted with buffer or 0.9% saline, and the heat resistance was determined in a thermoresistometer. Lyophilization had no effect on the heat resistance of the spores but did result in a slight decrease in population (≤0.3-logarithm reduction). The lyophilized spores maintained heat resistance and population levels over the test periods. The D-...

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

    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

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

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

    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 ...

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

    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

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

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

    2015-03-01

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

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

    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.

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

    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

  5. The Role of the Electrostatic Force in Spore Adhesion

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

    2010-01-01

    Electrostatic force is investigated as one of the components of the adhesion force between Bacillus thuringiensis (Bt) spores and planar surfaces. The surface potentials of a Bt spore and a mica surface are experimentally obtained using a combined atomic force microscopy (AFM)-scanning surface potential microscopy technique. On the basis of experimental information, the surface charge density of the spores is estimated at 0.03 {micro}C/cm{sup 2} at 20% relative humidity and decreases with increasing humidity. The Coulombic force is introduced for the spore-mica system (both charged, nonconductive surfaces), and an electrostatic image force is introduced to the spore-gold system because gold is electrically conductive. The Coulombic force for spore-mica is repulsive because the components are similarly charged, while the image force for the spore-gold system is attractive. The magnitude of both forces decreases with increasing humidity. The electrostatic forces are added to other force components, e.g., van der Waals and capillary forces, to obtain the adhesion force for each system. The adhesion forces measured by AFM are compared to the estimated values. It is shown that the electrostatic (Coulombic and image) forces play a significant role in the adhesion force between spores and planar surfaces.

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

    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

  7. IN VITRO PROPAGATION OF ANGIOPTERIS EVECT A USING SPORES

    DAMIEN CUPITT

    2001-01-01

    Full Text Available Techniques of e s t a b l i s h i n g Angiopleris evecta plants in vitro were studied. Soaking of A. evecta spores in water for 2 4 h ours mar kedly r edu ced s pore contamination . Soaking of the spores in 1 -2 % of sodi um hy po chlori te for less th an 5 minut es allo wed satisfa ctor y disinfes tation without affe cting spore v i a b i l i t y . Lower concentration of minerals (1/4 MS , p resence of charcoal in the me dium and exp osure of the spores to l i g h t were cr u c i a l for spore germination an d gainetophytc dev elopment of A. evec ta.

  8. Bacillus atrophaeus Outer Spore Coat Assembly and Ultrastructure

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

    2005-11-21

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

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

    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.

  10. Decontamination Options for Drinking Water Contaminated with Bacillus anthracis Spores

    Raber, E; Burklund, A

    2010-02-16

    Five parameters were evaluated with surrogates of Bacillus anthracis spores to determine effective decontamination options for use in a contaminated drinking water supply. The parameters were: (1) type of Bacillus spore surrogate (B. thuringiensis or B. atrophaeus); (2) spore concentration in suspension (10{sup 2} to 10{sup 6} spores/ml); (3) chemical characteristics of decontaminant [sodium dicholor-s-triazinetrione dihydrate (Dichlor), hydrogen peroxide, potassium peroxymonosulfate (Oxone), sodium hypochlorite, and VirkonS{reg_sign}]; (4) decontaminant concentration (0.01% to 5%); and (5) decontaminant exposure time (10 min to 24 hr). Results from 162 suspension tests with appropriate controls are reported. Hydrogen peroxide at a concentration of 5%, and Dichlor and sodium hypochlorite at a concentration of 2%, were effective at spore inactivation regardless of spore type tested, spore exposure time, or spore concentration evaluated. This is the first reported study of Dichlor as an effective decontaminant for B. anthracis spore surrogates. Dichlor's desirable characteristics of high oxidation potential, high level of free chlorine, and more neutral pH than that of other oxidizers evaluated appear to make it an excellent alternative. All three oxidizers were effective against B. atrophaeus spores in meeting EPA's biocide standard of greater than a 6 log kill after a 10-minute exposure time and at lower concentrations than typically reported for biocide use. Solutions of 5% VirkonS{reg_sign} and Oxone were less effective decontaminants than other options evaluated in this study and did not meet the EPA's efficacy standard for biocides. Differences in methods and procedures reported by other investigators make quantitative comparisons among studies difficult.

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

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

    2014-12-01

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

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

    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.

  13. Yeast Interacting Proteins Database: YDL239C, YPL124W [Yeast Interacting Proteins Database

    Full Text Available YDL239C ADY3 Protein required for spore wall formation, thought to mediate assembly...39C Bait ORF YDL239C Bait gene name ADY3 Bait description Protein required for spore wall formation, thoug...ht to mediate assembly of a Don1p-containing structure at the leading edge of the p

  14. Activity of essential oils against Bacillus subtilis spores.

    Lawrence, Hayley A; Palombo, Enzo A

    2009-12-01

    Alternative methods for controlling bacterial endospore contamination are desired in a range of industries and applications. Attention has recently turned to natural products, such as essential oils, which have sporicidal activity. In this study, a selection of essential oils was investigated to identify those with activity against Bacillus subtilis spores. Spores were exposed to thirteen essential oils, and surviving spores were enumerated. Cardamom, tea tree, and juniper leaf oils were the most effective, reducing the number of viable spores by 3 logs at concentrations above 1%. Sporicidal activity was enhanced at high temperatures (60 degrees C) or longer exposure times (up to one week). Gas chromatography-mass spectrometry analysis identified the components of the active essential oils. However, none of the major oil components exhibited equivalent activity to the whole oils. The fact that oil components, either alone or in combination, did not show the same level of sporicidal activity as the complete oils suggested that minor components may be involved, or that these act synergistically with major components. Scanning electron microscopy was used to examine spores after exposure to essential oils and suggested that leakage of spore contents was the likely mode of sporicidal action. Our data have shown that essential oils exert sporicidal activity and may be useful in applications where bacterial spore reduction is desired. PMID:20075624

  15. Quantification of spore-forming bacteria carried by dust particles

    Lin, Y.; Cholakian, T.; Gao, W.; Osman, S.; Barengoltz, J.

    An associated risk of space exploration is the potential of forward contamination of extraterrestrial environments with microorganisms originating on Earth Planetary protection seeks to minimize this risk by identifying and reducing sources of contamination during the spacecraft assembly process Bacterial endospores are of particular concern because their tolerance to a variety of hostile conditions which greatly increases their ability to tolerate outer space conditions and reach planetary bodies that may be capable of supporting life Spore-forming bacteria are ubiquitous in nature It is generally believed that airborne bacterial spores are transported into and within spacecraft assembly facilities by dust particles While the diversity and distribution of spore-forming bacteria in these facilities have been studied the level of bioburden by this mode of transport has not been quantified In order to establish a biological contamination transport model for predicting the cross contamination risk during spacecraft assembly and upon landing on Mars we conducted air and surface sampling in indoor outdoor and cleanroom environments to determine the ratio of spore forming bacteria to their dust particle carriers of different sizes The number of spore forming bacteria was determined from various size groups of particles in a given environment Our data also confirms the existence of multiple spores on a single particle and spore clumps This study will help in developing a better bio-contamination transport model which in turn will help in

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

    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

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

    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...

  18. New pressure and temperature effects on bacterial spores

    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

  19. New pressure and temperature effects on bacterial spores

    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

  20. New pressure and temperature effects on bacterial spores

    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

  1. Micromotors to capture and destroy anthrax simulant spores.

    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

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

    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

  3. Aerial Dissemination of Clostridium difficile spores

    Banfield Kathleen R

    2008-01-01

    Full Text Available Abstract Background Clostridium difficile-associated diarrhoea (CDAD is a frequently occurring healthcare-associated infection, which is responsible for significant morbidity and mortality amongst elderly patients in healthcare facilities. Environmental contamination is known to play an important contributory role in the spread of CDAD and it is suspected that contamination might be occurring as a result of aerial dissemination of C. difficile spores. However previous studies have failed to isolate C. difficile from air in hospitals. In an attempt to clarify this issue we undertook a short controlled pilot study in an elderly care ward with the aim of culturing C. difficile from the air. Methods In a survey undertaken during February (two days 2006 and March (two days 2007, air samples were collected using a portable cyclone sampler and surface samples collected using contact plates in a UK hospital. Sampling took place in a six bedded elderly care bay (Study during February 2006 and in March 2007 both the study bay and a four bedded orthopaedic bay (Control. Particulate material from the air was collected in Ringer's solution, alcohol shocked and plated out in triplicate onto Brazier's CCEY agar without egg yolk, but supplemented with 5 mg/L of lysozyme. After incubation, the identity of isolates was confirmed by standard techniques. Ribotyping and REP-PCR fingerprinting were used to further characterise isolates. Results On both days in February 2006, C. difficile was cultured from the air with 23 samples yielding the bacterium (mean counts 53 – 426 cfu/m3 of air. One representative isolate from each of these was characterized further. Of the 23 isolates, 22 were ribotype 001 and were indistinguishable on REP-PCR typing. C. difficile was not cultured from the air or surfaces of either hospital bay during the two days in March 2007. Conclusion This pilot study produced clear evidence of sporadic aerial dissemination of spores of a clone of C

  4. Determination of fungal spore release from wet building materials

    Kildesø, J.; Wurtz, H.; Nielsen, Kristian Fog; Kruse, P.; Wilkins, K.; Thrane, Ulf; Gravesen, S.; Nielsen, P.A.; Schneider, T.

    2003-01-01

    The release and transport of fungal spores from water-damaged building materials is a key factor for understanding the exposure to particles of fungal origin as a possible cause of adverse health effects associated to growth of fungi indoors. In this study, the release of spores from nine species...... of typical indoor fungi has been measured under controlled conditions. The fungi were cultivated for a period of 4-6 weeks on sterilized wet wallpapered gypsum boards at a relative humidity (RH) of approximately 97%. A specially designed small chamber (P-FLEC) was placed on the gypsum board. The...... release of fungal spores was induced by well-defined jets of air impacting from rotating nozzles. The spores and other particles released from the surface were transported by the air flowing from the chamber through a top outlet to a particle counter and sizer. For two of the fungi (Penicillium...

  5. Effect of Electrochemically Activated Water on Spore-Forming Bacteria

    Pankiv, Nataliya; Palianytsia, Liubov; Berezovska, Nataliya; Kosiv, Ruslana

    2013-01-01

    The effect of electrochemically activated water on the viability spore-forming bacteria Bacillus and Clostridium genera is investigated. It is established that the anolyte inhibits the growth of microorganisms, causing the death of 98% of the cells.

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

    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...

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

    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

  8. Fate of ingested Clostridium difficile spores in mice.

    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.

  9. Natural Dissemination of Bacillus anthracis Spores in Northern Canada

    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...

  10. The Silicon Layer Supports Acid Resistance of Bacillus cereus Spores

    Hirota, Ryuichi; Hata, Yumehiro; Ikeda, Takeshi; Ishida, Takenori; Kuroda, Akio

    2010-01-01

    Silicon (Si) is considered to be a “quasiessential” element for most living organisms. However, silicate uptake in bacteria and its physiological functions have remained obscure. We observed that Si is deposited in a spore coat layer of nanometer-sized particles in Bacillus cereus and that the Si layer enhances acid resistance. The novel acid resistance of the spore mediated by Si encapsulation was also observed in other Bacillus strains, representing a general adaptation enhancing survival u...

  11. Anthrax Spores Make an Essential Contribution to Vaccine Efficacy

    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...

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

    Zimmermann, Boris; Tkalčec, Zdenko; Mešić, Armin; Kohler, Achim

    2015-01-01

    Background 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. Methodology The study is based on measurement...

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

    WesleyTobyFraser; BarryHarveyLomax; PhillipEJardine; MarkASephton

    2014-01-01

    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...

  14. Difference in the homology of two nuclear nonhistone protein fractions as compared by polyacrylamide gel electrophoresis.

    Tsutsui, Ken; Tsutsui, Kimiko; Aoyama, Koji; Oda,Takuzo

    1985-01-01

    The extent of homology between two protein fractions was compared by simple electrophoretic analysis. Nuclear proteins of several rodent cells of different origins were fractionated into acid-soluble and acid-insoluble fractions. The two protein fractions were subjected to polyacrylamide gel electrophoresis in separate gel systems, and protein bands with identical mobilities were sought either in all possible combinational pairs of cell types or in all cell types. The paired and overall homol...

  15. Effect of Radiation and Heat on Bacterial Spore DNA

    Mild irradiation (administered first) is known to sensitize bacterial spores to subsequent heat injury. This project was concerned with the molecular changes underlying this type of synergistic enhancement of lethal effect. Using the alkaline sucrose gradient sedimentation technique it was found that ionizing radiation of 0.05-0.3 Mrad as well as heating at 90°C for 10-30 min (applied individually) induced single-strand breaks in the [3H] DNA of spores of B. subtilis 168 and C. botulinum 62A. In each case more DNA breaks were induced in the more sensitive strain. Combination treatments of radiation (administered first) followed by heating at 90°C showed a distinct synergistic enhancement effect in the observed number of single-strand breaks in the spore [3H] DNA. Depending on the particular treatment schedule, synergistic enhancement of DNA breakage reached up to 95%. The concurrent synergism in the inactivation of spores of B. subtilis under the conditions of this project was in excess of 500 000. It is clear that a combination of radiation and heat enhances both DNA breakage and spore inactivation. It is proposed that synergism may be due to the fact that lethal heat inactivates repair enzymes, while radiation sufficient to injure the spores leaves these enzymes virtually unharmed. (author)

  16. Quantification of Nonproteolytic Clostridium botulinum Spore Loads in Food Materials.

    Barker, Gary C; Malakar, Pradeep K; Plowman, June; Peck, Michael W

    2016-01-01

    We have produced data and developed analysis to build representations for the concentration of spores of nonproteolytic Clostridium botulinum in materials that are used during the manufacture of minimally processed chilled foods in the United Kingdom. Food materials are categorized into homogenous groups which include meat, fish, shellfish, cereals, fresh plant material, dairy liquid, dairy nonliquid, mushroom and fungi, and dried herbs and spices. Models are constructed in a Bayesian framework and represent a combination of information from a literature survey of spore loads from positive-control experiments that establish a detection limit and from dedicated microbiological tests for real food materials. The detection of nonproteolytic C. botulinum employed an optimized protocol that combines selective enrichment culture with multiplex PCR, and the majority of tests on food materials were negative. Posterior beliefs about spore loads center on a concentration range of 1 to 10 spores kg(-1). Posterior beliefs for larger spore loads were most significant for dried herbs and spices and were most sensitive to the detailed results from control experiments. Probability distributions for spore loads are represented in a convenient form that can be used for numerical analysis and risk assessments. PMID:26729721

  17. Tip-enhanced Raman scattering of bacillus subtilis spores

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

    2015-07-01

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

  18. Methods for Integrated Air Sampling and DNA Analysis for Detection of Airborne Fungal Spores

    Williams, Roger H.; Ward, Elaine; McCartney, H. Alastair

    2001-01-01

    Integrated air sampling and PCR-based methods for detecting airborne fungal spores, using Penicillium roqueforti as a model fungus, are described. P. roqueforti spores were collected directly into Eppendorf tubes using a miniature cyclone-type air sampler. They were then suspended in 0.1% Nonidet P-40, and counted using microscopy. Serial dilutions of the spores were made. Three methods were used to produce DNA for PCR tests: adding untreated spores to PCRs, disrupting spores (fracturing of s...

  19. Heat resistance of bacterial spores correlated with protoplast dehydration, mineralization, and thermal adaptation.

    Beaman, T C; Gerhardt, P

    1986-01-01

    Twenty-eight types of lysozyme-sensitive spores among seven Bacillus species representative of thermophiles, mesophiles, and psychrophiles were obtained spanning a 3,000-fold range in moist-heat resistance. The resistance within species was altered by demineralization of the native spores to protonated spores and remineralization of the protonated spores to calcified spores and by thermal adaptation at maximum, optimum, and minimum sporulation temperatures. Protoplast wet densities, and there...

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

    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...

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

    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...

  2. UV-Photobiology of bacterial spores in space

    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

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

    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

  4. Fungal Spores Viability on the International Space Station

    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. Concerted action of two avirulent spore effectors activates Reaction to Puccinia graminis 1 (Rpg1)-mediated cereal stem rust resistance.

    Nirmala, Jayaveeramuthu; Drader, Tom; Lawrence, Paulraj K; Yin, Chuntao; Hulbert, Scot; Steber, Camille M; Steffenson, Brian J; Szabo, Les J; von Wettstein, Diter; Kleinhofs, Andris

    2011-08-30

    The barley stem rust resistance gene Reaction to Puccinia graminis 1 (Rpg1), encoding a receptor-like kinase, confers durable resistance to the stem rust pathogen Puccinia graminis f. sp. tritici. The fungal urediniospores form adhesion structures with the leaf epidermal cells within 1 h of inoculation, followed by hyphae and haustorium formation. The RPG1 protein is constitutively expressed and not phosphorylated. On inoculation with avirulent urediniospores, it is phosphorylated in vivo within 5 min and subsequently degraded. Application of arginine-glycine-aspartic acid peptide loops prevented the formation of adhesion structures for spore attachment, the phosphorylation of RPG1, and germination of the viable spores. Arginine-glycine-aspartic acid affinity chromatography of proteins from the ungerminated avirulent rust spores led to the purification and identification of a protein with fibronectin type III and breast cancer type 1 susceptibility protein domains and a vacuolar protein sorting-associated protein 9 with a coupling of ubiquitin to endoplasmic reticulum degradation domain. Both proteins are required to induce in vivo phosphorylation and degradation of RPG1. Combined application of both proteins caused hypersensitive reaction on the stem rust-resistant cultivar Morex but not on the susceptible cultivar Steptoe. Expression studies indicated that mRNA of both genes are present in ungerminated urediniospores and are constitutively transcribed in sporelings, infected leaves, and haustoria in the investigated avirulent races. Evidence is presented that RPG1, in yeast, interacts with the two protein effectors from the urediniospores that activate cooperatively the stem rust resistance protein RPG1 long before haustoria formation. PMID:21873196

  6. 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

    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 .

  7. Pumilio Homologue from Saprolegnia parasitica Specifically Expressed in Undifferentiated Spore Cysts

    Andersson, M. Gunnar; Cerenius, Lage

    2002-01-01

    The expression of spore-specific marker transcripts at different stages of the asexual life cycle of Saprolegnia parasitica was analyzed. One of the markers, designated puf1, was found to be expressed transiently upon each of several cycles of zoospore encystment and reemergence. The transcript is induced immediately upon zoospore encystment and is rapidly lost when a cyst is triggered to germinate. In nongerminating cysts, puf1 is maintained until a time point when the cysts can no longer be triggered to germinate and thus have become determined for zoospore reemergence. The results show that the cyst stage has two phases, of about equal duration, which are physiologically and transcriptionally distinct and that the transcriptional machinery of oomycetes is also active in nongerminating spores. puf1 encodes a putative mRNA binding protein belonging to a conserved class of proteins including the Drosophila melanogaster Pumilio protein, Caenorhabditis elegans FBF, and Saccharomyces cerevisiae Puf5, all of which are involved in regulation of gene expression by posttranscriptional mechanisms. PMID:12455976

  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.

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

    Stewart, George C

    2015-12-01

    Much of what we know regarding bacterial spore structure and function has been learned from studies of the genetically well-characterized bacterium Bacillus subtilis. Molecular aspects of spore structure, assembly, and function are well defined. However, certain bacteria produce spores with an outer spore layer, the exosporium, which is not present on B. subtilis spores. Our understanding of the composition and biological functions of the exosporium layer is much more limited than that of other aspects of the spore. Because the bacterial spore surface is important for the spore's interactions with the environment, as well as being the site of interaction of the spore with the host's innate immune system in the case of spore-forming bacterial pathogens, the exosporium is worthy of continued investigation. Recent exosporium studies have focused largely on members of the Bacillus cereus family, principally Bacillus anthracis and Bacillus cereus. Our understanding of the composition of the exosporium, the pathway of its assembly, and its role in spore biology is now coming into sharper focus. This review expands on a 2007 review of spore surface layers which provided an excellent conceptual framework of exosporium structure and function (A. O. Henriques and C. P. Moran, Jr., Annu Rev Microbiol 61:555-588, 2007, http://dx.doi.org/10.1146/annurev.micro.61.080706.093224). That review began a process of considering outer spore layers as an integrated, multilayered structure rather than simply regarding the outer spore components as independent parts. PMID:26512126

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

    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

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

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

    2015-01-01

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

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

    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...

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

    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...

  14. Environmental Persistence of Bacillus anthracis and Bacillus subtilis Spores

    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

  15. Scanning Surface Potential Microscopy of Spore Adhesion on Surfaces

    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.

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

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

    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.

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

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

    1995-12-01

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

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

    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 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.

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

    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.

  1. The Fungal Spores Survival Under the Low-Temperature Plasma

    Soušková, Hana; Scholtz, V.; Julák, J.; Savická, D.

    This paper presents an experimental apparatus for the decontamination and sterilization of water suspension of fungal spores. The fungicidal effect of stabilized positive and negative corona discharges on four fungal species Aspergillus oryzae, Clacosporium sphaerospermum, Penicillium crustosum and Alternaria sp. was studied. Simultaneously, the slower growing of exposed fungal spores was observed. The obtained results are substantially different in comparison with those of the analogous experiments performed with bacteria. It may be concluded that fungi are more resistant to the low-temperature plasma.

  2. Flow-cytometric Analysis of Bacillus anthracis Spores

    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.

  3. Discrimination of Spore-Forming Bacilli Using spoIVA

    Venkateswaran, Kasthuri; LaDuc, Myron; Stuecker, Tara

    2009-01-01

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

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

    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...

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

    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

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

    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.

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

    Colas de la Noue, Alexandre; Peters, Judith; Gervais, Patrick; Martinez, Nicolas; Perrier-Cornet, Jean-Marie; Natali, Francesca

    2015-01-01

    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.

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

    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.

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

    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

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

    Spores of Bicillus cereus BIS-59, isolated in this laboratory from shrimps, exhibited an exponential gamma radiation survival curve with a d10 value of 400 krad as compared with a D10 value of 30 krad for the vegetative cells. The D10 value of DPA-depleted spores was also 400 krad indicating that DPA does not influence the radiation response of these spores. Maximum germination monitored with irradiated spores was 60 percent as compared with 80 percent in case of unirradiated spores. Radiation-induced inhibition of the germination processes was not dose dependent. Heat treatment (15 min at 80 C) to spores resulted in activation of the germination process; however, increase in heating time (30 min and 60 min) increased the germination lag period. DPA-depleted spores were less heat resistant than normal spores and exhibited biphasic exponential inactivation. (author)

  11. Global transcriptome analysis of spore formation in Myxococcus xanthus reveals a locus necessary for cell differentiation

    Treuner-Lange Anke

    2010-04-01

    Full Text Available Abstract Background Myxococcus xanthus is a Gram negative bacterium that can differentiate into metabolically quiescent, environmentally resistant spores. Little is known about the mechanisms involved in differentiation in part because sporulation is normally initiated at the culmination of a complex starvation-induced developmental program and only inside multicellular fruiting bodies. To obtain a broad overview of the sporulation process and to identify novel genes necessary for differentiation, we instead performed global transcriptome analysis of an artificial chemically-induced sporulation process in which addition of glycerol to vegetatively growing liquid cultures of M. xanthus leads to rapid and synchronized differentiation of nearly all cells into myxospore-like entities. Results Our analyses identified 1 486 genes whose expression was significantly regulated at least two-fold within four hours of chemical-induced differentiation. Most of the previously identified sporulation marker genes were significantly upregulated. In contrast, most genes that are required to build starvation-induced multicellular fruiting bodies, but which are not required for sporulation per se, were not significantly regulated in our analysis. Analysis of functional gene categories significantly over-represented in the regulated genes, suggested large rearrangements in core metabolic pathways, and in genes involved in protein synthesis and fate. We used the microarray data to identify a novel operon of eight genes that, when mutated, rendered cells unable to produce viable chemical- or starvation-induced spores. Importantly, these mutants displayed no defects in building fruiting bodies, suggesting these genes are necessary for the core sporulation process. Furthermore, during the starvation-induced developmental program, these genes were expressed in fruiting bodies but not in peripheral rods, a subpopulation of developing cells which do not sporulate

  12. Differentiation between spores of Bacillus anthracis and Bacillus cereus by a quantitative immunofluorescence technique.

    Phillips, A. P.; Martin, K L; Broster, M G

    1983-01-01

    A quantitative immunofluorescence assay based on fiber optic microscopy was used to measure the reaction of formalized spores of Bacillus anthracis and Bacillus cereus isolates with fluorescein conjugates prepared by hyperimmunization with B. anthracis Vollum spores. The spores of 11 of the 20 B. cereus strains reacted with the anti-anthrax conjugate to such an extent that they were indistinguishable from the spores of the several B. anthracis isolates tested. However, absorption of the conju...

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

    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...

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

    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...

  15. Stenocybe fragmenta, a new species of Mycocaliciaceae with fragmenting spores

    Peterson, E.B.; Rikkinen, Jouko

    1998-01-01

    The new species Stenocybe fragmenta (Ascomycota, Mycocaliciaceae) is described from western North America. The species was collected from twigs of Cercocarpus montanus and Rhamnus purshiana. Stenocybe fragmenta is characterized by 5-7 septate, 18-30 I?m long ascospores that fragment at maturity. This is the first report of spore fragmentation among the Mycocaliciaceae.

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

    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)

  17. Carboniferous and permian noeggerathialean plants and their spores; preliminary report

    Bek, Jiří; Wang, J.

    Prague : National Museum, 2006. ISBN 80-7036-198-0. [European Palaeobotany- Palynology Conference /7./. 06.09.2006-11.09.2006, Prague] R&D Projects: GA AV ČR IAA3013902 Institutional research plan: CEZ:AV0Z30130516 Keywords : Noeggerathiales * in situ spores * Palaeozoic-Permian Subject RIV: EF - Botanics

  18. Ferns of the Bohemian and their in situ spores

    Dašková, Jiřina; Kvaček, J.

    Prague : National Museum, 2006. s. 28-28. ISBN 80-7036-198-0. [European Palaeobotany- Palynology Conference /7./. 06.09.2006-11.09.2006, Prague] Institutional research plan: CEZ:AV0Z30130516 Keywords : fern * cenomanian * spores Subject RIV: EF - Botanics

  19. Sporicidal characteristics of heated dolomite powder against Bacillus subtilis spores.

    Yasue, Syogo; Sawai, Jun; Kikuchi, Mikio; Nakakuki, Takahito; Sano, Kazuo; Kikuchi, Takahide

    2014-01-01

    Dolomite is a double salt composed of calcium carbonate (CaCO3) and magnesium carbonate (MgCO3). The heat treatment of CaCO3 and MgCO3 respectively generates calcium oxide (CaO) and magnesium oxide (MgO), which have antimicrobial activity. In this study, heated dolomite powder (HDP) slurry was investigated for its sporicidal activity against Bacillus subtilis ATCC 6633 spores. The B. subtilis spores used in this study were not affected by acidic (pH 1) or alkaline (pH 13) conditions, indicating that they were highly resistant. However, dolomite powder heated to 1000℃ for 1 h could kill B. subtilis spores, even at pH 12.7. Sporicidal activity was only apparent when the dolomite powder was heated to 800℃ or higher, and sporicidal activity increased with increases in the heating temperature. This temperature corresponded to that of the generation of CaO. We determined that MgO did not contribute to the sporicidal activity of HDP. To elucidate the sporicidal mechanism of the HDP against B. subtilis spores, the generation of active oxygen from HDP slurry was examined by chemiluminescence analysis. The generation of active oxygen increased when the HDP slurry concentration rose. The results suggested that, in addition to its alkalinity, the active oxygen species generated from HDP were associated with sporicidal activity. PMID:25252642

  20. Changes in spore chemistry and appearance with increasing maturity

    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

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

    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.

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

    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

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

    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.

  4. Airway inflammation among compost workers exposed to actinomycetes spores

    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

  5. The Ice Nucleation Ability of Selected Atmospherically Abundant Fungal Spores

    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

  6. NanoSIMS analysis of Bacillus spores for forensics

    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

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

    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.

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

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

    2007-07-01

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

  9. Fighting Ebola through Novel Spore Decontamination Technologies for the Military

    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

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

    Irradiation, usually used in combination with other conventional methods of conservation, has been proven to be an efficient tool to ensure the safety of many types of foods by destroying pathogenic microorganisms and extending their shelf-lives. This work has investigated the efficacy of gamma irradiation to inactivate spores of the bacterium Paenibacillus larvae that causes the 'American foulbrood', a highly contagious disease still exotic in Brazil that kills bees and contaminates honey, preventing its commercialization and causing great economical losses. In this study, 60 g samples of two types of honey inoculated with 3.5x103 spores/mL of that bacterium were irradiated with doses of 0, 5, 7.5, 10, 12.5 and 15 kGy and counted. The analyses indicated a mean reduction of 97.5±0.7% in the number of viable spores exposed to 5 kGy. The application of doses of 7.5 kGy or higher yielded no viable spores above the detection threshold (10/mL). In addition the value of D10 (3.1±0.3 kGy) was estimated and the logarithm of the population of viable spores of Paenibacillus larvae subsp. larvae was determined as linear and quadratic polynomial functions of the radiation dose. The results indicated that the dose of 10 kGy could be insufficient to assure complete sterilization of honey in some cases while suggesting that 25 kGy would perform such task adequately. (author)

  11. Characterization of heavy ion-induced damage in Bacillus subtilis spores and their global transcriptional response during spore germination. First results

    The proposed research project is aimed to provide new insights on the spore resistance to heavy ions and the effects on different linear energy transfer (LET)-charged HZE particles. With this project, spores of Bacillus subtilis 168, (wild-type and several selected DNA repair-deficient strains) were used for studying the microbial response heavy ions irradiation. DNA repair and mutation induction events were investigated be the determination of the spore survivability, behavior to selected antibiotics, spore-specific protection mechanisms after irradiation. The activation of DNA repair genes were detected during germination by using DNA microarrays. For studying the DNA repair of treated spores during germination an integrated systems approach was used, id est (i.e.) all experiments were performed in a combination of various biochemical and molecular biological methods to study the spore resistance to heavy ion bombardment. (author)

  12. Liver mitochondria contain an ATP-dependent, vanadate-sensitive pathway for the degradation of proteins.

    Desautels, M; Goldberg, A L

    1982-01-01

    A large fraction (30-50%) of the various proteins synthesized within isolated rat liver mitochondria were degraded to amino acids within 60 min after synthesis. Incomplete mitochondrial polypeptides resulting from the incorporation of puromycin were degraded even more extensively (80% per hr). Protein breakdown was measured by the appearance of acid-soluble radioactivity and by the disappearance of labeled polypeptides detected on NaDodSO4/polyacrylamide gel electrophoresis. The amino acids g...

  13. Stingless bees (Hymenoptera, Meliponini feeding on stinkhorn spores (Fungi, Phallales: robbery or dispersal?

    Marcio L. Oliveira

    2000-09-01

    Full Text Available Records about stingless bee-fungi interaction are very rare. In Brazilian Amazonia, workers of Trigona crassipes (Fabricius, 1793 and Trigona fulviventris Guérin, 1835 visiting two stinkhorn species, Dictyophora sp. and Phallus sp., respectively, were observed. The workers licked the fungi gleba, a mucilaginous mass of spores covering the pileum. Neither gleba residue nor spores were found on the body surface of these bee workers. These observations indicate that these bee species include spores as a complement in their diet. On the other hand, they also suggest that these stingless bees can, at times, facilitale spore dispersal, in case intact spores are eliminated with the feces.

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

    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.

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

    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

  16. Beet necrotic yellow vein virus accumulates inside resting spores and zoosporangia of its vector Polymyxa betae BNYVV infects P. betae

    Payton Mark

    2007-04-01

    Full Text Available Abstract Background Plasmodiophorids and chytrids are zoosporic parasites of algae and land plant and are distributed worldwide. There are 35 species belonging to the order Plasmodiophorales and three species, Polymyxa betae, P. graminis, and Spongospora subterranea, are plant viral vectors. Plasmodiophorid transmitted viruses are positive strand RNA viruses belonging to five genera. Beet necrotic yellow vein virus (BNYVV and its vector, P. betae, are the causal agents for rhizomania. Results Evidence of BNYVV replication and movement proteins associating with P. betae resting spores was initially obtained using immunofluorescence labeling and well characterized antisera to each of the BNYVV proteins. Root cross sections were further examined using immunogold labeling and electron microscopy. BNYVV proteins translated from each of the four genomic and subgenomic RNAs accumulate inside P. betae resting spores and zoospores. Statistical analysis was used to determine if immunolabelling detected viral proteins in specific subcellular domains and at a level greater than in control samples. Conclusion Virus-like particles were detected in zoosporangia. Association of BNYVV replication and movement proteins with sporangial and sporogenic stages of P. betae suggest that BNYVV resides inside its vector during more than one life cycle stage. These data suggest that P. betae might be a host as well as a vector for BNYVV

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

    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.

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

    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.

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

    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.

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

    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. PMID:26742618

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

    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...

  2. Identifying experimental surrogates for Bacillus anthracis spores: a review

    Greenberg David L

    2010-09-01

    Full Text Available Abstract Bacillus anthracis, the causative agent of anthrax, is a proven biological weapon. In order to study this threat, a number of experimental surrogates have been used over the past 70 years. However, not all surrogates are appropriate for B. anthracis, especially when investigating transport, fate and survival. Although B. atrophaeus has been widely used as a B. anthracis surrogate, the two species do not always behave identically in transport and survival models. Therefore, we devised a scheme to identify a more appropriate surrogate for B. anthracis. Our selection criteria included risk of use (pathogenicity, phylogenetic relationship, morphology and comparative survivability when challenged with biocides. Although our knowledge of certain parameters remains incomplete, especially with regards to comparisons of spore longevity under natural conditions, we found that B. thuringiensis provided the best overall fit as a non-pathogenic surrogate for B. anthracis. Thus, we suggest focusing on this surrogate in future experiments of spore fate and transport modelling.

  3. Cytokine Response to Infection with Bacillus anthracis Spores

    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. Aerobic and anaerobic spore-forming bacteria in Sardinian honey.

    Farris, Giovanni Antonio; Fatichenti, Fabrizio; Deiana, Pietrino; Agostini, Franco

    1986-01-01

    Apart from an ubiquitous microflora, this investigation into 52 samples of honey revealed some undesirable spore-forming bacteria, Bacillus alvei and B. larvae which are bee pathogens. Bacillus cereus can cause spoilage and food poisoning. It is, therefore, considered essential that every country includes microbiological standards in its Food Safety Regulations for honey, so that the consumer is guaranteed as to the wholesomeness as well as the quality of the product.

  5. Quantification of Nonproteolytic Clostridium botulinum Spore Loads in Food Materials

    Barker, Gary C; Malakar, Pradeep K.; Plowman, June; Peck, Michael W.

    2016-01-01

    We have produced data and developed analysis to build representations for the concentration of spores of nonproteolytic Clostridium botulinum in materials that are used during the manufacture of minimally processed chilled foods in the United Kingdom. Food materials are categorized into homogenous groups which include meat, fish, shellfish, cereals, fresh plant material, dairy liquid, dairy nonliquid, mushroom and fungi, and dried herbs and spices. Models are constructed in a Bayesian framewo...

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

    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.

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

    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.

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

    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. PMID:26190468

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

    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.

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

    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...