Science.gov

Sample records for international fungus spore

  1. Fifth international fungus spore conference

    SciTech Connect

    Timberlake, W.E.

    1993-04-01

    This folio contains the proceedings of the Fifth International Fungal Spore Conference held August 17-21, 1991 at the Unicoi State Park at Helen, Georgia. The volume contains abstracts of each oral presentation as well as a collection of abstracts describing the poster sessions. Presentations were organized around the themes (1) Induction of Sporulation, (2) Nuclear Division, (3) Spore Formation, (4) Spore Release and Dispersal, and (4) Spore Germination.

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

    SciTech Connect

    Timberlake, W.E.

    1993-04-01

    This folio contains the proceedings of the Fifth International Fungal Spore Conference held August 17-21, 1991 at the Unicoi State Park at Helen, Georgia. The volume contains abstracts of each oral presentation as well as a collection of abstracts describing the poster sessions. Presentations were organized around the themes (1) Induction of Sporulation, (2) Nuclear Division, (3) Spore Formation, (4) Spore Release and Dispersal, and (4) Spore Germination.

  3. Spore dispersal of a resupinate ectomycorrhizal fungus, Tomentella sublilacina, via soil food webs.

    PubMed

    Lilleskov, Erik A; Bruns, Thomas D

    2005-01-01

    Patterns of fungal spore dispersal affect gene flow, population structure and fungal community structure. Many Basidiomycota produce resupinate (crust-like) basidiocarps buried in the soil. Although spores are actively discharged, they often do not appear to be well positioned for aerial dispersal. We investigated the potential spore dispersal mechanisms of one exemplar of this growth form, Tomentella sublilacina. It is a widespread ectomycorrhizal fungus that sporulates in the soil organic horizon, can establish from the spore bank shortly after disturbance, but also can be a dominant species in mature forest stands. We investigated whether its spores could be dispersed via spore-based food webs. We examined external surfaces, gut contents and feces from arthropod fungivores (mites, springtails, millipedes, beetles, fly larvae) and arthropod and vertebrate predators (centipedes, salamanders) from on and around T. sublilacina sporocarps. Spore densities were high in the guts of many individuals from all fungivore groups. Centipede gut contents, centipede feces and salamander feces contained undigested invertebrate exoskeletons and many apparently intact spores. DAPI staining of spores from feces of fungivores indicated that 7-73% of spores contained intact nuclei, whereas spores from predators had lower percentages of intact nuclei. The spiny spores often were lodged on invertebrate exoskeletons. To test the viability of spores that had passed through invertebrate guts we used fecal droppings of the millipede Harpaphe haydeniana to successfully inoculate seedlings of Pinus muricata (Bishop pine). These results indicate the potential for T. sublilacina spore dispersal via invertebrates and their predators in soil food webs and might help to explain the widespread distribution of this species. It is likely that this is a general mechanism of dispersal for fungi producing resupinate sporocarps, indicating a need to develop a fuller understanding of the linkages of

  4. Invasion of Spores of the Arbuscular Mycorrhizal Fungus Gigaspora decipiens by Burkholderia spp.

    PubMed Central

    Levy, Avram; Chang, Barbara J.; Abbott, Lynette K.; Kuo, John; Harnett, Gerry; Inglis, Timothy J. J.

    2003-01-01

    Burkholderia species are bacterial soil inhabitants that are capable of interacting with a variety of eukaryotes, in some cases occupying intracellular habitats. Pathogenic and nonpathogenic Burkholderia spp., including B. vietnamiensis, B. cepacia, and B. pseudomallei, were grown on germinating spores of the arbuscular mycorrhizal fungus Gigaspora decipiens. Spore lysis assays revealed that all Burkholderia spp. tested were able to colonize the interior of G. decipiens spores. Amplification of specific DNA sequences and transmission electron microscopy confirmed the intracellular presence of B. vietnamiensis. Twelve percent of all spores were invaded by B. vietnamiensis, with an average of 1.5 × 106 CFU recovered from individual infected spores. Of those spores inoculated with B. pseudomallei, 7% were invaded, with an average of 5.5 × 105 CFU recovered from individual infected spores. Scanning electron and fluorescence microscopy provided insights into the morphology of surfaces of spores and hyphae of G. decipiens and the attachment of bacteria. Burkholderia spp. colonized both hyphae and spores, attaching to surfaces in either an end-on or side-on fashion. Adherence of Burkholderia spp. to eukaryotic surfaces also involved the formation of numerous fibrillar structures. PMID:14532087

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

    PubMed

    Korsnes, Reinert; Westrum, Karin; Fløistad, Erling; Klingen, Ingeborg

    2016-01-01

    This contribution demonstrates an example of experimental automatic image analysis to detect spores prepared on microscope slides derived from trapping. The application is to monitor aerial spore counts of the entomopathogenic fungus Pandora neoaphidis which may serve as a biological control agent for aphids. Automatic detection of such spores can therefore play a role in plant protection. The present approach for such detection is a modification of traditional manual microscopy of prepared slides, where autonomous image recording precedes computerised image analysis. The purpose of the present image analysis is to support human visual inspection of imagery data - not to replace it. The workflow has three components:•Preparation of slides for microscopy.•Image recording.•Computerised image processing where the initial part is, as usual, segmentation depending on the actual data product. Then comes identification of blobs, calculation of principal axes of blobs, symmetry operations and projection on a three parameter egg shape space.

  6. Bioluminescence in the ghost fungus Omphalotus nidiformis does not attract potential spore dispersing insects.

    PubMed

    Weinstein, Philip; Delean, Steven; Wood, Tom; Austin, Andrew D

    2016-12-01

    Bioluminescence has been known from fungi since ancient times, but little work has been done to establish its potential role. There is evidence that some bioluminescent fungi differentially attract potential spore-dispersing insects, and we aimed to establish if this was the case for the ghost fungus, Omphalotus nidiformis (Agaricales,Marasmiaceae), a widespread Australian temperate zone species. We examined three corroborative lines of evidence: circadian rhythmicity of bioluminescence; field-recorded insect abundance at the time of basidiome production; and attractiveness of glowing fungi to flying insects. Basidiomes glowed continuously day and night, and were present in winter (June-July) when insect abundance was low. To assess attractiveness, we deployed sticky-traps in open woodland in the absence of light pollution, in Treatment (baited with fresh bioluminescent O. nidiformis) and Control pairs, for 480 trap-hours on moonless nights. There was no statistical difference in mean insect abundance between Treatment and Control traps (mean 0.33 and 0.54 individuals per trap night, respectively). To interpret these results, we provide a brief review of competing hypotheses for fungal bioluminescence, and conclude that for some fungi, bioluminescence may be an incidental by-product of metabolism rather than conferring any selective advantage. It is possible that the role of bioluminescence differs among evolutionary lineages of fungi and/or with attributes of their growth environments that could affect spore dispersal, such as wind and insect abundance.

  7. Fungal Spores Viability on the International Space Station

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

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

  8. Fungal Spores Viability on the International Space Station.

    PubMed

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

    2016-11-01

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

  9. The effects of triclosan on spore germination and hyphal growth of the arbuscular mycorrhizal fungus Glomus intraradices.

    PubMed

    Twanabasu, Bishnu R; Stevens, Kevin J; Venables, Barney J

    2013-06-01

    The effect of triclosan (5-chloro-2-[2,4-dichlorophenoxy]phenol; TCS), on spore germination, hyphal growth, and hyphal branching of the arbuscular mycorrhizal (AM) fungus, Glomus intraradices spores was evaluated at exposure concentrations of 0.4 and 4.0 μg/L in a static renewal exposure system. To determine if potential effects were mycotoxic or a consequence of impaired signaling between a host plant and the fungal symbiont, spores were incubated with and without the addition of a root exudate. Exposed spores were harvested at days 7, 14, and 21. AM spore germination, hyphal growth, and hyphal branching were significantly lower in both TCS concentrations compared to controls in non-root exudate treatments suggesting direct mycotoxic effects of TCS on AM development. Greater hyphal growth and hyphal branching in controls and 0.4μg/L TCS treatments with root exudate compared to non-root exudate treatments demonstrated growth stimulation by signaling chemicals present in the root exudate. This stimulatory effect was absent in the 4.0 μg/L TCS treatments indicating a direct effect on plant signaling compounds or plant signal response.

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

    PubMed Central

    2011-01-01

    Background Rust fungi are biotrophic basidiomycete plant pathogens that cause major diseases on plants and trees world-wide, affecting agriculture and forestry. Their biotrophic nature precludes many established molecular genetic manipulations and lines of research. The generation of genomic resources for these microbes is leading to novel insights into biology such as interactions with the hosts and guiding directions for breakthrough research in plant pathology. Results To support gene discovery and gene model verification in the genome of the wheat leaf rust fungus, Puccinia triticina (Pt), we have generated Expressed Sequence Tags (ESTs) by sampling several life cycle stages. We focused on several spore stages and isolated haustorial structures from infected wheat, generating 17,684 ESTs. We produced sequences from both the sexual (pycniospores, aeciospores and teliospores) and asexual (germinated urediniospores) stages of the life cycle. From pycniospores and aeciospores, produced by infecting the alternate host, meadow rue (Thalictrum speciosissimum), 4,869 and 1,292 reads were generated, respectively. We generated 3,703 ESTs from teliospores produced on the senescent primary wheat host. Finally, we generated 6,817 reads from haustoria isolated from infected wheat as well as 1,003 sequences from germinated urediniospores. Along with 25,558 previously generated ESTs, we compiled a database of 13,328 non-redundant sequences (4,506 singlets and 8,822 contigs). Fungal genes were predicted using the EST version of the self-training GeneMarkS algorithm. To refine the EST database, we compared EST sequences by BLASTN to a set of 454 pyrosequencing-generated contigs and Sanger BAC-end sequences derived both from the Pt genome, and to ESTs and genome reads from wheat. A collection of 6,308 fungal genes was identified and compared to sequences of the cereal rusts, Puccinia graminis f. sp. tritici (Pgt) and stripe rust, P. striiformis f. sp. tritici (Pst), and poplar

  11. THE EFFECTS OF ULTRAVIOLET RADIATION ON SPORES OF THE FUNGUS ASPERGILLUS NIGER

    PubMed Central

    Zahl, Paul A.; Koller, L. R.; Haskins, C. P.

    1939-01-01

    The survival ratio of Aspergillus spores exposed to ultraviolet radiation has been measured as a function of total incident energy for wave lengths of 2537 Å, 3022 Å, 3129 Å, and 3650 Å. The effect of humidity on killing of Aspergillus spores by ultraviolet radiation has been found to be negligible. A delay in germination as a result of irradiation has been found. The Bunsen-Roscoe reciprocity law has been found to hold within the limits of the radiation intensities studied. Certain morphological changes have been observed. PMID:19873127

  12. Spore Density Determines Infection Strategy by the Plant Pathogenic Fungus Plectosphaerella cucumerina1[OPEN

    PubMed Central

    2016-01-01

    Necrotrophic and biotrophic pathogens are resisted by different plant defenses. While necrotrophic pathogens are sensitive to jasmonic acid (JA)-dependent resistance, biotrophic pathogens are resisted by salicylic acid (SA)- and reactive oxygen species (ROS)-dependent resistance. Although many pathogens switch from biotrophy to necrotrophy during infection, little is known about the signals triggering this transition. This study is based on the observation that the early colonization pattern and symptom development by the ascomycete pathogen Plectosphaerella cucumerina (P. cucumerina) vary between inoculation methods. Using the Arabidopsis (Arabidopsis thaliana) defense response as a proxy for infection strategy, we examined whether P. cucumerina alternates between hemibiotrophic and necrotrophic lifestyles, depending on initial spore density and distribution on the leaf surface. Untargeted metabolome analysis revealed profound differences in metabolic defense signatures upon different inoculation methods. Quantification of JA and SA, marker gene expression, and cell death confirmed that infection from high spore densities activates JA-dependent defenses with excessive cell death, while infection from low spore densities induces SA-dependent defenses with lower levels of cell death. Phenotyping of Arabidopsis mutants in JA, SA, and ROS signaling confirmed that P. cucumerina is differentially resisted by JA- and SA/ROS-dependent defenses, depending on initial spore density and distribution on the leaf. Furthermore, in situ staining for early callose deposition at the infection sites revealed that necrotrophy by P. cucumerina is associated with elevated host defense. We conclude that P. cucumerina adapts to early-acting plant defenses by switching from a hemibiotrophic to a necrotrophic infection program, thereby gaining an advantage of immunity-related cell death in the host. PMID:26842622

  13. Total Dose Effects (TDE) of heavy ionizing radiation in fungus spores and plant seeds: Preliminary investigations

    NASA Technical Reports Server (NTRS)

    Kranz, A. R.; Zimmermann, M. W.; Stadler, R.; Gartenbach, K. E.; Pickert, M.

    1992-01-01

    The opportunity to compare cosmic radiation effects caused during long and short duration exposure flights in biological objects are limited until now, and data obtained so far are very rare and insufficient. Because of the very long exposure of the experiment during the Long Duration Exposure Facility (LDEF) mission (approximately 2000 days) structural changes of the hardware material can be expected which will influence its biocompatibility and, thus, will interact with the radiobiological effects. The aim of the experiment flown on LDEF was a detailed investigation of biological effects caused by cosmic radiation especially of particles of high atomic number Z and high energy. The flight hardware consisted of standard BIOSTACK containers; in these containers a special sandwich construction consisted of visual plastic detectors with seed rsp. spore layers interlocked.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  15. Single spore isolation and morphological characterization of local Malaysian isolates of rice blast fungus Magnoporthe grisea

    NASA Astrophysics Data System (ADS)

    Mishra, Ankitta; Ratnam, Wickneswari; Bhuiyan, Md Atiqur Rahman; Ponaya, Ariane; Jena, Khisord K.

    2015-09-01

    Rice blast is a destructive disease, caused by the fungal pathogen Magnaporthe grisea. It causes considerable damage to rice and leads to crop loss in rice growing regions worldwide. Although fungicides can be used to control rice blast, they generate additional cost in rice production and contamination of environment and food. Therefore, the use of resistant varieties is thought to be one of the most economically and environmentally efficient ways of crop protection from the disease. Six new local Malaysian isolates of M. grisea were isolated using single spore isolation method. Five isolates were from infected leaf samples collected from Kompleks Latihan MADA, Kedah and one was from Kelantan. These isolates were identified using morphological characteristics and microscopic studies and later confirmed by ITSequences. These isolates were induced to sporulate and used for greenhouse screening on two differential rice varieties: Mahsuri (susceptible) and Pongsu Seribu 2 (resistant). Among the 6 isolates, isolate number 3 was found to be the most virulent showing high sporulation while isolate number 4 was very slow growing, and the least virulent.

  16. Tilletia vankyi, a new species of reticulate-spored bunt fungus with non-conjugating basidiospores infecting species of Festuca and Lolium.

    PubMed

    Carris, Lori M; Castlebury, Lisa A; Huang, Guoming; Alderman, Steve C; Luo, Jiafeng; Bao, Xiaodong

    2007-12-01

    A bunt fungus, exhibiting a spore germination pattern unique to known reticulate-spored species of Tilletia was found infecting plants in seed production fields of Festuca rubra ssp. rubra (red fescue) and F. rubra ssp. fallax (Chewing's fescue) in Oregon, and in seed lots of Lolium perenne (perennial ryegrass) from Australia and Germany. Teliospores germinated to form 20-40 uninucleate, non-conjugating basidiospores, and colonies derived from single basidiospores produced teliospores in culture. In inoculation studies using single basidiospore colonies, perennial ryegrass and L. perenne ssp. multiflorum (Italian or annual ryegrass) were infected. A phylogenetic analysis, based on ITS region rDNA, eukaryotic translation elongation factor 1 alpha, and the second largest subunit of RNA polymerase II demonstrated that the fescue and ryegrass bunts are conspecific, and distinct from known species of Tilletia.

  17. Infectivity of resting spores of Massospora cicadina (Entomophthorales: Entomophthoraceae), an entomopathogenic fungus of periodical cicadas (Magicicada spp.) (Homoptera: Cicadidae).

    PubMed

    Duke, L; Steinkraus, D C; English, J E; Smith, K G

    2002-05-01

    Massospora cicadina Peck is a fungal pathogen of 13- and 17-year periodical cicadas (Magicicada spp.). In northwest Arkansas, during the spring 1998 emergence of the 13-year periodical cicada, Magicicada tredecassini (Brood XIX), <1% of emerging cicadas were infected with the conidial stage of M. cicadina, similar to data collected from the same population in 1985. However, in northwest Arkansas plots treated with M. cicadina resting spores collected from infected 17-year Magicicada septendecim cicadas (Brood IV) in 1997 from southern Iowa, 10 months prior to the 1998 emergence in Arkansas, conidial stage infections of M. cicadina in 13-year Arkansas M. tredecassini cicadas increased significantly to 10.6% (7.9% in males and 2.6% in females). These data suggest that M. cicadina resting spores do not require a dormancy of 13 or 17 years between cicada emergences. Instead M. cicadina resting spores appear to be capable of germinating and infecting periodical cicadas after less than 1 year. In addition, M. cicadina resting spores derived from one species (17-year M. septendecim cicadas) were infective for a second species (13-year M. tredecassini cicadas). A mean of 1.4 x 10(6)(SE = 1.8 x 10(5)) mature resting spores were produced per infected male M. septendecim.

  18. Spore behaviors reveal a category of mating-competent infertile heterokaryons in the offspring of the medicinal fungus Agaricus subrufescens.

    PubMed

    Rocha de Brito, Manuela; Foulongne-Oriol, Marie; Moinard, Magalie; Souza Dias, Eustáquio; Savoie, Jean-Michel; Callac, Philippe

    2016-01-01

    Strain breeding is much less advanced in the edible and medicinal species Agaricus subrufescens than in Agaricus bisporus, the button mushroom. Both species have a unifactorial system of sexual incompatibility, a mating type locus tightly linked to a centromere, and basidia producing both homokaryotic (n) and heterokaryotic (n + n) spores. In A. bisporus, breeding is mainly based on direct selection among the heterokaryotic offspring and on hybridization between homokaryotic offspring. The parental heterozygosity is highly maintained in the heterokaryotic offspring due to suppression of recombination and preferential pairing in the spores of nuclei, each one per second meiotic divisions; such "non-sister nuclei" heterokaryons are fertile. In A. subrufescens, recent studies revealed that recombination is not suppressed and that nuclei from the same second meiotic division can also be paired in a spore that give rise to a "sister nuclei" heterokaryon in which the nuclei bear the same mating type allele. The objective of the present work was to investigate the potential function of the different categories of spores in A. subrufescens and their possible use in a genetic breeding program. Using eight co-dominant molecular markers, we found that half of the offspring of the A. subrufescens strain WC837 were heterokaryotic, one quarter of them being sister nuclei heterokaryons. These heterokaryons were infertile and behaved like homokaryons, being even able to cross between each other. In contrast, non-sister nuclei heterokaryons could fruit but inconsistently due to inbreeding depression. Potential roles of these two categories of heterokaryons in nature and consequences for strain breeding are discussed.

  19. Fungus Amongus

    ERIC Educational Resources Information Center

    Wakeley, Deidra

    2005-01-01

    This role-playing simulation is designed to help teach middle level students about the typical lifecycle of a fungus. In this interactive simulation, students assume the roles of fungi, spores, living and dead organisms, bacteria, and rain. As they move around a playing field collecting food and water chips, they discover how the organisms…

  20. The CarO rhodopsin of the fungus Fusarium fujikuroi is a light-driven proton pump that retards spore germination

    PubMed Central

    García-Martínez, Jorge; Brunk, Michael; Avalos, Javier; Terpitz, Ulrich

    2015-01-01

    Rhodopsins are membrane-embedded photoreceptors found in all major taxonomic kingdoms using retinal as their chromophore. They play well-known functions in different biological systems, but their roles in fungi remain unknown. The filamentous fungus Fusarium fujikuroi contains two putative rhodopsins, CarO and OpsA. The gene carO is light-regulated, and the predicted polypeptide contains all conserved residues required for proton pumping. We aimed to elucidate the expression and cellular location of the fungal rhodopsin CarO, its presumed proton-pumping activity and the possible effect of such function on F. fujikuroi growth. In electrophysiology experiments we confirmed that CarO is a green-light driven proton pump. Visualization of fluorescent CarO-YFP expressed in F. fujikuroi under control of its native promoter revealed higher accumulation in spores (conidia) produced by light-exposed mycelia. Germination analyses of conidia from carO− mutant and carO+ control strains showed a faster development of light-exposed carO− germlings. In conclusion, CarO is an active proton pump, abundant in light-formed conidia, whose activity slows down early hyphal development under light. Interestingly, CarO-related rhodopsins are typically found in plant-associated fungi, where green light dominates the phyllosphere. Our data provide the first reliable clue on a possible biological role of a fungal rhodopsin. PMID:25589426

  1. The CarO rhodopsin of the fungus Fusarium fujikuroi is a light-driven proton pump that retards spore germination.

    PubMed

    García-Martínez, Jorge; Brunk, Michael; Avalos, Javier; Terpitz, Ulrich

    2015-01-15

    Rhodopsins are membrane-embedded photoreceptors found in all major taxonomic kingdoms using retinal as their chromophore. They play well-known functions in different biological systems, but their roles in fungi remain unknown. The filamentous fungus Fusarium fujikuroi contains two putative rhodopsins, CarO and OpsA. The gene carO is light-regulated, and the predicted polypeptide contains all conserved residues required for proton pumping. We aimed to elucidate the expression and cellular location of the fungal rhodopsin CarO, its presumed proton-pumping activity and the possible effect of such function on F. fujikuroi growth. In electrophysiology experiments we confirmed that CarO is a green-light driven proton pump. Visualization of fluorescent CarO-YFP expressed in F. fujikuroi under control of its native promoter revealed higher accumulation in spores (conidia) produced by light-exposed mycelia. Germination analyses of conidia from carO(-) mutant and carO(+) control strains showed a faster development of light-exposed carO(-) germlings. In conclusion, CarO is an active proton pump, abundant in light-formed conidia, whose activity slows down early hyphal development under light. Interestingly, CarO-related rhodopsins are typically found in plant-associated fungi, where green light dominates the phyllosphere. Our data provide the first reliable clue on a possible biological role of a fungal rhodopsin.

  2. A role for the asexual spores in infection of Fraxinus excelsior by the ash-dieback fungus Hymenoscyphus fraxineus

    PubMed Central

    Fones, Helen Nicola; Mardon, Charlotte; Gurr, Sarah Jane

    2016-01-01

    The invasive pathogen, ash dieback fungus Hymenoscyphus fraxineus, is spreading rapidly across Europe. It shows high levels of outcrossing and limited population structure, even at the epidemic front. The anamorphic (asexual) form produces prolific conidia, thought to function solely as spermatia (male gametes), facilitating gene flow between sympatric strains. Here, we show that conidia are capable of germination on ash leaves and in vitro, and can infect seedlings via leaves or soil. In leaves, germlings form structures resembling fruiting bodies. Additionally, H. fraxineus colonises ash debris and grows in soil in the absence of ash tissues. We propose an amended life-cycle in which wind-dispersed, insect-vectored or water-spread conidia infect ash and may sporulate in planta, as well as in forest debris. This amplifies inoculum levels of different strains in ash stands. In combination with their function as spermatia, conidia thus act to maximise gene flow between sympatric strains, including those originally present at low inoculum. Such mixing increases evolutionary potential, as well as enhancing the likelihood of gene introgression from closely-related strains or assimilation of further genetic diversity from parental Asian populations. This scenario increases the adaptability of H. fraxineus to new climates and, indeed, onto new host species. PMID:27694963

  3. Fungus Infections: Preventing Recurrence

    MedlinePlus

    ... place for these spores to collect is in shoes. Therefore, after effective treatment, a fungus may recur ... feet clean, cool and dry. Change socks. Wear shoes that "breathe" like leather, rather than plastic. Make ...

  4. Internal control for nucleic acid testing based on the use of purified Bacillus atrophaeus subsp. globigii spores.

    PubMed

    Picard, François J; Gagnon, Martin; Bernier, Marthe R; Parham, Nicholas J; Bastien, Martine; Boissinot, Maurice; Peytavi, Régis; Bergeron, Michel G

    2009-03-01

    Commonly used internal controls (ICs) to monitor the efficiency of nucleic acid testing (NAT) assays do not allow verification of nucleic acid extraction efficiency. Since microbial cells are often difficult to lyse, it is important to ensure that nucleic acids are efficiently extracted from any target organism. For this purpose, we developed a cellular IC based on the use of nonpathogenic Bacillus spores. Purified Bacillus atrophaeus subsp. globigii (referred to hereafter as simply B. atrophaeus) spores were added to vaginal and anal samples, which were then subjected to rapid DNA extraction and subsequent PCR amplification. The proof of concept of this cellular IC was made through the use of both manual and automated DNA extraction methods, using vaginal or anal samples spiked with B. atrophaeus spores, combined with a multiplex real-time PCR assay for the specific detection of group B streptococci (GBS) and B. atrophaeus. The performance of the cellular IC was compared to that of a standard IC plasmid added to PCRs. Approximately 500 B. atrophaeus spores per PCR was found to be optimal since this did not interfere significantly with GBS detection for either DNA extraction method and yielded reproducible amplification and/or detection of B. atrophaeus genomic DNA serving as an IC template. Performance of the cellular IC was comparable to that of the standard IC. This novel IC system using nonpathogenic and hard-to-lyse B. atrophaeus spores allowed validation of both the DNA extraction procedure and the amplification and detection process. Use of a spore-based control also provides a universal control for microbial cell lysis.

  5. Absence of Fungal Spore Internalization by Bronchial Epithelium in Mouse Models Evidenced by a New Bioimaging Approach and Transmission Electronic Microscopy.

    PubMed

    Rammaert, Blandine; Jouvion, Grégory; de Chaumont, Fabrice; Garcia-Hermoso, Dea; Szczepaniak, Claire; Renaudat, Charlotte; Olivo-Marin, Jean-Christophe; Chrétien, Fabrice; Dromer, Françoise; Bretagne, Stéphane

    2015-09-01

    Clinical data and experimental studies suggest that bronchial epithelium could serve as a portal of entry for invasive fungal infections. We therefore analyzed the interactions between molds and the bronchial/bronchiolar epithelium at the early steps after inhalation. We developed invasive aspergillosis (Aspergillus fumigatus) and mucormycosis (Lichtheimia corymbifera) murine models that mimic the main clinical risk factors for these infections. Histopathology studies were completed with a specific computer-assisted morphometric method to quantify bronchial and alveolar spores and with transmission electron microscopy. Morphometric analysis revealed a higher number of bronchial/bronchiolar spores for A. fumigatus than L. corymbifera. The bronchial/bronchiolar spores decreased between 1 and 18 hours after inoculation for both fungi, except in corticosteroid-treated mice infected with A. fumigatus, suggesting an effect of cortisone on bronchial spore clearance. No increase in the number of spores of any species was observed over time at the basal pole of the epithelium, suggesting the lack of transepithelial crossing. Transmission electron microscopy did not show spore internalization by bronchial epithelial cells. Instead, spores were phagocytized by mononuclear cells on the apical pole of epithelial cells. Early epithelial internalization of fungal spores in vivo cannot explain the bronchial/bronchiolar epithelium invasion observed in some invasive mold infections. The bioimaging approach provides a useful means to accurately enumerate and localize the fungal spores in the pulmonary tissues.

  6. Airborne mesophilic fungal spores in various residential environments

    NASA Astrophysics Data System (ADS)

    Pasanen, A.-L.

    In the present work viable fungal spore counts and flora of indoor air were compared in various residences. Total viable spore counts were lowest in the urban/suburban residences and highest in the rural residences. Moisture problems in the urban environment did not increase total viable spore count, but affected composition of fungal flora. In the rural environment, spore counts were much higher in the old houses than in the new ones. Penicillium was the most prevalent fungus in the air of all the residences studied. Airborne Aspergillus, Cladosporium spores and yeast cells were more common in the damp residences and the old rural houses than in the other residences.

  7. Viability and infectivity of fresh and cryopreserved Nosema ceranae spores.

    PubMed

    McGowan, Janine; De la Mora, Alvaro; Goodwin, Paul H; Habash, Marc; Hamiduzzaman, Mollah Md; Kelly, Paul G; Guzman-Novoa, Ernesto

    2016-12-01

    The microsporidium fungus Nosema ceranae is an intracellular parasite that infects the midgut of the honey bee, Apis mellifera. A major limitation of research on N. ceranae is that the fungus is non-culturable and thus studying it depends on the seasonal availability of Nosema spores. Also, spore viability and infectivity can vary considerably, and thus there is a need for reliable methods for determining those traits. This study examined different conditions for N. ceranae spore cryopreservation at -70°C, assessing spore viability and infectivity. Viability was determined by a staining procedure counting total spores numbers with bright field microscopy and un-viable spore numbers with the fluorescent dye, propidium iodide. Spore infectivity was determined with a dilution inoculation assay. Infectivity was dependent on the inoculum dose for the proportion of bees with detectable Nosema infections based on the number of spores per bee at 18days after inoculation; 4000 spores per bee or higher were needed to get approx. 100% of the inoculated bees infected. The median infective dose (ID50) was 149 spores per bee, and the minimum dose capable of causing a detectable infection was 1.28 spores. The proportion of N. ceranae infected bees correlated significantly with the number of spores per bee (r=0.98, P<0.0001). N. ceranae spores cryopreserved in water or 10% glycerol did not differ in viability compared to fresh spores, but lost infectivity when inoculated into bees. This study shows that while cryopreservation of N. ceranae spores can preserve viability, the spores can have reduced infectivity.

  8. Fun Microbiology: Using a Plant Pathogenic Fungus To Demonstrate Koch's Postulates.

    ERIC Educational Resources Information Center

    Mitchell, James K.; Orsted, Kathy M.; Warnes, Carl E.

    1997-01-01

    Describes an experiment using a plant pathogenic fungus in which students learn to follow aseptic techniques, grow and produce spores of a fungus, use a hemacytometer for enumerating spores, prepare serial dilutions, grow and inoculate plants, isolate a pure culture using agar streak plates, and demonstrate the four steps of Koch's postulates.…

  9. Mushrooms use convectively created airflows to disperse their spores

    PubMed Central

    Dressaire, Emilie; Yamada, Lisa; Song, Boya; Roper, Marcus

    2016-01-01

    Thousands of basidiomycete fungal species rely on mushroom spores to spread across landscapes. It has long been thought that spores depend on favorable winds 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 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 1 cm high and lift spores 10 cm or more into the air. This work reveals how mushrooms tolerate and even benefit from crowding and explains their high water needs. PMID:26929324

  10. Hydrazine vapor inactivates Bacillus spores

    NASA Astrophysics Data System (ADS)

    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.

  11. Spores Disperse, Too!

    ERIC Educational Resources Information Center

    Schumann, Donna N.

    1981-01-01

    Suggests the use of spores and spore-producing structures to show adaptations facilitating spore dispersal and dispersal to favorable environments. Describes several activities using horsetails, ferns, and mosses. Lists five safety factors related to use of mold spores in the classroom. (DS)

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    PubMed Central

    Burggraf, Larry W.; Xing, Yun

    2016-01-01

    ABSTRACT The study of structures and properties of bacterial spores is important to understanding spore formation and biological responses to environmental stresses. While significant progress has been made over the years in elucidating the multilayer architecture of spores, the mechanical properties of the spore interior are not known. Here, we present a thermal atomic force microscopy (AFM) study of the nanomechanical properties of internal structures of Bacillus anthracis spores. We developed a nanosurgical sectioning method in which a stiff diamond AFM tip was used to cut an individual spore, exposing its internal structure, and a soft AFM tip was used to image and characterize the spore interior on the nanometer scale. We observed that the elastic modulus and adhesion force, including their thermal responses at elevated temperatures, varied significantly in different regions of the spore section. Our AFM images indicated that the peptidoglycan (PG) cortex of Bacillus anthracis spores consisted of rod-like nanometer-sized structures that are oriented in the direction perpendicular to the spore surface. Our findings may shed light on the spore architecture and properties. IMPORTANCE A nanosurgical AFM method was developed that can be used to probe the structure and properties of the spore interior. The previously unknown ultrastructure of the PG cortex of Bacillus anthracis spores was observed to consist of nanometer-sized rod-like structures that are oriented in the direction perpendicular to the spore surface. The variations in the nanomechanical properties of the spore section were largely correlated with its chemical composition. Different components of the spore materials showed different thermal responses at elevated temperatures. PMID:26969703

  14. Allergenic airborne pollen and spores in Anchorage, Alaska

    SciTech Connect

    Anderson, J.H.

    1985-05-01

    Major aeroallergens in Anchorage are birch, alder, poplar, spruce, grass pollen, Cladosporium, and unspecified fungus spores. Lesser pollens are sorrel, willow, pine, juniper, sedge, lamb's-quarters, wormwood, plantain, and others. The aero-flora is discussed in terms of the frequency of allergenically significant events and within-season and year-to-year dynamics.

  15. Detection of 3-hydroxykynurenine in a plant pathogenic fungus.

    PubMed Central

    Wilson, T J Greer; Thomsen, Karl Kristian; Petersen, Bent O; Duus, Jens Ø; Oliver, Richard P

    2003-01-01

    A redox-active compound has been purified from the barley powdery mildew fungus Blumeria ( Erysiphe ) graminis f. sp. hordei. A combination of spectrophotometry, MS and NMR has identified it as 3-hydroxykynurenine (3OHKyn). This compound, never previously detected in any fungus or pathogen, is best known for its role in vertebrate cataracts. It is found abundantly in developing and germinating spores and also in runner hyphae. Two roles for 3OHKyn are discussed: first, the presence of active oxygen species would enable 3OHKyn to cross-link the spore chemically with the plant. Secondly, it may be acting as an UV protectant and an antioxidant. PMID:12556224

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

    PubMed

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

    2014-08-01

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

  17. Hydrazine inactivates bacillus spores

    NASA Technical Reports Server (NTRS)

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

    2005-01-01

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

  18. Fungus gnats and Pythium in the attack on greenhouse plants: conspirators or just cohabitants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Research conducted by collaborating Cornell University and USDA-ARS scientists investigated the potential for fungus gnats to vector Pythium root-rot pathogens. Fungus gnat larvae readily consumed Pythium oospores; the spores survived passage through the larval gut and, upon defecation, were able to...

  19. Fungal microcolonies on indoor surfaces — an explanation for the base-level fungal spore counts in indoor air

    NASA Astrophysics Data System (ADS)

    Pasanen, A.-L.; Heinonen-Tanski, H.; Kalliokoski, P.; Jantunen, M. J.

    In the subarctic winter, fungal spores are found in indoor air even when outdoor spore levels are very low. The results of this study support an explanation that some indoor airborne fungal spores are derived from unnoticeable fungal microcolonies, which may develop on temporarily wet surfaces. Laboratory experiments on Penicillium verrucosum indicated that the fungus germinated on new wallpaper very quickly (about half an hour) under moist conditions. Hyphal growth and sporulation of the fungus on moist wallpaper occured within one day of incubation. In gravity-settling tape samples from occasionally wet surfaces in a suburban home, large spore aggregates, hyphal fragments with some spores and spores in the germination stage were found, indicating fungal growth. These experiments showed that fungal microcolonies can develop within a week on occasionally wet indoor surfaces.

  20. Cryopreservation of fern spores

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  1. Malaria Mosquitoes Attracted by Fatal Fungus

    PubMed Central

    George, Justin; Jenkins, Nina E.; Blanford, Simon; Thomas, Matthew B.; Baker, Thomas C.

    2013-01-01

    Insect-killing fungi such as Beauveria bassiana are being evaluated as possible active ingredients for use in novel biopesticides against mosquito vectors that transmit malaria. Fungal pathogens infect through contact and so applications of spores to surfaces such as walls, nets, or other resting sites provide possible routes to infect mosquitoes in and around domestic dwellings. However, some insects can detect and actively avoid fungal spores to reduce infection risk. If true for mosquitoes, such behavior could render the biopesticide approach ineffective. Here we find that the spores of B. bassiana are highly attractive to females of Anopheles stephensi, a major anopheline mosquito vector of human malaria in Asia. We further find that An. stephensi females are preferentially attracted to dead and dying caterpillars infected with B. bassiana, landing on them and subsequently becoming infected with the fungus. Females are also preferentially attracted to cloth sprayed with oil-formulated B. bassiana spores, with 95% of the attracted females becoming infected after a one-minute visit on the cloth. This is the first report of an insect being attracted to a lethal fungal pathogen. The exact mechanisms involved in this behavior remain unclear. Nonetheless, our results indicate that biopesticidal formulations comprising B. bassiana spores will be conducive to attraction and on-source visitation by malaria vectors. PMID:23658757

  2. Internal transcribed spacer (ITS) evolution in populations of the hyperparasitic European mistletoe pathogen fungus, Sphaeropsis visci (Botryosphaeriaceae): The utility of ITS2 secondary structures.

    PubMed

    Poczai, Péter; Varga, Ildikó; Hyvönen, Jaakko

    2015-03-01

    We investigated patterns of nucleotide polymorphism in the internal transcribed spacer (ITS) region for Sphaeropsis visci, a hyperparasitic fungus that causes the leaf spot disease of the hemiparasite European mistletoe (Viscum album). Samples of S. visci were obtained from Hungary covering all major infected forest areas. For obtaining PCR products we used a fast and efficient direct PCR approach based on a high fidelity DNA polymerase. A total of 140 ITS sequences were subjected to an array of complementary sequence analyses, which included analyses of secondary structure stability, nucleotide polymorphism patterns, GC content, and presence of conserved motifs. Analysed sequences exhibited features of functional rRNAs. Overall, polymorphism was observed within less conserved motifs, such as loops and bulges, or, alternatively, as non-canonical G-U pairs within conserved regions of double stranded helices. The secondary structure of ITS2 provides new opportunities for obtaining further valuable information, which could be used in phylogenetic analyses, or at population level as demonstrated in our study. This is due to additional information provided by secondary structures and their models. The combined score matrix was used with the methods implemented in the programme 4SALE. Besides the pseudoprotein coding method of 4SALE, the molecular morphometric character coding also has potential for gaining further information for phylogenetic analyses based on the geometric features of the sub-structural elements of the ITS2 RNA transcript.

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

    PubMed Central

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

    2016-01-01

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

  4. Ambispora granatensis, a new arbuscular mycorrhizal fungus, associated with Asparagus officinalis in Andalucia (Spain).

    PubMed

    Palenzuela, Javier; Barea, José-Miguel; Ferrol, Nuria; Oehl, Fritz

    2011-01-01

    A new dimorphic fungal species in the arbuscular mycorrhiza-forming Glomeromycota, Ambispora granatensis, was isolated from an agricultural site in the province of Granada (Andalucía, Spain) growing in the rhizosphere of Asparagus officinalis. It was propagated in pot cultures with Trifolium pratense and Sorghum vulgare. The fungus also colonized Ri T-DNA transformed Daucus carota roots but did not form spores in these root organ cultures. The spores of the acaulosporoid morph are 90-150 μm diam and hyaline to white to pale yellow. They have three walls and a papillae-like rough irregular surface on the outer surface of the outer wall. The irregular surface might become difficult to detect within a few hours in lactic acid-based mountings but are clearly visible in water. The structural central wall layer of the outer wall is only 0.8-1.5 μm thick. The glomoid spores are formed singly or in small, loose spore clusters of 2-10 spores. They are hyaline to pale yellow, (25)40-70 μm diam and have a bilayered spore wall without ornamentation. Nearly full length sequences of the 18S and the ITS regions of the ribosomal gene place the new fungus in a separate clade next to Ambispora fennica and Ambispora gerdemannii. The acaulosporoid spores of the new fungus can be distinguished easily from all other spores in genus Ambispora by the conspicuous thin outer wall.

  5. The SPORES experiment of the EXPOSE-R mission: Bacillus subtilis spores in artificial meteorites

    NASA Astrophysics Data System (ADS)

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

    2015-01-01

    The experiment SPORES `Spores in artificial meteorites' was part of European Space Agency's EXPOSE-R mission, which exposed chemical and biological samples for nearly 2 years (March 10, 2009 to February 21, 2011) to outer space, when attached to the outside of the Russian Zvezda module of the International Space Station. The overall objective of the SPORES experiment was to address the question whether the meteorite material offers enough protection against the harsh environment of space for spores to survive a long-term journey in space by experimentally mimicking the hypothetical scenario of Lithopanspermia, which assumes interplanetary transfer of life via impact-ejected rocks. For this purpose, spores of Bacillus subtilis 168 were exposed to selected parameters of outer space (solar ultraviolet (UV) radiation at λ>110 or >200 nm, space vacuum, galactic cosmic radiation and temperature fluctuations) either as a pure spore monolayer or mixed with different concentrations of artificial meteorite powder. Total fluence of solar UV radiation (100-400 nm) during the mission was 859 MJ m-2. After retrieval the viability of the samples was analysed. A Mission Ground Reference program was performed in parallel to the flight experiment. The results of SPORES demonstrate the high inactivating potential of extraterrestrial UV radiation as one of the most harmful factors of space, especially UV at λ>110 nm. The UV-induced inactivation is mainly caused by photodamaging of the DNA, as documented by the identification of the spore photoproduct 5,6-dihydro-5(α-thyminyl)thymine. The data disclose the limits of Lithopanspermia for spores located in the upper layers of impact-ejected rocks due to access of harmful extraterrestrial solar UV radiation.

  6. Bacillus subtilis Spore Coat

    PubMed Central

    Driks, Adam

    1999-01-01

    In response to starvation, bacilli and clostridia undergo a specialized program of development that results in the production of a highly resistant dormant cell type known as the spore. A proteinacious shell, called the coat, encases the spore and plays a major role in spore survival. The coat is composed of over 25 polypeptide species, organized into several morphologically distinct layers. The mechanisms that guide coat assembly have been largely unknown until recently. We now know that proper formation of the coat relies on the genetic program that guides the synthesis of spore components during development as well as on morphogenetic proteins dedicated to coat assembly. Over 20 structural and morphogenetic genes have been cloned. In this review, we consider the contributions of the known coat and morphogenetic proteins to coat function and assembly. We present a model that describes how morphogenetic proteins direct coat assembly to the specific subcellular site of the nascent spore surface and how they establish the coat layers. We also discuss the importance of posttranslational processing of coat proteins in coat morphogenesis. Finally, we review some of the major outstanding questions in the field. PMID:10066829

  7. The role of mites in insect-fungus associations.

    PubMed

    Hofstetter, R W; Moser, J C

    2014-01-01

    The interactions among insects, mites, and fungi are diverse and complex but poorly understood in most cases. Associations among insects, mites, and fungi span an almost incomprehensible array of ecological interactions and evolutionary histories. Insects and mites often share habitats and resources and thus interact within communities. Many mites and insects rely on fungi for nutrients, and fungi benefit from them with regard to spore dispersal, habitat provision, or nutrient resources. Mites have important impacts on community dynamics, ecosystem processes, and biodiversity within many insect-fungus systems. Given that mites are understudied but highly abundant, they likely have bigger, more important, and more widespread impacts on communities than previously recognized. We describe mutualistic and antagonistic effects of mites on insect-fungus associations, explore the processes that underpin ecological and evolutionary patterns of these multipartite communities, review well-researched examples of the effects of mites on insect-fungus associations, and discuss approaches for studying mites within insect-fungus communities.

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

    PubMed Central

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

    2015-01-01

    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

  9. Spores do travel.

    PubMed

    Dam, Nico

    2013-01-01

    Model calculations are presented on the horizontal dispersal distance of basidiospores from their source (any typical agaric). The results are compared to old and recent experimental data obtained by sampling on sticky microscope slides placed on soil. I argue that such experimental data alone are insufficient to determine the dispersion kernel because of sampling paucity: Only a minor fraction of the released spores is sampled, and the fate of the rest is unknown. Spore dispersal is determined largely by wind, whereas deposition may be due predominantly to wash-out by rainfall.

  10. Factors affecting fungus-induced larval mortality in Anopheles gambiae and Anopheles stephensi

    PubMed Central

    2010-01-01

    Background Entomopathogenic fungi have shown great potential for the control of adult malaria vectors. However, their ability to control aquatic stages of anopheline vectors remains largely unexplored. Therefore, how larval characteristics (Anopheles species, age and larval density), fungus (species and concentration) and environmental effects (exposure duration and food availability) influence larval mortality caused by fungus, was studied. Methods Laboratory bioassays were performed on the larval stages of Anopheles gambiae and Anopheles stephensi with spores of two fungus species, Metarhizium anisopliae and Beauveria bassiana. For various larval and fungal characteristics and environmental effects the time to death was determined and survival curves established. These curves were compared by Kaplan Meier and Cox regression analyses. Results Beauveria bassiana and Metarhizium anisopliae caused high mortality of An. gambiae and An. stephensi larvae. However, Beauveria bassiana was less effective (Hazard ratio (HR) <1) compared to Metarhizium anisopliae. Anopheles stephensi and An. gambiae were equally susceptible to each fungus. Older larvae were less likely to die than young larvae (HR < 1). The effect of increase in fungus concentration on larval mortality was influenced by spore clumping. One day exposure to fungal spores was found to be equally effective as seven days exposure. In different exposure time treatments 0 - 4.9% of the total larvae, exposed to fungus, showed infection at either the pupal or adult stage. Mortality rate increased with increasing larval density and amount of available food. Conclusions This study shows that both fungus species have potential to kill mosquitoes in the larval stage, and that mortality rate depends on fungus species itself, larval stage targeted, larval density and amount of nutrients available to the larvae. Increasing the concentration of fungal spores or reducing the exposure time to spores did not show a proportional

  11. Morphological and molecular characterization of Magnaporthe oryzae (fungus) from infected rice leaf samples

    NASA Astrophysics Data System (ADS)

    Muni, Nurulhidayah Mat; Nadarajah, Kalaivani

    2014-09-01

    Magnaporthe oryzae is a plant-pathogenic fungus that causes a serious disease affecting rice called rice blast. Outbreaks of rice blast have been a threat to the global production of rice. This fungal disease is estimated to cause production losses of US55 million each year in South and Southeast Asia. It has been used as a primary model for elucidating various aspects of the host-pathogen interaction with its host. We have isolated five isolates of Magnaporthe oryzae from diseased leaf samples obtained from the field at Kompleks Latihan MADA, Kedah, Malaysia. We have identified the isolates using morphological and microscopic studies on the fungal spores and the lesions on the diseased leaves. Amplification of the internal transcribed spacer (ITS) was carried out with universal primers ITS1 and ITS4. The sequence of each isolates showed at least 99% nucleotide identity with the corresponding sequence in GenBank for Magnaporthe oryzae.

  12. Analysis of the antimicrobial effects of nonthermal plasma on fungal spores in ionic solutions.

    PubMed

    Kang, Min Ho; Hong, Young June; Attri, Pankaj; Sim, Geon Bo; Lee, Geon Joon; Panngom, Kamonporn; Kwon, Gi Chung; Choi, Eun Ha; Uhm, Han S; Park, Gyungsoon

    2014-07-01

    The antimicrobial efficiency of reactive species-based control strategies is significantly affected by the dynamics of reactive species in the biological environment. Atmospheric-pressure nonthermal plasma is an ionized gas in which various reactive species are produced. The various levels of antimicrobial activity may result from the dynamic interaction of the plasma-generated reactive species with the environment. However, the nature of the interaction between plasma and environments is poorly understood. In this study, we analyzed the influence of the ionic strength of surrounding solutions (environment) on the antimicrobial activity of plasma in relation to the plasma-generated reactive species using a model filamentous fungus, Neurospora crassa. Our data revealed that the presence of sodium chloride (NaCl) in the background solution attenuated the deleterious effects of plasma on germination, internal structure, and genomic DNA of fungal spores. The protective effects of NaCl were not explained exclusively by pH, osmotic stability, or the level of reactive species in the solution. These were strongly associated with the ionic strength of the background solution. The presence of ions reduced plasma toxicity, which might be due to a reduced access of reactive species to fungal spores, and fungal spores were inactivated by plasma in a background fluid of nonionic osmolytes despite the low level of reactive species. Our results suggest that the surrounding environment may affect the behavior of reactive species, which leads to different biological consequences regardless of their quantity. Moreover, the microbicidal effect of plasma can be synergistically regulated through control of the microenvironment.

  13. Anthrax Spores under a microscope

    NASA Technical Reports Server (NTRS)

    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.

  14. Biomarkers of Aspergillus spores

    NASA Astrophysics Data System (ADS)

    Sulc, Miroslav; Peslova, Katerina; Zabka, Martin; Hajduch, Marian; Havlicek, Vladimir

    2009-02-01

    We applied both matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometric and 1D sodium dodecylsulfate polyacrylamide gel electrophoretic (1D-PAGE) approaches for direct analysis of intact fungal spores of twenty four Aspergillus species. In parallel, we optimized various protocols for protein extraction from Aspergillus spores using acidic conditions, step organic gradient and variable sonication treatment. The MALDI-TOF mass spectra obtained from optimally prepared samples provided a reproducible fingerprint demonstrating the capability of the MALDI-TOF approach to type and characterize different fungal strains within the Aspergillus genus. Mass spectra of intact fungal spores provided signals mostly below 20 kDa. The minimum material amount represented 0.3 [mu]g (10,000 spores). Proteins with higher molecular weight were detected by 1D-PAGEE Eleven proteins were identified from three selected strains in the range 5-25 kDa by the proteomic approach. Hemolysin and hydrophobin have the highest relevance in host-pathogen interactions.

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

    SciTech Connect

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

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

    NASA Astrophysics Data System (ADS)

    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.

  17. Label-Free Detection of Soybean Rust Spores using Photonic Crystal Biosensors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Soybean rust, caused by the fungus Phakopsora pachyrhizi, is one of the most devastating foliar diseases affecting soybeans grown worldwide. The disease was reported for the first time in the United States in 2004. Early spore detection, prior to the appearance of visible symptoms, is critical to ef...

  18. Possible Contribution of Blast Spores to the Oxidative Burst in the Infection Droplet on Rice Leaf

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The infection-induced overproduction of reactive oxygen species (ROS) in resistant plants is usually ascribed to the host. Here we tested the possible contribution of the parasite, the rice blast fungus to ROS production. Droplets of spore suspensions or water were kept on rice leaves or on plastic....

  19. Pathogenicity of entomopathogenic fungus Metarhizium anisopliae (Deuteromycetes) to Ixodes scapularis (Acari: Ixodidae)

    USGS Publications Warehouse

    Zhioua, E.; Browning, M.; Johnson, P.W.; Ginsberg, H.S.; LeBrun, R.A.

    1997-01-01

    The entomopathogenic fungus Metarhizium anisopliae is highly pathogenic to the black-legged tick, Ixodes scapularis. Spore concentrations of 108/ml for engorged larvae and 107/ml for engorged females resulted in 100% tick mortality, 2 wk post-infection. The LC50 value for engorged larvae (concentration to kill 50% of ticks) was 107 spores/ml. Metarhizium anisopliae shows considerable potential as a microbial control agent for the management of Ixodes scapularis.

  20. Spore: Spawning Evolutionary Misconceptions?

    NASA Astrophysics Data System (ADS)

    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.

  1. Thermal Spore Exposure Vessels

    NASA Technical Reports Server (NTRS)

    Beaudet, Robert A.; Kempf, Michael; Kirschner, Larry

    2006-01-01

    Thermal spore exposure vessels (TSEVs) are laboratory containers designed for use in measuring rates of death or survival of microbial spores at elevated temperatures. A major consideration in the design of a TSEV is minimizing thermal mass in order to minimize heating and cooling times. This is necessary in order to minimize the number of microbes killed before and after exposure at the test temperature, so that the results of the test accurately reflect the effect of the test temperature. A typical prototype TSEV (see figure) includes a flat-bottomed stainless-steel cylinder 4 in. (10.16 cm) long, 0.5 in. (1.27 cm) in diameter, having a wall thickness of 0.010 plus or minus 0.002 in. (0.254 plus or minus 0.051 mm). Microbial spores are deposited in the bottom of the cylinder, then the top of the cylinder is closed with a sterile rubber stopper. Hypodermic needles are used to puncture the rubber stopper to evacuate the inside of the cylinder or to purge the inside of the cylinder with a gas. In a typical application, the inside of the cylinder is purged with dry nitrogen prior to a test. During a test, the lower portion of the cylinder is immersed in a silicone-oil bath that has been preheated to and maintained at the test temperature. Test temperatures up to 220 C have been used. Because the spores are in direct contact with the thin cylinder wall, they quickly become heated to the test temperature.

  2. Spore collection and elimination apparatus and method

    DOEpatents

    Czajkowski, Carl; Warren, Barbara Panessa

    2007-04-03

    The present invention is for a spore collection apparatus and its method of use. The portable spore collection apparatus includes a suction source, a nebulizer, an ionization chamber and a filter canister. The suction source collects the spores from a surface. The spores are activated by heating whereby spore dormancy is broken. Moisture is then applied to the spores to begin germination. The spores are then exposed to alpha particles causing extinction.

  3. Science hub spore data

    EPA Pesticide Factsheets

    Data set includes UV dose, and Bacillus pumilus spore plate counts in colony forming unitsThis dataset is associated with the following publication:Boczek , L., E. Rhodes , J. Cashdollar, J. Ryu, J. Popovici , J. Hoelle , M. Sivaganesan , S. Hayes , M. Rodgers , and H. Ryu. Applicability of UV resistant Bacillus pumilus endospores as a human adenovirus surrogate for evaluating the effectiveness of virus inactivation in low-pressure UV treatment systems. JOURNAL OF MICROBIOLOGICAL METHODS. Elsevier Science Ltd, New York, NY, USA, 122: 43-49, (2016).

  4. Slope aspect influences arbuscular mycorrhizal fungus communities in arid ecosystems of the Daqingshan Mountains, Inner Mongolia, North China.

    PubMed

    Liu, Min; Zheng, Rong; Bai, Shulan; Bai, Yv E; Wang, Jugang

    2017-04-01

    Arbuscular mycorrhizal (AM) symbiosis plays an important role in ecosystem functioning, particularly in fragile environments. Little is known, however, about how AM fungus community composition responds to slope aspect. Our objective was to compare the AM fungus communities between sunny and shady slopes and to detect factors that influenced the distributions of AM fungi in arid ecosystems of the Daqingshan Mountains, Inner Mongolia, North China. AM fungus communities were evaluated based on small subunit ribosomal RNA genes (SSUs) using Illumina MiSeq sequencing. AM fungus community composition differed significantly between slope aspects, and sunny slopes had significantly higher AM fungus diversity and richness as well as spore density, total root colonization, arbuscule abundance, vesicle abundance, and hyphal colonization than shady slopes. Structural equation modeling (SEM) illustrated that the effects of slope aspect on AM fungus richness likely were mediated by available phosphorus, soil organic carbon, plant cover, and plant diversity. Available phosphorus was the principal factor that influenced AM fungus species richness, and soil organic carbon was the principal factor influencing spore density and total root colonization, suggesting that these factors especially might be responsible for differences between the AM fungus communities of different slope aspects. These findings elucidate the influence of slope aspect on AM fungus communities and may inform use of AM fungi in protection and restoration of vegetation with different slope aspects in arid ecosystems.

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

    PubMed Central

    Sander, Sarah E.; Altizer, Sonia; de Roode, Jacobus C.; Davis, Andrew K.

    2013-01-01

    Monarch butterflies (Danaus plexippus) throughout the world are commonly infected by the specialist pathogen Ophryocystis elektroscirrha (OE). This protozoan is transmitted when larvae ingest infectious stages (spores) scattered onto host plant leaves by infected adults. Parasites replicate internally during larval and pupal stages, and adult monarchs emerge covered with millions of dormant spores on the outsides of their bodies. Across multiple monarch populations, OE varies in prevalence and virulence. Here, we examined geographic and genetic variation in OE spore morphology using clonal parasite lineages derived from each of four host populations (eastern and western North America, South Florida and Hawaii). Spores were harvested from experimentally inoculated, captive-reared adult monarchs. Using light microscopy and digital image analysis, we measured the size, shape and color of 30 replicate spores per host. Analyses examined predictors of spore morphology, including parasite source population and clone, parasite load, and the following host traits: family line, sex, wing area, and wing color (orange and black pigmentation). Results showed significant differences in spore size and shape among parasite clones, suggesting genetic determinants of morphological variation. Spore size also increased with monarch wing size, and monarchs with larger and darker orange wings tended to have darker colored spores, consistent with the idea that parasite development depends on variation in host quality and resources. We found no evidence for effects of source population on variation in spore morphology. Collectively, these results provide support for heritable variation in spore morphology and a role for host traits in affecting parasite development. PMID:26462429

  6. "Spore" and the Sociocultural Moment

    ERIC Educational Resources Information Center

    Meyer, W. Max

    2012-01-01

    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…

  7. PROPERTIES OF ELECTRODIALYZED BACTERIAL SPORES

    PubMed Central

    Harper, M. K.; Curran, H. R.; Pallansch, M. J.

    1964-01-01

    Harper, M. K. (U.S. Department of Agriculture, Washington, D.C.), H. R. Curran, and M. J. Pallansch. Properties of electrodialyzed bacterial spores. J. Bacteriol. 88:1338–1340. 1964.—Washed spores of Bacillus cereus, B. megaterium, and B. stearothermophilis suspended in distilled water were electrodialyzed at a potential of 250 v, 50 ma, for 6.5 hr, under conditions which precluded rise in temperature or shift in pH. Dipicolinic acid (DPA) was not released from the spores by electrodialysis, as indicated by essentially complete recovery of residual DPA from the treated spores. Uptake of stain, heat stability, and viability of the electrodialyzed spores were comparable to the nondialyzed controls. These findings are discussed in relation to those reported by Rode and Foster. PMID:14234790

  8. Studies on the air-borne fungal spores in Amritsar: their role in keratomycosis.

    PubMed

    Sandhu, D K; Randhawa, I S

    1979-08-31

    An aerial survey for fungal spores in Amritsar has been carried out by petri plate exposure method for a period of one year. A total of 23 fungi appeared in the plates. Out of these Aspergillus was the commonest fungus representing 21.69% of the total colony count followed by Alternaria, Curvularia and Fusarium. There was seasonal variation in the prevalence of fungal spores. A comparison of the prevalence of fungi in diseased and healthy eyes and the atmosphere of Amritsar appears to support the view that these fungi are transient residents in the eyes depending on their availability in the atmosphere.

  9. Fungus Infections: Tinea

    MedlinePlus

    ... Share: Yes No, Keep Private Fungus Infections Share | Tinea is the name given to a fungal skin ... Sometime the susceptibility will run in the family. Tinea Pedis (Athlete's foot) This is the most common ...

  10. Beyond animals and plants: dynamic maternal effects in the fungus Neurospora crassa.

    PubMed

    Zimmerman, K C K; Levitis, D A; Pringle, A

    2016-07-01

    Maternal effects are widely documented in animals and plants, but not in fungi or other eukaryotes. A principal cause of maternal effects is asymmetrical parental investment in a zygote, creating greater maternal vs. paternal influence on offspring phenotypes. Asymmetrical investments are not limited to animals and plants, but are also prevalent in fungi and groups including apicomplexans, dinoflagellates and red algae. Evidence suggesting maternal effects among fungi is sparse and anecdotal. In an experiment designed to test for maternal effects across sexual reproduction in the model fungus Neurospora crassa, we measured offspring phenotypes from crosses of all possible pairs of 22 individuals. Crosses encompassed reciprocals of 11 mating-type 'A' and 11 mating-type 'a' wild strains. After controlling for the genetic and geographic distances between strains in any individual cross, we found strong evidence for maternal control of perithecia (sporocarp) production, as well as maternal effects on spore numbers and spore germination. However, both parents exert equal influence on the percentage of spores that are pigmented and size of pigmented spores. We propose a model linking the stage-specific presence or absence of maternal effects to cellular developmental processes: effects appear to be mediated primarily through the maternal cytoplasm, and, after spore cell walls form, maternal influence on spore development is limited. Maternal effects in fungi, thus far largely ignored, are likely to shape species' evolution and ecologies. Moreover, the association of anisogamy and maternal effects in a fungus suggests maternal effects may also influence the biology of other anisogamous eukaryotes.

  11. Methyl Iodide Fumigation of Bacillus anthracis Spores.

    PubMed

    Sutton, Mark; Kane, Staci R; Wollard, Jessica R

    2015-09-01

    Fumigation techniques such as chlorine dioxide, vaporous hydrogen peroxide, and paraformaldehyde previously used to decontaminate items, rooms, and buildings following contamination with Bacillus anthracis spores are often incompatible with materials (e.g., porous surfaces, organics, and metals), causing damage or residue. Alternative fumigation with methyl bromide is subject to U.S. and international restrictions due to its ozone-depleting properties. Methyl iodide, however, does not pose a risk to the ozone layer and has previously been demonstrated as a fumigant for fungi, insects, and nematodes. Until now, methyl iodide has not been evaluated against Bacillus anthracis. Sterne strain Bacillus anthracis spores were subjected to methyl iodide fumigation at room temperature and at 550C. Efficacy was measured on a log-scale with a 6-log reduction in CFUs being considered successful compared to the U.S. Environmental Protection Agency biocide standard. Such efficacies were obtained after just one hour at 55 °C and after 12 hours at room temperature. No detrimental effects were observed on glassware, PTFE O-rings, or stainless steel. This is the first reported efficacy of methyl iodide in the reduction of Bacillus anthracis spore contamination at ambient and elevated temperatures.

  12. Microbial profile modification with spores

    SciTech Connect

    Bae, J.H.; Chambers, K.T.; Lee, H.O.

    1996-08-01

    To overcome the shortcomings of conventional, near-wellbore profile modification methods, a microbial profile modification (MPM) method with spores was investigated. A halotolerant, spore-forming mesophile was isolated and characterized. These biopolymer-producing spores propagate easily in Berea cores with permeabilities more than about 500 md. With a specifically formulated nutrient package, they are readily germinated and produce biofilm, which reduces the permeability of the rock. The depth of penetration and the degree of permeability reduction can be controlled by varying injection schemes.

  13. Spore Coat Architecture of Clostridium novyi-NT spores

    SciTech Connect

    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.

  14. Life cycle and spore resistance of spore-forming Bacillus atrophaeus.

    PubMed

    Sella, Sandra R B R; Vandenberghe, Luciana P S; Soccol, Carlos Ricardo

    2014-12-01

    Bacillus endospores have a wide variety of important medical and industrial applications. This is an overview of the fundamental aspects of the life cycle, spore structure and factors that influence the spore resistance of spore-forming Bacillus. Bacillus atrophaeus was used as reference microorganism for this review because their spores are widely used to study spore resistance and morphology. Understanding the mechanisms involved in the cell cycle and spore survival is important for developing strategies for spore killing; producing highly resistant spores for biodefense, food and pharmaceutical applications; and developing new bioactive molecules and methods for spore surface display.

  15. Bacillus anthracis spore movement does not require a carrier cell and is not affected by lethal toxin in human lung models.

    PubMed

    Booth, J Leland; Duggan, Elizabeth S; Patel, Vineet I; Langer, Marybeth; Wu, Wenxin; Braun, Armin; Coggeshall, K Mark; Metcalf, Jordan P

    2016-10-01

    The lung is the entry site for Bacillus anthracis in inhalation anthrax, the most deadly form of the disease. Spores escape from the alveolus to regional lymph nodes, germinate and enter the circulatory system to cause disease. The roles of carrier cells and the effects of B. anthracis toxins in this process are unclear. We used a human lung organ culture model to measure spore uptake by antigen presenting cells (APC) and alveolar epithelial cells (AEC), spore partitioning between these cells, and the effects of B. anthracis lethal toxin and protective antigen. We repeated the study in a human A549 alveolar epithelial cell model. Most spores remained unassociated with cells, but the majority of cell-associated spores were in AEC, not in APC. Spore movement was not dependent on internalization, although the location of internalized spores changed in both cell types. Spores also internalized in a non-uniform pattern. Toxins affected neither transit of the spores nor the partitioning of spores into AEC and APC. Our results support a model of spore escape from the alveolus that involves spore clustering with transient passage through intact AEC. However, subsequent transport of spores by APC from the lung to the lymph nodes may occur.

  16. Spore Size Comparison Between Several Bacillus Species

    DTIC Science & Technology

    2005-10-01

    Spore Size Comparison Between Several Bacillus Species Ruben O. Zandomeni1, Joseph E. Fitzgibbon2, Monica Carrera1, Edward Stuebing2, James E...OCT 2005 2. REPORT TYPE N/A 3. DATES COVERED - 4. TITLE AND SUBTITLE Spore Size Comparison Between Several Bacillus Species 5a. CONTRACT...Systematic comparison of the size of B.anthracis spores to size of other Bacillus spores (simulants/surrogates) - all spores produced under the same

  17. Direct probing of the surface ultrastructure and molecular interactions of dormant and germinating spores of Phanerochaete chrysosporium.

    PubMed

    Dufrêne, Y F; Boonaert, C J; Gerin, P A; Asther, M; Rouxhet, P G

    1999-09-01

    Atomic force microscopy (AFM) has been used to probe, under physiological conditions, the surface ultrastructure and molecular interactions of spores of the filamentous fungus Phanerochaete chrysosporium. High-resolution images revealed that the surface of dormant spores was uniformly covered with rodlets having a periodicity of 10 +/- 1 nm, which is in agreement with earlier freeze-etching measurements. In contrast, germinating spores had a very smooth surface partially covered with rough granular structures. Force-distance curve measurements demonstrated that the changes in spore surface ultrastructure during germination are correlated with profound modifications of molecular interactions: while dormant spores showed no adhesion with the AFM probe, germinating spores exhibited strong adhesion forces, of 9 +/- 2 nN magnitude. These forces are attributed to polysaccharide binding and suggested to be responsible for spore aggregation. This study represents the first direct characterization of the surface ultrastructure and molecular interactions of living fungal spores at the nanometer scale and offers new prospects for mapping microbial cell surface properties under native conditions.

  18. Identifying and Inactivating Bacterial Spores

    NASA Technical Reports Server (NTRS)

    Newcombe, David; Dekas, Anne; Venkateswaran, Kasthuri

    2009-01-01

    Problems associated with, and new strategies for, inactivating resistant organisms like Bacillus canaveralius (found at Kennedy Space Center during a survey of three NASA cleanrooms) have been defined. Identifying the particular component of the spore that allows its heightened resistance can guide the development of sterilization procedures that are targeted to the specific molecules responsible for resistance, while avoiding using unduly harsh methods that jeopardize equipment. The key element of spore resistance is a multilayered protein shell that encases the spore called the spore coat. The coat of the best-studied spore-forming microbe, B. subtilis, consists of at least 45 proteins, most of which are poorly characterized. Several protective roles for the coat are well characterized including resistance to desiccation, large toxic molecules, ortho-phthalaldehyde, and ultraviolet (UV) radiation. One important long-term specific goal is an improved sterilization procedure that will enable NASA to meet planetary protection requirements without a terminal heat sterilization step. This would support the implementation of planetary protection policies for life-detection missions. Typically, hospitals and government agencies use biological indicators to ensure the quality control of sterilization processes. The spores of B. canaveralius that are more resistant to osmotic stress would serve as a better biological indicator for potential survival than those in use currently.

  19. NASA Facts: SporeSat

    NASA Technical Reports Server (NTRS)

    Martinez, Andres; Cappuccio, Gelsomina; Tomko, David

    2013-01-01

    SporeSat is an autonomous, free-flying three-unit (3U) spacecraft that will be used to conduct scientific experiments to gain a deeper knowledge of the mechanisms of plant cell gravity sensing. SporeSat is being developed through a partnership between NASAs Ames Research Center and the Department of Agricultural and Biological Engineering at Purdue University. Amani Salim and Jenna L. Rickus are the Purdue University Principal Investigators. The SporeSat mission will be flown using a 3U nanosatellite weighing approximately 12 pounds and measuring 14 inches long by 4 inches wide by 4 inches tall. SporeSat will utilize flight-proven spacecraft technologies demonstrated on prior Ames nanosatellite missions such as PharmaSat and OrganismOrganic Exposure to Orbital Stresses (OOREOS) as well as upgrades that increase the hardware integration capabilities with SporeSat science instrumentation. In addition, the SporeSat science payload will serve as a technology platform to evaluate new microsensor technologies for enabling future fundamental biology missions.

  20. Effects of meteorological conditions on spore plumes

    NASA Astrophysics Data System (ADS)

    Burch, M.; Levetin, E.

    2002-05-01

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

  1. UV-Photobiology of bacterial spores in space

    NASA Astrophysics Data System (ADS)

    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

  2. Developmental modulation of DNA methylation in the fungus Phycomyces blakesleeanus.

    PubMed Central

    Antequera, F; Tamame, M; Vilanueva, J R; Santos, T

    1985-01-01

    DNA methylation is a rather sparse event among fungi. Phycomyces blakesleeanus seems to be one of the few exceptions in this context. 5-Methylcytosine represents 2.9% of the total cytosine in spore DNA and is located in approximately the same amount at any of the four CA, CT, CC or CG dinucleotides. A progressive and gradual drop in total 5-methylcytosine parallels the development of the fungus. This demethylation is non random but sequence specific and is not accounted for equally by the four different methylated dinucleotides, CG being much less affected (20% demethylated) than CA, CT and CC (more than 90% demethylated at the same time). "De novo" methylation to restore the initial pattern probably takes place during spore maturation. By using specific hybridization probes we have been able to show that the rRNA genes are not significantly methylated at any stage of development, regardless of their transcription status. Images PMID:2997714

  3. Ornithine and arginine decarboxylase activities and effect of some polyamine biosynthesis inhibitors on Gigaspora rosea germinating spores.

    PubMed

    Sannazzaro, Analía I; Alvarez, Cora L; Menéndez, Ana B; Pieckenstain, Fernando L; Albertó, Edgardo O; Ruiz, Oscar A

    2004-01-15

    The pathways for putrescine biosynthesis and the effects of polyamine biosynthesis inhibitors on the germination and hyphal development of Gigaspora rosea spores were investigated. Incubation of spores with different radioactive substrates demonstrated that both arginine and ornithine decarboxylase pathways participate in putrescine biosynthesis in G. rosea. Spermidine and spermine were the most abundant polyamines in this fungus. The putrescine biosynthesis inhibitors alpha-difluoromethylarginine and alpha-difluoromethylornithine, as well as the spermidine synthase inhibitor cyclohexylamine, slightly decreased polyamine levels. However, only the latter interfered with spore germination. The consequences of the use of putrescine biosynthesis inhibitors for the control of plant pathogenic fungi on the viability of G. rosea spores in soil are discussed.

  4. Measuring Total and Germinable Spore Populations

    NASA Technical Reports Server (NTRS)

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

    2011-01-01

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

  5. A study of spore identification from diffraction data

    NASA Astrophysics Data System (ADS)

    Le, Thanh; Cao, Yang; Fiddy, M. A.; Gardner, P.

    2007-04-01

    Much work has been reported on attempting to identify spores from their spectral signatures. Since spores are also complex scattering objects, with a layered internal refractive index structure, it makes sense to explore the possibility of making an identification simply from a scattering pattern or from anticipated scattering characteristics combined with a spectral signature. Models for scattering from simple geometrical coated shapes have been developed and recently Bragg spheres and onion-ring resonator-like scatterers in the Mie regime have received considerable attention driven by other applications. Also, our own group has recently advanced a method for inverting scattered field data from strongly scattering penetrable targets. We present here some very early considerations of the convergence of these possibilities and suggest some simple experiments that might advance our understanding of spore detection and identification.

  6. Fungal spores: hazardous to health?

    PubMed Central

    Sorenson, W G

    1999-01-01

    Fungi have long been known to affect human well being in various ways, including disease of essential crop plants, decay of stored foods with possible concomitant production of mycotoxins, superficial and systemic infection of human tissues, and disease associated with immune stimulation such as hypersensitivity pneumonitis and toxic pneumonitis. The spores of a large number of important fungi are less than 5 microm aerodynamic diameter, and therefore are able to enter the lungs. They also may contain significant amounts of mycotoxins. Diseases associated with inhalation of fungal spores include toxic pneumonitis, hypersensitivity pneumonitis, tremors, chronic fatigue syndrome, kidney failure, and cancer. PMID:10423389

  7. Spores of Aspergillus versicolor isolated from indoor air of a moisture-damaged building provoke acute inflammation in mouse lungs.

    PubMed

    Jussila, Juha; Komulainen, Hannu; Kosma, Veli-Matti; Nevalainen, Aino; Pelkonen, Jukka; Hirvonen, Maija-Riitta

    2002-12-01

    Microbial growth in moisture-damaged buildings has been associated with respiratory health effects, and the spores of the mycotoxin producing fungus Aspergillus versicolor are frequently present in the indoor air. To characterize the potential of these spores to cause harmful respiratory effects, mice were exposed via intratracheal instillation to a single dose of the spores of A. versicolor (1 x 10(5), 1 x 10(6), 5 x 10(6), 1 x 10(7), or 1 x 10(8) spores), isolated from the indoor air of a moisture-damaged building. Inflammation and toxicity in lungs were evaluated 24 h later by assessment of biochemical markers and histopathology. The time course of the effects was investigated with the dose of 5 x 10(6) spores for up to 28 days. The exposure to the spores increased transiently proinflammatory cytokine levels (tumor necrosis factor [TNF] alpha and interleukin [IL]-6) in bronchoalveolar lavage fluid (BALF). The cytokine responses were dose and time dependent. The highest cytokine concentrations were measured at 6 h after the dose, and they returned to the control level by 3 days. Moreover, the spores of A. versicolor recruited inflammatory cells into airways: Neutrophils peaked transiently at 24 h, macrophages at 3 days, and lymphocytes at 7 days after the dosing. The inflammatory cell response did not completely disappear during the subsequent 28 days, though no histopathological changes were seen at that time point. The spores did not induce expression of inducible nitric oxide synthase in lavaged cells. Only the highest spore dose (1 x 10(8)) markedly increased serum IL-6, increased vascular leakage, and caused cytotoxicity (i.e., increased levels of albumin, total protein, lactate dehydrogenase [LDH], and hemoglobin in BALF) in the airways. In summary, the spores of A. versicolor caused acute inflammation in mouse lungs. This indicates that they have potential to provoke adverse health effects in the occupants of moisture-damaged buildings.

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

    PubMed Central

    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

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

    PubMed

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

    2015-01-01

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

  10. Mechanisms of Resistance in Microbial Spores

    DTIC Science & Technology

    1990-12-20

    heat shock affects permeability and resistance of Bacillus stearotbermo2hilus spores; low heat resistance of )2. SQhaericus spores correlated with...DNA content in~· megaterium spores; compact structure of cortical peptidoglycans from bacterial spores. The titles of four published re-.view...among 8 Bacillus species spanning a 3,000-fold range in SHR, which was altered by acid demineralization and specific remineralization and also by

  11. Identification of Bacterial Spores using Statistical Analysis of Fourier Transform Infrared Photoacoustic Spectroscopy Data

    SciTech Connect

    Thompson, Sandra E.; Foster, Nancy S.; Johnson, Timothy J.; Valentine, Nancy B.; Amonette, James E.

    2003-08-28

    Fourier Transform Infrared Photoacoustic Spectroscopy (FTIR-PAS) has been applied for the first time to the identification and speciation of bacterial spores. With minimal preparation the spores were deposited into the photoacoustic sample cup and their spectra recorded. A total of 40 different samples of 5 different strains of Bacillus spores were analyzed: Bacillus subtilis ATCC 49760, Bacillus atrophaeus ATCC 49337, Bacillus subtilis 6051, Bacillus thuringiensis ssp. kurstaki, and Bacillus globigii Dugway. The statistical methods used included principal-component analysis (PCA), classification and regression trees (CART), and Mahalanobis-distance calculations. Internal cross-validation studies successfully classify the spores according to their bacterial strain in 38 of 40 cases (95%) and 36 of 40 (90%) in cross-validation. Analysis of fifteen blind samples, which included library and other spores, and nonbacterial materials, resulted in correct strain classification the blind samples that were members of the library and correct rejection of the nonbacterial samples.

  12. 1-Octanol, a self-inhibitor of spore germination in Penicillium camemberti.

    PubMed

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

    2016-08-01

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

  13. High gas pressure: an innovative method for the inactivation of dried bacterial spores.

    PubMed

    Colas de la Noue, A; Espinasse, V; Perrier-Cornet, J-M; Gervais, P

    2012-08-01

    In this article, an original non-thermal process to inactivate dehydrated bacterial spores is described. The use of gases such as nitrogen or argon as transmission media under high isostatic pressure led to an inactivation of over 2 logs CFU/g of Bacillus subtilis spores at 430 MPa, room temperature, for a 1 min treatment. A major requirement for the effectiveness of the process resided in the highly dehydrated state of the spores. Only a water activity below 0.3 led to substantial inactivation. The solubility of the gas in the lipid components of the spore and its diffusion properties was essential to inactivation. The main phenomenon involved seems to be the sorption of the gas under pressure by the spores' structures such as residual pores and plasma membranes, followed by a sudden drop in pressure. Observation by phase-contrast microscopy suggests that internal structures have been affected by the treatment. Some parallels with polymer permeability to gas and rigidity at various water activities offer a few clues about the behavior of the outer layers of spores in response to this parameter and provide a good explanation for the sensitivity of spores to high gas pressure discharge at low hydration levels. Specificity of microorganisms such as size, organization, and composition could help in understanding the differences between spores and yeast regarding the parameters required for inactivation, such as pressure or maintenance time.

  14. Protective effects of Ganoderma lucidum spore on cadmium hepatotoxicity in mice.

    PubMed

    Jin, Hai; Jin, Feng; Jin, Jia-Xing; Xu, Jie; Tao, Ting-Ting; Liu, Jie; Huang, Hou-Jin

    2013-02-01

    The medicinal fungus Ganoderma lucidum has been shown to have hepatoprotective effects. G. lucidum contains triterpenes and polysaccharides, and the Sporoderm-broken G. lucidum powder is particular beneficial. This study utilized G. lucidum spore to examine its effect on [Cd(II)]-induced hepatotoxicity in mice and the mechanism of the protection. Mice were pretreated with G. lucidum spore (0.1, 0.5, and 1.0 g/kg, po, for 7 days), and subsequently challenged with a hepatotoxic dose of Cd(II) (3.7 mg/kg, ip). Liver injury was evaluated 8h later. G. lucidum spore protected against Cd(II)-induced liver injury in a dose-dependent manner, as evidenced by serum alanine aminotransferase, aspartate aminotransferase and histopathology. To examine the mechanism of protection, subcellular distribution of Cd(II) was determined. G. lucidum spore decreased Cd(II) accumulation in hepatic nuclei, mitochondria, and microsomes, but increased Cd(II) distribution to the cytosol, where Cd(II) is sequestered by metallothionein, a protein against Cd(II) toxicity. Indeed, G. lucidum spore induced hepatic metallothionein-1 mRNA 8-fold, and also increased metallothionein protein as determined by the Cd(II)/hemoglobin assay. Cd(II)-induced oxidative stress was also decreased by G. lucidum spore, as evidenced by decreased formation of malondialdehyde. In summary, G. lucidum spore is effective in protection against Cd(II)-induced hepatotoxicity, and this effect is due, at least in part, to the induction of hepatic metallothionein to achieve beneficial effects.

  15. Optimizing Bacillus subtilis spore isolation and quantifying spore harvest purity.

    PubMed

    Harrold, Zoë R; Hertel, Mikaela R; Gorman-Lewis, Drew

    2011-12-01

    Investigating the biochemistry, resilience and environmental interactions of bacterial endospores often requires a pure endospore biomass free of vegetative cells. Numerous endospore isolation methods, however, neglect to quantify the purity of the final endospore biomass. To ensure low vegetative cell contamination we developed a quality control technique that enables rapid quantification of endospore harvest purity. This method quantifies spore purity using bright-field and fluorescence microscopy imaging in conjunction with automated cell counting software. We applied this method to Bacillus subtilis endospore harvests isolated using a two-phase separation method that utilizes mild chemicals. The average spore purity of twenty-two harvests was 88±11% (error is 1σ) with a median value of 93%. A spearman coefficient of 0.97 correlating automated and manual bacterial counts confirms the accuracy of software generated data.

  16. Efficient transformation of Rhizopus delemar by electroporation of germinated spores.

    PubMed

    Xu, Sha; Zhou, Zhengxiong; Du, Guocheng; Zhou, Jingwen; Chen, Jian

    2014-08-01

    High efficient transformation of mycelial fungi is essential to both metabolic engineering and physiological analysis of these industrially important microorganisms. However, transformation efficiencies for mycelial fungi are highly restricted by difficulties in colony formation and competent cell preparation. In this work, an innovative transformation procedure that could significantly improve the efficiency of colony formation and transformation process has been established for a typical mycelial fungus, Rhizopus delemar. Single colonies of R. delemar were obtained with the addition of sodium deoxycholate. Fresh germinated spores of R. delemar were successfully transformed by electroporation. In addition, by pretreatment of the germinated spores with 0.05M lithium acetate (LiAc) and 20mM dithiothreitol (DTT) before electroporation, the transformation efficiency was further improved by 9.5-fold. The final transformation efficiency at optimal conditions reached 1239 transformants/μg DNA. The method described here would facilitate more efficient metabolic engineering and investigation of physiological functions in R. delemar or other similar mycelial fungi.

  17. Differential Inactivation of Fungal Spores in Water and on Seeds by Ozone and Arc Discharge Plasma.

    PubMed

    Kang, Min Ho; Pengkit, Anchalee; Choi, Kihong; Jeon, Seong Sil; Choi, Hyo Won; Shin, Dong Bum; Choi, Eun Ha; Uhm, Han Sup; Park, Gyungsoon

    2015-01-01

    Seed sterilization is essential for preventing seed borne fungal diseases. Sterilization tools based on physical technologies have recently received much attention. However, available information is very limited in terms of efficiency, safety, and mode of action. In this study, we have examined antifungal activity of ozone and arc discharge plasma, potential tools for seed sterilization. In our results, ozone and arc discharge plasma have shown differential antifungal effects, depending on the environment associated with fungal spores (freely submerged in water or infected seeds). Ozone inactivates Fusarium fujikuroi (fungus causing rice bakanae disease) spores submerged in water more efficiently than arc discharge plasma. However, fungal spores associated with or infecting rice seeds are more effectively deactivated by arc discharge plasma. ROS generated in water by ozone may function as a powerful fungicidal factor. On the other hand, shockwave generated from arc discharge plasma may have greatly contributed to antifungal effects on fungus associated with rice seeds. In support of this notion, addition of ultrasonic wave in ozone generating water has greatly increased the efficiency of seed disinfection.

  18. Differential Inactivation of Fungal Spores in Water and on Seeds by Ozone and Arc Discharge Plasma

    PubMed Central

    Kang, Min Ho; Pengkit, Anchalee; Choi, Kihong; Jeon, Seong Sil; Choi, Hyo Won; Shin, Dong Bum; Choi, Eun Ha; Uhm, Han Sup; Park, Gyungsoon

    2015-01-01

    Seed sterilization is essential for preventing seed borne fungal diseases. Sterilization tools based on physical technologies have recently received much attention. However, available information is very limited in terms of efficiency, safety, and mode of action. In this study, we have examined antifungal activity of ozone and arc discharge plasma, potential tools for seed sterilization. In our results, ozone and arc discharge plasma have shown differential antifungal effects, depending on the environment associated with fungal spores (freely submerged in water or infected seeds). Ozone inactivates Fusarium fujikuroi (fungus causing rice bakanae disease) spores submerged in water more efficiently than arc discharge plasma. However, fungal spores associated with or infecting rice seeds are more effectively deactivated by arc discharge plasma. ROS generated in water by ozone may function as a powerful fungicidal factor. On the other hand, shockwave generated from arc discharge plasma may have greatly contributed to antifungal effects on fungus associated with rice seeds. In support of this notion, addition of ultrasonic wave in ozone generating water has greatly increased the efficiency of seed disinfection. PMID:26406468

  19. Compatibility of the entomopathogenic fungus Lecanicillium muscarium and insecticides for eradication of sweetpotato whitefly, Bemisia tabaci.

    PubMed

    Cuthbertson, Andrew G S; Walters, Keith F A; Deppe, Carola

    2005-08-01

    The compatibility of the entomopathogenic fungus Lecanicillium muscarium and chemical insecticides used to control the second instar stages of the sweetpotato whitefly, Bemisia tabaci, was investigated. The effect on spore germination of direct exposure for 24 h to the insecticides imidacloprid, buprofezin, teflubenzuron and nicotine was determined. Only exposure to buprofezin was followed by acceptable spore germination. However, all chemicals significantly reduced spore germination when compared to a water control. Infectivity of L. muscarium in the presence of dry residues of buprofezin, teflubenzuron and nicotine (imidacloprid is a systemic pesticide) on foliage were also investigated. No significant detrimental effects on the level of control of B. tabaci was recorded when compared with fungi applied to residue free foliage on either tomato or verbena plants. Fungi in combination with imidacloprid gave higher B. tabaci mortality on verbena foliage compared to either teflubenzuron or nicotine and fungi combinations. Use of these chemical insecticides with L. muscarium in integrated control programmes for B. tabaci is discussed.

  20. Ultraviolet-Resistant Bacterial Spores

    NASA Technical Reports Server (NTRS)

    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. The Glyoxylate Cycle in an Arbuscular Mycorrhizal Fungus. Carbon Flux and Gene Expression

    PubMed Central

    Lammers, Peter J.; Jun, Jeongwon; Abubaker, Jehad; Arreola, Raul; Gopalan, Anjali; Bago, Berta; Hernandez-Sebastia, Cinta; Allen, James W.; Douds, David D.; Pfeffer, Philip E.; Shachar-Hill, Yair

    2001-01-01

    The arbuscular mycorrhizal (AM) symbiosis is responsible for huge fluxes of photosynthetically fixed carbon from plants to the soil. Lipid, which is the dominant form of stored carbon in the fungal partner and which fuels spore germination, is made by the fungus within the root and is exported to the extraradical mycelium. We tested the hypothesis that the glyoxylate cycle is central to the flow of carbon in the AM symbiosis. The results of 13C labeling of germinating spores and extraradical mycelium with 13C2-acetate and 13C2-glycerol and analysis by nuclear magnetic resonance spectroscopy indicate that there are very substantial fluxes through the glyoxylate cycle in the fungal partner. Full-length sequences obtained by polymerase chain reaction from a cDNA library from germinating spores of the AM fungus Glomus intraradices showed strong homology to gene sequences for isocitrate lyase and malate synthase from plants and other fungal species. Quantitative real-time polymerase chain reaction measurements show that these genes are expressed at significant levels during the symbiosis. Glyoxysome-like bodies were observed by electron microscopy in fungal structures where the glyoxylate cycle is expected to be active, which is consistent with the presence in both enzyme sequences of motifs associated with glyoxysomal targeting. We also identified among several hundred expressed sequence tags several enzymes of primary metabolism whose expression during spore germination is consistent with previous labeling studies and with fluxes into and out of the glyoxylate cycle. PMID:11706207

  2. Reaerosolization of Fluidized Spores in Ventilation Systems▿

    PubMed Central

    Krauter, Paula; Biermann, Arthur

    2007-01-01

    This project examined dry, fluidized spore reaerosolization in a heating, ventilating, and air conditioning duct system. Experiments using spores of Bacillus atrophaeus, a nonpathogenic surrogate for Bacillus anthracis, were conducted to delineate the extent of spore reaerosolization behavior under normal indoor airflow conditions. Short-term (five air-volume exchanges), long-term (up to 21,000 air-volume exchanges), and cycled (on-off) reaerosolization tests were conducted using two common duct materials. Spores were released into the test apparatus in turbulent airflow (Reynolds number, 26,000). After the initial pulse of spores (approximately 1010 to 1011 viable spores) was released, high-efficiency particulate air filters were added to the air intake. Airflow was again used to perturb the spores that had previously deposited onto the duct. Resuspension rates on both steel and plastic duct materials were between 10−3 and 10−5 per second, which decreased to 10 times less than initial rates within 30 min. Pulsed flow caused an initial spike in spore resuspension concentration that rapidly decreased. The resuspension rates were greater than those predicted by resuspension models for contamination in the environment, a result attributed to surface roughness differences. There was no difference between spore reaerosolization from metal and that from plastic duct surfaces over 5 hours of constant airflow. The spores that deposited onto the duct remained a persistent source of contamination over a period of several hours. PMID:17293522

  3. Fungus Resistant XM205 Nonmetallic Cartridge Case,

    DTIC Science & Technology

    CARTRIDGE CASES, *FUNGICIDES, FUNGUS PROOFING, FUNGUS DETERIORATION, RESISTANCE, NITROCELLULOSE, POLYMERS, FIBERS, SYNTHETIC FIBERS, MATERIALS, ZINC COMPOUNDS, ORGANIC COMPOUNDS, ORGANIC SULFUR COMPOUNDS.

  4. On the fate of ingested Bacillus spores.

    PubMed

    Spinosa, M R; Braccini, T; Ricca, E; De Felice, M; Morelli, L; Pozzi, G; Oggioni, M R

    2000-06-01

    Spores of various Bacillus species, including B. subtilis, B. cereus and B. clausii, are used as probiotics, although they are generally absent from the normal microflora of man. We used two nonpathogenic Bacillus species, B. subtilis and B. clausii, to follow the fate of spores inoculated intragastrically in mice. We did not find detectable amounts of vegetative cells in intestinal samples, probably because of high toxicity of the conjugated bile salt taurodeoxycholic acid against Bacillus species. Both spores and cells were detected in the lymph nodes and spleen of one mouse. Our results indicate that Bacillus is present in the intestinal tract solely as spores and that nonpathogenic Bacillus spores may germinate in lymphoid organs, a finding reminiscent of B. anthracis germination in macrophages. These results indicate that any claimed probiotic effect of B. subtilis should be due to spores or, alternatively, to vegetative growth outside the intestine.

  5. Modeling Thermal Inactivation of Bacillus Spores

    DTIC Science & Technology

    2009-03-01

    models which theorize damage to Bacillus spores by various methods. These models use multiple Bacillus species such as anthracis, cereus , and subtilis...MODELING THERMAL INACTIVATION OF BACILLUS SPORES THESIS Emily A. Knight Captain, USAF AFIT/GAM/ENC/09-01 DEPARTMENT OF THE AIR FORCE AIR UNIVERSITY...United States Government. AFIT/GAM/ENC/09-01 MODELING THERMAL INACTIVATION OF BACILLUS SPORES THESIS Presented to the Faculty Department of Mathematics

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

    DTIC Science & Technology

    2008-01-01

    primarily on production of antibodies against the protective antigen component of the anthrax toxins, which are secreted by the bacilli. It has been... production . A spore-associated protein was identified that was specific to the B. cereus group of bacteria and referred to as spore opsonization-associated...in the Ames strain of B. anthracis appeared to increase the phagocytic uptake of the spores in the presence of anti-spore antibodies , since, unlike

  7. Hemp Sesbania (Sesbania exaltata) control in rice (Oryza sativa) with the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene formulated in an invert emulsion

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In greenhouse and field experiments, an invert emulsion (MSG 8.25) was tested with dried, formulated spores of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene, a highly virulent pathogen of the leguminous weed Aeschynomene virginica (northern jointvetch), but considered “...

  8. Riboflavin, overproduced during sporulation of Ashbya gossypii, protects its hyaline spores against ultraviolet light.

    PubMed

    Stahmann, K P; Arst, H N; Althöfer, H; Revuelta, J L; Monschau, N; Schlüpen, C; Gätgens, C; Wiesenburg, A; Schlösser, T

    2001-09-01

    Riboflavin (vitamin B2), essential in tiny amounts as a precursor for oxidoreductase coenzymes, is a yellow pigment. Although it causes cytotoxicity via photoinduced damage of macromolecules, several microorganisms are striking overproducers. A question, unanswered for decades, is whether riboflavin overproducers can benefit from this property. Here, we report an ultraviolet (UV) protective effect of riboflavin. The spores of Ashbya gossypii, a riboflavin-overproducing fungus, are more sensitive to UV than those of Aspergillus nidulans. The addition of riboflavin to suspensions improves the UV resistance of both spore types. Interestingly, we show that regulation of sporulation and riboflavin overproduction in A. gossypii are linked. In batch culture, both were elevated when growth ceased. At constant growth rates, obtained in a chemostat culture, neither was elevated. Supplementation of cultures by cAMP, a known stress signal, negatively affected sporulation as well as riboflavin overproduction, establishing a second, independent argument for the linkage.

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

    PubMed Central

    Cote, Christopher K.; Welkos, Susan L.

    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. PMID:26287244

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

    PubMed

    Cote, Christopher K; Welkos, Susan L

    2015-08-17

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

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

    NASA Astrophysics Data System (ADS)

    Chen, Xianliang; Liu, Xingcun; Sheng, Daping; Huang, Dake; Li, Weizu; Wang, Xin

    2012-11-01

    In this paper, FTIR microspectroscopy was used to identify broken cellular wall Ganoderma lucidum spores and G. lucidum spores. For IR spectra, broken cellular wall G. lucidum spores and G. lucidum spores were mainly different in the regions of 3000-2800, 1660-1600, 1400-1200 and 1100-1000 cm-1. For curve fitting, the results showed the differences in the protein secondary structures and the polysaccharide structures/content between broken cellular wall G. lucidum spores and G. lucidum spores. Moreover, the value of A1078/A1741 might be a potentially useful factor to distinguish broken cellular wall G. lucidum spores from G. lucidum spores. Additionally, FTIR microspectroscopy could identify broken cellular wall G. lucidum spores and G. lucidum spores accurately when it was combined with hierarchical cluster analysis. The result suggests FTIR microspectroscopy is very simple and efficient for distinction of broken cellular wall G. lucidum spores and G. lucidum spores. The result also indicates FTIR microspectroscopy may be useful for TCM identification.

  12. FAR1 and FAR2 Regulate the Expression of Genes Associated with Lipid Metabolism in the Rice Blast Fungus Magnaporthe oryzae

    PubMed Central

    bin Yusof, Mohammad Termizi; Kershaw, Michael J.; Soanes, Darren M.; Talbot, Nicholas J.

    2014-01-01

    The rice blast fungus Magnaporthe oryzae causes plant disease via specialised infection structures called appressoria. These dome-shaped cells are able to generate enormous internal pressure, which enables penetration of rice tissue by invasive hyphae. Previous studies have shown that mobilisation of lipid bodies and subsequent lipid metabolism are essential pre-requisites for successful appressorium-mediated plant infection, which requires autophagic recycling of the contents of germinated spores and germ tubes to the developing appressorium. Here, we set out to identify putative regulators of lipid metabolism in the rice blast fungus. We report the identification of FAR1 and FAR2, which encode highly conserved members of the Zn2-Cys6 family of transcriptional regulators. We generated Δfar1, Δfar2 and Δfar1Δfar2 double mutants in M. oryzae and show that these deletion mutants are deficient in growth on long chain fatty acids. In addition, Δfar2 mutants are also unable to grow on acetate and short chain fatty acids. FAR1 and FAR2 are necessary for differential expression of genes involved in fatty acid β-oxidation, acetyl-CoA translocation, peroxisomal biogenesis, and the glyoxylate cycle in response to the presence of lipids. Furthermore, FAR2 is necessary for expression of genes associated with acetyl-CoA synthesis. Interestingly, Δfar1, Δfar2 and Δfar1Δfar2 mutants show no observable delay or reduction in lipid body mobilisation during plant infection, suggesting that these transcriptional regulators control lipid substrate utilization by the fungus but not the mobilisation of intracellular lipid reserves during infection-related morphogenesis. PMID:24949933

  13. Shape characteristics of biological spores

    NASA Astrophysics Data System (ADS)

    Hahn, Daniel V.; Limsui, Diane; Joseph, Richard I.; Baldwin, Kevin C.; Boggs, Nathan T.; Carr, Alison K.; Carter, Christopher C.; Han, Timothy S.; Thomas, Michael E.

    2008-04-01

    Calculation of scattering properties of biological materials has classically been addressed using numerical calculations based on T-matrix theory. These calculations use bulk optical properties, particle size distribution, and a limited selection of shape descriptors to calculate the resulting aerosol properties. However, the most applicable shape available in T-matrix codes, the spheroid, is not the best descriptor of most biological materials. Based on imagery of the spores of Bacillus atrophaeus and Bacillus anthracis, capsule and egg shapes are mathematically described and programmed into the Amsterdam Discrete Dipole Approximation (ADDA). Spectrally dependent cross sections and depolarization ratios are calculated and a comparison made to spheroidal shapes of equivalent sizes.

  14. Spores

    MedlinePlus

    ... Schmucker R, Bryant K. Antibiotic-associated colitis. In: Cherry JD, Harrison GJ, Kaplan SL, Steinbach WJ, Hotez PJ, eds. Feigin and Cherry's Textbook of Pediatric Infectious Diseases . 7th ed. Philadelphia, ...

  15. Antimicrobial effects of interferon-inducible CXC chemokines against Bacillus anthracis spores and bacilli.

    PubMed

    Crawford, Matthew A; Zhu, Yinghua; Green, Candace S; Burdick, Marie D; Sanz, Patrick; Alem, Farhang; O'Brien, Alison D; Mehrad, Borna; Strieter, Robert M; Hughes, Molly A

    2009-04-01

    Based on previous studies showing that host chemokines exert antimicrobial activities against bacteria, we sought to determine whether the interferon-inducible Glu-Leu-Arg-negative CXC chemokines CXCL9, CXCL10, and CXCL11 exhibit antimicrobial activities against Bacillus anthracis. In vitro analysis demonstrated that all three CXC chemokines exerted direct antimicrobial effects against B. anthracis spores and bacilli including marked reductions in spore and bacillus viability as determined using a fluorometric assay of bacterial viability and CFU determinations. Electron microscopy studies revealed that CXCL10-treated spores failed to undergo germination as judged by an absence of cytological changes in spore structure that occur during the process of germination. Immunogold labeling of CXCL10-treated spores demonstrated that the chemokine was located internal to the exosporium in association primarily with the spore coat and its interface with the cortex. To begin examining the potential biological relevance of chemokine-mediated antimicrobial activity, we used a murine model of inhalational anthrax. Upon spore challenge, the lungs of C57BL/6 mice (resistant to inhalational B. anthracis infection) had significantly higher levels of CXCL9, CXCL10, and CXCL11 than did the lungs of A/J mice (highly susceptible to infection). Increased CXC chemokine levels were associated with significantly reduced levels of spore germination within the lungs as determined by in vivo imaging. Taken together, our data demonstrate a novel antimicrobial role for host chemokines against B. anthracis that provides unique insight into host defense against inhalational anthrax; these data also support the notion for an innovative approach in treating B. anthracis infection as well as infections caused by other spore-forming organisms.

  16. Optical Chromatography of Bacterial Spores

    NASA Astrophysics Data System (ADS)

    Sundbeck, Steven; Terray, Alex; Arnold, Jonathan; Leski, Tomasz; Hart, Sean

    2007-03-01

    The technique of optical chromatography uses a laser mildly focused against fluid flow in a microfluidic channel to trap microscopic particles. Particles in the channel near the focal point of the laser are drawn toward the beam axis and then accelerated via optical pressure against the fluid flow, reaching an equilibrium point when the optical and fluidic forces on the particle are balanced. This equilibrium point may occur at differing distances from the focal point for microscopic particles with differing properties, such as size, shape, morphology, and refractive index. Thus, identification and separation of particles may be achieved in the system. Optical chromatography may be used as a detection technique for biological particles of interest, either directly or as a means of concentrating and filtering a sample. Of particular interest would be reliable methods for detection of Bacillus anthracis, a common weaponized biological agent. In this work we present optical chromatography experiments on bacterial spores which may be environmentally present with B. anthracis spores and interfere with detection.

  17. Epicoccum allergy: skin reaction patterns and spore/mycelium disparities recognized by IgG and IgE ELISA inhibition.

    PubMed

    Portnoy, J; Chapman, J; Burge, H; Muilenberg, M; Solomon, W

    1987-07-01

    Comparable degrees of skin reactivity were observed towards spore and mycelium extracts from two isolates of Epicoccum and to one preparation of Alternaria in 35 rural and 120 university patients. The best experimental extracts detected Epicoccum sensitivity in 70% of the group tested while the commercial extract detected sensitivity in only 6%. Skin reaction correlations were greatest within isolates (eg, spore-A/mycelium-A), then for specific fungus parts (eg, spore-A/spore-B), then between isolates and parts (spore-A/mycelium-B). High correlations were found between individual IgG and IgE ELISA values for all antigens using serum from Epicoccum skin-reactive patients. ELISA inhibition results suggested that significant cross-reactivity exists between Epicoccum and Alternaria antigens recognized by IgG but not by IgE. ELISA inhibition cross-reaction patterns among Epicoccum antigens were comparable to skin reactions while IgG patterns showed little variability. Further characterization of spore/mycelium and interstrain recognition patterns among different immunoglobulin isotypes will be necessary before complete standardization of extracts from different parts of fungi will be possible. The use of spore material for skin testing and treatment of Epicoccum sensitivity appears to be both premature and unnecessary at this time.

  18. Sphagnum moss disperses spores with vortex rings.

    PubMed

    Whitaker, Dwight L; Edwards, Joan

    2010-07-23

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

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

    NASA Astrophysics Data System (ADS)

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

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

  20. Fern spore extracts can damage DNA

    PubMed Central

    Simán, S E; Povey, A C; Ward, T H; Margison, G P; Sheffield, E

    2000-01-01

    The carcinogenicity of the vegetative tissues of bracken fern (Pteridium) has long been established. More recently, the carcinogenic effects of the spores of bracken have also been recognized. Both vegetative tissues and spores of bracken can induce adducts in DNA in animal tissues, but the possible genotoxic or carcinogenic effects of spores from fern species other than bracken are unknown. The single-cell gel electrophoresis (‘comet’) assay was used to investigate whether fern spores can cause DNA damage in vitro. Extracts of spores from six fern species were administered to cultured human premyeloid leukaemia (K562) cells. Spore extracts of five fern species: Anemia phyllitidis, Dicksonia antarctica, Pteridium aquilinum, Pteris vittata and Sadleria pallida, induced significantly more DNA strand breaks than those in the control groups. Only in one species, Osmunda regalis, was the effect no different from that in the control groups. Using extracts from A. phyllitidis and P. vittata, the extent of DNA damage was increased by increasing the original dose 10 times, whereas an experiment in which exposure times were varied suggested that the highest levels of strand breaks appear after 2 h exposure. Simultaneous incubation with human S9 liver enzyme mix ablated the damaging effect of the extracts. Our data show that fern spore extracts can cause DNA damage in human cells in vitro. Considering the strong correlation between DNA damage and carcinogenic events, the observations made in this report may well have some implications for human health. © 2000 Cancer Research Campaign PMID:10883670

  1. Protection of Bacillus pumilus spores by catalases.

    PubMed

    Checinska, Aleksandra; Burbank, Malcolm; Paszczynski, Andrzej J

    2012-09-01

    Bacillus pumilus SAFR-032, isolated at spacecraft assembly facilities of the National Aeronautics and Space Administration Jet Propulsion Laboratory, is difficult to kill by the sterilization method of choice, which uses liquid or vapor hydrogen peroxide. We identified two manganese catalases, YjqC and BPUM_1305, in spore protein extracts of several B. pumilus strains by using PAGE and mass spectrometric analyses. While the BPUM_1305 catalase was present in six of the B. pumilus strains tested, YjqC was not detected in ATCC 7061 and BG-B79. Furthermore, both catalases were localized in the spore coat layer along with laccase and superoxide dismutase. Although the initial catalase activity in ATCC 7061 spores was higher, it was less stable over time than the SAFR-032 enzyme. We propose that synergistic activity of YjqC and BPUM_1305, along with other coat oxidoreductases, contributes to the enhanced resistance of B. pumilus spores to hydrogen peroxide. We observed that the product of the catalase reaction, gaseous oxygen, forms expanding vesicles on the spore surface, affecting the mechanical integrity of the coat layer, resulting in aggregation of the spores. The accumulation of oxygen gas and aggregations may play a crucial role in limiting further exposure of Bacilli spore surfaces to hydrogen peroxide or other toxic chemicals when water is present.

  2. Protection of Bacillus pumilus Spores by Catalases

    PubMed Central

    Checinska, Aleksandra; Burbank, Malcolm

    2012-01-01

    Bacillus pumilus SAFR-032, isolated at spacecraft assembly facilities of the National Aeronautics and Space Administration Jet Propulsion Laboratory, is difficult to kill by the sterilization method of choice, which uses liquid or vapor hydrogen peroxide. We identified two manganese catalases, YjqC and BPUM_1305, in spore protein extracts of several B. pumilus strains by using PAGE and mass spectrometric analyses. While the BPUM_1305 catalase was present in six of the B. pumilus strains tested, YjqC was not detected in ATCC 7061 and BG-B79. Furthermore, both catalases were localized in the spore coat layer along with laccase and superoxide dismutase. Although the initial catalase activity in ATCC 7061 spores was higher, it was less stable over time than the SAFR-032 enzyme. We propose that synergistic activity of YjqC and BPUM_1305, along with other coat oxidoreductases, contributes to the enhanced resistance of B. pumilus spores to hydrogen peroxide. We observed that the product of the catalase reaction, gaseous oxygen, forms expanding vesicles on the spore surface, affecting the mechanical integrity of the coat layer, resulting in aggregation of the spores. The accumulation of oxygen gas and aggregations may play a crucial role in limiting further exposure of Bacilli spore surfaces to hydrogen peroxide or other toxic chemicals when water is present. PMID:22752169

  3. Water absorption in a refractive index model for bacterial spores

    NASA Astrophysics Data System (ADS)

    Siegrist, K. M.; Thrush, E.; Airola, M.; Carr, A. K.; Limsui, D. M.; Boggs, N. T.; Thomas, M. E.; Carter, C. C.

    2009-05-01

    The complexity of biological agents can make it difficult to identify the important factors impacting scattering characteristics among variables such as size, shape, internal structure and biochemical composition, particle aggregation, and sample additives. This difficulty is exacerbated by the environmentally interactive nature of biological organisms. In particular, bacterial spores equilibrate with environmental humidity by absorption/desorption of water which can affect both the complex refractive index and the size/shape distributions of particles - two factors upon which scattering characteristics depend critically. Therefore accurate analysis of experimental data for determination of refractive index must take account of particle water content. First, spectral transmission measurements to determine visible refractive index done on suspensions of bacterial spores must account for water (or other solvent) uptake. Second, realistic calculations of aerosol scattering cross sections should consider effects of atmospheric humidity on particle water content, size and shape. In this work we demonstrate a method for determining refractive index of bacterial spores bacillus atropheus (BG), bacillus thuringiensis (BT) and bacillus anthracis Sterne (BAs) which accounts for these effects. Visible index is found from transmission measurements on aqueous and DMSO suspensions of particles, using an anomalous diffraction approximation. A simplified version of the anomalous diffraction theory is used to eliminate the need for knowledge of particle size. Results using this approach indicate the technique can be useful in determining the visible refractive index of particles when size and shape distributions are not well known but fall within the region of validity of anomalous dispersion theory.

  4. Mechanism by which contact with plant cuticle triggers cutinase gene expression in the spores of Fusarium solani f. sp. pisi

    SciTech Connect

    Woloshuk, C.P.; Kolattukudy, P.E.

    1986-03-01

    Spores of the phytopathogenic fungus Fusarium solani f. sp. pisi were shown to produce the extracellular enzyme, cutinase, only when cutin or cutin hydrolysate was added to the spore suspension. Dihydroxy-C/sub 16/ acid and trihydroxy-C/sub 18/ acid, which are unique cutin monomers, showed the greatest cutinase-inducing activity. Experiments with several compounds structurally related to these fatty acids suggested that both a omega-hydroxyl and a midchain hydroxyl are required for cutinase-inducing activity. Cutinase appeared in the medium 30-45 min after the addition of the inducers to the spore suspension, and the activity level increased for 6 hr. Addition of cycloheximide (5 ..mu..g/ml) completely inhibited cutinase production, suggesting that protein synthesis was involved in the increase of cutinase activity. Immunoblot analysis with rabbit antibodies prepared against cutinase showed that cutinase protein increased in parallel with the increase in enzyme activity. Measurement of cutinase-specific RNA levels by dot-blot hybridization with /sup 32/P-labeled cutinase cDNA showed that the cutinase gene transcripts could be detected within 15 min after addition of the inducers. Addition of exogenous cutinase greatly enhanced the level of cutinase gene transcripts induced by cutin. These results strongly suggest that the fungal spore senses that it is in contact with the plant by the production of small amounts of cutin monomers catalyzed by the low level of cutinase carried by the spore and that these monomers induce the synthesis of cutinase needed for penetration of the fungus into the plant.

  5. Micro-sonicator for spore lysis

    DOEpatents

    Miles, Robin R.; Belgrader, Phillip; Nasarabadi, Shanavaz L.

    2000-01-01

    A micro-sonicator for spore lysis. Using micromachining technology, the micro-sonicator uses ultrasonic excitation of spores to perform spore and cell lysis. The micro-sonicator comprises a container with a cavity therein for retaining the sample in an ultrasonic transmission medium, the cavity being closed by a silicon membrane to which an electrode and piezoelectric material are attached, with the electrode and piezoelectric material being electrically connected to an AC signal generator which causes the membrane to flex and vibrate at the frequency of the applied voltage.

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

    PubMed

    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

  7. Mechanisms of Bacterial Spore Germination and Its Heterogeneity

    DTIC Science & Technology

    2015-01-10

    SECURITY CLASSIFICATION OF: Great progress has been made in understanding the mechanisms of Bacillus and Clostridium spore germination and its...12211 Research Triangle Park, NC 27709-2211 Bacillus , Clostridium, spores, spore germination, germinant receptors, germination heterogeneity REPORT...made in understanding the mechanisms of Bacillus and Clostridium spore germination and its heterogeneity in the 5+ years of the MURI award. The

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

    PubMed

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

    2014-12-01

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

  9. Bacillus Spore Inactivation Methods Affect Detection Assays

    PubMed Central

    Dang, Jessica L.; Heroux, Karen; Kearney, John; Arasteh, Ameneh; Gostomski, Mark; Emanuel, Peter A.

    2001-01-01

    Detection of biological weapons is a primary concern in force protection, treaty verification, and safeguarding civilian populations against domestic terrorism. One great concern is the detection of Bacillus anthracis, the causative agent of anthrax. Assays for detection in the laboratory often employ inactivated preparations of spores or nonpathogenic simulants. This study uses several common biodetection platforms to detect B. anthracis spores that have been inactivated by two methods and compares those data to detection of spores that have not been inactivated. The data demonstrate that inactivation methods can affect the sensitivity of nucleic acid- and antibody-based assays for the detection of B. anthracis spores. These effects should be taken into consideration when comparing laboratory results to data collected and assayed during field deployment. PMID:11472945

  10. Bacterial Spores Survive Electrospray Charging and Desolvation

    NASA Astrophysics Data System (ADS)

    Pratt, Sara N.; Austin, Daniel E.

    2014-05-01

    The survivability of Bacillus subtilis spores and vegetative Escherichia coli cells after electrospray from aqueous suspension was tested using mobility experiments at atmospheric pressure. E. coli did not survive electrospray charging and desolvation, but B. subtilis did. Experimental conditions ensured that any surviving bacteria were de-agglomerated, desolvated, and electrically charged. Based on mobility measurements, B. subtilis spores survived even with 2,000-20,000 positive charges. B. subtilis was also found to survive introduction into vacuum after either positive or negative electrospray. Attempts to measure the charge distribution of viable B. subtilis spores using electrostatic deflection in vacuum were inconclusive; however, viable spores with low charge states (less than 42 positive or less than 26 negative charges) were observed.

  11. [Overview of study on Bacillus subtilis spores].

    PubMed

    Watabe, Kazuhito

    2013-01-01

    This review documents my research for the past 29 years in the work of bacterial sporulation. The Gram-positive bacterium Bacillus subtilis forms spores when conditions are unsuitable for growth. The mature spores remain for long periods of starvation and are resistant to harsh environment. This property is attributed mainly to the unique figures of spore's outer layers, spore coat. The protein composition of the spores was comprehensively analyzed by a combination of SDS-PAGE and LC-MS/MS. The total of 154 proteins were identified and 69 of them were novel. The expression of the genes encoding them was dependent on sporulation-specific sigma factors, σF, σE, σG and σK. The expression of a coat protein gene, cotS, was dependent on σK and GerE. CotE is essential for the assembly of CotS in the coat layer. Many coat genes were identified by reverse genetics and the regulation of the gene expression was studied in detail. Some cot genes are functioned in the resistance to heat and lysozyme, and some of the coat proteins are involved in the specificity of germinants. The yrbA is essential in spore development, yrbA deficient cells revealed abnormal figures of spore coat structure and changed the response to germinants. The location of 16 coat proteins was determined by the observation of fluorescence microscopy using fluorescence-labelled proteins. One protein was assigned to the cortex, nine to the inner coat, and four to the outer coat. In addition, CotZ and CgeA appeared in the outermost layer of the spore coat.

  12. Rapid onsite assessment of spore viability.

    SciTech Connect

    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.

  13. Draft Genome Sequence of Two Monosporidial Lines of the Karnal Bunt Fungus Tilletia indica Mitra (PSWKBGH-1 and PSWKBGH-2)

    PubMed Central

    Sharma, Pradeep; Saharan, M. S.; Sharma, Indu; Kumar, Jitender; Mishra, Shefali; Muthusamy, Senthilkumar K.; Gupta, R. K.; Jaiswal, Sarika; Iquebal, M. A.; Angadi, U. B.; Kumar, Neeraj; Fatma, Samar; Rai, Anil; Kumar, Dinesh

    2016-01-01

    Karnal bunt disease caused by the fungus Tilletia indica Mitra is a serious concern due to strict quarantines affecting international trade of wheat. We announce here the first draft assembly of two monosporidial lines, PSWKBGH-1 and -2, of this fungus, having approximate sizes of 37.46 and 37.21 Mbp, respectively. PMID:27634992

  14. Bacterial spores in silage and raw milk.

    PubMed

    te Giffel, M C; Wagendorp, A; Herrewegh, A; Driehuis, F

    2002-08-01

    Spore-forming bacteria can survive food-processing treatments. In the dairy industry, Bacillus and Clostridium species determine the shelf-life of a variety of heat-treated milk products, mainly if the level of post-process contamination is low. In order to minimize problems caused by bacterial spores in foods and food production processes a chain management approach, from raw materials, ingredients and environmental sources to final product storage conditions, is most effective. Silage is considered to be a significant source of contamination of raw milk with spores. PCR-RAPD fingerprinting and heat resistance studies of populations of aerobic spore-formers isolated from grass and maize silage and from raw milk confirmed this assumption. Prevention of outgrowth of aerobic spores in silage will contribute to reduction of the total spore load of raw milk. Therefore, it is important that the silage fermentation process is controlled. Application of cultures of lactic acid bacteria or chemical additives can aid silage fermentation and improve aerobic stability.

  15. Spore inoculum optimization to maximize cyclosporin A production in Tolypocladium niveum.

    PubMed

    Lee, Mi-Jin; Lee, Han-Na; Han, Kyubeom; Kim, Eung-Soo

    2008-05-01

    The cyclic undecapeptide, cyclosporin A (CyA), is one of the most commonly prescribed immunosuppressive drugs. It is generated nonribosomally from a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. In order to maximize the production of CyA by wild-type T. niveum (ATCC 34921), each of three culture stages (sporulation culture, growth culture, and production culture) were sequentially optimized. Among the three potential sporulation media, the SSMA medium generated the highest numbers of T. niveum spores. The SSM and SM media were then selected as the optimal growth and production culture media, respectively. The addition of valine and fructose to the SM production medium was also determined to be crucial for CyA biosynthesis. In this optimized three-stage culture system, 3% of the spore inoculum generated the highest level of CyA productivity in a 15-day T. niveum production culture, thereby implying that the determination of an appropriate size of T. niveum spore inoculum plays a critical role in the maximization of CyA production.

  16. Bacterial communities in termite fungus combs are comprised of consistent gut deposits and contributions from the environment.

    PubMed

    Otani, Saria; Hansen, Lars H; Sørensen, Søren J; Poulsen, Michael

    2016-01-01

    Fungus-growing termites (subfamily Macrotermitinae) mix plant forage with asexual spores of their plant-degrading fungal symbiont Termitomyces in their guts and deposit this blend in fungus comb structures, within which the plant matter is degraded. As Termitomyces grows, it produces nodules with asexual spores, which the termites feed on. Since all comb material passes through termite guts, it is inevitable that gut bacteria are also deposited in the comb, but it has remained unknown which bacteria are deposited and whether distinct comb bacterial communities are sustained. Using high-throughput sequencing of the 16S rRNA gene, we explored the bacterial community compositions of 33 fungus comb samples from four termite species (three genera) collected at four South African geographic locations in 2011 and 2013. We identified 33 bacterial phyla, with Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, and Candidate division TM7 jointly accounting for 92 % of the reads. Analyses of gut microbiotas from 25 of the 33 colonies showed that dominant fungus comb taxa originate from the termite gut. While gut communities were consistent between 2011 and 2013, comb community compositions shifted over time. These shifts did not appear to be due to changes in the taxa present, but rather due to differences in the relative abundances of primarily gut-derived bacteria within fungus combs. This indicates that fungus comb microbiotas are largely termite species-specific due to major contributions from gut deposits and also that environment affects which gut bacteria dominate comb communities at a given point in time.

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

    DTIC Science & Technology

    2013-02-01

    P. Setlow. Mechanism of killing of spores of Bacillus cereus and Bacillus megaterium by wet heat, Letters in Applied Microbiology, (02 2010...REPORT Inactivation of spores of Bacillus species by wet heat: studies on single spores using laser tweezers Taman spectroscopy (Final Report) 14...heterogeneity of single Bacillus spores during wet-heat treatment that are commonly used in spore killing and inactivation. Achievements include: (1

  18. Tilletia puccinelliae, a new species of reticulate-spored bunt fungus infecting Puccinellia distans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A shipment of seeds of Fults alkali grass (Puccinellia distans) grown in Washington state containing bunted seeds was intercepted by quarantine officials at China’s Tianjin Entry-Exit Quarantine and Inspection Bureau. The bunted seeds were filled with irregularly shaped, reticulately ornamented teli...

  19. Sensitive, Rapid Detection of Bacterial Spores

    NASA Technical Reports Server (NTRS)

    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.

  20. Ultrastructure of spore development in Scutellospora heterogama.

    PubMed

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

    2007-07-01

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

  1. The Influence of Sporulation Conditions on the Spore Coat Protein Composition of Bacillus subtilis Spores

    PubMed Central

    Abhyankar, Wishwas R.; Kamphorst, Kiki; Swarge, Bhagyashree N.; van Veen, Henk; van der Wel, Nicole N.; Brul, Stanley; de Koster, Chris G.; de Koning, Leo J.

    2016-01-01

    Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has also been shown that the more mature a spore is, the higher is its heat resistance most likely mediated, at least in part, by the progressive cross-linking of coat proteins. The current study for the first time assesses, at the proteomic level, the effect of two commonly used sporulation conditions on spore protein presence. 14N spores prepared on solid Schaeffer’s-glucose (SG) agar plates and 15N metabolically labeled spores prepared in shake flasks containing 3-(N-morpholino) propane sulfonic acid (MOPS) buffered defined liquid medium differ in their coat protein composition as revealed by LC-FT-MS/MS analyses. The former condition mimics the industrial settings while the latter conditions mimic the routine laboratory environment wherein spores are developed. As seen previously in many studies, the spores prepared on the solid agar plates show a higher thermal resistance than the spores prepared under liquid culture conditions. The 14N:15N isotopic ratio of the 1:1 mixture of the spore suspensions exposes that most of the identified inner coat and crust proteins are significantly more abundant while most of the outer coat proteins are significantly less abundant for the spores prepared on solid SG agar plates relative to the spores prepared in the liquid MOPS buffered defined medium. Sporulation condition-specific differences and variation in isotopic ratios between the tryptic peptides of expected cross-linked proteins suggest that the coat protein cross-linking may also be condition specific. Since the core dipicolinic acid content is found to be similar in both the spore populations, it appears that the difference in wet heat resistance is connected to the

  2. An appraisal of Impregon as a deterrent of domestic fungus growth.

    PubMed

    Burge, H A; Solomon, W R; Boise, J R

    1976-10-01

    Impregon brand of tetrachlorsalicylanilide (TCSA) has been profferred widely as a household fungistatic agent, although its value remains unproved. To assess its effects, this agent was used as a laundry and paint additive and as a treatment for burlap rug backing; after recommended applications of Impregon, coded replicate materials were inoculated with mixed suspensions of fungus particles. No difference was evident in fungus growth points on fabric swatches washed in tap water with and without Impregon. However, growth on both of these was significantly greater than on samples laundered in tap water using only a commercial soap or liquid detergent. Fungus soiling of burlap was not clearly diminished by prescribed applications of Impregon solution 3 mo previously. Similarly, the addition of this agent to paint did not suppress fungus growth on Masonite plaques to which it had been applied. However, comparable levels of Impregon incorporated into agar media substantially inhibited spore germination. These findings suggest that the bioavailability of TCSA is insufficient to provide desired household antifungal effects when Impregon is used in accord with current recommendations.

  3. Seasonal fungus prevalence inside and outside of domestic environments in the subtropical climate

    NASA Astrophysics Data System (ADS)

    Kuo, Yu-Mei; Li, Chin-Shan

    Airborne fungi were collected using the N6 Andersen sampler at 1-month intervals for I yr inside and outside of six apartments in Taipei. It was shown that seasonal variations of indoor and outdoor fungus number concentrations were remarkable and indoor and outdoor air spore counts varied considerably from residence to residence. The geometric mean concentrations of indoor and outdoor fungi were found to be higher than 1000 CFU m -3 during the summer months and abruptly decreased to below 100 CFU m -3 in the winter. A high correlation coefficient was found between fungus concentrations in living rooms and outdoors. Moreover, the ratios of indoor to outdoor fungus concentrations (0.21-3.81) were too low to indicate the presence of any indoor fungus sources. A large variety of mold genera was isolated, and Aspergillus, Penicillium, Cladosporium, and yeast were observed to be predominant. Indoors, Penicillium showed the highest concentrations in the summer and autumn months, while Asperyillus and Cladosporium were also observed frequently. The outside air was dominated by Asperyillus, Penicillium, and Cladosporium in spring, summer, and autumn, but by Penicillium and yeast during winter months. In addition, Cladosporium was found to be absent indoors and outdoors in the winter.

  4. FORMATION AND STRUCTURE OF THE SPORE OF BACILLUS COAGULANS

    PubMed Central

    Ohye, D. F.; Murrell, W. G.

    1962-01-01

    Spore formation in Bacillus coagulans has been studied by electron microscopy using an epoxy resin (Araldite) embedding technique. The developmental stages from the origin of the initial spore septum to the mature spore were investigated. The two forespore membranes developed from the double layer of cytoplasmic membrane. The cortex was progressively deposited between these two membranes. The inner membrane finally became the spore protoplasmic membrane, and the outer membrane part of the inner spore coat or the outer spore coat itself. In the mature spore the completed integuments around the spore protoplasm consisted of the cortex, a laminated inner coat, and a dense outer coat. No exosporium was observed. The method of formation of the cortex and the spore coats is discussed. PMID:14481435

  5. ROS Involves the Fungicidal Actions of Thymol against Spores of Aspergillus flavus via the Induction of Nitric Oxide

    PubMed Central

    Shen, Qingshan; Zhou, Wei; Li, Hongbo; Hu, Liangbin; Mo, Haizhen

    2016-01-01

    Aspergillus flavus is a well-known pathogenic fungus for both crops and human beings. The acquisition of resistance to azoles by A. flavus is leading to more failures occurring in the prevention of infection by A. flavus. In this study, we found that thymol, one of the major chemical constituents of the essential oil of Monarda punctate, had efficient fungicidal activity against A. flavus and led to sporular lysis. Further studies indicated that thymol treatment induced the generation of both ROS and NO in spores, whereas NO accumulation was far later than ROS accumulation in response to thymol. By blocking ROS production with the inhibitors of NADPH oxidase, NO generation was also significantly inhibited in the presence of thymol, which indicated that ROS induced NO generation in A. flavus in response to thymol treatment. Moreover, the removal of either ROS or NO attenuated lysis and death of spores exposed to thymol. The addition of SNP (exogenous NO donor) eliminated the protective effects of the inhibitors of NADPH oxidase on thymol-induced lysis and death of spores. Taken together, it could be concluded that ROS is involved in spore death induced by thymol via the induction of NO. PMID:27196096

  6. Immunolocalization of an Alternative Respiratory Chain in Antonospora (Paranosema) locustae Spores: Mitosomes Retain Their Role in Microsporidial Energy Metabolism ▿

    PubMed Central

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

    2011-01-01

    Microsporidia are a group of fungus-related intracellular parasites with severely reduced metabolic machinery. They lack canonical mitochondria, a Krebs cycle, and a respiratory chain but possess genes encoding glycolysis enzymes, a glycerol phosphate shuttle, and ATP/ADP carriers to import host ATP. The recent finding of alternative oxidase genes in two clades suggests that microsporidial mitosomes may retain an alternative respiratory pathway. We expressed the fragments of mitochondrial chaperone Hsp70 (mitHsp70), mitochondrial glycerol-3-phosphate dehydrogenase (mitG3PDH), and alternative oxidase (AOX) from the microsporidium Antonospora (Paranosema) locustae in Escherichia coli. Immunoblotting with antibodies against recombinant polypeptides demonstrated specific accumulation of both metabolic enzymes in A. locustae spores. At the same time comparable amounts of mitochondrial Hsp70 were found in spores and in stages of intracellular development as well. Immunoelectron microscopy of ultrathin cryosections of spores confirmed mitosomal localization of the studied proteins. Small amounts of enzymes of an alternative respiratory chain in merogonial and early sporogonial stages, alongside their accumulation in mature spores, suggest conspicuous changes in components and functions of mitosomes during the life cycle of microsporidia and the important role of these organelles in parasite energy metabolism, at least at the final stages of sporogenesis. PMID:21296913

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

    PubMed

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

    2011-04-01

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

  8. Pulmonary Injury after Combined Exposures to Low-Dose Low-LET Radiation and Fungal Spores

    PubMed Central

    Marples, B.; Downing, L.; Sawarynski, K. E.; Finkelstein, J. N.; Williams, J. P.; Martinez, A. A.; Wilson, G. D.; Sims, M. D.

    2013-01-01

    Exposure to infectious microbes is a likely confounder after a nuclear terrorism event. In combination with radiation, morbidity and mortality from an infection may increase significantly. Pulmonary damage after low-dose low-LET irradiation is characterized by an initial diffuse alveolar inflammation. By contrast, inhaled fungal spores produce localized damage around pulmonary bronchioles. In the present study, we assessed lung injury in C57BL/6 mice after combined exposures to whole-body X radiation and inhaled fungal spores. Either animals were exposed to Aspergillus spores and immediately irradiated with 2 Gy, or the inoculation and irradiation were separated by 8 weeks. Pulmonary injury was assessed at 24 and 48 h and 1, 2, 4, 8, and 24 weeks later using standard H&E-stained sections and compared with sham-treated age-matched controls. Immunohistochemistry for invasive inflammatory cells (macrophages, neutrophils and B and T lymphocytes) was performed. A semi-quantitative assessment of pulmonary injury was made using three distinct parameters: local infiltration of inflammatory cells, diffuse inflammation, and thickening and distortion of alveolar architecture. Radiation-induced changes in lung architecture were most evident during the first 2 weeks postexposure. Fungal changes were seen over the first 4 weeks. Simultaneous combined exposures significantly increased the duration of acute pulmonary damage up to 24 weeks (P < 0.01). In contrast, administration of the fungus 8 weeks after irradiation did not produce enhanced levels of acute pulmonary damage. These data imply that the inhalation of fungal spores at the time of a radiation exposure alters the susceptibility of the lungs to radiation-induced injury. PMID:21275606

  9. Transcriptome sequencing and characterization of ungerminated and germinated spores of Nosema bombycis

    PubMed Central

    Liu, Han; Li, Mingqian; He, Xinyi; Cai, Shunfeng; He, Xiangkang; Lu, Xingmeng

    2016-01-01

    Nosema bombycis is an obligate intracellular parasitic fungus that utilizes a distinctive mechanism to infect Bombyx mori. Germination, an indispensible process through which microsporidia infect the host cells, is regarded as a key developmental turning point for microsporidia from dormant state to reproduction state. Thus, elucidating the transcriptome changes before and after germination is crucial for parasite control. However, the molecular basis of germination of microsporidia remains unknown. To investigate this germination process, the transcriptome of N. bombycis ungerminated spores and germinated spores were sequenced and analyzed. More than 60 million high-quality transcript reads were generated from these two groups using RNA-Seq technology. After assembly, 2756 and 2690 unigenes were identified, respectively, and subsequently annotated based on known proteins. After analysis of differentially expressed genes, 66 genes were identified to be differentially expressed (P ≤ 0.05) between these two groups. A protein phosphatase-associated gene was first identified to be significantly up-regulated as determined by RNA-Seq and immunoblot analysis, indicating that dephosphorylation might potentially contribute to microsporidia germination. The DEGs that encode proteins involved in glycometabolism, spore wall proteins and ricin B lectin of N. bombycis were also analyzed. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed genes responsible for some specific biological functions and processes. The datasets generated in this study provide a basic characterization of the transcriptome changes in N. bombycis during germination. The analysis of transcriptome data and identification of certain functional genes which are robust candidate genes related to germination will help to provide a deep understanding of spore germination and invasion. PMID:26837419

  10. Method for collecting spores from a mold

    DOEpatents

    Au, Frederick H. F.; Beckert, Werner F.

    1977-01-01

    A technique and apparatus used therewith for determining the uptake of plutonium and other contaminants by soil microorganisms which, in turn, gives a measure of the plutonium and/or other contaminants available to the biosphere at that particular time. A measured quantity of uncontaminated spores of a selected mold is added to a moistened sample of the soil to be tested. The mixture is allowed to sit a predetermined number of days under specified temperature conditions. An agar layer is then applied to the top of the sample. After three or more days, when spores of the mold growing in the sample have formed, the spores are collected by a miniature vacuum collection apparatus operated under preselected vacuum conditions, which collect only the spores with essentially no contamination by mycelial fragments or culture medium. After collection, the fungal spores are dried and analyzed for the plutonium and/or other contaminants. The apparatus is also suitable for collection of pollen, small insects, dust and other small particles, material from thin-layer chromatography plates, etc.

  11. Aerodynamics of puffball mushroom spore dispersal

    NASA Astrophysics Data System (ADS)

    Amador, Guillermo; Barberie, Alex; Hu, David

    2012-11-01

    Puffball mushrooms Lycoperdon are spherical fungi that release a cloud of spores in response to raindrop impacts. In this combined experimental and theoretical study, we elucidate the aerodynamics of this unique impact-based spore-dispersal. We characterize live puffball ejections by high speed video, the geometry and elasticity of their shells by cantilever experiments, and the packing fraction and size of their spores by scanning electron microscope. We build a dynamically similar puffball mimic composed of a tied-off latex balloon filled with baby powder and topped with a 1-cm slit. A jet of powder is elicited by steady lateral compression of the mimic between two plates. The jet height is a bell-shaped function of force applied, with a peak of 18 cm at loads of 45 N. We rationalize the increase in jet height with force using Darcy's Law: the applied force generates an overpressure maintained by the air-tight elastic membrane. Pressure is relieved as the air travels through the spore interstitial spaces, entrains spores, and exits through the puffball orifice. This mechanism demonstrates how powder-filled elastic shells can generate high-speed jets using energy harvested from rain.

  12. Multigeneration Cross Contamination of Mail with Bacillus Species Spores by Tumbling ▿

    PubMed Central

    Edmonds, Jason; Clark, Paul; Williams, Leslie; Lindquist, H. D. Alan; Martinez, Kenneth; Gardner, Warren; Shadomy, Sean; Hornsby-Myers, Jennifer

    2010-01-01

    In 2001, envelopes loaded with Bacillus anthracis spores were mailed to Senators Daschle and Leahy as well as to the New York Post and NBC News buildings. Additional letters may have been mailed to other news agencies because there was confirmed anthrax infection of employees at these locations. These events heightened the awareness of the lack of understanding of the mechanism(s) by which objects contaminated with a biological agent might spread disease. This understanding is crucial for the estimation of the potential for exposure to ensure the appropriate response in the event of future attacks. In this study, equipment to simulate interactions between envelopes and procedures to analyze the spread of spores from a “payload” envelope (i.e., loaded internally with a powdered spore preparation) onto neighboring envelopes were developed. Another process to determine whether an aerosol could be generated by opening contaminated envelopes was developed. Subsequent generations of contaminated envelopes originating from a single payload envelope showed a consistent two-log decrease in the number of spores transferred from one generation to the next. Opening a tertiary contaminated envelope resulted in an aerosol containing 103 B. anthracis spores. We developed a procedure for sampling contaminated letters by a nondestructive method aimed at providing information useful for consequence management while preserving the integrity of objects contaminated during the incident and preserving evidence for law enforcement agencies. PMID:20511424

  13. Multigeneration cross contamination of mail with Bacillus species spores by tumbling.

    PubMed

    Edmonds, Jason; Clark, Paul; Williams, Leslie; Lindquist, H D Alan; Martinez, Kenneth; Gardner, Warren; Shadomy, Sean; Hornsby-Myers, Jennifer

    2010-07-01

    In 2001, envelopes loaded with Bacillus anthracis spores were mailed to Senators Daschle and Leahy as well as to the New York Post and NBC News buildings. Additional letters may have been mailed to other news agencies because there was confirmed anthrax infection of employees at these locations. These events heightened the awareness of the lack of understanding of the mechanism(s) by which objects contaminated with a biological agent might spread disease. This understanding is crucial for the estimation of the potential for exposure to ensure the appropriate response in the event of future attacks. In this study, equipment to simulate interactions between envelopes and procedures to analyze the spread of spores from a "payload" envelope (i.e., loaded internally with a powdered spore preparation) onto neighboring envelopes were developed. Another process to determine whether an aerosol could be generated by opening contaminated envelopes was developed. Subsequent generations of contaminated envelopes originating from a single payload envelope showed a consistent two-log decrease in the number of spores transferred from one generation to the next. Opening a tertiary contaminated envelope resulted in an aerosol containing 10(3) B. anthracis spores. We developed a procedure for sampling contaminated letters by a nondestructive method aimed at providing information useful for consequence management while preserving the integrity of objects contaminated during the incident and preserving evidence for law enforcement agencies.

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

    SciTech Connect

    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

  15. Bacterial spores and chemical sporicidal agents.

    PubMed Central

    Russell, A D

    1990-01-01

    Bacterial spores are among the most resistant of all living cells to biocides, although the response depends on the stage of sporulation. The development of resistance to some agents such as chlorhexidine occurs much earlier in sporulation than does resistance to glutaraldehyde, which is a very late event. During germination or outgrowth or both, resistance is lost and the cells become as susceptible to biocides as nonsporulating bacteria. Mechanisms of spore resistance to, and the action of, biocides are discussed, and possible means of enhancing antispore activity are considered. The clinical and other uses of sporicidal and sporostatic chemical agents are described. Images PMID:2187595

  16. Factors affecting spore germination in algae - review.

    PubMed

    Agrawal, S C

    2009-01-01

    This review surveys whatever little is known on the influence of different environmental factors like light, temperature, nutrients, chemicals (such as plant hormones, vitamins, etc.), pH of the medium, biotic factors (such as algal extracellular substances, algal concentration, bacterial extracellular products, animal grazing and animal extracellular products), water movement, water stress, antibiotics, UV light, X-rays, gamma-rays, and pollution on the spore germination in algae. The work done on the dormancy of algal spores and on the role of vegetative cells in tolerating environmental stress is also incorporated.

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

    PubMed Central

    Francis, Michael B.; Allen, Charlotte A.

    2015-01-01

    ABSTRACT Bacterial spore germination is a process whereby a dormant spore returns to active, vegetative growth, and this process has largely been studied in the model organism Bacillus subtilis. In B. subtilis, the initiation of germinant receptor-mediated spore germination is divided into two genetically separable stages. Stage I is characterized by the release of dipicolinic acid (DPA) from the spore core. Stage II is characterized by cortex degradation, and stage II is activated by the DPA released during stage I. Thus, DPA release precedes cortex hydrolysis during B. subtilis spore germination. Here, we investigated the timing of DPA release and cortex hydrolysis during Clostridium difficile spore germination and found that cortex hydrolysis precedes DPA release. Inactivation of either the bile acid germinant receptor, cspC, or the cortex hydrolase, sleC, prevented both cortex hydrolysis and DPA release. Because both cortex hydrolysis and DPA release during C. difficile spore germination are dependent on the presence of the germinant receptor and the cortex hydrolase, the release of DPA from the core may rely on the osmotic swelling of the core upon cortex hydrolysis. These results have implications for the hypothesized glycine receptor and suggest that the initiation of germinant receptor-mediated C. difficile spore germination proceeds through a novel germination pathway. IMPORTANCE Clostridium difficile infects antibiotic-treated hosts and spreads between hosts as a dormant spore. In a host, spores germinate to the vegetative form that produces the toxins necessary for disease. C. difficile spore germination is stimulated by certain bile acids and glycine. We recently identified the bile acid germinant receptor as the germination-specific, protease-like CspC. CspC is likely cortex localized, where it can transmit the bile acid signal to the cortex hydrolase, SleC. Due to the differences in location of CspC compared to the Bacillus subtilis germinant

  18. Bacillus anthracis spore interactions with mammalian cells: Relationship between germination state and the outcome of in vitro

    PubMed Central

    2011-01-01

    Background During inhalational anthrax, internalization of Bacillus anthracis spores by host cells within the lung is believed to be a key step for initiating the transition from the localized to disseminated stages of infection. Despite compelling in vivo evidence that spores remain dormant within the bronchioalveolar spaces of the lungs, and germinate only after uptake into host cells, most in vitro studies of infection have been conducted under conditions that promote rapid germination of spores within the culture medium. Results Using an in vitro model of infection, we evaluated the influence of the germination state of B. anthracis spores, as controlled by defined culture conditions, on the outcome of infection. Spores prepared from B. anthracis Sterne 7702 germinated in a variety of common cell culture media supplemented with fetal bovine serum (FBS) while, in the absence of FBS, germination was strictly dependent on medium composition. RAW264.7 macrophage-like cells internalized spores to the same extent in either germinating or non-germinating media. However, significantly more viable, intracellular B. anthracis were recovered from cells infected under non-germinating conditions compared to germinating conditions. At the same time, RAW264.7 cells demonstrated a significant loss in viability when infected under non-germinating conditions. Conclusions These results suggest that the outcome of host cell infection is sensitive to the germination state of spores at the time of uptake. Moreover, this study demonstrates the efficacy of studying B. anthracis spore infection of host cells within a defined, non-germinating, in vitro environment. PMID:21356113

  19. Production and counting of spores of Clostridium chauvoei.

    PubMed

    Bagadi, H O

    1977-06-01

    The concentration and viability of spores produced by four different strains of Clostridium chauvoei (C. feseri) grown in a modified medium for 18 days are described. The medium yielded enough viable spores for experimental work.

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

    PubMed

    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.

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

    DTIC Science & Technology

    2015-01-14

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

  2. Growth of ferns from spores in axenic culture.

    PubMed

    Ford, M V; Fay, M F

    1990-01-01

    In this chapter, a method by which many fern species can be successfully grown from spores in axenic culture will be described. Unlike the conventional method of sowing the spores on compost, this method allows spore populations free from contamination by spores of other species to be sown. The method can be used for the production of mature sporophytes or to provide a controllable system for biosystematic studies of, or experimentation with, fern gametophytes (1,2).

  3. Targeted Disruption of Melanin Biosynthesis Genes in the Human Pathogenic Fungus Lomentospora prolificans and Its Consequences for Pathogen Survival.

    PubMed

    Al-Laaeiby, Ayat; Kershaw, Michael J; Penn, Tina J; Thornton, Christopher R

    2016-03-24

    The dematiaceous (melanised) fungus Lomentospora (Scedosporium) prolificans is a life-threatening opportunistic pathogen of immunocompromised humans, resistant to anti-fungal drugs. Melanin has been shown to protect human pathogenic fungi against antifungal drugs, oxidative killing and environmental stresses. To determine the protective role of melanin in L. prolificans to oxidative killing (H₂O₂), UV radiation and the polyene anti-fungal drug amphotericin B, targeted gene disruption was used to generate mutants of the pathogen lacking the dihydroxynaphthalene (DHN)-melanin biosynthetic enzymes polyketide synthase (PKS1), tetrahydroxynapthalene reductase (4HNR) and scytalone dehydratase (SCD1). Infectious propagules (spores) of the wild-type strain 3.1 were black/brown, whereas spores of the PKS-deficient mutant ΔLppks1::hph were white. Complementation of the albino mutant ΔLppks1::hph restored the black-brown spore pigmentation, while the 4HNR-deficient mutant ΔLp4hnr::hph and SCD-deficient mutant ΔLpscd1::hph both produced orange-yellow spores. The mutants ΔLppks1::hph and ΔLp4hnr::hph showed significant reductions in spore survival following H₂O₂ treatment, while spores of ΔLpscd1::hph and the ΔLppks1::hph complemented strain ΔLppks1::hph:PKS showed spore survivals similar to strain 3.1. Spores of the mutants ΔLp4hnr::hph and ΔLpscd1::hph and complemented strain ΔLppks1::hph:PKS showed spore survivals similar to 3.1 following exposure to UV radiation, but survival of ΔLppks1::hph spores was significantly reduced compared to the wild-type strain. Strain 3.1 and mutants ΔLp4hnr::hph and ΔLppks1::hph:PKS were resistant to amphotericin B while, paradoxically, the PKS1- and SCD1-deficient mutants showed significant increases in growth in the presence of the antifungal drug. Taken together, these results show that while melanin plays a protective role in the survival of the pathogen to oxidative killing and UV radiation, melanin does not

  4. Targeted Disruption of Melanin Biosynthesis Genes in the Human Pathogenic Fungus Lomentospora prolificans and Its Consequences for Pathogen Survival

    PubMed Central

    Al-Laaeiby, Ayat; Kershaw, Michael J.; Penn, Tina J.; Thornton, Christopher R.

    2016-01-01

    The dematiaceous (melanised) fungus Lomentospora (Scedosporium) prolificans is a life-threatening opportunistic pathogen of immunocompromised humans, resistant to anti-fungal drugs. Melanin has been shown to protect human pathogenic fungi against antifungal drugs, oxidative killing and environmental stresses. To determine the protective role of melanin in L. prolificans to oxidative killing (H2O2), UV radiation and the polyene anti-fungal drug amphotericin B, targeted gene disruption was used to generate mutants of the pathogen lacking the dihydroxynaphthalene (DHN)-melanin biosynthetic enzymes polyketide synthase (PKS1), tetrahydroxynapthalene reductase (4HNR) and scytalone dehydratase (SCD1). Infectious propagules (spores) of the wild-type strain 3.1 were black/brown, whereas spores of the PKS-deficient mutant ΔLppks1::hph were white. Complementation of the albino mutant ΔLppks1::hph restored the black-brown spore pigmentation, while the 4HNR-deficient mutant ΔLp4hnr::hph and SCD-deficient mutant ΔLpscd1::hph both produced orange-yellow spores. The mutants ΔLppks1::hph and ΔLp4hnr::hph showed significant reductions in spore survival following H2O2 treatment, while spores of ΔLpscd1::hph and the ΔLppks1::hph complemented strain ΔLppks1::hph:PKS showed spore survivals similar to strain 3.1. Spores of the mutants ΔLp4hnr::hph and ΔLpscd1::hph and complemented strain ΔLppks1::hph:PKS showed spore survivals similar to 3.1 following exposure to UV radiation, but survival of ΔLppks1::hph spores was significantly reduced compared to the wild-type strain. Strain 3.1 and mutants ΔLp4hnr::hph and ΔLppks1::hph:PKS were resistant to amphotericin B while, paradoxically, the PKS1- and SCD1-deficient mutants showed significant increases in growth in the presence of the antifungal drug. Taken together, these results show that while melanin plays a protective role in the survival of the pathogen to oxidative killing and UV radiation, melanin does not contribute to

  5. 9 CFR 113.66 - Anthrax Spore Vaccine-Nonencapsulated.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Anthrax Spore Vaccine-Nonencapsulated... REQUIREMENTS Live Bacterial Vaccines § 113.66 Anthrax Spore Vaccine—Nonencapsulated. Anthrax Spore Vaccine.... All serials of vaccine shall be prepared from the first through the fifth passage from the Master...

  6. 9 CFR 113.66 - Anthrax Spore Vaccine-Nonencapsulated.

    Code of Federal Regulations, 2010 CFR

    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.... All serials of vaccine shall be prepared from the first through the fifth passage from the Master...

  7. 9 CFR 113.66 - Anthrax Spore Vaccine-Nonencapsulated.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Anthrax Spore Vaccine-Nonencapsulated... REQUIREMENTS Live Bacterial Vaccines § 113.66 Anthrax Spore Vaccine—Nonencapsulated. Anthrax Spore Vaccine.... All serials of vaccine shall be prepared from the first through the fifth passage from the Master...

  8. 9 CFR 113.66 - Anthrax Spore Vaccine-Nonencapsulated.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Anthrax Spore Vaccine-Nonencapsulated... REQUIREMENTS Live Bacterial Vaccines § 113.66 Anthrax Spore Vaccine—Nonencapsulated. Anthrax Spore Vaccine.... All serials of vaccine shall be prepared from the first through the fifth passage from the Master...

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

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Anthrax Spore Vaccine-Nonencapsulated... REQUIREMENTS Live Bacterial Vaccines § 113.66 Anthrax Spore Vaccine—Nonencapsulated. Anthrax Spore Vaccine.... All serials of vaccine shall be prepared from the first through the fifth passage from the Master...

  10. Classification of Streptomyces Spore Surfaces into Five Groups

    PubMed Central

    Dietz, Alma; Mathews, John

    1971-01-01

    Streptomyces spores surfaces have been classified into five groups, smooth, warty, spiny, hairy, and rugose, by examination of carbon replicas of spores with the transmission electron microscope and by direct examination of spores with the scanning electron microscope. Images PMID:4928607

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

    SciTech Connect

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

    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.

  12. Use of molecular methods for the detection of fungal spores.

    PubMed

    Ward, Elaine

    2009-01-01

    Traditional methods for the isolation and identification of fungal spores can be time-consuming and laborious. DNA-based methods for fungal detection can be used to detect the spores of plant-pathogenic fungi. Air borne spores can be collected and identified by PCR allowing identification of the species.

  13. Airborne myxomycete spores: detection using molecular techniques

    NASA Astrophysics Data System (ADS)

    Kamono, Akiko; Kojima, Hisaya; Matsumoto, Jun; Kawamura, Kimitaka; Fukui, Manabu

    2009-01-01

    Myxomycetes are organisms characterized by a life cycle that includes a fruiting body stage. Myxomycete fruiting bodies contain spores, and wind dispersal of the spores is considered important for this organism to colonize new areas. In this study, the presence of airborne myxomycetes and the temporal changes in the myxomycete composition of atmospheric particles (aerosols) were investigated with a polymerase chain reaction (PCR)-based method for Didymiaceae and Physaraceae. Twenty-one aerosol samples were collected on the roof of a three-story building located in Sapporo, Hokkaido Island, northern Japan. PCR analysis of DNA extracts from the aerosol samples indicated the presence of airborne myxomycetes in all the samples, except for the one collected during the snowfall season. Denaturing gradient gel electrophoresis (DGGE) analysis of the PCR products showed seasonally varying banding patterns. The detected DGGE bands were subjected to sequence analyses, and four out of nine obtained sequences were identical to those of fruiting body samples collected in Hokkaido Island. It appears that the difference in the fruiting period of each species was correlated with the seasonal changes in the myxomycete composition of the aerosols. Molecular evidence shows that newly formed spores are released and dispersed in the air, suggesting that wind-driven dispersal of spores is an important process in the life history of myxomycetes. This study is the first to detect airborne myxomycetes with the use of molecular ecological analyses and to characterize their seasonal distribution.

  14. Real time viability detection of bacterial spores

    DOEpatents

    Vanderberg, Laura A.; Herdendorf, Timothy J.; Obiso, Richard J.

    2003-07-29

    This invention relates to a process for detecting the presence of viable bacterial spores in a sample and to a spore detection system, the process including placing a sample in a germination medium for a period of time sufficient for commitment of any present viable bacterial spores to occur, mixing the sample with a solution of a lanthanide capable of forming a fluorescent complex with dipicolinic acid, and, measuring the sample for the presence of dipicolinic acid, and the system including a germination chamber having inlets from a sample chamber, a germinant chamber and a bleach chamber, the germination chamber further including an outlet through a filtering means, the outlet connected to a detection chamber, the detection chamber having an inlet from a fluorescence promoting metal chamber and the detection chamber including a spectral excitation source and a means of measuring emission spectra from a sample, the detection chamber further connected to a waste chamber. A germination reaction mixture useful for promoting commitment of any viable bacterial spores in a sample including a combination of L-alanine, L-asparagine and D-glucose is also described.

  15. Pan-European Distribution of White-Nose Syndrome Fungus (Geomyces destructans) Not Associated with Mass Mortality

    PubMed Central

    Korn, Vanessa; Fuller, Hubert; Forget, Frédéric; Mühldorfer, Kristin; Kurth, Andreas; Bogdanowicz, Wieslaw; Borel, Christophe; Bosch, Thijs; Cherezy, Thomas; Drebet, Mikhail; Görföl, Tamás; Haarsma, Anne-Jifke; Herhaus, Frank; Hallart, Guénael; Hammer, Matthias; Jungmann, Christian; Le Bris, Yann; Lutsar, Lauri; Masing, Matti; Mulkens, Bart; Passior, Karsten; Starrach, Martin; Wojtaszewski, Andrzej; Zöphel, Ulrich; Teeling, Emma C.

    2011-01-01

    Background The dramatic mass mortalities amongst hibernating bats in Northeastern America caused by “white nose-syndrome” (WNS) continue to threaten populations of different bat species. The cold-loving fungus, Geomyces destructans, is the most likely causative agent leading to extensive destruction of the skin, particularly the wing membranes. Recent investigations in Europe confirmed the presence of the fungus G. destructans without associated mass mortality in hibernating bats in six countries but its distribution remains poorly known. Methodology/Principal Findings We collected data on the presence of bats with white fungal growth in 12 countries in Europe between 2003 and 2010 and conducted morphological and genetic analysis to confirm the identity of the fungus as Geomyces destructans. Our results demonstrate the presence of the fungus in eight countries spanning over 2000 km from West to East and provide compelling photographic evidence for its presence in another four countries including Romania, and Turkey. Furthermore, matching prevalence data of a hibernaculum monitored over two consecutive years with data from across Europe show that the temporal occurrence of the fungus, which first becomes visible around February, peaks in March but can still be seen in some torpid bats in May or June, is strikingly similar throughout Europe. Finally, we isolated and cultured G. destructans from a cave wall adjacent to a bat with fungal growth. Conclusions/Significance G. destructans is widely found over large areas of the European continent without associated mass mortalities in bats, suggesting that the fungus is native to Europe. The characterisation of the temporal variation in G. destructans growth on bats provides reference data for studying the spatio-temporal dynamic of the fungus. Finally, the presence of G. destructans spores on cave walls suggests that hibernacula could act as passive vectors and/or reservoirs for G. destructans and therefore, might

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

    DTIC Science & Technology

    2014-01-01

    SECURITY CLASSIFICATION OF: The germination of multiple individual Bacillus subtilis spores by a high pressure (HP) of 140-150 (unless noted...public release; distribution is unlimited. Monitoring Rates and Heterogeneity of High-Pressure Germination of Bacillus Spores by Phase-Contrast Microscopy...ADDRESS (ES) U.S. Army Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 Bacillus , spores, spore germination, high pressure, pressure

  17. Genome Diversity of Spore-Forming Firmicutes

    PubMed Central

    Galperin, Michael Y.

    2015-01-01

    Summary Formation of heat-resistant endospores is a specific property of the members of the phylum Firmicutes (low-G+C Gram-positive bacteria). It is found in representatives of four different classes of Firmicutes: Bacilli, Clostridia, Erysipelotrichia, and Negativicutes, which all encode similar sets of core sporulation proteins. Each of these classes also includes non-spore-forming organisms that sometimes belong to the same genus or even species as their spore-forming relatives. This chapter reviews the diversity of the members of phylum Firmicutes, its current taxonomy, and the status of genome sequencing projects for various subgroups within the phylum. It also discusses the evolution of the Firmicutes from their apparently spore-forming common ancestor and the independent loss of sporulation genes in several different lineages (staphylococci, streptococci, listeria, lactobacilli, ruminococci) in the course of their adaptation to the saprophytic lifestyle in nutrient-rich environment. It argues that systematics of Firmicutes is a rapidly developing area of research that benefits from the evolutionary approaches to the ever-increasing amount of genomic and phenotypic data and allows arranging these data into a common framework. Later the Bacillus filaments begin to prepare for spore formation. In their homogenous contents strongly refracting bodies appear. From each of these bodies develops an oblong or shortly cylindrical, strongly refracting, dark-rimmed spore. Ferdinand Cohn. 1876. Untersuchungen über Bacterien. IV. Beiträge zur Biologie der Bacillen. Beiträge zur Biologie der Pflanzen, vol. 2, pp. 249–276. (Studies on the biology of the bacilli. In: Milestones in Microbiology: 1546 to 1940. Translated and edited by Thomas D. Brock. Prentice-Hall, Englewood Cliffs, NJ, 1961, pp. 49–56). PMID:26184964

  18. Effects of Major Spore-Specific DNA Binding Proteins on Bacillus subtilis Sporulation and Spore Properties

    PubMed Central

    Setlow, Barbara; McGinnis, Kelly A.; Ragkousi, Katerina; Setlow, Peter

    2000-01-01

    Sporulation of a Bacillus subtilis strain (termed α− β−) lacking the majority of the α/β-type small, acid-soluble spore proteins (SASP) that are synthesized in the developing forespore and saturate spore DNA exhibited a number of differences from that of the wild-type strain, including delayed forespore accumulation of dipicolinic acid, overexpression of forespore-specific genes, and delayed expression of at least one mother cell-specific gene turned on late in sporulation, although genes turned on earlier in the mother cell were expressed normally in α− β− strains. The sporulation defects in α− β− strains were corrected by synthesis of chromosome-saturating levels of either of two wild-type, α/β-type SASP but not by a mutant SASP that binds DNA poorly. Spores from α− β− strains also exhibited less glutaraldehyde resistance and slower outgrowth than did wild-type spores, but at least some of these defects in α− β− spores were abolished by the synthesis of normal levels of α/β-type SASP. These results indicate that α/β-type SASP may well have global effects on gene expression during sporulation and spore outgrowth. PMID:11092849

  19. Protective role of spore structural components in determining Bacillus subtilis spore resistance to simulated mars surface conditions.

    PubMed

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

    2012-12-01

    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.

  20. Cytological and Proteomic Analyses of Osmunda cinnamomea Germinating Spores Reveal Characteristics of Fern Spore Germination and Rhizoid Tip Growth.

    PubMed

    Suo, Jinwei; Zhao, Qi; Zhang, Zhengxiu; Chen, Sixue; Cao, Jian'guo; Liu, Guanjun; Wei, Xing; Wang, Tai; Yang, Chuanping; Dai, Shaojun

    2015-09-01

    Fern spore is a good single-cell model for studying the sophisticated molecular networks in asymmetric cell division, differentiation, and polar growth. Osmunda cinnamomea L. var. asiatica is one of the oldest fern species with typical separate-growing trophophyll and sporophyll. The chlorophyllous spores generated from sporophyll can germinate without dormancy. In this study, the spore ultrastructure, antioxidant enzyme activities, as well as protein and gene expression patterns were analyzed in the course of spore germination at five typical stages (i.e. mature spores, rehydrated spores, double-celled spores, germinated spores, and spores with protonemal cells). Proteomic analysis revealed 113 differentially expressed proteins, which were mainly involved in photosynthesis, reserve mobilization, energy supplying, protein synthesis and turnover, reactive oxygen species scavenging, signaling, and cell structure modulation. The presence of multiple proteoforms of 25 differentially expressed proteins implies that post-translational modification may play important roles in spore germination. The dynamic patterns of proteins and their encoding genes exhibited specific characteristics in the processes of cell division and rhizoid tip growth, which include heterotrophic and autotrophic metabolisms, de novo protein synthesis and active protein turnover, reactive oxygen species and hormone (brassinosteroid and ethylene) signaling, and vesicle trafficking and cytoskeleton dynamic. In addition, the function skew of proteins in fern spores highlights the unique and common mechanisms when compared with evolutionarily divergent spermatophyte pollen. These findings provide an improved understanding of the typical single-celled asymmetric division and polar growth during fern spore germination.

  1. Maturation of Released Spores Is Necessary for Acquisition of Full Spore Heat Resistance during Bacillus subtilis Sporulation ▿

    PubMed Central

    Sanchez-Salas, Jose-Luis; Setlow, Barbara; Zhang, Pengfei; Li, Yong-qing; Setlow, Peter

    2011-01-01

    The first ∼10% of spores released from sporangia (early spores) during Bacillus subtilis sporulation were isolated, and their properties were compared to those of the total spores produced from the same culture. The early spores had significantly lower resistance to wet heat and hypochlorite than the total spores but identical resistance to dry heat and UV radiation. Early and total spores also had the same levels of core water, dipicolinic acid, and Ca and germinated similarly with several nutrient germinants. The wet heat resistance of the early spores could be increased to that of total spores if early spores were incubated in conditioned sporulation medium for ∼24 h at 37°C (maturation), and some hypochlorite resistance was also restored. The maturation of early spores took place in pH 8 buffer with Ca2+ but was blocked by EDTA; maturation was also seen with early spores of strains lacking the CotE protein or the coat-associated transglutaminase, both of which are needed for normal coat structure. Nonetheless, it appears to be most likely that it is changes in coat structure that are responsible for the increased resistance to wet heat and hypochlorite upon early spore maturation. PMID:21821751

  2. Cytological and Proteomic Analyses of Osmunda cinnamomea Germinating Spores Reveal Characteristics of Fern Spore Germination and Rhizoid Tip Growth*

    PubMed Central

    Suo, Jinwei; Zhao, Qi; Zhang, Zhengxiu; Chen, Sixue; Cao, Jian'guo; Liu, Guanjun; Wei, Xing; Wang, Tai; Yang, Chuanping; Dai, Shaojun

    2015-01-01

    Fern spore is a good single-cell model for studying the sophisticated molecular networks in asymmetric cell division, differentiation, and polar growth. Osmunda cinnamomea L. var. asiatica is one of the oldest fern species with typical separate-growing trophophyll and sporophyll. The chlorophyllous spores generated from sporophyll can germinate without dormancy. In this study, the spore ultrastructure, antioxidant enzyme activities, as well as protein and gene expression patterns were analyzed in the course of spore germination at five typical stages (i.e. mature spores, rehydrated spores, double-celled spores, germinated spores, and spores with protonemal cells). Proteomic analysis revealed 113 differentially expressed proteins, which were mainly involved in photosynthesis, reserve mobilization, energy supplying, protein synthesis and turnover, reactive oxygen species scavenging, signaling, and cell structure modulation. The presence of multiple proteoforms of 25 differentially expressed proteins implies that post-translational modification may play important roles in spore germination. The dynamic patterns of proteins and their encoding genes exhibited specific characteristics in the processes of cell division and rhizoid tip growth, which include heterotrophic and autotrophic metabolisms, de novo protein synthesis and active protein turnover, reactive oxygen species and hormone (brassinosteroid and ethylene) signaling, and vesicle trafficking and cytoskeleton dynamic. In addition, the function skew of proteins in fern spores highlights the unique and common mechanisms when compared with evolutionarily divergent spermatophyte pollen. These findings provide an improved understanding of the typical single-celled asymmetric division and polar growth during fern spore germination. PMID:26091698

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

    PubMed Central

    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. PMID:23064347

  4. Apparatus and method for automated monitoring of airborne bacterial spores

    NASA Technical Reports Server (NTRS)

    Ponce, Adrian (Inventor)

    2009-01-01

    An apparatus and method for automated monitoring of airborne bacterial spores. The apparatus is provided with an air sampler, a surface for capturing airborne spores, a thermal lysis unit to release DPA from bacterial spores, a source of lanthanide ions, and a spectrometer for excitation and detection of the characteristic fluorescence of the aromatic molecules in bacterial spores complexed with lanthanide ions. In accordance with the method: computer-programmed steps allow for automation of the apparatus for the monitoring of airborne bacterial spores.

  5. Decrease in optical density as a results of germination of Alicyclobacillus acidoterrestris spores under high hydrostatic pressure

    NASA Astrophysics Data System (ADS)

    Porębska, I.; Rutkowska, M.; Sokołowska, B.

    2015-01-01

    Alicyclobacillus acidoterrestris is a spore-forming bacterium, causing spoilage of juices. The spores of these bacteria have the ability to survive in the typical conditions used for thermal pasteurization. Therefore, the use of other techniques such as high hydrostatic pressure is considered for their inactivation. The effect of hydrostatic pressure of 200-500 MPa, at temperatures 4-50 °C for 15 min, on the dynamics of germination of A. acidoterrestris spores in apple juice and pH 4 buffer was studied. To estimate the share of germinated spores, the method of determining the optical density at a wavelength of 660 nm (OD660) was used. Parameters of hydrostatic pressure treatment used in this work affected the dynamics of germination of A. acidoterrestris spores in apple juice, and the temperature had the greatest effect. The results indicate that nutrients present in apple juice can promote the germination of A. acidoterrestris spores. This paper was presented at the 8th International Conference on High Pressure Bioscience & Biotechnology (HPBB 2014) in Nantes (France) 15-18 July 2014.

  6. Flavonoids induce germination of basidiospores of the ectomycorrhizal fungus Suillus bovinus.

    PubMed

    Kikuchi, Kensuke; Matsushita, Norihisa; Suzuki, Kazuo; Hogetsu, Taizo

    2007-10-01

    Under laboratory conditions, spores of ectomycorrhizal fungi usually germinate very poorly or not at all. In a previous study, we showed that spores of the ectomycorrhizal fungus Suillus bovinus germinated through the combination of activated charcoal treatment of media and co-culture with seedlings of Pinus densiflora, which suggested that some substances contained in root exudates induced the germination. Among the compounds reported from root exudates, flavonoids have been elucidated to play various and substantial roles in plant-microbe interactions; we therefore investigated the effects of flavonoids on basidiospore germination of S. bovinus by the diffusion gradient assay on water agar plates pretreated with charcoal powder. Seven out of the 11 flavonoids tested, hesperidin, morin, rutin, quercitrin, naringenin, genistein, and chrysin, had greater effects than controls, whereas flavone, biochanin A, luteolin, and quercetin showed no positive effects. The effective concentration presumably corresponded to several micromolar levels, which was equivalent to those effective for pollen development, nod gene induction, and spore germination of F. solani f. sp. pisi and AM fungi. The results suggest that flavonoids play a role as signaling molecules in symbiotic relationships between woody plants and ectomycorrhizal fungi.

  7. Identification by Quantitative Carrier Test of Surrogate Spore-Forming Bacteria To Assess Sporicidal Chemicals for Use against Bacillus anthracis▿

    PubMed Central

    Majcher, Miles R.; Bernard, Kathryn A.; Sattar, Syed A.

    2008-01-01

    The spores of six strains of Bacillus anthracis (four virulent and two avirulent) were compared with those of four other types of spore-forming bacteria for their resistance to four liquid chemical sporicides (sodium hypochlorite at 5,000 ppm available chlorine, 70,000 ppm accelerated H2O2, 1,000 ppm chlorine dioxide, and 3,000 ppm peracetic acid). All test bacteria were grown in a 1:10 dilution of Columbia broth (with manganese) incubated at 37°C for 72 h. The spore suspensions, heat treated at 80°C for 10 min to rid them of any viable vegetative cells, contained 1 × 108 to 3 × 108 CFU/ml. The second tier of the quantitative carrier test (QCT-2), a standard of ASTM International, was used to assess for sporicidal activity, with disks (1 cm in diameter) of brushed and magnetized stainless steel as spore carriers. Each carrier, with 10 μl (≥106 CFU) of the test spore suspension in a soil load, was dried and then overlaid with 50 μl of the sporicide being evaluated. The contact time at room temperature ranged from 5 to 20 min, and the arbitrarily set criterion for acceptable sporicidal activity was a reduction of ≥106 in viable spore count. Each test was repeated at least three times. In the final analysis, the spores of Bacillus licheniformis (ATCC 14580T) and Bacillus subtilis (ATCC 6051T) proved to be generally more resistant than the spores of the strains of B. anthracis tested. The use of one or both of the safe and easy-to-handle surrogates identified here should help in developing safer and more-effective sporicides and also in evaluating the field effectiveness of existing and newer formulations in the decontamination of objects and surfaces suspected of B. anthracis contamination. PMID:18083869

  8. Detection of chlorophylls in spores of seven ferns.

    PubMed

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

    2017-03-01

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

  9. Sources of Variability in the Measurement of Fungal Spore Yields

    PubMed Central

    Smith, C. S.; Slade, S. J.; Nordheim, E. V.; Cascino, J. J.; Harris, R. F.; Andrews, J. H.

    1988-01-01

    Variability in the production of fungal spores and in the measurement of spore yields was investigated in four species of fungi: Colletotrichum gloeosporioides, Colletotrichum coccodes, Colletotrichum phomoides, and Acremonium strictum. When the fungi were grown on solid medium in microplates and spore yields were measured by counting the subsamples with a hemacytometer, the variability among hemacytometer squares was always the largest source of variation, accounting for 51 to 91% of the total variation. Variability among replicate cultures and results of repeat experiments were generally also significant. The effect of square-to-square variability on the precision of spore yield measurement was minimized by counting a moderate number (ca. 30) of squares per culture. Culture-to-culture variability limited the practical precision of spore production measurements to a 95% confidence interval of approximately the mean ± 25%. We provide guidelines for determining the number of replicate cultures required to attain this or other degrees of precision. Particle counter-derived spore counts and counts based on spore weights were much less variable than were hemacytometer counts, but they did not improve spore production estimates very much because of culture-to-culture variability. Results obtained by both of these methods differed from those obtained with a hemacytometer; particle counter measurements required a correction for spore pairs, while the relationship between spore weights and spore counts changed as the cultures aged. PMID:16347653

  10. Inactivation of Clostridium difficile spores by microwave irradiation.

    PubMed

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

    2016-04-01

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

  11. Fern Spore Longevity in Saline Water: Can Sea Bottom Sediments Maintain a Viable Spore Bank?

    PubMed Central

    de Groot, G. Arjen; During, Heinjo

    2013-01-01

    Freshwater and marine sediments often harbor reservoirs of plant diaspores, from which germination and establishment may occur whenever the sediment falls dry. Therewith, they form valuable records of historical inter- and intraspecific diversity, and are increasingly exploited to facilitate diversity establishment in new or restored nature areas. Yet, while ferns may constitute a considerable part of a vegetation’s diversity and sediments are known to contain fern spores, little is known about their longevity, which may suffer from inundation and - in sea bottoms - salt stress. We tested the potential of ferns to establish from a sea or lake bottom, using experimental studies on spore survival and gametophyte formation, as well as a spore bank analysis on sediments from a former Dutch inland sea. Our experimental results revealed clear differences among species. For Asplenium scolopendrium and Gymnocarpium dryopteris, spore germination was not affected by inundated storage alone, but decreased with rising salt concentrations. In contrast, for Asplenium trichomanes subsp. quadrivalens germination decreased following inundation, but not in response to salt. Germination rates decreased with time of storage in saline water. Smaller and less viable gametophytes were produced when saline storage lasted for a year. Effects on germination and gametophyte development clearly differed among genotypes of A. scolopendrium. Spore bank analyses detected no viable spores in marine sediment layers. Only two very small gametophytes (identified as Thelypteris palustris via DNA barcoding) emerged from freshwater sediments. Both died before maturation. We conclude that marine, and likely even freshwater sediments, will generally be of little value for long-term storage of fern diversity. The development of any fern vegetation on a former sea floor will depend heavily on the deposition of spores onto the drained land by natural or artificial means of dispersal. PMID:24223951

  12. Fern spore longevity in saline water: can sea bottom sediments maintain a viable spore bank?

    PubMed

    de Groot, G Arjen; During, Heinjo

    2013-01-01

    Freshwater and marine sediments often harbor reservoirs of plant diaspores, from which germination and establishment may occur whenever the sediment falls dry. Therewith, they form valuable records of historical inter- and intraspecific diversity, and are increasingly exploited to facilitate diversity establishment in new or restored nature areas. Yet, while ferns may constitute a considerable part of a vegetation's diversity and sediments are known to contain fern spores, little is known about their longevity, which may suffer from inundation and--in sea bottoms--salt stress. We tested the potential of ferns to establish from a sea or lake bottom, using experimental studies on spore survival and gametophyte formation, as well as a spore bank analysis on sediments from a former Dutch inland sea. Our experimental results revealed clear differences among species. For Asplenium scolopendrium and Gymnocarpium dryopteris, spore germination was not affected by inundated storage alone, but decreased with rising salt concentrations. In contrast, for Asplenium trichomanes subsp. quadrivalens germination decreased following inundation, but not in response to salt. Germination rates decreased with time of storage in saline water. Smaller and less viable gametophytes were produced when saline storage lasted for a year. Effects on germination and gametophyte development clearly differed among genotypes of A. scolopendrium. Spore bank analyses detected no viable spores in marine sediment layers. Only two very small gametophytes (identified as Thelypteris palustris via DNA barcoding) emerged from freshwater sediments. Both died before maturation. We conclude that marine, and likely even freshwater sediments, will generally be of little value for long-term storage of fern diversity. The development of any fern vegetation on a former sea floor will depend heavily on the deposition of spores onto the drained land by natural or artificial means of dispersal.

  13. Do fungicides used to control Rhizoctonia solani impact the non-target arbuscular mycorrhizal fungus Rhizophagus irregularis?

    PubMed

    Buysens, Catherine; Dupré de Boulois, Hervé; Declerck, Stéphane

    2015-05-01

    There is growing evidence that the application of biocontrol organisms (e.g., Pseudomonas and Bacillus spp., arbuscular mycorrhizal fungi-AMF) is a feasible option to reduce incidence of plant pathogens in an integrated control strategy. However, the utilization of these microorganisms, in particular AMF, may be threatened by the application of fungicides, a widely-used measure to control Rhizoctonia solani in various crops among which potato. Prior to their application, it is thus important to determine the impact of fungicides on AMF. The present study investigated, under in vitro controlled conditions, the impact of azoxystrobin (a systemic broad-spectrum fungicide), flutolanil (a systemic Basidiomycota-specific fungicide), and pencycuron (a contact Rhizoctonia-specific fungicide) and their respective formulations (Amistar, Monarch, and Monceren) on the growth and development of the AMF Rhizophagus irregularis MUCL 41833 (spore germination, root colonization, extraradical mycelium development, and spore production) at doses used to control R. solani. Results demonstrated that azoxystrobin and its formulation Amistar, at threshold values for R. solani control (estimated by the half maximal inhibitory concentration, IC50, on a dry weight basis), did not affect spore germination and potato root colonization by R. irregularis, while the development of extra-radical mycelium and spore production was reduced at 10 times the threshold value. Flutolanil and its formulation Monarch at threshold value did not affect spore germination or extra-radical development but decreased root colonization and arbuscule formation. At threshold value, pencycuron and its formulation Monceren, did not affect spore germination and intra- or extraradical development of R. irregularis. These results suggest that azoxystrobin and pencycuron do not affect the AMF at threshold concentrations to control R. solani in vitro, while flutolanil (as formulation) impacts the intraradical phase of the

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

    PubMed

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

    2013-09-02

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

  15. Intra and Inter-Spore Variability in Rhizophagus irregularis AOX Gene

    PubMed Central

    Nogales, Amaia; Svensson, Jan; Lopez-Ráez, Juan Antonio; Pozo, María José; Nobre, Tânia; Schneider, Carolin; Arnholdt-Schmitt, Birgit

    2015-01-01

    Arbuscular mycorrhizal fungi (AMF) are root-inhabiting fungi that form mutualistic symbioses with their host plants. AMF symbiosis improves nutrient uptake and buffers the plant against a diversity of stresses. Rhizophagus irregularis is one of the most widespread AMF species in the world, and its application in agricultural systems for yield improvement has increased over the last years. Still, from the inoculum production perspective, a lack of consistency of inoculum quality is referred to, which partially may be due to a high genetic variability of the fungus. The alternative oxidase (AOX) is an enzyme of the alternative respiratory chain already described in different taxa, including various fungi, which decreases the damage caused by oxidative stress. Nevertheless, virtually nothing is known on the involvement of AMF AOX on symbiosis establishment, as well on the existence of AOX variability that could affect AMF effectiveness and consequently plant performance. Here, we report the isolation and characterisation of the AOX gene of R. irregularis (RiAOX), and show that it is highly expressed during early phases of the symbiosis with plant roots. Phylogenetic analysis clustered RiAOX sequence with ancient fungi, and multiple sequence alignment revealed the lack of several regulatory motifs which are present in plant AOX. The analysis of RiAOX polymorphisms in single spores of three different isolates showed a reduced variability in one spore relatively to a group of spores. A high number of polymorphisms occurred in introns; nevertheless, some putative amino acid changes resulting from non-synonymous variants were found, offering a basis for selective pressure to occur within the populations. Given the AOX relatedness with stress responses, differences in gene variants amongst R. irregularis isolates are likely to be related with its origin and environmental constraints and might have a potential impact on inoculum production. PMID:26540237

  16. Changes in Atmospheric CO2 Influence the Allergenicity of Aspergillus fumigatus fungal spore

    NASA Astrophysics Data System (ADS)

    Lang-Yona, N.; Levin, Y.; Dannemoller, K. C.; Yarden, O.; Peccia, J.; Rudich, Y.

    2013-12-01

    Increased allergic susceptibility has been documented without a comprehensive understanding for its causes. Therefore understanding trends and mechanisms of allergy inducing agents is essential. In this study we investigated whether elevated atmospheric CO2 levels can affect the allergenicity of Aspergillus fumigatus, a common allergenic fungal species. Both direct exposure to changing CO2 levels during fungal growth, and indirect exposure through changes in the C:N ratios in the growth media were inspected. We determined the allergenicity of the spores through two types of immunoassays, accompanied with genes expression analysis, and proteins relative quantification. We show that fungi grown under present day CO2 levels (392 ppm) exhibit 8.5 and 3.5 fold higher allergenicity compared to fungi grown at preindustrial (280 ppm) and double (560 ppm) CO2 levels, respectively. A corresponding trend is observed in the expression of genes encoding for known allergenic proteins and in the major allergen Asp f1 concentrations, possibly due to physiological changes such as respiration rates and the nitrogen content of the fungus, influenced by the CO2 concentrations. Increased carbon and nitrogen levels in the growth medium also lead to a significant increase in the allergenicity, for which we propose two different biological mechanisms. We suggest that climatic changes such as increasing atmospheric CO2 levels and changes in the fungal growth medium may impact the ability of allergenic fungi such as Aspergillus fumigatus to induce allergies. The effect of changing CO2 concentrations on the total allergenicity per 10^7 spores of A. fumigatus (A), the major allergen Asp f1 concentration in ng per 10^7 spores (B), and the gene expression by RT-PCR (C). The error bars represent the standard error of the mean.

  17. CLOSTRIDIUM SPORE ATTACHMENT TO HUMAN CELLS

    SciTech Connect

    PANESSA-WARREN,B.; TORTORA,G.; WARREN,J.

    1997-08-10

    This paper uses high resolution scanning electron microscopy (SEM) with a LaB6 gun and the newest commercial field emission guns, to obtain high magnification images of intact clostridial spores throughout the activation/germination/outgrowth process. By high resolution SEM, the clostridial exosporial membrane can be seen to produce numerous delicate projections (following activation), that extend from the exosporial surface to a nutritive substrate (agar), or cell surface when anaerobically incubated in the presence of human cells (embryonic fibroblasts and colon carcinoma cells). Magnifications of 20,000 to 200,000Xs at accelerating voltages low enough to minimize or eliminate specimen damage (1--5 kV) have permitted the entire surface of C.sporogenes and C.difficile endospores to be examined during all stages of germination. The relationships between the spore and the agar or human cell surface were also clearly visible.

  18. Viability of bacterial spores exposed to hydrazine

    NASA Astrophysics Data System (ADS)

    Schubert, W.; Plett, G.; Yavrouian, A.; Barengoltz, J.

    For the purposes of planetary protection a series of experiments were performed to answer a long-standing question about the potential of bacterial contamination of interplanetary spacecraft from liquid hydrazine Spores of Bacillus atrophaeus ATCC No 9372 also known as Bacillus subtilis var niger and BSN were exposed to hydrazine for various durations Then the survivors were enumerated using the NASA standard planetary protection pour plate assay It is important to note that in these experiments the hydrazine was removed prior to the assay This eliminated the possibility that the presence of hydrazine rather than a prior exposure was inhibiting germination and or reproduction Populations of 10 6 spores were eliminated within 30 minutes These results indicate that bulk hydrazine rocket propellant may be considered free of living bacterial cells for planetary protection compliance

  19. EFFECT OF THE FILAMENTOUS FUNGUS MUCOR CIRCINELLOIDES ON THE DEVELOPMENT OF EGGS OF THE RUMEN FLUKE CALICOPHORON DAUBNEYI (PARAMPHISTOMIDAE).

    PubMed

    Arroyo, Fabià  N; Hernà  Ndez, Josà  à  Ngel; Cazapal-Monteiro, Cristiana Filipa; Pedreira, Josà Â; Sanchà  S, Jaime; Romasanta, à  Ngel; Sã  Nchez-Andrade, Rita; Paz-Silva, Adolfo; Arias, Marà  A Sol

    2017-02-06

    Ruminants infected by Paramphistomidae flukes shed eggs in the feces, which pass through different stages in the environment until the infective stages (metacercariae) are reached. The activity of the soil fungus Mucor circinelloides on the development of eggs of the rumen fluke Calicophoron daubneyi was presently tested with 3 probes, i.e., in Petri plates, feces and an aqueous environment (tubes). The effect of the fungus was assessed by recording the numbers of undeveloped, non-viable and embryonated eggs. Non-viable eggs were considered when vacuolization occurred, the inner structures were not clearly observed, the egg-shell was broken, and/or the embryo inside was destroyed. By considering the ability of hyphae of M. circinelloides to develop in the presence of C. daubneyi eggs, attach to their surface, penetrate and destroy the inner embryo, this ovicidal effect was classified as type 3. After a period of 50 days, the percentage of undeveloped eggs in the feces of infected cattle was 40%; furthermore, 27% eggs were non-viable, and 33% were embryonated (1 miracidium inside). The addition of 4 doses of M. circinelloides spores directly onto the feces resulted in 9-31% undeveloped eggs, 38-60% non-viable eggs, and 9-21% embryonated eggs, and no statistical significances were obtained among the different doses. Placing the eggs of C. daubneyi into an aqueous solution containing 107 spores M. circinelloides/ml for 29 days resulted in 43% undeveloped eggs, 40% non-viable eggs, and 17% embryonated eggs, whereas in the controls, the percentages were 48%, 12%, and 40%, respectively. These data demonstrate the usefulness of the spores of the fungus M. circinelloides in limiting the development of the eggs of the trematode Calicophoron daubneyi.

  20. Mechanisms of Resistance in Microbial Spores.

    DTIC Science & Technology

    1986-11-14

    characterization of forespores isolated from Bacillus meqaterium ATCC 19213. J. Bacteriol. 153:436-442. Isolated stage III forespores of Bacillus megaterium ...other factors is complex. At Michigan State University, four morphotypes of B. megaterium spores, obtained by progressive divestment of the integument...permeating medium. Thereby, the PWC was determined with 28 types among 7 Bacillus species spanning a 3,OCO-fold range in heat resistance, which was

  1. Morphogenesis of the Bacillus anthracis Spore

    DTIC Science & Technology

    2007-02-01

    layers in B. subtilis is unknown. Unlike B. subtilis, in B. anthracis, Bacillus megaterium , and other species, the spore is surrounded by an additional...nonpathogenic species including B. megaterium and Bacillus odysseyi (45, 85), suggesting that their primary role need not be in disease. Nonetheless, the exospo...S. 1994. Prime time for Bacillus megaterium . Microbiology 140:1001– 1013. 86. Warth, A. D., D. F. Ohye, and W. G. Murrell. 1963. The composition and

  2. Viability of bacterial spores exposed to hydrazine

    NASA Astrophysics Data System (ADS)

    Schubert, W.; Plett, G.; Yavrouian, A.; Barengoltz, J.

    2008-09-01

    For the purposes of planetary protection, a series of experiments were performed to answer a long-standing question about the potential of bacterial contamination of interplanetary spacecraft from liquid hydrazine. Spores of Bacillus atrophaeus (ATCC No. 9372, also known as Bacillus subtilis var. niger, and BSN) were exposed to hydrazine and survivors were enumerated using the NASA standard planetary protection pour plate assay. Results indicate that bulk hydrazine rocket propellant may be considered free of living bacterial cells for planetary protection compliance.

  3. Association of Fidaxomicin with C. difficile Spores: Effects of Persistence on Subsequent Spore Recovery, Outgrowth and Toxin Production

    PubMed Central

    Crowther, Grace S.; Ashwin, Helen; Longshaw, Chris M.; Wilcox, Mark H.

    2016-01-01

    Background We have previously shown that fidaxomicin instillation prevents spore recovery in an in-vitro gut model, whereas vancomycin does not. The reasons for this are unclear. Here, we have investigated persistence of fidaxomicin and vancomycin on C. difficile spores, and examined post-antibiotic exposure spore recovery, outgrowth and toxin production. Methods Prevalent UK C. difficile ribotypes (n = 10) were incubated with 200mg/L fidaxomicin, vancomycin or a non-antimicrobial containing control for 1 h in faecal filtrate or Phosphate Buffered Saline. Spores were washed three times with faecal filtrate or phosphate buffered saline, and residual spore-associated antimicrobial activity was determined by bioassay. For three ribotypes (027, 078, 015), antimicrobial-exposed, faecal filtrate-washed spores and controls were inoculated into broth. Viable vegetative and spore counts were enumerated on CCEYL agar. Percentage phase bright spores, phase dark spores and vegetative cells were enumerated by phase contrast microscopy at 0, 3, 6, 24 and 48 h post-inoculation. Toxin levels (24 and 48h) were determined by cell cytotoxicity assay. Results Fidaxomicin, but not vancomycin persisted on spores of all ribotypes following washing in saline (mean = 10.1mg/L; range = 4.0-14mg/L) and faecal filtrate (mean = 17.4mg/L; 8.4–22.1mg/L). Outgrowth and proliferation rates of vancomycin-exposed spores were similar to controls, whereas fidaxomicin-exposed spores showed no vegetative cell growth after 24 and 48 h. At 48h, toxin levels averaged 3.7 and 3.3 relative units (RU) in control and vancomycin-exposed samples, respectively, but were undetectable in fidaxomicin-exposed samples. Conclusion Fidaxomicin persists on C. difficile spores, whereas vancomycin does not. This persistence prevents subsequent growth and toxin production in vitro. This may have implications on spore viability, thereby impacting CDI recurrence and transmission rates. PMID:27556739

  4. Airborne spores of Basidiomycetes in Mérida (SW Spain).

    PubMed

    Hernández Trejo, Fernando; Muñoz Rodríguez, Adolfo F; Tormo Molina, Rafael; Silva Palacios, Inmaculada

    2013-01-01

    The aim of this work was to detect the presence of Basidiomycetes spores (basidiospores, teliospores, uredospores and aeciospores) in Mérida (SW Spain) and assess the influence of weather parameters. Air was sampled continuously with a volumetric seven-day Burkard spore trap for two years. Fungi spores were identified and counted at x1,000 microscope resolution. Daily and weekly meteorological data and airborne spore concentration were analysed. Twenty-three spores types were identified, including basidiospores (Amanita, Agrocybe, Cortinarius, Coprinus -2 types-, Boletus, Bovista, Calvatia, Entoloma, Ganoderma, Inocybe, Russula, Scleroderma, Telephora), teliospores (Phragmidium, Tilletia, Ustillago -4 types-), uredospores, and aeciospores (2 types), all of these types of spores included different taxa. Average concentration was of 616 spores/m(3), with maximum concentration in autumn (October), and a second concentration in spring (May-June); however, some spore types were more frequent in summer (Bovista, Ganoderma) or even in winter (Entoloma, Calvatia). The Amanita type was the most frequent (white-hyaline basidiospores); the second were teliospores of Ustilago, the third spore type was basidiospores of Coprinus (blackish basidiospores) and Agrocybe type (smoothed light to dark coloured basidiospores). Basidiospore concentration was positively correlated with temperature and negatively with relative humidity in most cases, and Ustilago teliospores concentration was positively correlated with wind speed. Differences in monthly rain were probably the origin between years. Airborne spores of Basidiomycetes may be separated into more than 20 types, and their seasonal concentration depended on meteorology as well as whether they were saprotrophic or parasitic.

  5. Bryophyte spore germinability is inhibited by peatland substrates

    NASA Astrophysics Data System (ADS)

    Bu, Zhao-Jun; Li, Zhi; Liu, Li-Jie; Sundberg, Sebastian; Feng, Ya-Min; Yang, Yun-He; Liu, Shuang; Song, Xue; Zhang, Xing-Lin

    2017-01-01

    Bryophyte substrates and species may affect spore germination through allelopathy. Polytrichum strictum is currently expanding in peatlands in north-eastern China - is this an effect of its superior spore germinability or do its gametophytes have a stronger allelopathic effect than do Sphagnum? We conducted a spore burial experiment to test the effect of species identity, substrate and water table depth (WTD) on spore germinability and bryophyte allelopathic effect with P. strictum and two Sphagnum species (S. palustre and S. magellanicum). After 5 months of burial during a growing season, the spores were tested for germinability. Allelopathic effect of bryophyte substrates was assessed by the difference between spore germinability after being stored inside or outside the substrates. After burial, more than 90% of the spores lost their germinability across all three species due to ageing and allelopathy. Spore germinability differed among species, where the spores in S. palustre had a higher germination frequency than those in P. strictum. The three bryophytes maintained a higher germinability in Sphagnum than in Polytrichum hummocks, probably due to a stronger allelopathic effect of P. strictum. Water table drawdown by 10 cm increased germinability by more than 60% across the three species. The study indicates that P. strictum does not possess an advantage regarding spore germination but rather its gametophytes have a stronger allelopathic effect. Due to the weaker inhibitive effect of Sphagnum gametophytes, P. strictum may have a potential establishment superiority over Sphagnum in peatlands, in addition to a better drought tolerance, which may explain its current expansion.

  6. The scope for nuclear selection within Termitomyces fungi associated with fungus-growing termites is limited

    PubMed Central

    2014-01-01

    Background We investigate the scope for selection at the level of nuclei within fungal individuals (mycelia) of the mutualistic Termitomyces cultivated by fungus-growing termites. Whereas in most basidiomycete fungi the number and kind of nuclei is strictly regulated to be two per cell, in Termitomyces mycelia the number of nuclei per cell is highly variable. We hypothesised that natural selection on these fungi not only occurs between mycelia, but also at the level of nuclei within the mycelium. We test this hypothesis using in vitro tests with five nuclear haplotypes of a Termitomyces species. Results First, we studied the transition from a mixture of five homokaryons (mycelia with identical nuclei) each with a different nuclear haplotype to heterokaryons (mycelia with genetically different nuclei). In vitro cultivation of this mixture for multiple asexual transfers led to the formation of multiple heterokaryotic mycelia, and a reduction of mycelial diversity over time. All heterokaryotic mycelia contained exactly two types of nucleus. The success of a heterokaryon during in vitro cultivation was mainly determined by spore production and to a lesser extent by mycelial growth rate. Second, heterokaryons invariably produced more spores than homokaryons implying that homokaryons will be outcompeted. Third, no homokaryotic ‘escapes’ from a heterokaryon via the formation of homokaryotic spores were found, despite extensive spore genotyping. Fourth, in contrast to most studied basidiomycete fungi, in Termitomyces sp. no nuclear migration occurs during mating, limiting the scope for nuclear competition within the mycelium. Conclusions Our experiments demonstrate that in this species of Termitomyces the scope for selection at the level of the nucleus within an established mycelium is limited. Although ‘mate choice’ of a particular nuclear haplotype is possible during mating, we infer that selection primarily occurs between mycelia with two types of nucleus

  7. A study of Ganoderma lucidum spores by FTIR microspectroscopy

    NASA Astrophysics Data System (ADS)

    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.

  8. Architecture and Assembly of the Bacillus subtilis Spore Coat

    DTIC Science & Technology

    2014-09-26

    arrows). EM of ruthenium red stained B. subtilis spores demonstrated the presence of an outermost glycoprotein layer, and it was suggested that this layer...Rather the amorphous layer likely corresponds to the outer crust layer of B. subtilis spores that stains with ruthenium red and is glycoprotein rich...RL (2004) Ruthenium red staining for ultrastructural visualization of a glycoprotein layer surrounding the spore of Bacillus anthracis and Bacillus

  9. Thermal Inactivation of Bacillus anthracis Spores Using Rapid Resistive Heating

    DTIC Science & Technology

    2016-03-24

    microbiological study sought to obtain a correlation between exposure time, temperature , and spore viability. This information is invaluable when...of the spores were found using rapid resistive heating at short duration exposure times from 0.26 to 7 seconds at temperatures ranging from 73.5 to...ranging from 0.1 to 10 seconds. Higher temperatures were needed to thermally inactivate the B.a. spores as exposure times decreased. vi

  10. Temporaly germinating rhythms of moss Funaria hygrometrica Hedw. spores

    NASA Astrophysics Data System (ADS)

    Pundiak, O.; Demkiv, O.

    The process of an organism development is regular and gradual. These characteristics of the development are especially evident in archegonial plants. It was shown that spores of moss Funar ia hygrometrica Hedw. in Knop's nutrient medium with 0,2% glucose in the dark in vertical orientation of Petry dishes, germinated polarly depending on gravity direction. At the begining, the primary rhyziod developed being usually directed downwards and then after 24 hours primary chloronema developed growing usually upwards. The amyloplasts sedimentation was shown before the rhyzoid and chloronema formation. It determines not only the time, but spatial orientation of the primary rhyzoid and chloronema (Pundjak at al., 2001). EGTA in concentration of 510- 5 M inhibited the initiation of the primary rhyzoid. The primary chloronema developed as usual in 48 h after the spores sowing. Temporary cooling caused analogical effect. Basing on these results we drew the conclusion that the primary rhyzoid and chloronema differently react on the action of EGTA and the cooling. The primary chloronema was more tolerant then the rhyzoid and maintained its usual gravisensitivy. Thus, we can think that EGTA and the cooling stop the development of primary rhyzoid, but it does not disturb physiological rhythm which underlais in the base of the function of the biological clock. The stability of biological rhythms and their indeterminism in respect of described above external and internal factors is real thanks to dissipation, which makes considerable interval of uncertainties of distributions of distances between segments of biopolymers and thus, of their fermentative activities (Pundjak,2001). Therefore the rise of biological clocks of each organism is in certain sense transcendental.

  11. Transcriptomic responses of germinating Bacillus subtilis spores exposed to 1.5 years of space and simulated martian conditions on the EXPOSE-E experiment PROTECT.

    PubMed

    Nicholson, Wayne L; Moeller, Ralf; Horneck, Gerda

    2012-05-01

    Because of their ubiquity and resistance to spacecraft decontamination, bacterial spores are considered likely potential forward contaminants on robotic missions to Mars. Thus, it is important to understand their global responses to long-term exposure to space or martian environments. As part of the PROTECT experiment, spores of B. subtilis 168 were exposed to real space conditions and to simulated martian conditions for 559 days in low-Earth orbit mounted on the EXPOSE-E exposure platform outside the European Columbus module on the International Space Station. Upon return, spores were germinated, total RNA extracted, fluorescently labeled, and used to probe a custom Bacillus subtilis microarray to identify genes preferentially activated or repressed relative to ground control spores. Increased transcript levels were detected for a number of stress-related regulons responding to DNA damage (SOS response, SPβ prophage induction), protein damage (CtsR/Clp system), oxidative stress (PerR regulon), and cell envelope stress (SigV regulon). Spores exposed to space demonstrated a much broader and more severe stress response than spores exposed to simulated martian conditions. The results are discussed in the context of planetary protection for a hypothetical journey of potential forward contaminant spores from Earth to Mars and their subsequent residence on Mars.

  12. Heat killing of bacterial spores analyzed by differential scanning calorimetry.

    PubMed

    Belliveau, B H; Beaman, T C; Pankratz, H S; Gerhardt, P

    1992-07-01

    Thermograms of the exosporium-lacking dormant spores of Bacillus megaterium ATCC 33729, obtained by differential scanning calorimetry, showed three major irreversible endothermic transitions with peaks at 56, 100, and 114 degrees C and a major irreversible exothermic transition with a peak at 119 degrees C. The 114 degrees C transition was identified with coat proteins, and the 56 degrees C transition was identified with heat inactivation. Thermograms of the germinated spores and vegetative cells were much alike, including an endothermic transition attributable to DNA. The ascending part of the main endothermic 100 degrees C transition in the dormant-spore thermograms corresponded to a first-order reaction and was correlated with spore death; i.e., greater than 99.9% of the spores were killed when the transition peak was reached. The maximum death rate of the dormant spores during calorimetry, calculated from separately measured D and z values, occurred at temperatures above the 73 degrees C onset of thermal denaturation and was equivalent to the maximum inactivation rate calculated for the critical target. Most of the spore killing occurred before the release of most of the dipicolinic acid and other intraprotoplast materials. The exothermic 119 degrees C transition was a consequence of the endothermic 100 degrees C transition and probably represented the aggregation of intraprotoplast spore components. Taken together with prior evidence, the results suggest that a crucial protein is the rate-limiting primary target in the heat killing of dormant bacterial spores.

  13. Quantitative and sensitive RNA based detection of Bacillus spores

    PubMed Central

    Osmekhina, Ekaterina; Shvetsova, Antonina; Ruottinen, Maria; Neubauer, Peter

    2014-01-01

    The fast and reliable detection of bacterial spores is of great importance and still remains a challenge. Here we describe a direct RNA-based diagnostic method for the specific detection of viable bacterial spores which does not depends on an enzymatic amplification step and therefore is directly appropriate for quantification. The procedure includes the following steps: (i) heat activation of spores, (ii) germination and enrichment cultivation, (iii) cell lysis, and (iv) analysis of 16S rRNA in crude cell lysates using a sandwich hybridization assay. The sensitivity of the method is dependent on the cultivation time and the detection limit; it is possible to detect 10 spores per ml when the RNA analysis is performed after 6 h of enrichment cultivation. At spore concentrations above 106 spores per ml the cultivation time can be shortened to 30 min. Total analysis times are in the range of 2–8 h depending on the spore concentration in samples. The developed procedure is optimized at the example of Bacillus subtilis spores but should be applicable to other organisms. The new method can easily be modified for other target RNAs and is suitable for specific detection of spores from known groups of organisms. PMID:24653718

  14. Effect of Lipid Materials on Heat Resistance of Bacterial Spores

    PubMed Central

    Molin, N.; Snygg, B. G.

    1967-01-01

    The apparent heat resistance of spores of Bacillus megaterium, B. subtilis, B. cereus, B. stearothermophilus, and Clostridium botulinum type E in lipids was investigated and compared with the resistance of the spores in phosphate buffer solution. The most pronounced increase in heat resistance was noted for B. subtilis and C. botulinum type E, the increase varying with the type of lipid used. A high water content of the lipids used as heating menstruum lowered the heat resistance of the spores. Possible explanations for the high heat resistance of spores in lipids are discussed. PMID:16349757

  15. Spore germination promoter of Dictyostelium discoideum excreted by Aerobacter aerogenes.

    PubMed

    Hashimoto, Y; Tanaka, Y; Yamada, T

    1976-07-01

    The nutrient medium in which Aerobacter aerogenes was grown, contains a spore germination promoter (SGP) for the cellular slime mould Dictyostelium discoideum. SGP can cuase synchronous spore germination in a short time, and triggers the germination process in just a few minutes. Germination-promoting capacity of SGP decreases as it comes in contact with increasing number of spores. When spores activated by SGP are stored at 4 degrees C, they gradually return to the dormant state. SGP is comparatively heat-stable, but is unstable at pH above 10 or under 3.

  16. The physical state of water in bacterial spores

    PubMed Central

    Sunde, Erik P.; Setlow, Peter; Hederstedt, Lars; Halle, Bertil

    2009-01-01

    The bacterial spore, the hardiest known life form, can survive in a metabolically dormant state for many years and can withstand high temperatures, radiation, and toxic chemicals. The molecular basis of spore dormancy and resistance is not understood, but the physical state of water in the different spore compartments is thought to play a key role. To characterize this water in situ, we recorded the water 2H and 17O spin relaxation rates in D2O-exchanged Bacillus subtilis spores over a wide frequency range. The data indicate high water mobility throughout the spore, comparable with binary protein–water systems at similar hydration levels. Even in the dense core, the average water rotational correlation time is only 50 ps. Spore dormancy therefore cannot be explained by glass-like quenching of molecular diffusion but may be linked to dehydration-induced conformational changes in key enzymes. The data demonstrate that most spore proteins are rotationally immobilized, which may contribute to heat resistance by preventing heat-denatured proteins from aggregating irreversibly. We also find that the water permeability of the inner membrane is at least 2 orders of magnitude lower than for model membranes, consistent with the reported high degree of lipid immobilization in this membrane and with its proposed role in spore resistance to chemicals that damage DNA. The quantitative results reported here on water mobility and transport provide important clues about the mechanism of spore dormancy and resistance, with relevance to food preservation, disease prevention, and astrobiology. PMID:19892742

  17. Dispersal of fungal spores on a cooperatively generated wind

    PubMed Central

    Roper, Marcus; Seminara, Agnese; Bandi, M. M.; Cobb, Ann; Dillard, Helene R.; Pringle, Anne

    2010-01-01

    Because of their microscopic size, the forcibly ejected spores of ascomycete fungi are quickly brought to rest by drag. Nonetheless some apothecial species, including the pathogen Sclerotinia sclerotiorum, disperse with astonishing rapidity between ephemeral habitats. Here we show that by synchronizing the ejection of thousands of spores, these fungi create a flow of air that carries spores through the nearly still air surrounding the apothecium, around intervening obstacles, and to atmospheric currents and new infection sites. High-speed imaging shows that synchronization is self-organized and likely triggered by mechanical stresses. Although many spores are sacrificed to produce the favorable airflow, creating the potential for conflict among spores, the geometry of the spore jet physically targets benefits of the airflow to spores that cooperate maximally in its production. The ability to manipulate a local fluid environment to enhance spore dispersal is a previously overlooked feature of the biology of fungal pathogens, and almost certainly shapes the virulence of species including S. sclerotiorum. Synchronous spore ejection may also provide a model for the evolution of stable, self-organized behaviors. PMID:20880834

  18. Defining the natural habitat of Bacillus spore-formers.

    PubMed

    Hong, Huynh A; To, Ellen; Fakhry, Saad; Baccigalupi, Loredana; Ricca, Ezio; Cutting, Simon M

    2009-01-01

    Our understanding of the genetics and physiology of the spore-forming genus Bacillus is remarkable. On the other hand, though, where these Gram-positive bacteria live and grow is far from clear. The soil, once considered their habitat, may simply serve as a reservoir. A growing number of studies show that Bacillus spores can be found in the intestinal tracts of animals, raising the question of whether this could be where they live and grow. In this study, we have conducted the first evaluation of Bacillus spore formers in soil and in human faeces. Our aim is simply to determine the abundance of aerobic spore-formers. Our results show that soil carries approximately approximately 10(6)spores/g while human faeces an average of up to 10(4)spores/g. The numbers of spores found in faeces, we reason, is too high to be accounted for principally by ingestion of food contaminated with spores from soil. This provides further evidence that Bacillus spore formers may have adapted to survival within the intestinal tract of insects and other animals that ingest them; if so they may well be hitherto undiscovered gut commensals.

  19. Entomology: A Bee Farming a Fungus.

    PubMed

    Oldroyd, Benjamin P; Aanen, Duur K

    2015-11-16

    Farming is done not only by humans, but also by some ant, beetle and termite species. With the discovery of a stingless bee farming a fungus that provides benefits to its larvae, bees can be added to this list.

  20. Spore-Forming Bacteria that Resist Sterilization

    NASA Technical Reports Server (NTRS)

    LaDuc, Myron; Venkateswaran, Kasthuri

    2003-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

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

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

    NASA Astrophysics Data System (ADS)

    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. Utilizing DART Mass Spectrometry to Pinpoint Halogenated Metabolites from a Marine Invertebrate-Derived Fungus

    PubMed Central

    Watts, Katharine R.; Loveridge, Steven T.; Tenney, Karen; Media, Joseph; Valeriote, Frederick A.; Crews, Phillip

    2011-01-01

    Prenylated indole alkaloids are a diverse group of fungal secondary metabolites and represent an important biosynthetic class. In this study we have identified new halogenated prenyl-indole alkaloids from an invertebrate-derived Malbranchea graminicola strain. Using Direct Analysis in Real Time (DART) Mass Spectrometry, these compounds were initially detected from spores of the fungus grown on agar plates, without the need for any organic extraction. Subsequently, the metabolites were isolated from liquid culture in artificial seawater. The structures of two novel chlorinated metabolites, named (−)-spiromalbramide and (+)-isomalbrancheamide B, provide additional insights into the assembly of the malbrancheamide compound family. Remarkably, two new brominated analogs, (+)-malbrancheamide C and (+)-isomalbrancheamide C, were produced by enriching the growth medium with bromine salts. PMID:21682275

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Rust fungi are biotrophic basidiomycete plant pathogens that cause major diseases on plants and trees world-wide, affecting agriculture and forestry. Their biotrophic nature precludes many established molecular genetic manipulations and lines of research. The generation of genomic resour...

  5. The Role of the Electrostatic Force in Spore Adhesion

    SciTech Connect

    Chung, Eunhyea; Yiacoumi, Sotira; Lee, Ida; Tsouris, Costas

    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. Mushrooms as Rainmakers: How Spores Act as Nuclei for Raindrops

    PubMed Central

    2015-01-01

    Millions of tons of fungal spores are dispersed in the atmosphere every year. These living cells, along with plant spores and pollen grains, may act as nuclei for condensation of water in clouds. Basidiospores released by mushrooms form a significant proportion of these aerosols, particularly above tropical forests. Mushroom spores are discharged from gills by the rapid displacement of a droplet of fluid on the cell surface. This droplet is formed by the condensation of water on the spore surface stimulated by the secretion of mannitol and other hygroscopic sugars. This fluid is carried with the spore during discharge, but evaporates once the spore is airborne. Using environmental electron microscopy, we have demonstrated that droplets reform on spores in humid air. The kinetics of this process suggest that basidiospores are especially effective as nuclei for the formation of large water drops in clouds. Through this mechanism, mushroom spores may promote rainfall in ecosystems that support large populations of ectomycorrhizal and saprotrophic basidiomycetes. Our research heightens interest in the global significance of the fungi and raises additional concerns about the sustainability of forests that depend on heavy precipitation. PMID:26509436

  7. Enumerating Spore-Forming Bacteria Airborne with Particles

    NASA Technical Reports Server (NTRS)

    Lin, Ying; Barengoltz, Jack

    2006-01-01

    A laboratory method has been conceived to enable the enumeration of (1) Cultivable bacteria and bacterial spores that are, variously, airborne by themselves or carried by, parts of, or otherwise associated with, other airborne particles; and (2) Spore-forming bacteria among all of the aforementioned cultivable microbes.

  8. Inhibition of Bacillus anthracis Spore Outgrowth by Nisin▿

    PubMed Central

    Gut, Ian M.; Prouty, Angela M.; Ballard, Jimmy D.; van der Donk, Wilfred A.; Blanke, Steven R.

    2008-01-01

    The lantibiotic nisin has previously been reported to inhibit the outgrowth of spores from several Bacillus species. However, the mode of action of nisin responsible for outgrowth inhibition is poorly understood. By using B. anthracis Sterne 7702 as a model, nisin acted against spores with a 50% inhibitory concentration (IC50) and an IC90 of 0.57 μM and 0.90 μM, respectively. Viable B. anthracis organisms were not recoverable from cultures containing concentrations of nisin greater than the IC90. These studies demonstrated that spores lose heat resistance and become hydrated in the presence of nisin, thereby ruling out a possible mechanism of inhibition in which nisin acts to block germination initiation. Rather, germination initiation is requisite for the action of nisin. This study also revealed that nisin rapidly and irreversibly inhibits growth by preventing the establishment of oxidative metabolism and the membrane potential in germinating spores. On the other hand, nisin had no detectable effects on the typical changes associated with the dissolution of the outer spore structures (e.g., the spore coats, cortex, and exosporium). Thus, the action of nisin results in the uncoupling of two critical sequences of events necessary for the outgrowth of spores: the establishment of metabolism and the shedding of the external spore structures. PMID:18809941

  9. Binding Affinity of Glycoconjugates to BACILLUS Spores and Toxins

    NASA Astrophysics Data System (ADS)

    Rasol, Aveen; Eassa, Souzan; Tarasenko, Olga

    2010-04-01

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

  10. Microalgae harvesting via co-culture with filamentous fungus

    NASA Astrophysics Data System (ADS)

    Gultom, Sarman Oktovianus

    Microalgae harvesting is a labor- and energy-intensive process. For instance, classical harvesting technologies such as chemical addition and mechanical separation are economically prohibiting for biofuel production. Newer approaches to harvest microalgae have been developed in order to decrease costs. Among these new methods, fungal co-pelletization seems to be a promising technology. By co-culturing filamentous fungi with microalgae, it is possible to form pellets, which can easily be separated. In this study, different parameters for the cultivation of filamentous fungus (Aspergillus niger) and microalgae (Chlorella vulgaris) to efficiently form cell pellets were evaluated under heterotrophic and phototrophic conditions, including organic carbon source (glucose, glycerol and sodium acetate) concentration, pH, initial concentration of fungal spores, initial concentration of microalgal cells, concentration of ionic strength (Calcium and Magnesium) and concentration of salinity (NaCl). In addition, zeta-potential measurements were carried out in order to get a better understanding of the mechanism of attraction. It was found that 2 g/L of glucose, a fungus to microalgae ratio of 1:300, and uncontrolled pH (around 7) are the best culturing conditions for co-pelletization. Under these conditions, it was possible to achieve a high harvesting performance (>90%). In addition, it was observed that most pellets formed in the co-culture were spherical with an average diameter of 3.5 mm and in concentrations of about 5 pellets per mL of culture media. Under phototrophic conditions, co-pelletization required the addition of glucose as organic carbon source to sustain the growth of fungi and to allow the harvesting of microalgae. Zeta-potential measurements indicated that (i) both microalgae and fungi have low zeta-potential values regardless of the pH on the bulk (i.e. <-10 mV) (ii) fungi can have a positive electric charge at low pH (ie. pH=3). These values suggest that it

  11. Adenylyl cyclase G, an osmosensor controlling germination of Dictyostelium spores.

    PubMed

    van Es, S; Virdy, K J; Pitt, G S; Meima, M; Sands, T W; Devreotes, P N; Cotter, D A; Schaap, P

    1996-09-27

    Dictyostelium cells express a G-protein-coupled adenylyl cyclase, ACA, during aggregation and an atypical adenylyl cyclase, ACG, in mature spores. The ACG gene was disrupted by homologous recombination. acg- cells developed into normal fruiting bodies with viable spores, but spore germination was no longer inhibited by high osmolarity, a fairly universal constraint for spore and seed germination. ACG activity, measured in aca-/ACG cells, was strongly stimulated by high osmolarity with optimal stimulation occurring at 200 milliosmolar. RdeC mutants, which display unrestrained protein kinase A (PKA) activity and a cell line, which overexpresses PKA under a prespore specific promoter, germinate very poorly, both at high and low osmolarity. These data indicate that ACG is an osmosensor controlling spore germination through activation of protein kinase A.

  12. Removal of dissolved heavy metals and radionuclides by microbial spores

    SciTech Connect

    Revis, N.W.; Hadden, C.T.; Edenborn, H.

    1997-11-01

    Microbial systems have been shown to remove specific heavy metals from contaminated aqueous waste to levels acceptable to EPA for environmental release. However, systems capable of removing a variety of heavy metals from aqueous waste to environmentally acceptable levels remain to be reported. The present studies were performed to determine the specificity of spores of the bacterium Bacillus megaterium for the adsorption of dissolved metals and radionuclides from aqueous waste. The spores effectively adsorbed eight heavy metals from a prepared metal mix and from a plating rinse waste to EPA acceptable levels for waste water. These results suggest that spores have multiple binding sites for the adsorption of heavy metals. Spores were also effective in adsorbing the radionuclides {sup 85}strontium and {sup 197}cesium. The presence of multiple sites in spores for the adsorption of heavy metals and radionuclides makes this biosorbent a good candidate for the treatment of aqueous wastes associated with the plating and nuclear industries. 17 refs., 4 tabs.

  13. Bacillus atrophaeus Outer Spore Coat Assembly and Ultrastructure

    SciTech Connect

    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.

  14. Quantification of Spore-forming Bacteria Carried by Dust Particles

    NASA Technical Reports Server (NTRS)

    Lin, Ying; Cholakian, Tanya; Gao, Wenming; Osman, Shariff; Barengoltz, Jack

    2006-01-01

    In order to establish a biological contamination transport model for predicting the cross contamination risk during spacecraft assembly and upon landing on Mars, it is important to understand the relationship between spore-forming bacteria and their carrier particles. 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 determining forward contamination risks for future missions.

  15. Bacillus atrophaeus outer spore coat assembly and ultrastructure.

    PubMed

    Plomp, Marco; Leighton, Terrance J; Wheeler, Katherine E; Pitesky, Maurice E; Malkin, Alexander J

    2005-11-08

    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 approximately 11 nm thick rodlets, having a periodicity of approximately 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.

  16. Surface Sampling Methods for Bacillus anthracis Spore Contamination

    PubMed Central

    Hein, Misty J.; Taylor, Lauralynn; Curwin, Brian D.; Kinnes, Gregory M.; Seitz, Teresa A.; Popovic, Tanja; Holmes, Harvey T.; Kellum, Molly E.; McAllister, Sigrid K.; Whaley, David N.; Tupin, Edward A.; Walker, Timothy; Freed, Jennifer A.; Small, Dorothy S.; Klusaritz, Brian; Bridges, John H.

    2002-01-01

    During an investigation conducted December 17–20, 2001, we collected environmental samples from a U.S. postal facility in Washington, D.C., known to be extensively contaminated with Bacillus anthracis spores. Because methods for collecting and analyzing B. anthracis spores have not yet been validated, our objective was to compare the relative effectiveness of sampling methods used for collecting spores from contaminated surfaces. Comparison of wipe, wet and dry swab, and HEPA vacuum sock samples on nonporous surfaces indicated good agreement between results with HEPA vacuum and wipe samples. However, results from HEPA vacuum sock and wipe samples agreed poorly with the swab samples. Dry swabs failed to detect spores >75% of the time they were detected by wipe and HEPA vacuum samples. Wipe samples collected after HEPA vacuum samples and HEPA vacuum samples after wipe samples indicated that neither method completely removed spores from the sampled surfaces. PMID:12396930

  17. Isolation and in vitro cultivation of Auriculoscypha anacardiicola D.A. Reid et Manim., an insect-associated and potentially medicinal fungus from India.

    PubMed

    Kumar, T K Arun; Chandran, Jisha K; Sreekandan, Jisha E; Manimohan, Patinjareveettil

    2011-01-01

    Auriculoscypha anacardiicola, an obligate insect-associate and a potential medicinal fungus, is isolated and studied in vitro. Suitable methods for isolation and cultivation of the fungus have been developed. Incubating spore deposits made to fall from basidiomata on tap water agar seems to be the best method for developing cultures. Successful isolations were also accomplished from infected coccids. Cultures could not be developed from single basidiospores and from tissues of the basidiomata. Although production of ballistospores and blastospores as well as germ tube formation were observed at the time of germination of basidiospores, budding blastospores alone produced mycelial cultures. Observations such as the inability of single basidiospores to germinate, emergence of mycelium from a spore deposit, and the apparent conjugation of yeast cells indicate that dikaryotization resulting from fusion of compatible yeast cells is essential for development of mycelium in A. anacardiicola. The fungus grew well on all complex media tested. It seems that a purely synthetic medium devoid of any growth factors cannot support the growth ofA. anacardiicola and yeast extract seems to provide the required growth factors.

  18. Characterizing Aeroallergens by Infrared Spectroscopy of Fungal Spores and Pollen

    PubMed Central

    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 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. Results 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. PMID:25867755

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

    SciTech Connect

    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.

  20. Functional characterization of Clostridium difficile spore coat proteins.

    PubMed

    Permpoonpattana, Patima; Phetcharaburanin, Jutarop; Mikelsone, Anna; Dembek, Marcin; Tan, Sisareuth; Brisson, Marie-Clémence; La Ragione, Roberto; Brisson, Alain R; Fairweather, Neil; Hong, Huynh A; Cutting, Simon M

    2013-04-01

    Spores of Clostridium difficile play a key role in the dissemination of this important human pathogen, and until recently little has been known of their functional characteristics. Genes encoding six spore coat proteins (cotA, cotB, cotCB, cotD, cotE, and sodA) were disrupted by ClosTron insertional mutagenesis. Mutation of one gene, cotA, presented a major structural defect in spore assembly, with a clear misassembly of the outermost layers of the spore coat. The CotA protein is most probably subject to posttranslational modification and could play a key role in stabilizing the spore coat. Surprisingly, mutation of the other spore coat genes did not affect the integrity of the spore, although for the cotD, cotE, and sodA mutants, enzyme activity was reduced or abolished. This could imply that these enzymatic proteins are located in the exosporium or alternatively that they are structurally redundant. Of the spore coat proteins predicted to carry enzymatic activity, three were confirmed to be enzymes using both in vivo and in vitro methods, the latter using recombinant expressed proteins. These were a manganese catalase, encoded by cotD, a superoxide dismutase (SOD), encoded by sodA, and a bifunctional enzyme with peroxiredoxin and chitinase activity, encoded by cotE. These enzymes being exposed on the spore surface would play a role in coat polymerization and detoxification of H2O2. Two additional proteins, CotF (a tyrosine-rich protein and potential substrate for SodA) and CotG (a putative manganese catalase) were shown to be located at the spore surface.

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

    NASA Astrophysics Data System (ADS)

    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.

  2. Fungus-insect gall of Phlebopus portentosus.

    PubMed

    Zhang, Chun-Xia; He, Ming-Xia; Cao, Yang; Liu, Jing; Gao, Feng; Wang, Wen-Bing; Ji, Kai-Ping; Shao, Shi-Cheng; Wang, Yun

    2015-01-01

    Phlebopus portentosus is a popular edible wild mushroom found in the tropical Yunnan, China, and northern Thailand. In its natural habitats, a gall often has been found on some plant roots, around which fungal fruiting bodies are produced. The galls are different from common insect galls in that their cavity walls are not made from plant tissue but rather from the hyphae of P. portentosus. Therefore we have termed this phenomenon "fungus-insect gall". Thus far six root mealy bug species in the family Pseudococcidae that form fungus-insect galls with P. portentosus have been identified: Formicococcus polysperes, Geococcus satellitum, Planococcus minor, Pseudococcus cryptus, Paraputo banzigeri and Rastrococcus invadens. Fungus-insect galls were found on the roots of more than 21 plant species, including Delonix regia, Citrus maxima, Coffea arabica and Artocarpus heterophyllus. Greenhouse inoculation trials showed that fungus-insect galls were found on the roots of A. heterophyllus 1 mo after inoculation. The galls were subglobose to globose, fulvous when young and became dark brown at maturation. Each gall harbored one or more mealy bugs and had a chimney-like vent for ventilation and access to the gall. The cavity wall had three layers. Various shaped mealy bug wax deposits were found inside the wall. Fungal hyphae invaded the epidermis of plant roots and sometimes even the cortical cells during the late stage of gall development. The identity of the fungus inside the cavity was confirmed by molecular methods.

  3. Studies on Bacterial Spore Ultraviolet Light Resistance and Regulation of the Activity of a Spore Protease

    DTIC Science & Technology

    1993-12-08

    SASP) of bacteria, FEBS Lett. 3M5, 115-120 (1992). Popham, D. and P. Setlow, The cortical peptidoglycan from spores of Bacillus megaterium and Bacillus ...vitro. We have shown that purified W/O3-type SASP from both Clostridium and Bacillus species interact similarly with and have the same effects on DNA...sequenced, and the proteins coded for were extremely homologous to those of Bacillus species. This work significantly extended the evolutionary time

  4. Methods for neutralizing anthrax or anthrax spores

    DOEpatents

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

    2013-02-26

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

  5. Optimization of nutritional requirements for mycelial growth and sporulation of entomogenous fungus Aschersonia aleyrodis Webber

    PubMed Central

    Zhu, Yanping; Pan, Jieru; Qiu, Junzhi; Guan, Xiong

    2008-01-01

    The objective of the present study was to investigate the optimal nutritional requirements for mycelial growth and sporulation of entomopathogenic fungus Aschersonia aleyrodis Webber by orthogonal layout methods. Herein the order of effects of nutrient components on mycelial growth was tryptone > Ca2+ > soluble starch > folacin, corresponding to the following optimal concentrations: 1.58% Soluble Starch, 3.16% Tryptone, 0.2 mmol l-1 Ca2+ and 0.005% Folacin. The optimal concentration of each factors for sporulation was 1.16% lactose, 0.394% tryptone, 0.4 mmol l-1 Fe2+ and 0.00125% VB1, and the effects of medium components on sporulation were found to be in the order lactose > VB1 > Fe2+ > tryptone. Under the optimal culture conditions, the maximum production of mycelial growth achieved 20.05 g l-1 after 7 days of fermentation, while the maximum spore yield reached 5.23 ×1010 spores l-1 after 22 days of cultivation. This is the first report on optimization of nutritional requirements and design of simplified semi-synthetic media for mycelial growth and sporulation of A. aleyrodis. PMID:24031305

  6. Optimization of nutritional requirements for mycelial growth and sporulation of entomogenous fungus Aschersonia aleyrodis Webber.

    PubMed

    Zhu, Yanping; Pan, Jieru; Qiu, Junzhi; Guan, Xiong

    2008-10-01

    The objective of the present study was to investigate the optimal nutritional requirements for mycelial growth and sporulation of entomopathogenic fungus Aschersonia aleyrodis Webber by orthogonal layout methods. Herein the order of effects of nutrient components on mycelial growth was tryptone > Ca(2+) > soluble starch > folacin, corresponding to the following optimal concentrations: 1.58% Soluble Starch, 3.16% Tryptone, 0.2 mmol l(-1) Ca(2+) and 0.005% Folacin. The optimal concentration of each factors for sporulation was 1.16% lactose, 0.394% tryptone, 0.4 mmol l(-1) Fe(2+) and 0.00125% VB1, and the effects of medium components on sporulation were found to be in the order lactose > VB1 > Fe(2+) > tryptone. Under the optimal culture conditions, the maximum production of mycelial growth achieved 20.05 g l(-1) after 7 days of fermentation, while the maximum spore yield reached 5.23 ×10(10) spores l(-1) after 22 days of cultivation. This is the first report on optimization of nutritional requirements and design of simplified semi-synthetic media for mycelial growth and sporulation of A. aleyrodis.

  7. Peroxisome dynamics during development of the fungus Podospora anserina.

    PubMed

    Takano-Rojas, Harumi; Zickler, Denise; Peraza-Reyes, Leonardo

    2016-01-01

    Peroxisomes are versatile and dynamic organelles that are required for the development of diverse eukaryotic organisms. We demonstrated previously that in the fungus Podospora anserina different peroxisomal functions are required at distinct stages of sexual development, including the initiation and progression of meiocyte (ascus) development and the differentiation and germination of sexual spores (ascospores). Peroxisome assembly during these processes relies on the differential activity of the protein machinery that drives the import of proteins into the organelle, indicating a complex developmental regulation of peroxisome formation and activity. Here we demonstrate that peroxisome dynamics is also highly regulated during development. We show that peroxisomes in P. anserina are highly dynamic and respond to metabolic and environmental cues by undergoing changes in size, morphology and number. In addition, peroxisomes of vegetative and sexual cell types are structurally different. During sexual development peroxisome number increases at two stages: at early ascus differentiation and during ascospore formation. These processes are accompanied by changes in peroxisome structure and distribution, which include a cell-polarized concentration of peroxisomes at the beginning of ascus development, as well as a morphological transition from predominantly spherical to elongated shapes at the end of the first meiotic division. Further, the mostly tubular peroxisomes present from second meiotic division to early ascospore formation again become rounded during ascospore differentiation. Ultimately the number of peroxisomes dramatically decreases upon ascospore maturation. Our results reveal a precise regulation of peroxisome dynamics during sexual development and suggest that peroxisome constitution and function during development is defined by the coordinated regulation of the proteins that control peroxisome assembly and dynamics.

  8. Long-Distance Wind-Dispersal of Spores in a Fungal Plant Pathogen: Estimation of Anisotropic Dispersal Kernels from an Extensive Field Experiment

    PubMed Central

    Rieux, Adrien; Soubeyrand, Samuel; Bonnot, François; Klein, Etienne K.; Ngando, Josue E.; Mehl, Andreas; Ravigne, Virginie; Carlier, Jean; de Lapeyre de Bellaire, Luc

    2014-01-01

    Given its biological significance, determining the dispersal kernel (i.e., the distribution of dispersal distances) of spore-producing pathogens is essential. Here, we report two field experiments designed to measure disease gradients caused by sexually- and asexually-produced spores of the wind-dispersed banana plant fungus Mycosphaerella fijiensis. Gradients were measured during a single generation and over 272 traps installed up to 1000 m along eight directions radiating from a traceable source of inoculum composed of fungicide-resistant strains. We adjusted several kernels differing in the shape of their tail and tested for two types of anisotropy. Contrasting dispersal kernels were observed between the two types of spores. For sexual spores (ascospores), we characterized both a steep gradient in the first few metres in all directions and rare long-distance dispersal (LDD) events up to 1000 m from the source in two directions. A heavy-tailed kernel best fitted the disease gradient. Although ascospores distributed evenly in all directions, average dispersal distance was greater in two different directions without obvious correlation with wind patterns. For asexual spores (conidia), few dispersal events occurred outside of the source plot. A gradient up to 12.5 m from the source was observed in one direction only. Accordingly, a thin-tailed kernel best fitted the disease gradient, and anisotropy in both density and distance was correlated with averaged daily wind gust. We discuss the validity of our results as well as their implications in terms of disease diffusion and management strategy. PMID:25116080

  9. Alternariol 9-methyl ether from the endophytic fungus Alternaria sp. Samif01 and its bioactivities

    PubMed Central

    Lou, Jingfeng; Yu, Ruiting; Wang, Xiaohan; Mao, Ziling; Fu, Linyun; Liu, Yang; Zhou, Ligang

    2016-01-01

    One bioactive compound, identified as alternariol 9-methyl ether, was isolated from the crude extract of the endophytic fungus Alternaria sp. Samif01 residing in the roots of Salvia miltiorrhiza Bunge. Alternariol 9-methyl ether was active against bacteria with minimum inhibitory concentration values ranging from 25 to 75 μg/mL and median inhibitory concentration (IC50) values ranging from 16.00 to 38.27 μg/mL. The IC50 value of alternariol 9-methyl ether against spore germination of Magnaporthe oryzae was 87.18 μg/mL. Alternariol 9-methyl ether also showed antinematodal activity against Bursaphelenchus xylophilus and Caenorhabditis elegans with IC50 values of 98.17 μg/mL and 74.62 μg/mL, respectively. This work is the first report on alternariol 9-methyl ether and its biological activities from the endophytic fungus Alternaria sp. Samif01 derived from S. miltiorrhiza Bunge. The results indicate the potential of Alternaria sp. Samif01 as a source of alternariol 9-methyl ether and also support that alternariol 9-methyl ether is a natural compound with high potential bioactivity against microorganisms. PMID:26887231

  10. Alternariol 9-methyl ether from the endophytic fungus Alternaria sp. Samif01 and its bioactivities.

    PubMed

    Lou, Jingfeng; Yu, Ruiting; Wang, Xiaohan; Mao, Ziling; Fu, Linyun; Liu, Yang; Zhou, Ligang

    2016-01-01

    One bioactive compound, identified as alternariol 9-methyl ether, was isolated from the crude extract of the endophytic fungus Alternaria sp. Samif01 residing in the roots of Salvia miltiorrhiza Bunge. Alternariol 9-methyl ether was active against bacteria with minimum inhibitory concentration values ranging from 25 to 75μg/mL and median inhibitory concentration (IC50) values ranging from 16.00 to 38.27μg/mL. The IC50 value of alternariol 9-methyl ether against spore germination of Magnaporthe oryzae was 87.18μg/mL. Alternariol 9-methyl ether also showed antinematodal activity against Bursaphelenchus xylophilus and Caenorhabditis elegans with IC50 values of 98.17μg/mL and 74.62μg/mL, respectively. This work is the first report on alternariol 9-methyl ether and its biological activities from the endophytic fungus Alternaria sp. Samif01 derived from S. miltiorrhiza Bunge. The results indicate the potential of Alternaria sp. Samif01 as a source of alternariol 9-methyl ether and also support that alternariol 9-methyl ether is a natural compound with high potential bioactivity against microorganisms.

  11. Knock down of chitosanase expression in phytopathogenic fungus Fusarium solani and its effect on pathogenicity.

    PubMed

    Liu, Huaiwei; Zhang, Bo; Li, Changsong; Bao, Xiaoming

    2010-06-01

    Chitosanases are lytic enzymes involved in the degradation of chitosan, a component of fungal cell walls. The phytopathogenic fungus Fusarium solani produces an extracellular chitosanase, CSN1, the role of which in the physiology and virulence of the fungus remains to be expounded. Here, we studied the expression of the CSN1 gene through gene silencing and examined its effect on fungal pathogenicity. A vector construct encoding a hairpin RNA (hpRNA) of CSN1 was constructed and introduced into the F. solani 0114 strain. The results revealed that majority of the transformants exhibited a significant reduction in chitosanase activity compared with the wild-type strain. Further, transformants with silenced CSN1 exhibited no change in mycelial growth and spore formation. However, pea pod and seedling bioassays indicated that transformants with silenced CSN1 were more virulent compared with the wild-type strain, and in sharp contrast to strains in which overexpression of the CSN1 gene resulted in virulence reduction. Although the mechanism remains unclear, our findings did suggest that F. solani chitosanase has a negative effect on fungal pathogenicity.

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

    NASA Astrophysics Data System (ADS)

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

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

  13. Coat and enterotoxin-related proteins in Clostridium perfringens spores.

    PubMed

    Ryu, S; Labbe, R G

    1989-11-01

    Coat proteins from mature spores of two enterotoxin-positive (Ent+) and two enterotoxin-negative (Ent-) strains of Clostridium perfringens were solubilized using 50 mM-dithiothreitol and 1% sodium dodecyl sulphate at pH 9.7, and alkylated using 110 mM-iodoacetamide to prevent aggregation. The coat proteins and C. perfringens type A enterotoxin (CPE) were separated by SDS-PAGE and analysed by Western blotting using anti-CPE antibody. As previously reported, CPE aggregated in the presence of SDS, but no aggregation occurred at concentrations below 15 micrograms CPE ml-1. Two CPE-related proteins (34 and 48 kDa) were found in the solubilized spore coat protein of Ent+ strains while only the 48 kDa CPE-related protein was found in the spore coat fraction of Ent- strains. CPE-related proteins comprised 2.7% and 0.8% of the total solubilized coat protein of Ent+ and Ent- strains respectively. CPE-related proteins could be extracted from the spores with 1% SDS alone. They could also be released by disruption of whole spores, indicating that the CPE-related proteins may be in the spore core or trapped between the core and coat layers. The results suggest that CPE is not a major structural component of the coat fraction of C. perfringens spores.

  14. Activity of essential oils against Bacillus subtilis spores.

    PubMed

    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.

  15. Nutritionally defined conditions for germination of Streptomyces viridochromogenes spores.

    PubMed Central

    Hirsch, C F; Ensign, J C

    1976-01-01

    Spores of Streptomyces viridochromogenes were removed from the surface of solid media with glass beads and suspended in a buffer-detergent solution. Addition of yeast extract and glucose resulted in rapid loss of refractility of the spores. Appearance of germ tubes followed. Germination was accompanied by a decrease in the optical density (OD) of the suspension. The OD decrease was used as an assay for germination. A defined germination medium (DGM) comprised of L-alanine, L-glutamic acid, adenosine, para-aminobenzoic acid, and calcium and magnesium ions provided a germination rate nearly equal to that of complex media. The germination rate was essentially the same if D-alanine and D-glutamate replaced the L-isomers. The optimum pH and temperature for germination were 7.0 and 35 C. Germination was absolutely dependent on the presence of CO2. Spores harvested after growth for longer periods than the usual time (10 days) became less germinable in DGM. The same was observed for spores grown at 37 C as compared with 30 C. Spores incubated in DGM for various time periods before being transferred to a buffer solution did not continue to germinate. Spores harvested after growth of eight species of Streptomyces did not show a decrease in OD when incubated in yeast extract medium. Another strain of S. viridochromogenes did exhibit an OD decrease in the medium. Comparative properties of spores of streptomycetes, fungi, and bacilli are discussed. Images PMID:4421

  16. Immunogenicity and protective efficacy of Clostridium difficile spore proteins.

    PubMed

    Ghose, Chandrabali; Eugenis, Ioannis; Edwards, Adrianne N; Sun, Xingmin; McBride, Shonna M; Ho, David D

    2016-02-01

    Clostridium difficile is a spore-forming, anaerobic, Gram-positive organism that is the leading cause of antibiotic-associated infectious diarrhea, commonly known as C. difficile infection (CDI). C. difficile spores play an important role in the pathogenesis of CDI. Spore proteins, especially those that are surface-bound may play an essential role in the germination, colonization and persistence of C. difficile in the human gut. In our current study, we report the identification of two surface-bound spore proteins, CdeC and CdeM that may be utilized as immunization candidates against C. difficile. These spore proteins are immunogenic in mice and are able to protect mice against challenge with C. difficile UK1, a clinically-relevant 027/B1/NAP1 strain. These spore proteins are also able to afford high levels of protection against challenge with C. difficile 630Δerm in golden Syrian hamsters. This unprecedented study shows the vaccination potential of C. difficile spore exosporium proteins.

  17. Water Behavior in Bacterial Spores by Deuterium NMR Spectroscopy

    PubMed Central

    2015-01-01

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

  18. Infrared signatures to discriminate viability of autoclaved Bacillus spores

    NASA Astrophysics Data System (ADS)

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

    2011-11-01

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

  19. Bacillus subtilis spores as adjuvants for DNA vaccines.

    PubMed

    Aps, Luana R M M; Diniz, Mariana O; Porchia, Bruna F M M; Sales, Natiely S; Moreno, Ana Carolina R; Ferreira, Luís C S

    2015-05-11

    Recently, Bacillus subtilis spores were shown to be endowed with strong adjuvant capacity when co-administered with purified antigenic proteins. In the present study we assessed whether spores possess adjuvant properties when combined with DNA vaccines. We showed that B. subtilis spores promoted the activation of dendritic cells in vitro and induced migration of pro-inflammatory cells after parenteral administration to mice. Likewise, co-administration of spores with a DNA vaccine encoding the human papillomavirus type 16 (HPV-16) E7 protein enhanced the activation of antigen-specific CD8(+) T cell responses in vivo. Mice immunized with the DNA vaccine admixed with spores presented a protective immunity increase to previously implanted tumor cells, capable of expressing HPV-16 oncoproteins. Finally, we observed that the adjuvant effect can vary accordingly to the number of co-administered spores which may be ascribed with the ability to induce. Collectively, the present results demonstrate for the first time that B. subtilis spores can also confer adjuvant effects to DNA vaccines.

  20. Immunomagnetic capture of Bacillus anthracis spores from food.

    PubMed

    Shields, Michael J; Hahn, Kristen R; Janzen, Timothy W; Goji, Noriko; Thomas, Matthew C; Kingombe, Cesar Bin I; Paquet, Chantal; Kell, Arnold J; Amoako, Kingsley K

    2012-07-01

    Food is a vulnerable target for potential bioterrorist attacks; therefore, a critical mitigation strategy is needed for the rapid concentration and detection of biothreat agents from food matrices. Magnetic beads offer a unique advantage in that they have a large surface area for efficient capture of bacteria. We have demonstrated the efficient capture and concentration of Bacillus anthracis (Sterne) spores using immunomagnetic beads for a potential food application. Magnetic beads from three different sources, with varying sizes and surface chemistries, were functionalized with monoclonal antibodies and polyclonal antibodies from commercial sources and used to capture and concentrate anthrax spores from spiked food matrices, including milk, apple juice, bagged salad, processed meat, and bottled water. The results indicated that the Pathatrix beads were more effective in the binding and capture of anthrax spores than the other two bead types investigated. Furthermore, it was observed that the use of polyclonal antibodies resulted in a more efficient recovery of anthrax spores than the use of monoclonal antibodies. Three different magnetic capture methods, inversion, the Pathatrix Auto system, and the new i CropTheBug system, were investigated. The i CropTheBug system yielded a much higher recovery of spores than the Pathatrix Auto system. Spore recoveries ranged from 80 to 100% for the i CropTheBug system when using pure spore preparations, whereas the Pathatrix Auto system had recoveries from 20 to 30%. Spore capture from food samples inoculated at a level of 1 CFU/ml resulted in 80 to 100% capture for milk, bottled water, and juice samples and 60 to 80% for processed meat and bagged salad when using the i CropTheBug system. This efficient capture of anthrax spores at very low concentrations without enrichment has the potential to enhance the sensitivity of downstream detection technologies and will be a useful method in a foodborne bioterrorism response.

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

    PubMed

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

    2015-11-05

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

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

    PubMed

    Semenyuk, Ekaterina G; Laning, Michelle L; Foley, Jennifer; Johnston, Pehga F; Knight, Katherine L; Gerding, Dale N; Driks, Adam

    2014-01-01

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

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

    PubMed Central

    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

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

    PubMed Central

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

    2016-01-01

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

  5. Dentigerumycin: a bacterial mediator of an ant-fungus symbiosis.

    PubMed

    Oh, Dong-Chan; Poulsen, Michael; Currie, Cameron R; Clardy, Jon

    2009-06-01

    Fungus-growing ants engage in mutualistic associations with both the fungus they cultivate for food and actinobacteria (Pseudonocardia spp.) that produce selective antibiotics to defend that fungus from specialized fungal parasites. We have analyzed one such system at the molecular level and found that the bacterium associated with the ant Apterostigma dentigerum produces dentigerumycin, a cyclic depsipeptide with highly modified amino acids, to selectively inhibit the associated parasitic fungus (Escovopsis sp.).

  6. New pressure and temperature effects on bacterial spores

    NASA Astrophysics Data System (ADS)

    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

  7. Activity of Pera Safe(Trademark) Against Bacillus Anthracis Spores

    DTIC Science & Technology

    2002-01-01

    peroxide and peracetic acid . J.Appl. Bacteriol. 1983,54,417-23. 2. Dietz P., Böhm R.: Results of an experimental study on testing disinfectants with spores...Bacteriol. 1980, 48, 161-90. 5. Hussaini S.N., Ruby K.R.: Sporicidal activity of peracetic acid against Bacillus anthracis spores. Vet. Rec. 1976, 98, 257-9. ...challenging task. There exist a variety of disinfectants that can inactivate Bacillus anthracis spores; however, most of them have negative side effects

  8. Heterologous expression of VHb can improve the yield and quality of biocontrol fungus Paecilomyces lilacinus, during submerged fermentation.

    PubMed

    Zhang, Shumeng; Wang, Jieping; Wei, Yale; Tang, Qing; Ali, Maria Kanwal; He, Jin

    2014-10-10

    Paecilomyces lilacinus is an egg-parasitic fungus which is effective against plant-parasitic nematodes and it has been successfully commercialized for the control of many plant-parasitic nematodes. However, during the large-scale industrial fermentation process of the filamentous fungus, the dissolved oxygen supply is a limiting factor, which influences yield, product quality and production cost. To solve this problem, we intended to heterologously express VHb in P. lilacinus ACSS. After optimizing the vgb gene, we fused it with a selection marker gene nptII, a promoter PgpdA and a terminator TtrpC. The complete expression cassette PgpdA-nptII-vgb-TtrpC was transferred into P. lilacinus ACSS by Agrobacterium tumefaciens-mediated transformation. Consequently, we successfully screened an applicable fungus strain PNVT8 which efficiently expressed VHb. The submerged fermentation experiments demonstrated that the expression of VHb not only increased the production traits of P. lilacinus such as biomass and spore production, but also improved the beneficial product quality and application value, due to the secretion of more protease and chitinase. It can be speculated that the recombinant strain harboring vgb gene will have a growth advantage over the original strain under anaerobic conditions in soil and therefore will possess higher biocontrol efficiency against plant-parasitic nematodes.

  9. Effects of High-Pressure Treatment on Spores of Clostridium Species

    PubMed Central

    Doona, Christopher J.; Feeherry, Florence E.; Setlow, Barbara; Wang, Shiwei; Li, William; Nichols, Frank C.; Talukdar, Prabhat K.; Sarker, Mahfuzur R.; Li, Yong-Qing; Shen, Aimee

    2016-01-01

    ABSTRACT This work analyzes the high-pressure (HP) germination of spores of the food-borne pathogen Clostridium perfringens (with inner membrane [IM] germinant receptors [GRs]) and the opportunistic pathogen Clostridium difficile (with no IM GRs), which has growing implications as an emerging food safety threat. In contrast to those of spores of Bacillus species, mechanisms of HP germination of clostridial spores have not been well studied. HP treatments trigger Bacillus spore germination through spores' IM GRs at ∼150 MPa or through SpoVA channels for release of spores' dipicolinic acid (DPA) at ≥400 MPa, and DPA-less spores have lower wet heat resistance than dormant spores. We found that C. difficile spores exhibited no germination events upon 150-MPa treatment and were not heat sensitized. In contrast, 150-MPa-treated unactivated C. perfringens spores released DPA and became heat sensitive, although most spores did not complete germination by fully rehydrating the spore core, but this treatment of heat-activated spores led to almost complete germination and greater heat sensitization. Spores of both clostridial organisms released DPA during 550-MPa treatment, but C. difficile spores did not complete germination and remained heat resistant. Heat-activated 550-MPa-HP-treated C. perfringens spores germinated almost completely and became heat sensitive. However, unactivated 550-MPa-treated C. perfringens spores did not germinate completely and were less heat sensitive than spores that completed germination. Since C. difficile and C. perfringens spores use different mechanisms for sensing germinants, our results may allow refinement of HP methods for their inactivation in foods and other applications and may guide the development of commercially sterile low-acid foods. IMPORTANCE Spores of various clostridial organisms cause human disease, sometimes due to food contamination by spores. Because of these spores' resistance to normal decontamination regimens, there

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

    NASA Astrophysics Data System (ADS)

    Tychinsky, V. P.; Mulyukin, A. L.; Lisovskii, V. V.; Nikolaev, Yu. A.; Kretushev, A. V.; Vyshenskaya, T. V.; Suzina, N. E.; Duda, V. I.; El-Registan, G. I.

    One of the challenging tasks in monitoring studies is to estimate heterogeneity of microbial populations by the physiological state and potential viability of individual cells especially with regard of their ability to withstand various environmental assaults Previously we described some approaches based on electron microscopy methods to discriminate vegetative dormant and dead cells in both aged microbial cultures and environmental samples including permafrost In this communication we propose to extend the arsenal of microscopy methods for monitoring studies by a new non-invasive and informative method - dynamic phase microscopy DPM The substantial advantage of DPM is that it gives quantitative digitized data of un-destroyed living microscopic objects exemplified in our work by Bacillus licheniformis spores Using DPM made it possible to record interference images of objects spores and to produce picture of their phase thickness PT that is the optical path difference in nm Thus it was demonstrated the remarkable difference in the PT of spores at different physiological states dormant germinating and heat-killed spores had PT values of 80 nm 40-50 nm and 20 nm respectively The other found criterion to distinguish between spores was the PT fluctuations In contrast to dormant and killed spores the PT of germinating spores oscillated with amplitude of up to 7 nm with typical frequencies of 1 3 and 3 4 Hz A combination of the recorded PT values and PT fluctuations gave a key to detect viable and dead cells Under the conditions that did not

  11. 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.

    PubMed

    Moeller, Ralf; Setlow, Peter; Horneck, Gerda; Berger, Thomas; Reitz, Günther; Rettberg, Petra; Doherty, Aidan J; Okayasu, Ryuichi; Nicholson, Wayne L

    2008-02-01

    The role of DNA repair by nonhomologous end joining (NHEJ), homologous recombination, spore photoproduct lyase, and DNA polymerase I and genome protection via alpha/beta-type small, acid-soluble spore proteins (SASP) in Bacillus subtilis spore resistance to accelerated heavy ions (high-energy charged [HZE] particles) and X rays has been studied. Spores deficient in NHEJ and alpha/beta-type SASP were significantly more sensitive to HZE particle bombardment and X-ray irradiation than were the recA, polA, and splB mutant and wild-type spores, indicating that NHEJ provides an efficient DNA double-strand break repair pathway during spore germination and that the loss of the alpha/beta-type SASP leads to a significant radiosensitivity to ionizing radiation, suggesting the essential function of these spore proteins as protectants of spore DNA against ionizing radiation.

  12. A guide to the recent literature on aspergillosis as caused by Aspergillus fumigatus, a fungus frequently found in self-heating organic matter.

    PubMed

    Marsh, P B; Millner, P D; Kla, J M

    1979-11-30

    Spores of Aspergillus fumigatus have been found to be abundantly present in the outdoor air at a site where large scale experimental composting of sewage sludge is in progress at Beltsville, Maryland. The health significance of this finding, for that site and for others in the future, is still only incompletely understood. Further studies are in progress to characterize absolute concentrations of the spores of the fungus in air at the site, spore dispersal by air from composting operations, and background environmental spore levels in air. The present paper contains a list of references to papers on health effects of A. fumigatus, many published in the past ten years, along with a review of the same designed to assist the reader in finding information on particular aspects of the subject in the literature. It is intended primarily as an aid to individuals interested in sludge composting and wishing to attain an insight into the A. fumigatus-composting situation, but it may also interest others concerned with other substrates which become moldy at 40--50 C. A. fumigatus has been found in great numbers in naturally and artificially heated environments such as decaying leaves, compost heaps, solar heated sloughs, cooling canals for nuclear power generators, silos, grain storage bins, boiler rooms, detritus around steam turbines and sauna baths. The evident practical merits of sludge composting have been described elsewhere; the information presented here has its main significance in respect to requirements for choice of locations for composting sites and to process and design criteria.

  13. Mechanism of Bacillus subtilis Spore Inactivation by and Resistance to Supercritical CO2 plus Peracetic Acid

    PubMed Central

    Setlow, Barbara; Korza, George; Blatt, Kelly M.S.; Fey, Julien P.; Setlow, Peter

    2015-01-01

    Aims Determine how supercritical CO2 (scCO2) plus peracetic acid (PAA) inactivates Bacillus subtilis spores, factors important in spore resistance to scCO2-PAA, and if spores inactivated by scCO2-PAA are truly dead. Methods and Results Spores of wild-type B. subtilis and isogenic mutants lacking spore protective proteins were treated with scCO2-PAA in liquid or dry at 35°C. Wild-type wet spores (aqueous suspension) were more susceptible than dry spores. Treated spores were examined for viability (and were truly dead), dipicolinic acid (DPA), mutations, permeability to nucleic acid stains, germination under different conditions, energy metabolism and outgrowth. ScCO2-PAA-inactivated spores retained DPA, and survivors had no notable DNA damage. However, DPA was released from inactivated spores at a normally innocuous temperature (85°C), and colony formation from treated spores was salt sensitive. The inactivated spores germinated but did not outgrow, and these germinated spores had altered plasma membrane permeability and defective energy metabolism. Wet or dry coat-defective spores had increased scCO2-PAA sensitivity, and dry spores but not wet spores lacking DNA protective proteins were more scCO2-PAA sensitive. Conclusions These findings suggest that scCO2-PAA inactivates spores by damaging spores’ inner membrane. The spore coat provided scCO2-PAA resistance for both wet and dry spores. DNA protective proteins provided scCO2-PAA resistance only for dry spores. Significance and Impact of Study These results provide information on mechanisms of spore inactivation of and resistance to scCO2-PAA, an agent with increasing use in sterilization applications. PMID:26535794

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  15. Alicyclobacillus acidoterrestris: new methods for inhibiting spore germination.

    PubMed

    Bevilacqua, A; Sinigaglia, M; Corbo, M R

    2008-07-15

    For a long period the thermal processing has been considered as the only way to reduce the initial spore number of Alicyclobacillus acidoterrestris and prevent the spoilage of acidic beverage. New methods, however, were proposed by the literature to control spore germination both in laboratory media and in real systems. After a brief introduction on the impact of A. acidoterrestris in food microbiology and a description of enumeration methods and heat processing applied by the juices manufactures, a review of innovative approaches to inhibit and/or control spore germination is proposed. In particular, this paper focuses on two different topics; the 1st is the use of some natural compounds (monolaurin, lysozyme, nisin and essential oils) or some chemicals, conventional (like sodium-benzoate, organic acids, surfactants and chlorine dioxide) or not conventional (chlorine dioxide as gas). The 2nd topic is a description of some innovative methods to reduce the initial spore number (high hydrostatic and homogenisation pressures, radiation and microwaves).

  16. Evaluation of Surface Sampling for Bacillus Spores Using ...

    EPA Pesticide Factsheets

    Report The primary objectives of this project were to evaluate the Aggressive Air Sampling (AAS) method compared to currently used surface sampling methods and to determine if AAS is a viable option for sampling Bacillus anthracis spores.

  17. Enhanced photocatalytic inactivation of bacterial spores on surfaces in air.

    PubMed

    Vohra, Amit; Goswami, D Y; Deshpande, D A; Block, S S

    2005-08-01

    TiO(2) photocatalysis with ultraviolet (UV-A) light has proven to be a highly effective process for complete inactivation of airborne microbes. However, the overall efficiency of the technology needs to be improved to make it more attractive as a defense against bio-terrorism. The present research investigates the enhancement in the rate of destruction of bacterial spores on metal (aluminum) and fabric (polyester) substrates with metal (silver)-doped titanium dioxide and compares it to conventional photocatalysis (TiO(2) P25/+UV-A) and UV-A photolysis. Bacillus cereus bacterial spores were used as an index to demonstrate the enhanced disinfection efficiency. The results indicate complete inactivation of B. cereus spores with the enhanced photocatalyst. The enhanced spore destruction rate may be attributed to the highly oxidizing radicals generated by the doped TiO(2).

  18. Inactivation of Bacillus anthracis Spores in Soil Matrices with ...

    EPA Pesticide Factsheets

    Report This report documents the results of a laboratory study designed to better understand the effectiveness of chlorine dioxide (ClO2) gas to decontaminate soil materials contaminated with Bacillus anthracis spores.

  19. Environmental Stresses on Spore Populations of Bacillus stearothermophilus1

    PubMed Central

    Fields, M. L.

    1964-01-01

    Heat-shocking spores at 110 C in 20% sucrose solutions decreased the percentage of the rough variant in a mixed population (rough and smooth variants) of strain M. Heat-shocking spores of the rough variant of strain NCA 1518 in 20% sucrose produced a decline in the number which germinated, whereas the smooth variant of strain NCA 1518 increased in the number which germinated. By the use of phase microscopy and plate counts, from the same incubated spore suspension in distilled water, heat-induced dormancy was demonstrated at 52 C. Dormancy also occurred in 20% sucrose solutions when held at room temperatures. Heat-shocking spores of strain M in 20% sucrose solutions and plating immediately after 24- and 48-hr holding periods at 25 C produced a decline in the total population with the percentage of rough variant increasing with time. A second heat shock produced only an increase in the rough variant. PMID:14215969

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

    PubMed Central

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

    2015-01-01

    Dormant bacterial spores are encased in a thick protein shell, the ‘coat', which contains ∼70 different proteins. The coat protects the spore from environmental insults, and is among the most durable static structures in biology. Owing to extensive cross-linking among coat proteins, this structure has been recalcitrant to detailed biochemical analysis, so molecular details of how it assembles are largely unknown. Here, we reconstitute the basement layer of the coat atop spherical membranes supported by silica beads to create artificial spore-like particles. We report that these synthetic spore husk-encased lipid bilayers (SSHELs) assemble and polymerize into a static structure, mimicking in vivo basement layer assembly during sporulation in Bacillus subtilis. In addition, we demonstrate that SSHELs may be easily covalently modified with small molecules and proteins. We propose that SSHELs may be versatile display platforms for drugs and vaccines in clinical settings, or for enzymes that neutralize pollutants for environmental remediation. PMID:25854653

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

    NASA Technical Reports Server (NTRS)

    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

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

    PubMed

    Wang; Li

    2000-08-01

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

  3. VUV absorption spectroscopy of bacterial spores and DNA components

    NASA Astrophysics Data System (ADS)

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

    2017-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2014-01-01

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

  5. Identifying bacterial spores and anthrax hoax materials by Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Farquharson, Stuart; Brouillette, Carl R.; Smith, Wayne

    2004-12-01

    The distribution of Bacillus anthracis spores through the US postal system in the autumn of 2001, initiated a secondary form of terror, the mailing of hoax materials. In the past three years nearly 20,000 letters containing harmless powders have been mailed, creating additional anxiety. Thus, there is a need for analyzers that can not only identify anthrax-causing spores to save lives, but also identify hoax materials to eliminate time-consuming and costly shutdowns. Recently, we established that Raman spectroscopy has the ability to identify both Bacilli endospores and hoax materials. Here we present Raman spectra of several Bacilli spores along with the dipicolinate salts, to further define the abilities of this technology to not only identify hoax materials, but also identify spores at the genus and species level.

  6. Solanapyrone analogues from a Hawaiian fungicolous fungus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four new solanayrone analogues (solanapyrones J-M; 1-4) have been isolated from an unidentified fungicolous fungus collected in Hawaii. The structures and relative configurations of these compounds were determined by analysis of ID NMR, 2D NMR, and MS data. Solanapyrone J(1) showed antifungal acti...

  7. The fission yeast spore is coated by a proteinaceous surface layer comprising mainly Isp3

    PubMed Central

    Fukunishi, Kana; Miyakubi, Kana; Hatanaka, Mitsuko; Otsuru, Natsumi; Hirata, Aiko; Shimoda, Chikashi; Nakamura, Taro

    2014-01-01

    The spore is a dormant cell that is resistant to various environmental stresses. As compared with the vegetative cell wall, the spore wall has a more extensive structure that confers resistance on spores. In the fission yeast Schizosaccharomyces pombe, the polysaccharides glucan and chitosan are major components of the spore wall; however, the structure of the spore surface remains unknown. We identify the spore coat protein Isp3/Meu4. The isp3 disruptant is viable and executes meiotic nuclear divisions as efficiently as the wild type, but isp3∆ spores show decreased tolerance to heat, digestive enzymes, and ethanol. Electron microscopy shows that an electron-dense layer is formed at the outermost region of the wild-type spore wall. This layer is not observed in isp3∆ spores. Furthermore, Isp3 is abundantly detected in this layer by immunoelectron microscopy. Thus Isp3 constitutes the spore coat, thereby conferring resistance to various environmental stresses. PMID:24623719

  8. Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes

    DTIC Science & Technology

    2014-10-30

    2010 31-May-2014 Approved for Public Release; Distribution Unlimited Final Report: (Life Science Division/Biochemistry) Inactivation of Bacillus ...S) AND ADDRESS (ES) U.S. Army Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 Bacillus Anthracis, Spores, Biofilm, Inhibition...Biochemistry) Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes Report Title The Specific Aims of the project were to investigate: 1) the

  9. Biochemical Changes and their Regulation during Spore Formation and Germination.

    DTIC Science & Technology

    1980-04-09

    or completed. Achievements in the three year period are given below. Most of the work utilized Bacillus megaterium , but other species have given...and the major low moTecular-weighfttol1/disulfide identified was Coenzyme A (CoA). Dormant spores of Bacillus megaterium were found to contain...x 10- M, respectively. Phosphoglycerate phosphomutase was purified to homogeneity from vegetative cells and germinated spores of Bacillus megaterium

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

    DTIC Science & Technology

    1983-01-01

    occurred with, for example, urea/mercaptoethanol (UME) treatment of Bacillus megaterium (Vary, 1973). alkaline dithioerythritol/sodium dodecyl sulphate...cetylmuremides of Bacillus subtilis YT 25. Agr. Biol. Chem. 38 2357-65. Accepted 15 June 1979 91 Germination of C perfringns spores Journal qf General Microbiology...spore-lytic enzymes of Bacillus cereus have been isolated and extensively studied by Strange & Dark (1957). Gould et al. (1966), Warth (1972) and Brown et

  11. Decontamination Of Bacterial Spores by a Peptide-Mimic

    DTIC Science & Technology

    2006-11-01

    germination. Rode and Foster (1961) studied dodecylamine induced germination of Bacillus megaterium by following optical density changes in spore...from the four Bacillus organisms is shown at four different temperatures and various times. 5 Bacillus megaterium 30 60 90 120 150 180 210 240 Fr... Bacillus anthracis, has called urgent attention to detailed studies of bacterial spores, especially from the point of view of their decontamination

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

    DTIC Science & Technology

    2006-05-31

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

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

    PubMed

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

    2012-07-01

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

  14. Tip-enhanced Raman scattering of bacillus subtilis spores

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

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

    2009-09-01

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

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

    PubMed

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

    2016-01-04

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

  17. Air sampling of mold spores by slit impactors: yield comparison.

    PubMed

    Pityn, Peter J; Anderson, James

    2013-01-01

    The performance of simple slit impactors for air sampling of mold contamination was compared under field conditions. Samples were collected side-by-side, outdoors in quadruplicates with Burkhard (ambient sampler) and Allergenco MK3 spore traps and with two identical Allergenco slit cassettes operated at diverse flow rates of 5 and 15 L/min, respectively. The number and types of mold spores in each sample were quantified by microscopy. Results showed all four single-stage slit impactors produced similar spore yields. Moreover, paired slit cassettes produced similar outcomes despite a three-fold difference in their sampling rate. No measurable difference in the amount or mix of mold spores per m(3)of air was detected. The implications for assessment of human exposures and interpretation of indoor/outdoor fungal burden are discussed. These findings demonstrate that slit cassettes capture most small spores, effectively and without bias, when operated at a range of flow rates including the lower flow rates used for personal sampling. Our findings indicate sampling data for mold spores correlate for different single stage impactor collection methodologies and that data quality is not deteriorated by operating conditions deviating from manufacturers' norms allowing such sampling results to be used for scientific, legal, investigative, or property insurance purposes. The same conclusion may not be applied to other particle sampling instruments and mulit-stage impactors used for ambient particulate sampling, which represent an entirely different scenario. This knowledge may help facilitate comparison between scientific studies where methodological differences exist.

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

    PubMed Central

    Barker, Gary C.; Malakar, Pradeep K.; Plowman, June

    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

  19. Display of native proteins on Bacillus subtilis spores.

    PubMed

    Pan, Jae-Gu; Choi, Soo-Keun; Jung, Heung-Chae; Kim, Eui-Joong

    2014-09-01

    In principle, protein display is enabled by fusing target proteins to naturally secreted, surface-anchored protein motifs. In this work, we developed a method of native protein display on the Bacillus spore surface that obviates the need to construct fusion proteins to display a motif. Spore coat proteins are expressed in the mother cell compartment and are subsequently assembled and deposited on the surface of spores. Therefore, target proteins overexpressed in the mother cell compartment during the late sporulation phase were expected to be targeted and displayed on the spore surface. As a proof of principle, we demonstrated the display of carboxymethylcellulase (CMCase) in its native form on the spore surface. The target protein, CMCase, was expressed under the control of the cry1Aa promoter, which is controlled by σ(E) and σ(K) and is expressed in the mother cell compartment. The correct display was confirmed using enzyme activity assays, flow cytometry, and immunogold electron microscopy. In addition, we demonstrated the display of a β-galactosidase tetramer and confirmed its correct display using enzyme activity assays and protein characterization. This native protein display system, combined with the robust nature of Bacillus spores, will broaden the range of displayable target proteins. Consequently, the applications of display technology will be expanded, including high-throughput screening, vaccines, biosensors, biocatalysis, bioremediation, and other innovative bioprocesses.

  20. [Electron microscopy of the surfaces of bacillary spores].

    PubMed

    Smirnova, T A; Zubasheva, M V; Shevliagina, N V; Nikolaenko, M A; Azizbekian, R R

    2013-01-01

    The surface structures of the spores of Bacillus. cereus, B. thuringiensis, and Brevibacillus laterosporus were studied by transmission and scanning electron microscopy. Platinum deposition and negative staining with uranyl acetate revealed appendages and exosporium in B. thuringiensis and B. cereus. The exosporium structure was visualized by negative staining and ultrathin sectioning. For staining the exosporium polysaccharide, Alcian blue was used during fixation. The results obtained show the differences in structural organization of appendages and exosporium in different strains. Canoe-shaped inclusions were revealed in all Br. laterosporus strains, while strain IGM16-92 had a fibrillar capsule as well. Electron microscopy using a dual beam scanning electron microscope Quanta 200 3D provided the information of the spore surface relief without sample treatment (fixation and dehydration). The spores of Br. laterosporus strains had folded surface, unlike the smooth surface of B. cereus and B. thuringiensis spores. Diversity of external spore structures was shown within a species, which may be used for detection of bacteria at the strain level. Optimized procedures for visualization of spore surface by different electron microscopic techniques were discussed.

  1. Infrared Signatures to Discriminate Viability of Autoclaved Bacillus Spores

    SciTech Connect

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

    2011-10-06

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

  2. Small acid soluble proteins for rapid spore identification.

    SciTech Connect

    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.

  3. Interaction of antimicrobial cyclic lipopeptides from Bacillus subtilis influences their effect on spore germination and membrane permeability in fungal plant pathogens.

    PubMed

    Liu, Jiajie; Hagberg, Ingrid; Novitsky, Laura; Hadj-Moussa, Hanane; Avis, Tyler J

    2014-11-01

    Bacillus subtilis cyclic lipopeptides are known to have various antimicrobial effects including different types of interactions with the cell membranes of plant pathogenic fungi. The various spectra of activities of the three main lipopeptide families (fengycins, iturins, and surfactins) seem to be linked to their respective mechanisms of action on the fungal biomembrane. Few studies have shown the combined effect of more than one family of lipopeptides on fungal plant pathogens. In an effort to understand the effect of producing multiple lipopeptide families, sensitivity and membrane permeability of spores from four fungal plant pathogens (Alternaria solani, Fusarium sambucinum, Rhizopus stolonifer, and Verticillium dahliae) were assayed in response to lipopeptides, both individually and as combined treatments. Results showed that inhibition of spores was highly variable depending on the tested fungus-lipopeptide treatment. Results also showed that inhibition of the spores was closely associated with SYTOX stain absorption suggesting effects of efficient treatments on membrane permeability. Combined lipopeptide treatments revealed additive, synergistic or sometimes mutual inhibition of beneficial effects.

  4. IMMUNOCYTOCHEMICAL LOCALIZATION OF STACHYLYSIN IN STACHYBOTRYS CHARTARUM SPORES AND SPORE-IMPACTED MOUSE AND RAT LUNG TISSUES

    EPA Science Inventory

    Stachylysin is a proteinaceous hemolytic agent that is producted by S. chartarum. Stachylysin was found, using immunohistochemistical and immunocytochemical methods, to be localized in S. chartarum spores/mycelia primarily in the inner wall suggesting that it is constitutively ...

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

    PubMed Central

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

    2016-01-01

    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 × 107 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

  6. Ant-fungus species combinations engineer physiological activity of fungus gardens.

    PubMed

    Seal, J N; Schiøtt, M; Mueller, U G

    2014-07-15

    Fungus-gardening insects are among the most complex organisms because of their extensive co-evolutionary histories with obligate fungal symbionts and other microbes. Some fungus-gardening insect lineages share fungal symbionts with other members of their lineage and thus exhibit diffuse co-evolutionary relationships, while others exhibit little or no symbiont sharing, resulting in host-fungus fidelity. The mechanisms that maintain this symbiont fidelity are currently unknown. Prior work suggested that derived leaf-cutting ants in the genus Atta interact synergistically with leaf-cutter fungi (Attamyces) by exhibiting higher fungal growth rates and enzymatic activities than when growing a fungus from the sister-clade to Attamyces (so-called 'Trachymyces'), grown primarily by the non-leaf cutting Trachymyrmex ants that form, correspondingly, the sister-clade to leaf-cutting ants. To elucidate the enzymatic bases of host-fungus specialization in leaf-cutting ants, we conducted a reciprocal fungus-switch experiment between the ant Atta texana and the ant Trachymyrmex arizonensis and report measured enzymatic activities of switched and sham-switched fungus gardens to digest starch, pectin, xylan, cellulose and casein. Gardens exhibited higher amylase and pectinase activities when A. texana ants cultivated Attamyces compared with Trachymyces fungi, consistent with enzymatic specialization. In contrast, gardens showed comparable amylase and pectinase activities when T. arizonensis cultivated either fungal species. Although gardens of leaf-cutting ants are not known to be significant metabolizers of cellulose, T. arizonensis were able to maintain gardens with significant cellulase activity when growing either fungal species. In contrast to carbohydrate metabolism, protease activity was significantly higher in Attamyces than in Trachymyces, regardless of the ant host. Activity of some enzymes employed by this symbiosis therefore arises from complex interactions between the

  7. Virulence Plasmids of Spore-Forming Bacteria.

    PubMed

    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.

  8. Mechanisms of induction of germination of Bacillus subtilis spores by high pressure.

    PubMed

    Paidhungat, Madan; Setlow, Barbara; Daniels, William B; Hoover, Dallas; Papafragkou, Efstathia; Setlow, Peter

    2002-06-01

    Spores of Bacillus subtilis lacking all germinant receptors germinate >500-fold slower than wild-type spores in nutrients and were not induced to germinate by a pressure of 100 MPa. However, a pressure of 550 MPa induced germination of spores lacking all germinant receptors as well as of receptorless spores lacking either of the two lytic enzymes essential for cortex hydrolysis during germination. Complete germination of spores either lacking both cortex-lytic enzymes or with a cortex not attacked by these enzymes was not induced by a pressure of 550 MPa, but treatment of these mutant spores with this pressure caused the release of dipicolinic acid. These data suggest the following conclusions: (i) a pressure of 100 MPa induces spore germination by activating the germinant receptors; and (ii) a pressure of 550 MPa opens channels for release of dipicolinic acid from the spore core, which leads to the later steps in spore germination.

  9. Spores of Bacillus subtilis: their resistance to and killing by radiation, heat and chemicals.

    PubMed

    Setlow, P

    2006-09-01

    A number of mechanisms are responsible for the resistance of spores of Bacillus species to heat, radiation and chemicals and for spore killing by these agents. Spore resistance to wet heat is determined largely by the water content of spore core, which is much lower than that in the growing cell protoplast. A lower core water content generally gives more wet heat-resistant spores. The level and type of spore core mineral ions and the intrinsic stability of total spore proteins also play a role in spore wet heat resistance, and the saturation of spore DNA with alpha/beta-type small, acid-soluble spore proteins (SASP) protects DNA against wet heat damage. However, how wet heat kills spores is not clear, although it is not through DNA damage. The alpha/beta-type SASP are also important in spore resistance to dry heat, as is DNA repair in spore outgrowth, as Bacillus subtilis spores are killed by dry heat via DNA damage. Both UV and gamma-radiation also kill spores via DNA damage. The mechanism of spore resistance to gamma-radiation is not well understood, although the alpha/beta-type SASP are not involved. In contrast, spore UV resistance is due largely to an alteration in spore DNA photochemistry caused by the binding of alpha/beta-type SASP to the DNA, and to a lesser extent to the photosensitizing action of the spore core's large pool of dipicolinic acid. UV irradiation of spores at 254 nm does not generate the cyclobutane dimers (CPDs) and (6-4)-photoproducts (64PPs) formed between adjacent pyrimidines in growing cells, but rather a thymidyl-thymidine adduct termed spore photoproduct (SP). While SP is formed in spores with approximately the same quantum efficiency as that for generation of CPDs and 64PPs in growing cells, SP is repaired rapidly and efficiently in spore outgrowth by a number of repair systems, at least one of which is specific for SP. Some chemicals (e.g. nitrous acid, formaldehyde) again kill spores by DNA damage, while others, in particular

  10. At-line determining spore germination of Penicillium chrysogenum bioprocesses in complex media.

    PubMed

    Ehgartner, Daniela; Fricke, Jens; Schröder, Andreas; Herwig, Christoph

    2016-10-01

    Spore inoculum quality in filamentous bioprocesses is a critical parameter associated with viable spore concentration (1) and spore germination (2). It influences pellet morphology and, consequently, process performance. The state-of-the-art method to measure viable spore concentration is tedious, associated with significant inherent bias, and not applicable in real-time. Therefore, it is not usable as process analytical technology (PAT). Spore germination has so far been monitored using image analysis, which is hampered by complex medium background often observed in filamentous bioprocesses. The method presented here is based on the combination of viability staining and large-particle flow cytometry which enables measurements in real-time and hence aims to be applicable as a PAT tool. It is compatible with the complex media background and allows the quantification of metabolically active spores and the monitoring of spore germination. A distinction of germinated spores and not germinated spores was based on logistic regression, using multiparameteric data from flow cytometry. In a first step, a significant correlation between colony-forming unit (CFU) counts and viable spore concentration (1) in an industrially relevant model bioprocess was found. Spore germination (2) was followed over the initial process phase with close temporal resolution. The validation of the method showed an error below 5 %. Differences in spore germination for various spore inocula ages and spore inoculum concentrations were monitored. The real-time applicability of the method suggests the implementation as a PAT tool in filamentous bioprocesses.

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

    PubMed Central

    2015-01-01

    SUMMARY 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

  12. Germination and infectivity of ectomycorrhizal fungal spores in relation to their ecological traits during primary succession.

    PubMed

    Ishida, Takahide A; Nara, Kazuhide; Tanaka, Megumi; Kinoshita, Akihiko; Hogetsu, Taizo

    2008-01-01

    The spores of ectomycorrhizal fungi (EMF) play critical roles in the population and community development of EMF. Here, the germination and infectivity of EMF spores are examined with reference to the ecological traits of the EMF species. Spores were collected from 12 EMF species, whose successional patterns have been studied in the volcanic desert on Mount Fuji, Japan. Spore germination experiments were conducted with host plants (Salix reinii), with nonhost plants (Polygonum cuspidatum), and without plants. The mycorrhizal formation ability of spores was also examined in seven EMF using spore inoculation experiments. To determine the effects of the spore preservation period, both experiments were repeated up to 1 yr after spore collection. Spore germination was very low in the absence of host plants. In the presence of hosts, even 30 d after spore collection, spore germination was significantly enhanced in all pioneer EMF (c. 20%) but less so in late-stage EMF (< 5%), except in Hebeloma species. Mycorrhizal formation from spores was also greater in pioneer EMF but was significantly reduced by 1 yr of spore preservation. High spore germination and infectivity of pioneer EMF should enable these species to colonize disturbed and isolated areas in accordance with their ecological traits.

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

    PubMed

    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.

  14. 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.

    PubMed

    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.

  15. Environmental Persistence of Bacillus anthracis and Bacillus subtilis Spores

    PubMed Central

    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

  16. Scanning Surface Potential Microscopy of Spore Adhesion on Surfaces

    SciTech Connect

    Lee, Ida; Chung, Eunhyea; Kweon, Hyojin; Yiacoumi, Sotira; Tsouris, Costas

    2012-01-01

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

  17. Environmental Persistence of Bacillus anthracis and Bacillus subtilis Spores.

    PubMed

    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.

  18. STREPTOMYCES SPECIES COMPRISING THE BLUE-SPORE SERIES

    PubMed Central

    Trejo, W. H.; Bennett, R. E.

    1963-01-01

    Trejo, W. H. (Squibb Institute for Medical Research, New Brunswick, N.J.) and R. E. Bennett. Streptomyces species comprising the blue-spore series. J. Bacteriol. 85:676–690. 1963.—The objective of this study was to define and delimit the streptomycetes of the blue-spored (Viridochromogenes) series. The series, as defined in this study, includes 11 blue and blue-green species. The green-spored species were excluded on the basis of morphology as well as color. It was proposed that NRRL B-1511 be designated as the neotype strain of Streptomyces viridochromogenes (Krainsky) Waksman and Henrici, and that IMRU 3761 be designated as the neotype for Streptomyces cyaneus (Krassilnikov) Waksman. Evidence was presented to show that physiological criteria cannot be used to differentiate these organisms below the series level. The major characteristics of the Viridochromogenes series are blue to blue-green spores borne in spirals, and chromogenicity (melanin-positive). Reverse color and spore morphology provide a basis for separation below the series level. Images PMID:14042949

  19. Detection of Bacterial Spores with Lanthanide-Macrocycle Binary Complexes

    PubMed Central

    Cable, Morgan L.; Kirby, James P.; Levine, Dana J.; Manary, Micah J.; Gray, Harry B.; Ponce, Adrian

    2009-01-01

    The detection of bacterial spores via dipicolinate-triggered lanthanide luminescence has been improved in terms of detection limit, stability, and susceptibility to interferents by use of lanthanide-macrocycle binary complexes. Specifically, we compared the effectiveness of Sm, Eu, Tb and Dy complexes with the macrocycle 1,4,7,10-tetraazacyclododecane-1,7-diacetate (DO2A) to the corresponding lanthanide aquo ions. The Ln(DO2A)+ binary complexes bind dipicolinic acid (DPA), a major constituent of bacterial spores, with greater affinity and demonstrate significant improvement in bacterial spore detection. Of the four luminescent lanthanides studied, the terbium complex exhibits the greatest dipicolinate binding affinity (100-fold greater than Tb3+ alone, and 10-fold greater than other Ln(DO2A)+ complexes) and highest quantum yield. Moreover, the inclusion of DO2A extends the pH range over which Tb-DPA coordination is stable, reduces the interference of calcium ions nearly 5-fold, and mitigates phosphate interference 1000-fold compared to free terbium alone. In addition, detection of Bacillus atrophaeus bacterial spores was improved by the use of Tb(DO2A)+, yielding a 3-fold increase in the signal-to-noise ratio over Tb3+. Out of the eight cases investigated, the Tb(DO2A)+ binary complex is best for the detection of bacterial spores. PMID:19537757

  20. Detection of bacterial spores with lanthanide-macrocycle binary complexes.

    PubMed

    Cable, Morgan L; Kirby, James P; Levine, Dana J; Manary, Micah J; Gray, Harry B; Ponce, Adrian

    2009-07-15

    The detection of bacterial spores via dipicolinate-triggered lanthanide luminescence has been improved in terms of detection limit, stability, and susceptibility to interferents by use of lanthanide-macrocycle binary complexes. Specifically, we compared the effectiveness of Sm, Eu, Tb, and Dy complexes with the macrocycle 1,4,7,10-tetraazacyclododecane-1,7-diacetate (DO2A) to the corresponding lanthanide aquo ions. The Ln(DO2A)(+) binary complexes bind dipicolinic acid (DPA), a major constituent of bacterial spores, with greater affinity and demonstrate significant improvement in bacterial spore detection. Of the four luminescent lanthanides studied, the terbium complex exhibits the greatest dipicolinate binding affinity (100-fold greater than Tb(3+) alone, and 10-fold greater than other Ln(DO2A)(+) complexes) and highest quantum yield. Moreover, the inclusion of DO2A extends the pH range over which Tb-DPA coordination is stable, reduces the interference of calcium ions nearly 5-fold, and mitigates phosphate interference 1000-fold compared to free terbium alone. In addition, detection of Bacillus atrophaeus bacterial spores was improved by the use of Tb(DO2A)(+), yielding a 3-fold increase in the signal-to-noise ratio over Tb(3+). Out of the eight cases investigated, the Tb(DO2A)(+) binary complex is best for the detection of bacterial spores.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  3. Zombie bugs? The fungus Purpureocillium cf. lilacinum may manipulate the behavior of its host bug Edessa rufomarginata.

    PubMed

    Eberhard, William; Pacheco-Esquivel, Jessica; Carrasco-Rueda, Farah; Christopher, Yuliana; Gonzalez, Cely; Ramos, Daniel; Urbina, Hector; Blackwell, Meredith

    2014-01-01

    Just before dying, Edessa rufomarginata (Hemiptera, Pentotomidae) individuals that are infected with the fungus Purpureocillium cf. lilacinum (Ascomycota: Ophiocordycipitaceae) move from the leaves onto the stems of their Solanum sp. host and firmly grasp the stems in ways seldom employed by uninfected bugs. These alterations in host behavior probably improve the chances that the subsequently produced fungal spores will be dispersed aerially. Purpureocillium cf. lilacinum is a member of the Ophiocordycipitaceae, a group in which other species also modify the behavior of their hosts. As in the case of newly distinguished relatives of Ophiocordyceps unilateralis associated with "zombie ants" the discovery of P. cf. lilacinum infecting bugs reveals that P. lilacinum may be more diverse than previously appreciated.

  4. An extracellular glucoamylase produced by endophytic fungus EF6.

    PubMed

    Tangngamsakul, P; Karnchanatat, A; Sihanonth, P; Sangvanich, P

    2011-01-01

    A strain of endophytic fungus EF6 isolated from Thai medicinal plants was found to produce higher levels of extracellular glucoamylase. This strain produced glucoamylase of culture filtrate when grown on 1% soluble starch. The enzyme was purified and characterized. Purification steps involved (NH4)2SO4 precipitation, anion exchange, and gel filtration chromatography. Final purification fold was 14.49 and the yield obtained was 9.15%. The enzyme is monomeric with a molecular mass of 62.2 kDa as estimated by SDS-PAGE, and with a molecular mass of 62.031 kDa estimated by MALDI-TOF spectrometry. The temperature for maximum activity was 60 degrees C. After 30 min for incubation, glucoamylase was found to be stable lower than 50 degrees C. The activity decrease rapidly when residual activity was retained about 45% at 55 degrees C. The pH optimum of the enzyme activity was 6.0, and it was stable over a pH range of 4.0-7.0 at 50 degrees C. The activity of glucoamylase was stimulated by Ca2+, Co2+, Mg2+, Mn2+, glycerol, DMSO, DTT and EDTA, and strongly inhibited by Hg2+. Various types of starch were test, soluble starch proved to be the best substrate for digestion process. The enzyme catalyzes the hydrolysis of soluble starch and maltose as the substrate, the enzyme had Km values of 2.63, and 1.88 mg/ml and Vmax, values of 1.25, and 2.54 U/min/mg protein, and Vmax/Km values of 0.48 and 1.35, respectively. The internal amino acid sequences of endophytic fungus EF6 glucoamylase; RALAN HKQVV DSFRS have similarity to the sequence of the glucoamylase purified form Thermomyces lanuginosus. From all results indicated that this enzyme is a glucoamylase (1,4-alpha-D-glucan glucanohydrolase).

  5. Adenosine Monophosphate-Based Detection of Bacterial Spores

    NASA Technical Reports Server (NTRS)

    Kern, Roger G.; Chen, Fei; Venkateswaran, Kasthuri; Hattori, Nori; Suzuki, Shigeya

    2009-01-01

    A method of rapid detection of bacterial spores is based on the discovery that a heat shock consisting of exposure to a temperature of 100 C for 10 minutes causes the complete release of adenosine monophosphate (AMP) from the spores. This method could be an alternative to the method described in the immediately preceding article. Unlike that method and related prior methods, the present method does not involve germination and cultivation; this feature is an important advantage because in cases in which the spores are those of pathogens, delays involved in germination and cultivation could increase risks of infection. Also, in comparison with other prior methods that do not involve germination, the present method affords greater sensitivity. At present, the method is embodied in a laboratory procedure, though it would be desirable to implement the method by means of a miniaturized apparatus in order to make it convenient and economical enough to encourage widespread use.

  6. Plasma effects on bacterial spores in a wet environment

    NASA Astrophysics Data System (ADS)

    Kuo, Spencer P.; Tarasenko, Olga; Nourkbash, Said; Bakhtina, Assya; Levon, Kalle

    2006-03-01

    An arc-seed microwave plasma torch, which can run stably at low airflow rate (e.g., 0.393 l s-1) and produces an abundance of reactive atomic oxygen in its plasma effluent, is applied for studying the effects of atomic oxygen on bacterial spores in solution. Bacillus cereus was chosen as the biological agent. The experimental results show that the plasma effluent can penetrate into water to kill B. cereus spores. The kill time (i.e., 10-fold reduction time) is about 10 s at an exposure distance of 3 cm, 24 s at 4 cm, and 31 s at 5 cm. Morphological studies are performed via scanning electron and atomic force microscopes, which take two- and three-dimensional images of spores to record the changes in their morphological structures and shapes caused by the plasma effluent. The loss of appendages and exosporium in the structure as well as flattened cell shapes are observed.

  7. Differentiating bacterial spores from hoax materials by Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Farquharson, Stuart; Smith, Wayne W.

    2004-03-01

    The bioterrorism of October 2001 caused by the distribution of anthrax through the U.S. postal system was compounded by the significant delay associated with positive identification of the Bacillus anthracis spores and the unknown extent of their distribution along the eastern seaboard. In the ensuing two years, literally thousands of hoaxes, letters containing harmless powders, have been mailed creating additional anxiety. Thus, there is a need for instruments and/or methods that can not only identify anthrax-causing spores to save lives, but also identify hoax materials to eliminate costly shutdowns. Here we present Raman spectra of Bacillus cereus spores, an anthrax surrogate, as well as of 30 common substances that might be used as hoax materials. We also examine the choice of laser excitation, 785 nm or 1064 nm, and its impact on the ability to measure visible particles in 5 minutes or less, and to provide a complete answer to the question of suspicious material identity.

  8. Enhanced Spore Biomarker Detection Following Laser Induced Lysis

    SciTech Connect

    Wunschel, David S.; Beck, Kenneth M.; Wahl, Karen L.

    2002-12-01

    Matrix assisted laser desorption/ionization (MALDI) has grown in popularity as a means to rapidly analyze proteins directly from bacterial cells. This method provides identifying information by generating protein ?fingerprints? for each organism. However, generating rich protein fingerprints from spores, such as from the genus Bacillus, has proven difficult. We have examined the use of laser energy to induce spore lysis and increase the protein signature complexity. As a measure of lysis, the ions from calcium and dipicolinic acid (DPA) were monitored along with the higher m/z protein ions. DPA is a known marker of eubacterial spores usually as a complex with calcium. This is in contrast to the abundant geogenic calcium complexes with carbonate among other forms. A combination of general bacterial markers, DPA and calcium, and protein fingerprints can be used to provide complementary biomarkers from a single sample preparation.

  9. Role of dipicolinic acid in the germination, stability, and viability of spores of Bacillus subtilis.

    PubMed

    Magge, Anil; Granger, Amanda C; Wahome, Paul G; Setlow, Barbara; Vepachedu, Venkata R; Loshon, Charles A; Peng, Lixin; Chen, De; Li, Yong-Qing; Setlow, Peter

    2008-07-01

    Spores of Bacillus subtilis spoVF strains that cannot synthesize dipicolinic acid (DPA) but take it up during sporulation were prepared in medium with various DPA concentrations, and the germination and viability of these spores as well as the DPA content in individual spores were measured. Levels of some other small molecules in DPA-less spores were also measured. These studies have allowed the following conclusions. (i) Spores with no DPA or low DPA levels that lack either the cortex-lytic enzyme (CLE) SleB or the receptors that respond to nutrient germinants could be isolated but were unstable and spontaneously initiated early steps in spore germination. (ii) Spores that lacked SleB and nutrient germinant receptors and also had low DPA levels were more stable. (iii) Spontaneous germination of spores with no DPA or low DPA levels was at least in part via activation of SleB. (iv) The other redundant CLE, CwlJ, was activated only by the release of high levels of DPA from spores. (v) Low levels of DPA were sufficient for the viability of spores that lacked most alpha/beta-type small, acid-soluble spore proteins. (vi) DPA levels accumulated in spores prepared in low-DPA-containing media varied greatly between individual spores, in contrast to the presence of more homogeneous DPA levels in individual spores made in media with high DPA concentrations. (vii) At least the great majority of spores of several spoVF strains that contained no DPA also lacked other major spore small molecules and had gone through some of the early reactions in spore germination.

  10. Spore Yield and Microcycle Conidiation of Colletotrichum gloeosporioides in Liquid Culture

    PubMed Central

    Cascino, J. J.; Harris, R. F.; Smith, C. S.; Andrews, J. H.

    1990-01-01

    The effect of V8 juice concentration (5 to 40%, vol/vol), spore inoculum density (105 and 107 spores per ml), and liquid batch or fed-batch culture condition on mycelium and spore production by Colletotrichum gloeosporioides was evaluated. The amount of mycelium produced, the time required for initiation of sporulation following attainment of maximum mycelium, and the time for attainment of maximum spore concentration increased with increasing V8 juice concentration in batch culture. Cultures containing V8 juice at >10% achieved a similar spore density (apparent spore-carrying capacity) of about 0.8 mg of spores per ml (1 × 107 to 2 × 107 spores per ml) independent of inoculum density and V8 juice concentration. The relative spore yield decreased from a high of 64% of the total biomass for the low-inoculum 5% V8 culture, through 13% for the analogous 40% V8 culture, to a low of 2% for the high-inoculum 27% V8 culture. Fed-batch cultures were used to establish conditions of high spore density and low substrate availability but high substrate flux. The rate of addition of V8 juice was adjusted to approximate the rate of substrate utilization by the (increasing) biomass. The final spore concentration was about four times higher (3.0 mg of spores per ml) than the apparent spore-carrying capacity in batch culture. This high spore yield was obtained at the expense of greatly reduced mycelium, resulting in a high relative spore yield (62% of the total biomass). Microcycle conidiation occurred in the fed-batch but not batch systems. These data indicate that substrate-limited, fed-batch culture can be used to increase the amount and efficiency of spore production by C. gloeosporioides by maintaining microcycle conidiation conditions favoring allocation of nutrients to spore rather than mycelium production. PMID:16348245

  11. Expression of proteins and protein kinase activity during germination of aerial spores of Streptomyces granaticolor.

    PubMed

    Mikulík, Karel; Bobek, Jan; Bezousková, Silvia; Benada, Oldrich; Kofronová, Olga

    2002-11-29

    Dormant aerial spores of Streptomyces granaticolor contain pre-existing pool of mRNA and active ribosomes for rapid translation of proteins required for earlier steps of germination. Activated spores were labeled for 30 min with [35S]methionine/cysteine in the presence or absence of rifamycin (400 microg/ml) and resolved by two-dimensional electrophoresis. About 320 proteins were synthesized during the first 30 min of cultivation at the beginning of swelling, before the first DNA replication. Results from nine different experiments performed in the presence of rifamycin revealed 15 protein spots. Transition from dormant spores to swollen spores is not affected by the presence of rifamycin but further development of spores is stopped. To support existence of pre-existing pool of mRNA in spores, cell-free extract of spores (S30 fraction) was used for in vitro protein synthesis. These results indicate that RNA of spores possesses mRNA functionally competent and provides templates for protein synthesis. Cell-free extracts isolated from spores, activated spores, and during spore germination were further examined for in vitro protein phosphorylation. The analyses show that preparation from dormant spores catalyzes phosphorylation of only seven proteins. In the absence of phosphatase inhibitors, several proteins were partially dephosphorylated. The activation of spores leads to a reduction in phosphorylation activity. Results from in vitro phosphorylation reaction indicate that during germination phosphorylation/dephosphorylation of proteins is a complex function of developmental changes.

  12. The Bacillus subtilis spore coat provides "eat resistance" during phagocytic predation by the protozoan Tetrahymena thermophila.

    PubMed

    Klobutcher, Lawrence A; Ragkousi, Katerina; Setlow, Peter

    2006-01-03

    Bacillus spores are highly resistant to many environmental stresses, owing in part to the presence of multiple "extracellular" layers. Although the role of some of these extracellular layers in resistance to particular stresses is known, the function of one of the outermost layers, the spore coat, is not completely understood. This study sought to determine whether the spore coat plays a role in resistance to predation by the ciliated protozoan Tetrahymena, which uses phagocytosis to ingest and degrade other microorganisms. Wild-type dormant spores of Bacillus subtilis were efficiently ingested by the protozoan Tetrahymena thermophila but were neither digested nor killed. However, spores with various coat defects were killed and digested, leaving only an outer shell termed a rind, and supporting the growth of Tetrahymena. A similar rind was generated when coat-defective spores were treated with lysozyme alone. The sensitivity of spores with different coat defects to predation by T. thermophila paralleled the spores' sensitivities to lysozyme. Spore killing by T. thermophila was by means of lytic enzymes within the protozoal phagosome, not by initial spore germination followed by killing. These findings suggest that a major function of the coat of spores of Bacillus species is to protect spores against predation. We also found that indigestible rinds were generated even from spores in which cross-linking of coat proteins was greatly reduced, implying the existence of a coat structure that is highly resistant to degradative enzymes.

  13. Influence of transition metals added during sporulation on heat resistance of Clostridium botulinum 113B spores.

    PubMed Central

    Kihm, D J; Hutton, M T; Hanlin, J H; Johnson, E A

    1990-01-01

    Sporulation of Clostridium botulinum 113B in a complex medium supplemented with certain transition metals (Fe, Mn, Cu, or Zn) at 0.01 to 1.0 mM gave spores that were increased two to sevenfold in their contents of the added metals. The contents of calcium, magnesium, and other metals in the purified spores were relatively unchanged. Inclusion of sodium citrate (3 g/liter) in the medium enhanced metal accumulation and gave consistency in the transition metal contents of independent spore crops. In citrate-supplemented media, C. botulinum formed spores with very high contents of Zn (approximately 1% of the dry weight). Spores containing an increased content of Fe (0.1 to 0.2%) were more susceptible to thermal killing than were native spores or spores containing increased Zn or Mn. The spores formed with added Fe or Cu also appeared less able to repair heat-induced injuries than the spores with added Mn or Zn. Fe-increased spores appeared to germinate and outgrow at a higher frequency than did native and Mn-increased spores. This study shows that C. botulinum spores can be sensitized to increased thermal destruction by incorporation of Fe in the spores. PMID:2180370

  14. 32P-post-labelling analysis of DNA adducts formed in the upper gastrointestinal tissue of mice fed bracken extract or bracken spores.

    PubMed Central

    Povey, A. C.; Potter, D.; O'Connor, P. J.

    1996-01-01

    Bracken toxicity to both domestic and laboratory animals is well established and tumours are formed when rodents are treated with either bracken extracts or bracken spores. In this study we have administered bracken spores and extract to mice in order to investigate whether such exposure leads to the formation of DNA adducts. DNA, isolated from the upper gastrointestinal tract and liver, was digested to 3'-nucleotides. Adducts were extracted with butanol, 32P-post-labelled, separated by thin layer chromatography (TLC) and visualised and quantified using storage-phosphor technology. A cluster of adducts was clearly seen in the DNA of the upper gastrointestinal tract, but not liver, 5 and 24 h after treatment with bracken extract or bracken spores. These adducts were not observed in DNA extracted from vehicle-treated animals. Whereas, after 5 h adduct levels in extract and spore-treated animals were similar, after 24 h adduct levels in the extract-treated animals had diminished by > 75%, but levels in spore-treated animals remained similar to those found after 5 h. This suggests that the DNA-reactive compounds were being released slowly from the spores, even though the spores had been sonicated before administration. Adducts were also quantified after the addition of an internal standard (deoxyinosine 3'-monophosphate) by comparing the amount of label incorporated into the adducts with that found in a known amount of the internal standard. Adduct levels using this internal standard approach were similar to those found by direct measurement of radioactivity incorporated into the adduct, indicating that the labelling of adducts was quantitative. We have tried, unsuccessfully, to synthesise ptaquiloside, the principal carcinogenic component present within bracken. However, similar patterns of adducts were observed when two other compounds, (1-(4-chlorophenyl sulphonyl)-l-cyclopropane carbonitrile and 3-cyclopropylindeno [1,2-c] pyrazol-4-(O-methyl)oxime), which both

  15. Inactivation of Bacillus subtilis spores with ozone and monochloramine.

    PubMed

    Larson, Matthew A; Mariñas, Benito J

    2003-02-01

    The inactivation kinetics of Bacillus subtilis spores with ozone and monochloramine was characterized by a lag phase followed by a pseudo-first-order rate of inactivation. The lag phase decreased and the post-lag phase rate constant increased with increasing temperature within the range investigated (1-30 degrees C for ozone, 1-20 degrees C for monochloramine). The corresponding activation energies were 46820 J/mol for ozone and 79640 J/mol for monochloramine. The CT concept was found to be valid within the concentration range investigated of 0.44-4.8 mg/l for ozone, and 3.8-7.7 mg/l as Cl(2) for monochloramine. The inactivation kinetics of B. subtilis spores with both ozone and monochloramine varied with pH within the range of pH 6-10 investigated. The fastest ozone and monochloramine inactivation rates were observed at pH 10 and 6, respectively. Different stocks of the same strain of B. subtilis spores had different resistance to ozone and monochloramine mainly because of discrepancies in the extent of the lag phase. B. subtilis spores might not be conservative surrogates for C. parvum oocysts for ozone disinfection at relatively low temperature mainly due to the spores having a lower activation energy compared to that for the oocysts. In contrast, the activation energy for monochloramine was comparable for both microorganisms but differences in the extent of the lag phase might result in the spores being overly conservative surrogates for the oocysts at relatively low temperature.

  16. Two Novel Transcriptional Regulators Are Essential for Infection-related Morphogenesis and Pathogenicity of the Rice Blast Fungus Magnaporthe oryzae

    PubMed Central

    Yan, Xia; Li, Ya; Yue, Xiaofeng; Wang, Congcong; Que, Yawei; Kong, Dandan; Ma, Zhonghua; Talbot, Nicholas J.; Wang, Zhengyi

    2011-01-01

    The cyclic AMP-dependent protein kinase A signaling pathway plays a major role in regulating plant infection by the rice blast fungus Magnaporthe oryzae. Here, we report the identification of two novel genes, MoSOM1 and MoCDTF1, which were discovered in an insertional mutagenesis screen for non-pathogenic mutants of M. oryzae. MoSOM1 or MoCDTF1 are both necessary for development of spores and appressoria by M. oryzae and play roles in cell wall differentiation, regulating melanin pigmentation and cell surface hydrophobicity during spore formation. MoSom1 strongly interacts with MoStu1 (Mstu1), an APSES transcription factor protein, and with MoCdtf1, while also interacting more weakly with the catalytic subunit of protein kinase A (CpkA) in yeast two hybrid assays. Furthermore, the expression levels of MoSOM1 and MoCDTF1 were significantly reduced in both Δmac1 and ΔcpkA mutants, consistent with regulation by the cAMP/PKA signaling pathway. MoSom1-GFP and MoCdtf1-GFP fusion proteins localized to the nucleus of fungal cells. Site-directed mutagenesis confirmed that nuclear localization signal sequences in MoSom1 and MoCdtf1 are essential for their sub-cellular localization and biological functions. Transcriptional profiling revealed major changes in gene expression associated with loss of MoSOM1 during infection-related development. We conclude that MoSom1 and MoCdtf1 functions downstream of the cAMP/PKA signaling pathway and are novel transcriptional regulators associated with cellular differentiation during plant infection by the rice blast fungus. PMID:22144889

  17. Discrimination of Spore-Forming Bacilli Using spoIVA

    NASA Technical Reports Server (NTRS)

    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

  18. Physical determinants of radiation sensitivity in bacterial spores

    SciTech Connect

    Powers, E.L.

    1982-01-01

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

  19. Spore Photoproduct Lyase: The Known, the Controversial, and the Unknown*

    PubMed Central

    Yang, Linlin; Li, Lei

    2015-01-01

    Spore photoproduct lyase (SPL) repairs 5-thyminyl-5,6-dihydrothymine, a thymine dimer that is also called the spore photoproduct (SP), in germinating endospores. SPL is a radical S-adenosylmethionine (SAM) enzyme, utilizing the 5′-deoxyadenosyl radical generated by SAM reductive cleavage reaction to revert SP to two thymine residues. Here we review the current progress in SPL mechanistic studies. Protein radicals are known to be involved in SPL catalysis; however, how these radicals are quenched to close the catalytic cycle is under debate. PMID:25477522

  20. Enteric spore-forming opportunistic parasites in HIV / AIDS.

    PubMed

    Chawla, Rohit; Ichhpujani, R L

    2011-01-01

    Human Immunodeficiency Virus (HIV) infection causes progressive damage to both limbs of the immune system, which results in a plethora of opportunistic infections. Among the various opportunistic infections, gastrointestinal infections are very common in HIV / Acquired Immunodeficiency Syndrome (AIDS). Opportunistic spore-forming protozoal parasites, namely, Cryptosporidium parvum, Isospora belli, Cyclospora cayetanensis, and Microsporidia, play a major role in causing chronic diarrhea, accompanied with weight loss, in patients with HIV / AIDS. The purpose of this review is to discuss the salient microbiological, clinical, and diagnostic aspects of important enteric spore-forming opportunistic parasites in HIV / AIDS.

  1. Atmospheric transport of mold spores in clouds of desert dust.

    PubMed

    Shinn, Eugene A; Griffin, Dale W; Seba, Douglas B

    2003-08-01

    Fungal spores can be transported globally in clouds of desert dust. Many species of fungi (commonly known as molds) and bacteria--including some that are human pathogens--have characteristics suited to long-range atmospheric transport. Dust from the African desert can affect air quality in Africa, Europe, the Middle East, and the Americas. Asian desert dust can affect air quality in Asia, the Arctic, North America, and Europe. Atmospheric exposure to mold-carrying desert dust may affect human health directly through allergic induction of respiratory stress. In addition, mold spores within these dust clouds may seed downwind ecosystems in both outdoor and indoor environments.

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

    SciTech Connect

    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

  3. Spore-displayed streptavidin: A live diagnostic tool in biotechnology

    SciTech Connect

    Kim, June-Hyung; Lee, Chang-Soo; Kim, Byung-Gee . E-mail: byungkim@snu.ac.kr

    2005-05-27

    Streptavidin, which is one of the most widely used proteins in biotechnological application field and is active only in tetrameric form, was surface expressed on the surface of Bacillus subtilis spore. Spore coat protein of B. subtilis, CotG, was used as an anchoring motif to display streptavidin. FACS using anti-streptavidin antibody was used for the verification of surface localization of expressed CotG-streptavidin fusion protein. FACS and dot-blot were used for the verification of biological activity of displayed streptavidin with FITC-labeled biotin.

  4. Atmospheric transport of mold spores in clouds of desert dust

    USGS Publications Warehouse

    Shinn, E.A.; Griffin, Dale W.; Seba, D.B.

    2003-01-01

    Fungal spores can be transported globally in clouds of desert dust. Many species of fungi (commonly known as molds) and bacteria--including some that are human pathogens--have characteristics suited to long-range atmospheric transport. Dust from the African desert can affect air quality in Africa, Europe, the Middle East, and the Americas. Asian desert dust can affect air quality in Asia, the Arctic, North America, and Europe. Atmospheric exposure to mold-carrying desert dust may affect human health directly through allergic induction of respiratory stress. In addition, mold spores within these dust clouds may seed downwind ecosystems in both outdoor and indoor environments.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed Central

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

    2016-01-01

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

  7. Effect of synthetic detergents on germination of fern spores

    SciTech Connect

    Devi, Y.; Devi, S.

    1986-12-01

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

  8. Atmospheric pollen and fungal spores in Hamilton in 1972 estimated by the Hirst automatic volumetric spore trap.

    PubMed

    Chatterjee, J; Hargreave, F E

    1974-03-16

    A knowledge of the atmospheric pollen and fungal spores is necessary for the diagnosis and management of extrinsic rhinitis and asthma. The Hirst automatic volumetric spore trap has been used for the first time in Canada to identify the quantitative and seasonal incidence of these particles. The trap is easy to operate and has several advantages over the previously used gravity samplers. Tree, grass and ragweed pollens occurred in short, well-defined seasons. Fungal spores greatly outnumbered pollen by 120 to one, and occurred in long, ill-defined seasons. They included large numbers of small basidiospores and ascospores which have previously not been detected in Canada. The latter have not been considered as potential allergens; their clinical importance requires investigation.

  9. Atmospheric pollen and fungal spores in Hamilton in 1972 estimated by the Hirst automatic volumetric spore trap

    PubMed Central

    Chatterjee, J.; Hargreave, F. E.

    1974-01-01

    A knowledge of the atmospheric pollen and fungal spores is necessary for the diagnosis and management of extrinsic rhinitis and asthma. The Hirst automatic volumetric spore trap has been used for the first time in Canada to identify the quantitative and seasonal incidence of these particles. The trap is easy to operate and has several advantages over the previously used gravity samplers. Tree, grass and ragweed pollens occurred in short, well-defined seasons. Fungal spores greatly outnumbered pollen by 120 to one, and occurred in long, ill-defined seasons. They included large numbers of small basidiospores and ascospores which have previously not been detected in Canada. The latter have not been considered as potential allergens; their clinical importance requires investigation. ImagesFIG. 1 PMID:4817211

  10. The basidiomycete ganoderma and asthma: collection, quantitation and immunogenicity of the spores.

    PubMed

    Cutten, A E; Hasnain, S M; Segedin, B P; Bai, T R; McKay, E J

    1988-06-08

    Ganoderma fungal spores are a major component of the Auckland air-spora. Previous studies of ganoderma involvement in allergic asthma and rhinitis were extended by locating the sporophores (fruiting bodies) in the Auckland area and systematically collecting the ejected spores. Maximum production by one sporophore was 5 gram dry weight of spores in one week, equivalent to 11 billion spores. We have estimated that between 400 and 1200 sporophores would account for previously reported levels of ganoderma spores collected from the air by Burkhard spore traps. Both whole spores and extracts of spores were strongly immunogenic in rabbits. Of the 115 asthma patients who were skin prick tested with a variety of fungal extracts, 32 (28%) were positive to one or more fungi. Of these, 18 (16%) reacted positively to ganoderma extracts. A theory proposing how ganoderma might contribute to allergic hyperreactivity in susceptible patients is developed.

  11. Reversal of radiation-dependent heat sensitization of Clostridium perfringens spores.

    PubMed Central

    Gomez, R F; Gombas, D E; Herrero, A

    1980-01-01

    The effect of solute concentration on the sensitization of Clostridium perfringens spores to heat by ionizing radiation was investigated. As we have shown previously, spores of C. perfringens treated with gamma radiation are now sensitive to subsequent heat treatments than are spores that receive no radiation treatment. When gamma-irradiated spores were heated in the presence of increasing concentrations of glycerol or sucrose, the heat sensitivity induced by irradiation was progressively decreased. The magnitude of the increase in heat resistance induced by extracellular solutes was greater in gamma-irradiated spores than in nonirradiated spores. Based on these observations, it is proposed that the induction of heat sensitivity in spores by radiation is related to the loss of osmoregulatory or dehydrating mechanisms in irradiated spores. PMID:6247972

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

    EPA Science Inventory

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

  13. Major Transcriptome Reprogramming Underlies Floral Mimicry Induced by the Rust Fungus Puccinia monoica in Boechera stricta

    PubMed Central

    Haugen, Riston H.; Saunders, Diane G. O.; Leonelli, Lauriebeth; MacLean, Dan; Hogenhout, Saskia A.; Kamoun, Sophien

    2013-01-01

    Pucciniamonoica is a spectacular plant parasitic rust fungus that triggers the formation of flower-like structures (pseudoflowers) in its Brassicaceae host plant Boecherastricta. Pseudoflowers mimic in shape, color, nectar and scent co-occurring and unrelated flowers such as buttercups. They act to attract insects thereby aiding spore dispersal and sexual reproduction of the rust fungus. Although much ecological research has been performed on P. monoica-induced pseudoflowers, this system has yet to be investigated at the molecular or genomic level. To date, the molecular alterations underlying the development of pseudoflowers and the genes involved have not been described. To address this, we performed gene expression profiling to reveal 256 plant biological processes that are significantly altered in pseudoflowers. Among these biological processes, plant genes involved in cell fate specification, regulation of transcription, reproduction, floral organ development, anthocyanin (major floral pigments) and terpenoid biosynthesis (major floral volatile compounds) were down-regulated in pseudoflowers. In contrast, plant genes involved in shoot, cotyledon and leaf development, carbohydrate transport, wax biosynthesis, cutin transport and L-phenylalanine metabolism (pathway that results in phenylethanol and phenylacetaldehyde volatile production) were up-regulated. These findings point to an extensive reprogramming of host genes by the rust pathogen to induce floral mimicry. We also highlight 31 differentially regulated plant genes that are enriched in the biological processes mentioned above, and are potentially involved in the formation of pseudoflowers. This work illustrates the complex perturbations induced by rust pathogens in their host plants, and provides a starting point for understanding the molecular mechanisms of pathogen-induced floral mimicry. PMID:24069397

  14. Major transcriptome reprogramming underlies floral mimicry induced by the rust fungus Puccinia monoica in Boechera stricta.

    PubMed

    Cano, Liliana M; Raffaele, Sylvain; Haugen, Riston H; Saunders, Diane G O; Leonelli, Lauriebeth; MacLean, Dan; Hogenhout, Saskia A; Kamoun, Sophien

    2013-01-01

    Pucciniamonoica is a spectacular plant parasitic rust fungus that triggers the formation of flower-like structures (pseudoflowers) in its Brassicaceae host plant Boecherastricta. Pseudoflowers mimic in shape, color, nectar and scent co-occurring and unrelated flowers such as buttercups. They act to attract insects thereby aiding spore dispersal and sexual reproduction of the rust fungus. Although much ecological research has been performed on P. monoica-induced pseudoflowers, this system has yet to be investigated at the molecular or genomic level. To date, the molecular alterations underlying the development of pseudoflowers and the genes involved have not been described. To address this, we performed gene expression profiling to reveal 256 plant biological processes that are significantly altered in pseudoflowers. Among these biological processes, plant genes involved in cell fate specification, regulation of transcription, reproduction, floral organ development, anthocyanin (major floral pigments) and terpenoid biosynthesis (major floral volatile compounds) were down-regulated in pseudoflowers. In contrast, plant genes involved in shoot, cotyledon and leaf development, carbohydrate transport, wax biosynthesis, cutin transport and L-phenylalanine metabolism (pathway that results in phenylethanol and phenylacetaldehyde volatile production) were up-regulated. These findings point to an extensive reprogramming of host genes by the rust pathogen to induce floral mimicry. We also highlight 31 differentially regulated plant genes that are enriched in the biological processes mentioned above, and are potentially involved in the formation of pseudoflowers. This work illustrates the complex perturbations induced by rust pathogens in their host plants, and provides a starting point for understanding the molecular mechanisms of pathogen-induced floral mimicry.

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

    SciTech Connect

    Nicholson, W.L.; Setlow, B.; Setlow, P. )

    1991-10-01

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

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

    DTIC Science & Technology

    2005-06-01

    Protamine was reported to be inhibitory to the growth of Bacillus subtilis and B. licheniformis spores at concentrations ranging from 10- to 50-pug/ml...for rapid inactivation of Bacillus anthracis spores in aqueous suspension. Treatment of spores at the onset of germination with 10-, 100- and 1000-,ug...materials contaminated with the spore form of B. anthracis. 15. SUBJECT TERMS Chemical Defense, Biological Defense, CBD, Water Sampling, Bacillus Anthracis

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

    NASA Astrophysics Data System (ADS)

    Kumar, Ajay; Attri, Arun K.

    2016-10-01

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

  18. Water and Small-Molecule Permeation of Dormant Bacillus subtilis Spores

    PubMed Central

    Cermak, Nathan; Feijó Delgado, Francisco; Setlow, Barbara; Setlow, Peter

    2015-01-01

    ABSTRACT We use a suspended microchannel resonator to characterize the water and small-molecule permeability of Bacillus subtilis spores based on spores' buoyant mass in different solutions. Consistent with previous results, we found that the spore coat is not a significant barrier to small molecules, and the extent to which small molecules may enter the spore is size dependent. We have developed a method to directly observe the exchange kinetics of intraspore water with deuterium oxide, and we applied this method to wild-type spores and a panel of congenic mutants with deficiencies in the assembly or structure of the coat. Compared to wild-type spores, which exchange in approximately 1 s, several coat mutant spores were found to have relatively high water permeability with exchange times below the ∼200-ms temporal resolution of our assay. In addition, we found that the water permeability of the spore correlates with the ability of spores to germinate with dodecylamine and with the ability of TbCl3 to inhibit germination with l-valine. These results suggest that the structure of the coat may be necessary for maintaining low water permeability. IMPORTANCE Spores of Bacillus species cause food spoilage and disease and are extremely resistant to standard decontamination methods. This hardiness is partly due to spores' extremely low permeability to chemicals, including water. We present a method to directly monitor the uptake of molecules into B. subtilis spores by weighing spores in fluid. The results demonstrate the exchange of core water with subsecond resolution and show a correlation between water permeability and the rate at which small molecules can initiate or inhibit germination in coat-damaged spores. The ability to directly measure the uptake of molecules in the context of spores with known structural or genetic deficiencies is expected to provide insight into the determinants of spores' extreme resistance. PMID:26483518

  19. Verification of Commercial Decontamination Technologies in Bench-Scale Studies Using Bacillus anthracis Spores

    DTIC Science & Technology

    2004-11-17

    12980) • Spore Strips – Bacillus atrophaeus (ATCC 9372) Biological Indicator Spore Strip BUSINESS SENSITIVE Organisms Biological Indicators: SEM Images...BUSINESS SENSITIVE Verification of Commercial Decontamination Technologies in Bench-Scale Studies Using Bacillus anthracis Spores M.L. Taylor, J.V...Commercial Decontamination Technologies in Bench-Scale Studies Using Bacillus anthracis Spores 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM

  20. Calcineurin orchestrates dimorphic transitions, antifungal drug responses, and host-pathogen interactions of the pathogenic mucoralean fungus Mucor circinelloides

    PubMed Central

    Lee, Soo Chan; Li, Alicia; Calo, Silvia; Inoue, Makoto; Tonthat, Nam K.; Bain, Judith M.; Louw, Johanna; Shinohara, Mari L.; Erwig, Lars P.; Schumacher, Maria A.; Ko, Dennis C.; Heitman, Joseph

    2015-01-01

    Summary Calcineurin plays essential roles in virulence and growth of pathogenic fungi and is a target of the natural products FK506 and Cyclosporine A. In the pathogenic mucoralean fungus Mucor circinelloides, calcineurin mutation or inhibition confers a yeast-locked phenotype indicating that calcineurin governs the dimorphic transition. Genetic analysis in this study reveals that two calcineurin A catalytic subunits (out of three) are functionally diverged. Homology modeling illustrates modes of resistance resulting from amino substitutions in the interface between each calcineurin subunit and the inhibitory drugs. In addition, we show how the dimorphic transition orchestrated by calcineurin programs different outcomes during host-pathogen interactions. For example, when macrophages phagocytose Mucor yeast, subsequent phagosomal maturation occurs, indicating host cells respond appropriately to control the pathogen. On the other hand, upon phagocytosis of spores, macrophages fail to form mature phagosomes. Cytokine production from immune cells differs following exposure to yeast vs. spores (which germinate into hyphae). Thus, the morphogenic transition can be targeted as an efficient treatment option against Mucor infection. In addition, genetic analysis (including gene disruption and mutational studies) further strengthens the understanding of calcineurin and provides a foundation to develop antifungal agents targeting calcineurin to deploy against Mucor and other pathogenic fungi. PMID:26010100

  1. 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

    NASA Technical Reports Server (NTRS)

    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.

  2. Electron microscopic examination of the dormant spore and the sporulation of Paenibacillus motobuensis strain MC10.

    PubMed

    Iida, Ken-ichiro; Amako, Kazunobu; Takade, Akemi; Ueda, Yasuichi; Yoshida, Shin-ichi

    2007-01-01

    We previously reported a new species Paenibacillus motobuensis. The type strain MC10 was stained gram-negative, but had a gram-positive cell wall structure and its spore had a characteristic star shape. The spore and sporulation process of P. motobuensis strain MC10 were examined by electron microscopy using the technique of freeze-substitution in thin sectioning. The structure of the dormant spore was basically the same as that of the other Bacillus spp. The core of the spore was enveloped with two main spore components, the cortex and the spore coat. In thin section, the spore showed a star-shaped image, which was derived from the structure of the spore coat, which is composed of three layers, namely the inner, middle and outer spore coat. The middle coat was an electron-dense thick layer and had a characteristic ridge. By scanning electron microscopic observation, the ridges were seen running parallel to the long axis of the oval-shaped spore. The process of sporulation was essentially the same as that of the other Bacillus spp. The forespore was engulfed by the mother cell membrane, then the spore coat and the cortex were accumulated in the space between the mother cell membrane and forespore membrane. The mother cell membrane seemed to participate in the synthesis of the spore coat. MC10 strain showed almost identical heat resistance to that of B. subtilis.

  3. Anthrax surrogate spores are destroyed by PDT mediated by phenothiazinium dyes

    NASA Astrophysics Data System (ADS)

    Demidova, Tatiana N.; Hamblin, Michael R.

    2005-04-01

    Some Gram-positive bacteria (including the causative agent of anthrax - Bacillus anthracis) survive conditions of stress and starvation by producing dormant stage spores. The spore"s multilayered capsule consists of inner and outer membranes, cortex, proteinaceous spore coat, and in some species an exosporium. These outer layers enclose dehydrated and condensed DNA, saturated with small, acid-soluble proteins. These protective structures make spores highly resistant to damage by heat, radiation, and commonly employed anti-bacterial agents. Previously Bacillus spores have been shown to be resistant to photodynamic inactivation (PDI) using dyes and light that easily destroy the corresponding vegetative bacteria, but recently we have discovered that they are susceptible to PDI. Photoinactivation, however, is only possible if phenothiazinium dyes are used. Dimethylmethylene blue, methylene blue, new methylene blue and toluidine blue O are all effective photosensitizers. Alternative photosensitizers such as Rose Bengal, polylysine chlorin(e6) conjugate, a tricationic porphyrin and benzoporphyrin derivative are ineffective against spores even though they can easily kill vegetative cells. Spores of B. cereus and B. thuringiensis are most susceptible, B. subtilis and B. atrophaeus are also killed, while B. megaterium is resistant. Photoinactivation is most effective when excess dye is washed from the spores showing that the dye binds to the spores and that excess dye in solution can quench light delivery. The relatively mild conditions needed for spore killing could have applications for treating wounds contaminated by anthrax spores and for which conventional sporicides would have unacceptable tissue toxicity.

  4. Heat Resistance of Native and Demineralized Spores of Bacillus subtilis Sporulated at Different Temperatures

    PubMed Central

    Palop, Alfredo; Sala, Francisco J.; Condón, Santiago

    1999-01-01

    Demineralization reduced heat resistance of B. subtilis spores, but the pattern and magnitude of the reduction depended on sporulation temperature and on heating menstruum pH. The differences in heat resistance of native spores caused by sporulation temperature almost disappeared after demineralization. Demineralized spores were still susceptible to the heat-sensitizing effect of acidic pH. PMID:10049900

  5. Early silurian spore tetrads from new york: earliest new world evidence for vascular plants?

    PubMed

    Gray, J; Boucot, A J

    1971-09-03

    Several taxa of abundant cutinized trilete spores from earliest Silurian shale in New York predate by almost an entire period vascular land plant megafossils. Paleoecological evidence suggests that these spores may represent vascular land or semiaquatic plants but a bryophytic origin cannot be precluded on the basis of spore characters. An algal origin is considered unlikely.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  7. Evaluating the spore genome sizes of ferns and lycophytes: a flow cytometry approach.

    PubMed

    Kuo, Li-Yaung; Huang, Yi-Jia; Chang, JenYu; Chiou, Wen-Liang; Huang, Yao-Moan

    2017-03-01

    Ferns and lycophytes produce spores to initiate the gametophyte stage for sexual reproduction. Approximately 10% of these seedless vascular plants are apomictic, and produce genomic unreduced spores. Genome size comparisons between spores and leaves are a reliable, and potentially easier way to determine their reproductive mode compared to traditional approaches. However, estimation of the spore genome sizes of these plants has not been attempted. We attempted to evaluate the spore genome sizes of ferns and lycophytes using flow cytometry, collected spores from selected species representing different spore physical properties and taxonomic groups, and sought to optimize bead-vortexing conditions. By evaluating the spore and sporophyte genome sizes, we examined whether reproductive modes could be ascertained from these flow cytometry results. We proposed two separate sets of optimized bead-vortexing conditions for the nuclear extraction of green and nongreen spores. We further successfully extracted spore nuclei of 19 families covering most orders, and the qualities and quantities of these extractions satisfied the C-value criteria. These evaluated genome sizes further supported the reproductive modes reported previously. In the current study, flow cytometry was used for the first time to evaluate the spore genome sizes of ferns and lycophytes. This use of spore flow cytometry provides a new, efficient approach to ascertaining the reproductive modes of these plants.

  8. FACTORS RELATING TO THE RELEASE OF STACHYBOTRYS CHARTARUM SPORES FROM CONTAMINATED SOURCES

    EPA Science Inventory

    The paper describes preliminary results of a research project to determine the factors that control the release of S. chartarum spores from a contaminated source and test ways to reduce spore release and thus exposure. As anticipated, S. chartarum spore emissions from gypsum boar...

  9. Taphonomic bias in pollen and spore record: a review

    SciTech Connect

    Fisk, L.H.

    1986-05-01

    The high dispersibility and ease of pollen and spore transport have led researchers to conclude erroneously that fossil pollen and spore floras are relatively complete and record unbiased representations of the regional vegetation extant at the time of sediment deposition. That such conclusions are unjustified is obvious when the authors remember that polynomorphs are merely organic sedimentary particles and undergo hydraulic sorting not unlike clastic sedimentary particles. Prior to deposition in the fossil record, pollen and spores can be hydraulically sorted by size, shape, and weight, subtly biasing relative frequencies in fossil assemblages. Sorting during transport results in palynofloras whose composition is environmentally dependent. Therefore, depositional environment is an important consideration to make correct inferences on the source vegetation. Sediment particle size of original rock samples may contain important information on the probability of a taphonomically biased pollen and spore assemblage. In addition, a reasonable test of hydraulic sorting is the distribution of pollen grain sizes and shapes in each assemblage. Any assemblage containing a wide spectrum of grain sizes and shapes has obviously not undergone significant sorting. If unrecognized, taphonomic bias can lead to paleoecologic, paleoclimatic, and even biostratigraphic misinterpretations.

  10. Evaluation of Surface Sampling for Bacillus Spores Using ...

    EPA Pesticide Factsheets

    Journal Article In this study, commercially-available domestic cleaning robots were evaluated for spore surface sampling efficiency on common indoor surfaces. The current study determined the sampling efficiency of each robot, without modifying the sensors, algorithms, or logics set by the manufacturers.

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

    PubMed

    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.

  12. Evaluation of tools for environmental sampling of Bacillus anthracis spores.

    PubMed

    Fujinami, Yoshihito; Hosokawa-Muto, Junji; Mizuno, Natsuko

    2015-12-01

    This study describes the validation of sampling techniques used to detect biological warfare agents used in terror attacks. For this purpose, we tested the efficiencies of different sampling media and extraction solutions for the recovery of bacterial pathogens. We first used Bacillus cereus ATCC 4342 spores as a surrogate for highly pathogenic B. anthracis to compare recovery efficiencies of spores from four different surfaces. We used three different types of sampling swabs and four different solutions to extract spores from the swabs. The most effective sampling method employed rayon swabs moistened with water. The efficencies of the four extraction solutions did not differ significantly, although yields were highest using phosphate-buffered saline containing Tween 80 (PBS-T). Using rayon swabs and sterile water, we recovered B. cereus ATCC 4342 and B. anthracis spores with equivalent efficiencies. These findings indicate that because of its reduced pathogenicity and relative ease in handling (Biosafety Level 1), use of B. cereus ATCC 4342 will facilitate further optimization of techniques to detect B. anthracis.

  13. Isolation of the Paenibacillus phoenicis, a Spore-Forming Bacterium

    NASA Technical Reports Server (NTRS)

    Benardini, James N.; Vaishampayan, Parag A.; Venkateswaran, Kasthuri J.; Osman, Shariff; Satomi, Masataka

    2010-01-01

    A microorganism was isolated from the surfaces of the cleanroom facility in which the Phoenix lander was assembled. The isolated bacterial strain was subjected to a comprehensive polyphasic analysis to characterize its taxonomic position. Both phenotypic and phylogenetic analyses clearly indicate that this isolate belongs to the genus Paenibacillus and represents a novel species. Bacillus spores have been utilized to assess the degree and level of microbiological contamination on spacecraft and their associated spacecraft assembly facilities. Spores of Bacillus species are of particular concern to planetary protection due to the extreme resistance of some members of the genus to space environmental conditions such as UV and gamma radiation, vacuum, oxidation, and temperature fluctuation. These resistive spore phenotypes have enhanced potential for transfer, and subsequent proliferation, of terrestrial microbes on another solar body. Due to decreased nutrient conditions within spacecraft assembly facility clean rooms, the vegetative cells of Bacillus species and other spore-forming Paenibacillus species are induced to sporulate, thereby enhancing their survivability of bioreduction

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

    PubMed

    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.

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

    PubMed Central

    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

  16. 14C Analysis of protein extracts from Bacillus spores.

    PubMed

    Cappuccio, Jenny A; Falso, Miranda J Sarachine; Kashgarian, Michaele; Buchholz, Bruce A

    2014-07-01

    Investigators of bioagent incidents or interdicted materials need validated, independent analytical methods that will allow them to distinguish between recently made bioagent samples versus material drawn from the archives of a historical program. Heterotrophic bacteria convert the carbon in their food sources, growth substrate or culture media, into the biomolecules they need. The F(14)C (fraction modern radiocarbon) of a variety of media, Bacillus spores, and separated proteins from Bacillus spores was measured by accelerator mass spectrometry (AMS). AMS precisely measures F(14)C values of biological materials and has been used to date the synthesis of biomaterials over the bomb pulse era (1955 to present). The F(14)C of Bacillus spores reflects the radiocarbon content of the media in which they were grown. In a survey of commercial media we found that the F(14)C value indicated that carbon sources for the media were alive within about a year of the date of manufacture and generally of terrestrial origin. Hence, bacteria and their products can be dated using their (14)C signature. Bacillus spore samples were generated onsite with defined media and carbon free purification and also obtained from archived material. Using mechanical lysis and a variety of washes with carbon free acids and bases, contaminant carbon was removed from soluble proteins to enable accurate (14)C bomb-pulse dating. Since media is contemporary, (14)C bomb-pulse dating of isolated soluble proteins can be used to distinguish between historical archives of bioagents and those produced from recent media.

  17. Stem rust spores elicit rapid RPG1 phosphorylation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    SciTech Connect

    Chung, Eunhyea; Kweon, Hyojin; Yiacoumi, Sotira; Lee, Ida; Joy, David Charles; Palumbo, Anthony Vito; Tsouris, Costas

    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.

  19. Microwave inactivation of Bacillus atrophaeus spores in healthcare waste.

    PubMed

    Oliveira, E A; Nogueira, N G P; Innocentini, M D M; Pisani, R

    2010-11-01

    Public healthcare wastes from the region of Ribeirão Preto, Brazil, pre-sterilized in an autoclave, were inoculated with spores of Bacillus atrophaeus for microwave processing on a laboratory scale. The influence of waste moisture (40%, 50% and 60% wet basis), presence of surfactant, power per unit mass of waste (100, 150 and 200 W/kg) and radiation exposure time (from 5 to 40 min) on the heating curves was investigated. The most favorable conditions for waste heating with respect to moisture and use of surfactant were then applied in an experimental analysis of the degree of inactivation of B. atrophaeus spores as a function of time and power per unit mass of waste. Based on Chick's and Arrhenius laws, the experimental results were adjusted by the least squares method to determine the activation energies (9203-5782 J/mol) and the Arrhenius pre-exponential factor (0.23 min(-1)). The kinetic parameters thus obtained enabled us to predict the degree of inactivation achieved for B. atrophaeus spores in typical healthcare waste. The activation energy was found to decrease as the power per waste mass increased, leading to the conclusion that, in addition to the thermal effect on the inactivation of B. atrophaeus spores, there was an effect inherent to radiation.

  20. Lethal effect of microwaves on spores of Bacillus spp.

    PubMed

    Wang, Jinn-Chyi; Hu, Shu-Hui; Lin, Chin-Yang

    2003-04-01

    Strains representing four types of common heat-resistant spores of Bacillus spp., B. cereus CCRC 14655, B. coagulans CCRC10606, B. licheniformis CCRC14693, and B. subtilis CCRC14199, were heated with microwaves at different power levels and under different conditions in salt solutions, starch solutions, and containers. The results of this study showed that B. licheniformis spores had the highest microwave tolerance at a power level of 100% for different incubation times. B. coagulans spores showed the lowest microwave tolerance in salt solutions with water activity values of 0.6, 0.7, 0.8, and 0.9, and B. licheniformis spores were the most resistant in the tested salt solutions at different incubation times. An analysis of the effect of the viscosity of the medium revealed that the bacteria had the lowest microwave resistance in a medium containing <0.8% starch in solution. The microwave resistance levels of the test microorganisms were the lower in glass containers than in polypropylene containers and aluminum foil-enclosed pouches. Of the four species of bacilli, B. licheniformis had the highest microwave tolerance (P < 0.05) under all conditions.

  1. FgFlbD regulates hyphal differentiation required for sexual and asexual reproduction in the ascomycete fungus Fusarium graminearum.

    PubMed

    Son, Hokyoung; Kim, Myung-Gu; Chae, Suhn-Kee; Lee, Yin-Won

    2014-11-01

    Fusarium graminearum is a filamentous fungal plant pathogen that infects major cereal crops. The fungus produces both sexual and asexual spores in order to endure unfavorable environmental conditions and increase their numbers and distribution across plants. In a model filamentous fungus, Aspergillus nidulans, early induction of conidiogenesis is orchestrated by the fluffy genes. The objectives of this study were to characterize fluffy gene homologs involved in conidiogenesis and their mechanism of action in F. graminearum. We characterized five fluffy gene homologs in F. graminearum and found that FlbD is the only conserved regulator for conidiogenesis in A. nidulans and F. graminearum. Deletion of fgflbD prevented hyphal differentiation and the formation of perithecia. Successful interspecies complementation using A. nidulans flbD demonstrated that the molecular mechanisms responsible for FlbD functions are conserved in F. graminearum. Moreover, abaA-wetA pathway is positively regulated by FgFlbD during conidiogenesis in F. graminearum. Deleting fgflbD abolished morphological effects of abaA overexpression, which suggests that additional factors for FgFlbD or an AbaA-independent pathway for conidiogenesis are required for F. graminearum conidiation. Importantly, this study led to the construction of a genetic pathway of F. graminearum conidiogenesis and provides new insights into the genetics of conidiogenesis in fungi.

  2. Effects of anthracene on development of an arbuscular mycorrhizal fungus and contribution of the symbiotic association to pollutant dissipation.

    PubMed

    Verdin, A; Lounès-Hadj Sahraoui, A; Fontaine, J; Grandmougin-Ferjani, A; Durand, R

    2006-09-01

    The influence of anthracene, a low molecular weight polycyclic aromatic hydrocarbon (PAH), on chicory root colonization by Glomus intraradices and the effect of the root colonization on PAH degradation were investigated in vitro. The fungus presented a reduced development of extraradical mycelium and a decrease in sporulation, root colonization, and spore germination when exposed to anthracene. Mycorrhization improved the growth of the roots in the medium supplemented containing 140 mg l(-1) anthracene, suggesting a positive contribution of G. intraradices to the PAH tolerance of roots. Anthracene disappearance from the culture medium was quantified; results suggested that nonmycorrhizal chicory roots growing in vitro were able to contribute to anthracene dissipation, and in addition, that mycorrhization significantly enhanced anthracene dissipation. These monoxenic experiments demonstrated a positive contribution of the symbiotic association to anthracene dissipation in the absence of other microorganisms. In addition to anthracene dissipation, intracellular accumulation of anthracene was detected in lipid bodies of plant cells and fungal hyphae, indicating intracellular storage capacity of the pollutant by the roots and the mycorrhizal fungus.

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

    NASA Astrophysics Data System (ADS)

    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

  4. NanoSIMS analysis of Bacillus spores for forensics

    SciTech Connect

    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

  5. Expanding Distribution of Lethal Amphibian Fungus Batrachochytrium salamandrivorans in Europe

    PubMed Central

    Spitzen-van der Sluijs, Annemarieke; Martel, An; Asselberghs, Johan; Bales, Emma K.; Beukema, Wouter; Bletz, Molly C.; Dalbeck, Lutz; Goverse, Edo; Kerres, Alexander; Kinet, Thierry; Kirst, Kai; Laudelout, Arnaud; Marin da Fonte, Luis F.; Nöllert, Andreas; Ohlhoff, Dagmar; Sabino-Pinto, Joana; Schmidt, Benedikt R.; Speybroeck, Jeroen; Spikmans, Frank; Steinfartz, Sebastian; Veith, Michael; Vences, Miguel; Wagner, Norman; Pasmans, Frank

    2016-01-01

    Emerging fungal diseases can drive amphibian species to local extinction. During 2010–2016, we examined 1,921 urodeles in 3 European countries. Presence of the chytrid fungus Batrachochytrium salamandrivorans at new locations and in urodeles of different species expands the known geographic and host range of the fungus and underpins its imminent threat to biodiversity. PMID:27070102

  6. Metacridamides A and B from the biocontrol fungus metarhizium acridum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Metarhizium acridum, an entomopathogenic fungus, has been commercialized and used successfully for biocontrol of grasshopper pests in Africa and Australia. As part of an effort to catalog the secondary metabolites of this fungus we discovered that its conidia produce two novel 17-membered macrocycl...

  7. The effect of ionizing radiation on amino acids and bacterial spores in different geo- and cosmochemical environments

    NASA Astrophysics Data System (ADS)

    Kminek, Gerhard

    In this thesis I have investigated the impact of ionizing radiation from the environment on the stability of bacterial spores and amino acids. I measured the radiolysis constant of amino acids and the inactivation constant of bacterial spores. To put these results in the context of a natural setting, I have selected four different cases and calculated the radiation environment for meteorites, the Martian subsurface, terrestrial halite fluid inclusions, and fossil bones. Bacterial spores exhibit a remarkable resistance to adverse environments and are the best example for the long-term survival of life forms. On a molecular level, amino acids are of particular interest because of their importance in biochemistry and their stability in the environment. The significance of amino acids, however, goes back to a time before life existed. The exogenous delivery of amino acids by meteorites might have been essential to provide the required supply of organic molecules for the origin of life on the Earth. There is one common threat, however, to the preservation of amino acids and bacterial spores in all known terrestrial and extraterrestrial environments: ionizing radiation. Amino acids in meteorites are exposed to radiation from internal radioactivity and space radiation. I show that this radiation decomposes substantial amounts of amino acids over time, indicating a higher exogenous delivery of amino acids to the early Earth. The total radiodecomposition since the synthesis of amino acids is between 23 and 68%. Radiodecomposition induces a certain fractionation in favor of smaller amino acids. Fossil bones show a post-mortem uranium uptake. My results suggest a substantial radiodecomposition of amino acids on a 10 million year time scale. Age determination based on racemization of amino acids will be affected in fossil bones that are older than 1--30 million years. My results on the stability of bacterial spores in halite fluid inclusions and on Mars suggest that radiation

  8. Molecular dissection of Neurospora Spore killer meiotic drive elements.

    PubMed

    Hammond, Thomas M; Rehard, David G; Xiao, Hua; Shiu, Patrick K T

    2012-07-24

    Meiotic drive is a non-Mendelian inheritance phenomenon in which certain selfish genetic elements skew sexual transmission in their own favor. In some cases, progeny or gametes carrying a meiotic drive element can survive preferentially because it causes the death or malfunctioning of those that do not carry it. In Neurospora, meiotic drive can be observed in fungal spore killing. In a cross of Spore killer (Sk) × WT (Sk-sensitive), the ascospores containing the Spore killer allele survive, whereas the ones with the sensitive allele degenerate. Sk-2 and Sk-3 are the most studied meiotic drive elements in Neurospora, and they each theoretically contain two essential components: a killer element and a resistance gene. Here we report the identification and characterization of the Sk resistance gene, rsk (resistant to Spore killer). rsk seems to be a fungal-specific gene, and its deletion in a killer strain leads to self-killing. Sk-2, Sk-3, and naturally resistant isolates all use rsk for resistance. In each killer system, rsk sequences from an Sk strain and a resistant isolate are highly similar, suggesting that they share the same origin. Sk-2, Sk-3, and sensitive rsk alleles differ from each other by their unique indel patterns. Contrary to long-held belief, the killer targets not only late but also early ascospore development. The WT RSK protein is dispensable for ascospore production and is not a target of the spore-killing mechanism. Rather, a resistant version of RSK likely neutralizes the killer element and prevents it from interfering with ascospore development.

  9. Hazardous waste treatment using fungus enters marketplace

    SciTech Connect

    Illman, D.L.

    1993-07-01

    When the announcement was made eight years ago that a common fungus had been found that could degrade a variety of environmental pollutants, the news stirred interest in the scientific community, the private sector, and the general public. Here was the promise of a new technology that might be effective and economical in treating hazardous waste, especially the most recalcitrant of toxic pollutants. Today, commercialization is beginning amid a mixture of optimism and skepticism. The organism in question is white rot fungus, or Phanerochaete chrysosporium, and it belongs to a family of woodrotting fungi common all over North America. The fungi secrete enzymes that break down lignin in wood to carbon dioxide and water--a process called mineralization. These lignin-degrading enzymes are not very discriminating, however. The white rot fungi have been shown to degrade such materials as DDT, the herbicide (2,4,5-trichlorophenoxy)acetic acid (2,4,5-T), 2,4,6-trinitrotoluene (TNT), pentachlorophenol (PCP), creosote, coal tars, and heavy fuels, in many cases mineralizing these pollutants to a significant extent.

  10. Bacterial farming by the fungus Morchella crassipes.

    PubMed

    Pion, Martin; Spangenberg, Jorge E; Simon, Anaele; Bindschedler, Saskia; Flury, Coralie; Chatelain, Auriel; Bshary, Redouan; Job, Daniel; Junier, Pilar

    2013-12-22

    The interactions between bacteria and fungi, the main actors of the soil microbiome, remain poorly studied. Here, we show that the saprotrophic and ectomycorrhizal soil fungus Morchella crassipes acts as a bacterial farmer of Pseudomonas putida, which serves as a model soil bacterium. Farming by M. crassipes consists of bacterial dispersal, bacterial rearing with fungal exudates, as well as harvesting and translocation of bacterial carbon. The different phases were confirmed experimentally using cell counting and (13)C probing. Common criteria met by other non-human farming systems are also valid for M. crassipes farming, including habitual planting, cultivation and harvesting. Specific traits include delocalization of food production and consumption and separation of roles in the colony (source versus sink areas), which are also found in human agriculture. Our study evidences a hitherto unknown mutualistic association in which bacteria gain through dispersal and rearing, while the fungus gains through the harvesting of an additional carbon source and increased stress resistance of the mycelium. This type of interaction between fungi and bacteria may play a key role in soils.

  11. Bacterial farming by the fungus Morchella crassipes

    PubMed Central

    Pion, Martin; Spangenberg, Jorge E.; Simon, Anaele; Bindschedler, Saskia; Flury, Coralie; Chatelain, Auriel; Bshary, Redouan; Job, Daniel; Junier, Pilar

    2013-01-01

    The interactions between bacteria and fungi, the main actors of the soil microbiome, remain poorly studied. Here, we show that the saprotrophic and ectomycorrhizal soil fungus Morchella crassipes acts as a bacterial farmer of Pseudomonas putida, which serves as a model soil bacterium. Farming by M. crassipes consists of bacterial dispersal, bacterial rearing with fungal exudates, as well as harvesting and translocation of bacterial carbon. The different phases were confirmed experimentally using cell counting and 13C probing. Common criteria met by other non-human farming systems are also valid for M. crassipes farming, including habitual planting, cultivation and harvesting. Specific traits include delocalization of food production and consumption and separation of roles in the colony (source versus sink areas), which are also found in human agriculture. Our study evidences a hitherto unknown mutualistic association in which bacteria gain through dispersal and rearing, while the fungus gains through the harvesting of an additional carbon source and increased stress resistance of the mycelium. This type of interaction between fungi and bacteria may play a key role in soils. PMID:24174111

  12. General metabolism of the dimorphic and pathogenic fungus Paracoccidioides brasiliensis.

    PubMed

    Arraes, Fabrício B M; Benoliel, Bruno; Burtet, Rafael T; Costa, Patrícia L N; Galdino, Alexandro S; Lima, Luanne H A; Marinho-Silva, Camila; Oliveira-Pereira, Luciana; Pfrimer, Pollyanna; Procópio-Silva, Luciano; Reis, Viviane Castelo-Branco; Felipe, Maria Sueli S

    2005-06-30

    Annotation of the transcriptome of the dimorphic fungus Paracoccidioides brasiliensis has set the grounds for a global understanding of its metabolism in both mycelium and yeast forms. This fungus is able to use the main carbohydrate sources, including starch, and it can store reduced carbons in the form of glycogen and trehalose; these provide energy reserves that are relevant for metabolic adaptation, protection against stress and infectivity mechanisms. The glyoxylate cycle, which is also involved in pathogenicity, is present in this fungus. Classical pathways of lipid biosynthesis and degradation, including those of ketone body and sterol production, are well represented in the database of P. brasiliensis. It is able to synthesize de novo all nucleotides and amino acids, with the sole exception of asparagine, which was confirmed by the fungus growth in minimal medium. Sulfur metabolism, as well as the accessory synthetic pathways of vitamins and co-factors, are likely to exist in this fungus.

  13. KINETICS OF DRY RUPTURE OF BACTERIAL SPORES IN THE PRESENCE OF SALT.

    PubMed

    SACKS, L E; PERCELL, P B; THOMAS, R S; BAILEY, G F

    1964-04-01

    Sacks, L. E. (U.S. Department of Agriculture, Albany, Calif.), Peter B. Percell, Richard S. Thomas, and Glen F. Bailey. Kinetics of dry rupture of bacterial spores in the presence of salt. J. Bacteriol. 87:952-960. 1964.-The kinetics of breaking spores in the dry state by use of an excess of sodium chloride and a steel ball in a shaking device were investigated. Under most conditions, disruption is a first-order process. The disruption-rate constant varies directly with the weight of the ball and inversely with the weight of the capsule contents (spores plus salt). Different spore batches differ somewhat in susceptibility to dry rupture. The dry-rupture process is highly reproducible and it is relatively simple to obtain preparations in which exactly 50%, or 90%, of the spores are broken. The procedure is uniquely suited to the disruption of small (5 to 20 mg) samples, but 150 mg of spores have been handled with conventional equipment. Apparently, the chief function of the salt is to separate the spores from one another with a relatively hard, energy-nonabsorbing matrix, preventing aggregation and consequent cushioning of the ball's impact. However, under certain conditions (small ball, high salt, large crystals) appreciable breakage results from collisions of spores with the salt crystals. The minimal salt-spore ratio for efficient breakage depends on the spore batch, but is usually greater than 3:1. Fine glass beads or inorganic salts other than sodium chloride will also serve as the matrix. Electron micrographs of the spores in various stages of disruption are shown, as are electron micrographs of the spore coats of Bacillus macerans, B. megaterium, B. cereus, B. coagulans, and Clostridium bifermentans. Prolonged agitation disintegrates spore coats. The spore coats of B. macerans exhibit a characteristic ribbed structure, previously detected only by carbon replicas of intact spores. Possible application to other biological materials is considered.

  14. Influence of Spore Moisture Content on the Dry-Heat Resistance of Bacillus subtilis var. niger

    PubMed Central

    Angelotti, Robert; Maryanski, James H.; Butler, Thomas F.; Peeler, James T.; Campbell, Jeptha E.

    1968-01-01

    The dry-heat resistance of Bacillus subtilis var. niger spores located in or on various materials was determined as D and z values in the range of 105 through 160 C. The systems tested included spores located on steel and paper strips, spores located between stainless-steel washers mated together under 150 inch-lb and 12 inch-lb of torque, and spores encapsulated in methylmethacrylate and epoxy plastics. D values for a given temperature varied with the test system. High D values were observed for the systems in which spores were encapsulated or under heavy torque, whereas lower D values were observed for the steel and paper strip systems and the lightly torqued system. Similar z values were obtained for the plastic and steel strip systems (zD = 21 C), but an unusually low z for spores on paper (zD = 12.9 C) and an unusually high z for spores on steel washers mated at 150 inch-lb of torque (zD = 32 C) were observed. The effect of spore moisture content on the D value of spores encapsulated in water-impermeable plastic was determined, and maximal resistance was observed for spores with a water activity (aw) of 0.2 to 0.4. Significantly decreased D values were observed for spores with moisture contents below aw 0.2 or above aw 0.4. The data indicate that the important factors to be considered when measuring the dry heat resistance of spores are (i) the initial moisture content of the spore, (ii) the rate of spore desiccation during heating, (iii) the water retention capacity of the material in or on which spores are located, and (iv) the relative humidity of the system at the test temperature. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 7 PMID:4968962

  15. Agent-Based Modeling of Physical Factors That May Control the Growth of Coccidioides immitis (Valley Fever Fungus) in Soils

    NASA Astrophysics Data System (ADS)

    Gettings, M. E.; Fisher, F. S.

    2003-12-01

    A model of the spread and survival of the fungus Coccidioides immitis in soil via wind-borne spore transport has been completed using public domain agent-based modeling software. The hypothetical model posits that for a successful new site to become established, four factors must be simultaneously satisfied. 1) There must be transport of spores from a source site to sites with favorable soil geology, texture, topographic aspect, and lack of biomass competition. 2) There must be sufficient moisture for fungal growth. 3) Temperature of the surface and soil must be favorable for growth. Finally, 4) the temperature and moisture must remain in favorable ranges for a long enough time interval for the fungus to grow down to depths at which spores will survive subsequent heat, aridity, and ultraviolet radiation of the hot, dry season typical of the Southwest U.S. climate. Using agent-based modeling software, a model was built so that the effects of combinations of these controlling factors could be evaluated using realistic temperature, rain and wind models. The rain probability and amount, temperature annual and diurnal variation, and wind direction and intensity were based on the weather records at Tucson, Arizona for the 107-year period from 1894 to 2001. Favorable ground was defined using a fractal tree algorithm that emulates a drainage network in accordance with observations that favorable sites are often adjacent to drainage channels. Numerous model runs produced the following five conclusions. 1) If any property is not isotropic, for example wind direction or narrow paths of rainstorms, parts of the favorable areas will never become colonized no matter how long the model runs. 2)The spread of sites is extremely sensitive to moisture duration. The amount of wind and temperature after a rain control the length of time before a site becomes too dry. 3) The distribution of wind and rainstorm direction relative to that of the favorable sites is a strong control on the

  16. Protection against UV disinfection of E. coli bacteria and B. subtilis spores ingested by C. elegans nematodes.

    PubMed

    Bichai, Françoise; Barbeau, Benoit; Payment, Pierre

    2009-08-01

    Nematodes, which occur abundantly in granular media filters of drinking water treatment plants and in distribution systems, can ingest and transport pathogenic bacteria and provide them protection against chemical disinfectants. However, protection against UV disinfection had not been investigated to date. In this study, Caenorhabditis elegans nematodes (wild-type strain N2) were allowed to feed on Escherichia coli OP50 and Bacillus subtilis spores before being exposed to 5 and 40 mJ/cm(2) UV fluences, using a collimated beam apparatus (LP, 254 nm). Sonication (15 W, 60s) was used to extract bacteria from nematode guts following UV exposure in order to assess the amount of ingested bacteria that resisted the UV treatment using a standard culture method. Bacteria located inside the gut of C. elegans were shown to benefit from a significant protection against UV. Approximately 15% of the applied UV fluence of 40 mJ/cm(2) (as typically used in WTP) was found to reach the bacteria located inside nematode guts based on the inactivation of recovered bacteria (2.7 log reduction of E. coli bacteria and 0.7 log reduction of B. subtilis spores at 40 mJ/cm(2)). To our knowledge, this study is the first demonstration of the protection effect of bacterial internalization by higher organisms against UV treatment, using the specific case of E. coli and B. subtilis spores ingested by C. elegans.

  17. Lipid droplet dynamics during Schizosaccharomyces pombe sporulation and their role in spore survival

    PubMed Central

    Yang, Hui-Ju; Osakada, Hiroko; Kojidani, Tomoko; Haraguchi, Tokuko

    2017-01-01

    ABSTRACT Upon nitrogen starvation, the fission yeast Schizosaccharomyces pombe forms dormant spores; however, the mechanisms by which a spore sustains life without access to exogenous nutrients remain unclear. Lipid droplets are reservoirs of neutral lipids that act as important cellular energy resources. Using live-cell imaging analysis, we found that the lipid droplets of mother cells redistribute to their nascent spores. Notably, this process was actin polymerization-dependent and facilitated by the leading edge proteins of the forespore membrane. Spores lacking triacylglycerol synthesis, which is essential for lipid droplet formation, failed to germinate. Our results suggest that the lipid droplets are important for the sustenance of life in spores. PMID:28011631

  18. Isolation, characterization and application of a chitosan-degrading fungus from soil.

    PubMed

    Wei, Xinlin; Chen, Wei; Xiao, Ming; Xiao, Jianbo; Wang, Yuanfeng

    2010-07-01

    A chitosan-degrading fungus, BSF114, was isolated from soil. The culture preparation showed strong chitosanolytic enzyme activity at optimum pH 4.0 and optimum temperature of 60 degrees C after 36-40 h fermentation. The rapid decreased viscosity of chitosan solutions at early stage of reaction suggested an endo-type cleavage of the polymeric chitosan chains. To identify the isolated fungus, molecular biological and morphological methods were used. The fungal internal transcribed spacer (ITS) region 1 was amplified, sequenced, and then compared with related sequences in the GenBank database using BLAST. The phylogenetic relationships were then analyzed, and the results showed that the fungus belongs to Aspergillus fumigatus. Morphological observations were also used to confirm the above conclusion. The chitooligosaccharides (COS) obtained through hydrolyzing the colloidal chitosan showed A. fumigatus BSF114 is suitable for degrading chitosan and producing chitooligosaccharides on a large scale. High concentrations of the COS (1000 and 500 microg/mL) significantly proliferated mice marrow cells.

  19. Penicillium menonorum: A Novel Fungus to Promote Growth and Nutrient Management in Cucumber Plants

    PubMed Central

    Babu, Anam Giridhar; Kim, Sang Woo; Yadav, Dil Raj; Hyum, Umyong; Adhikari, Mahesh

    2015-01-01

    The present study is the first report on the isolation of Penicillium menonorum from rhizosphere soil in Korea and its identification based on morphological characteristics and internal transcribed spacer gene sequence. The fungal isolate was named KNU-3 and was found to exhibit plant growth-promoting (PGP) activity through indole acetic acid (IAA) and siderophore production, as well as P solubilization. KNU-3 produced 9.7 mg/L IAA and solubilized 408 mg of Ca3PO4/L, and inoculation with the isolate significantly (p < 0.05) increased the dry biomass of cucumber roots (57%) and shoots (52%). Chlorophyll, starch, protein, and P contents were increased by 16%, 45%, 22%, and 14%, respectively, compared to plants grown in uninoculated soil. The fungus also increased soil dehydrogenase (30%) and acid phosphatase (19%) activities. These results demonstrate that the isolate KNU-3 has potential PGP attributes, and therefore it can be considered as a new fungus to enhance soil fertility and promote plant growth. Moreover, the discovery of PGP ability and traits of this fungus will open new aspects of research and investigations. In this study, plant growth promotion by P. menonorum KNU-3 is reported for the first time in Korea after its original description. PMID:25892915

  20. Virulence determinants of the human pathogenic fungus Aspergillus fumigatus protect against soil amoeba predation.

    PubMed

    Hillmann, Falk; Novohradská, Silvia; Mattern, Derek J; Forberger, Tilmann; Heinekamp, Thorsten; Westermann, Martin; Winckler, Thomas; Brakhage, Axel A

    2015-08-01

    Filamentous fungi represent classical examples for environmentally acquired human pathogens whose major virulence mechanisms are likely to have emerged long before the appearance of innate immune systems. In natural habitats, amoeba predation could impose a major selection pressure towards the acquisition of virulence attributes. To test this hypothesis, we exploited the amoeba Dictyostelium discoideum to study its interaction with Aspergillus fumigatus, two abundant soil inhabitants for which we found co-occurrence in various sites. Fungal conidia were efficiently taken up by D. discoideum, but ingestion was higher when conidia were devoid of the green fungal spore pigment dihydroxynaphtalene melanin, in line with earlier results obtained for immune cells. Conidia were able to survive phagocytic processing, and intracellular germination was initiated only after several hours of co-incubation which eventually led to a lethal disruption of the host cell. Besides phagocytic interactions, both amoeba and fungus secreted cross inhibitory factors which suppressed fungal growth or induced amoeba aggregation with subsequent cell lysis, respectively. On the fungal side, we identified gliotoxin as the major fungal factor killing Dictyostelium, supporting the idea that major virulence attributes, such as escape from phagocytosis and the secretion of mycotoxins are beneficial to escape from environmental predators.

  1. Characterization of the glyoxalase I gene from the vascular wilt fungus Verticillium dahliae.

    PubMed

    Klimes, A; Neumann, M J; Grant, S J; Dobinson, K F

    2006-09-01

    A glyoxalase I gene homologue (VdGLO1) was identified in the vascular wilt fungus Verticillium dahliae by sequence tag analysis of genes expressed during resting structure development. The results of the current study show that the gene encodes a putative 345 amino acid protein with high similarity to glyoxalase I, which produces S-D-lactoylglutathione from the toxic metabolic by-product methylglyoxal (MG). Disruption of the V. dahliae gene by Agrobacterium tumefaciens-mediated transformation resulted in enhanced sensitivity to MG. Mycelial growth of disruption mutants was severely reduced in the presence of 5 mmol/L MG. In contrast, spore production in liquid medium was abolished at 1 mmol/L MG, although not at physiologically relevant concentrations of

  2. Comparison of the toxicity of reference mycotoxins and spore extracts of common indoor moulds.

    PubMed

    Schulz, Torsten; Senkpiel, Klaus; Ohgke, Helge

    2004-07-01

    There is an unclear endangering potential by toxic influences of inhaled conidiospores and therefore the conidia of indoor mould species were cultured and toxicologically examined after their mechanical disintegration. For this purpose high-performance liquid chromatography (HPLC) and three colorimetric bioassays, the PTGT (pollen tube growth test), the MB (methylene blue) and the MTT (methylthiazoltetrazolium) assay were applied. The sensitivity of the biological methods was evaluated by using 12 reference mycotoxins and 3 structural cell wall components. Only in one extract of disintegrated spores (Aspergillus fumigatus) a mycotoxin (0.22 microg gliotoxin/6.2 x 10(8) spores) was determined. All nine spore extracts, however, turned out to be cytotoxic and in this case the MTT assay was remarkably more sensitive than the two other test methods. The IC50 values of six different spore extracts determined by the MTT assay were lower than 10(6) spores/well (well = 0.2 ml) whereas the IC50 values determined by the MB assay and PTGT were higher than 10(6) spores per 0.2 ml for each spore extract. An examination of four spore extracts, which were fractionated depending on their polarity by HPLC, showed that single substances as well as synergistic effects contribute to the toxic properties of the spores. The results of this work indicate a health hazard due to toxic effects after the inhalation of extremely high spore concentrations of indoor moulds. This risk will also exist if the spores do not contain any mycotoxins.

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

    SciTech Connect

    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.

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

    PubMed

    Park, Hyun; Ka, Kang-Hyeon

    2010-09-01

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

  5. Activity of bleach, ethanol and two commercial disinfectants against spores of Encephalitozoon cuniculi.

    PubMed

    Jordan, Carly N; Dicristina, Jennifer A; Lindsay, David S

    2006-03-31

    Encephalitozoon cuniculi is a small protist parasite in the phylum Microspora. Hosts are infected by ingestion or inhalation of spores passed in the urine or feces. Infection with E. cuniculi is usually asymptomatic, except in young or immunocompromised hosts. This study examined the effects of various disinfectants on in vitro infectivity of E. cuniculi spores. Spores of E. cuniculi were exposed to several dilutions of commercial bleach, 70% ethanol and dilutions of commercial disinfectants HiTor and Roccal for 10 min and then loaded onto human fibroblast cells (Hs68 cells). Ten minutes of exposure to these disinfectants was lethal to E. cuniculi spores. Additional exposure time studies were done using dilutions of bleach at 0.1, 1 and 10%, and 70% ethanol. Exposure of E. cuniculi spores to 1 or 10% bleach for 30s rendered them non-infectious for Hs68 cells. Growth of E. cuniculi was observed in Hs68 cells inoculated with spores treated with 0.1% bleach for 30s or 1, 3 and 5 min, but not with spores treated for 7 min or longer. Exposure of E. cuniculi spores to 70% ethanol for 30s rendered them non-infectious for Hs68 cells. Spores of E. cuniculi are more sensitive to disinfectants than are coccidial oocysts and other parasite cysts. The relatively short contact time needed to kill spores indicates that disinfection of animal housing may be a viable means to reduce exposure of animals to E. cuniculi spores.

  6. The Clostridium botulinum GerAB germination protein is located in the inner membrane of spores.

    PubMed

    Alberto, François; Botella, Lucien; Carlin, Fréderic; Nguyen-The, Christophe; Broussolle, Véronique

    2005-12-15

    Clostridium botulinum dormant spores germinate in presence of l-alanine via a specific receptor composed of GerAA, GerAB and GerAC proteins. In Bacillus subtilis spores, GerAA and GerAC proteins were located in the inner membrane of the spore. We studied the location of the GerAB protein in C. botulinum spore fractions by Western-blot analysis, using an antipeptidic antibody. The protein GerAB was in vitro translated and used to confirm the specificity of the antibodies. GerAB was not present in a coat and spore outer membrane fraction but was present in a fraction of decoated spores containing inner membrane. These results strongly suggest that the protein GerAB is located in the inner membrane of the spore.

  7. Germination and growth from spores: variability and uncertainty in the assessment of food borne hazards.

    PubMed

    Barker, G C; Malakar, P K; Peck, M W

    2005-04-15

    We have developed a model for the variability of spore lag times and shown that variability has an important role in the quantitative assessment of risks associated with spore forming bacteria in food. The model includes two sequential independent delay times that contribute to the lag time for a single spore. We have shown that a population of variable spores also has a variable lag time, and we have emphasised the significance of this variability in quantitative representations of population dynamics for small populations. We have made a Bayesian estimate for the extent of the variability in spore lag times and made a comparison with direct microscopic observations of individual spores of nonproteolytic Clostridium botulinum. We conclude that Bayesian inference is a practical method for quantifying variability and hence a significant element in the development of quantitative risk assessments for hazards associated with spore forming bacteria.

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

    SciTech Connect

    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.

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

    SciTech Connect

    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.

  10. Ultrastructure and extreme heat resistance of spores from thermophilic Clostridium species.

    PubMed Central

    Hyun, H H; Zeikus, J G; Longin, R; Millet, J; Ryter, A

    1983-01-01

    The heat resistance and ultrastructural features of spore suspensions prepared from Clostridium thermocellum LQRI, Clostridium thermosulfurogenes 4B, and Clostridium thermohydrosulfuricum 39E were compared as a function of decimal reduction time. The decimal reduction times at 121 degrees C for strains LQRI, 4B, and 39E were 0.5, 2.5, and 11 min. The higher degree of spore heat resistance was associated with a spore architecture displaying a thicker cortex layer. Heat resistance of these spores was proportional to the ratio of spore cortex volume to cytoplasmic volume. These ratios for spores of strains LQRI, 4B, and 39E were 1.4, 1.6, and 6.6, respectively. The extreme heat resistance and autoclavable nature of C. thermohydrosulfuricum spores under routine sterilization procedures is suggested as a common cause of laboratory contamination with pure cultures of thermophilic, saccharide-fermenting anaerobes. Images PMID:6643392

  11. Purification of an Inducible DNase from a Thermophilic Fungus

    PubMed Central

    Landry, Kyle S.; Vu, Andrea; Levin, Robert E.

    2014-01-01

    The ability to induce an extracellular DNase from a novel thermophilic fungus was studied and the DNAse purified using both traditional and innovative purification techniques. The isolate produced sterile hyphae under all attempted growing conditions, with an average diameter of 2 μm and was found to have an optimal temperature of 45 °C and a maximum of 65 °C. Sequencing of the internal transcribed region resulted in a 91% match with Chaetomium sp., suggesting a new species, but further clarification on this point is needed. The optimal temperature for DNase production was found to be 55 °C and was induced by the presence of DNA and/or deoxyribose. Static growth of the organism resulted in significantly higher DNase production than agitated growth. The DNase was purified 145-fold using a novel affinity membrane purification system with 25% of the initial enzyme activity remaining. Electrophoresis of the purified enzyme resulted in a single protein band, indicating DNase homogeneity. PMID:24447923

  12. Acrophialophora, a Poorly Known Fungus with Clinical Significance

    PubMed Central

    Sandoval-Denis, Marcelo; Sutton, Deanna A.; Wiederhold, Nathan P.; Guarro, Josep

    2015-01-01

    Acrophialophora fusispora is an emerging opportunistic fungus capable of causing human infections. The taxonomy of the genus is not yet resolved and, in order to facilitate identification of clinical specimens, we have studied a set of clinical and environmental Acrophialophora isolates by morphological and molecular analyses. This set included the available type strains of Acrophialophora species and similar fungi, some of which were considered by various authors to be synonyms of A. fusispora. Sequence analysis of the large subunit (LSU) and internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA and a fragment of the β-tubulin (Tub) gene revealed that Acrophialophora belongs in the family Chaetomiaceae and comprises three different species, i.e., A. fusispora, Acrophialophora levis, and Acrophialophora seudatica; the latter was previously included in the genus Ampullifera. The most prevalent species among clinical isolates was A. levis (72.7%), followed by A. fusispora (27.3%), both of which were isolated mostly from respiratory specimens (72.7%), as well as subcutaneous and corneal tissue samples. In general, of the eight antifungal drugs tested, voriconazole had the greatest in vitro activity, while all other agents showed poor in vitro activity against these fungi. PMID:25716450

  13. Acrophialophora, a poorly known fungus with clinical significance.

    PubMed

    Sandoval-Denis, Marcelo; Gené, Josepa; Sutton, Deanna A; Wiederhold, Nathan P; Guarro, Josep

    2015-05-01

    Acrophialophora fusispora is an emerging opportunistic fungus capable of causing human infections. The taxonomy of the genus is not yet resolved and, in order to facilitate identification of clinical specimens, we have studied a set of clinical and environmental Acrophialophora isolates by morphological and molecular analyses. This set included the available type strains of Acrophialophora species and similar fungi, some of which were considered by various authors to be synonyms of A. fusispora. Sequence analysis of the large subunit (LSU) and internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA and a fragment of the β-tubulin (Tub) gene revealed that Acrophialophora belongs in the family Chaetomiaceae and comprises three different species, i.e., A. fusispora, Acrophialophora levis, and Acrophialophora seudatica; the latter was previously included in the genus Ampullifera. The most prevalent species among clinical isolates was A. levis (72.7%), followed by A. fusispora (27.3%), both of which were isolated mostly from respiratory specimens (72.7%), as well as subcutaneous and corneal tissue samples. In general, of the eight antifungal drugs tested, voriconazole had the greatest in vitro activity, while all other agents showed poor in vitro activity against these fungi.

  14. Chemical composition of metapleural gland secretions of fungus-growing and non-fungus-growing ants.

    PubMed

    Vieira, Alexsandro S; Morgan, E David; Drijfhout, Falko P; Camargo-Mathias, Maria I

    2012-10-01

    The metapleural gland is exclusive to ants, and unusual among exocrine glands in having no mechanism for closure and retention of secretion. As yet, no clear conclusion has been reached as to the function of metapleural gland secretion. Metapleural gland secretions were investigated for fungus-growing ants representing the derived attines Trachymyrmex fuscus, Atta laevigata, and Acromyrmex coronatus, the basal attines Apterostigma pilosum and Mycetarotes parallelus, and non-fungus-growing ants of the tribes Ectatommini (Ectatomma brunneum) and Myrmicini (Pogonomyrmex naegeli). Our results showed that the secretions of leaf-cutting ants (A. laevigata and A. coronatus) and the derived attine, T. fuscus, contain a greater variety and larger quantities of volatile compounds than those of myrmicine and ectatommine ants. The most abundant compounds found in the metapleural glands of A. laevigata and A. coronatus were hydroxyacids, and phenylacetic acid (only in A. laevigata). Indole was present in all groups examined, while skatole was found in large quantities only in attines. Ketones and aldehydes are present in the secretion of some attines. Esters are present in the metapleural gland secretion of all species examined, although mainly in A. laevigata, A. coronatus, and T. fuscus. Compared with basal attines and non-fungus-growing ants, the metapleural glands of leaf-cutting ants produce more acidic compounds that may have an antibiotic or antifungal function.

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

    PubMed Central

    2010-01-01

    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. PMID:21092338

  16. 14C Analysis of Protein Extracts from Bacillus Spores

    PubMed Central

    Cappucio, Jenny A.; Sarachine Falso, Miranda J.; Kashgarian, Michaele; Buchholz, Bruce A.

    2014-01-01

    Investigators of bioagent incidents or interdicted materials need validated, independent analytical methods that will allow them to distinguish between recently made bioagent samples versus material drawn from the archives of a historical program. Heterotrophic bacteria convert the carbon in their food sources, growth substrate or culture media, into the biomolecules they need. The F14C (fraction modern radiocarbon) of a variety of media, Bacillus spores, and separated proteins from Bacillus spores was measured by accelerator mass spectrometry (AMS). AMS precisely measures F14C values of biological materials and has been used to date the synthesis of biomaterials over the bomb pulse era (1955 to present). The F14C of Bacillus spores reflects the radiocarbon content of the media in which they were grown. In a survey of commercial media we found that the F14C value indicated that carbon sources for the media were alive within about a year of the date of manufacture and generally of terrestrial origin. Hence, bacteria and their products can be dated using their 14C signature. Bacillus spore samples were generated onsite with defined media and carbon free purification and also obtained from archived material. Using mechanical lysis and a variety of washes with carbon free acids and bases, contaminant carbon was removed from soluble proteins to enable accurate 14C bomb-pulse dating. Since media is contemporary, 14C bomb-pulse dating of isolated soluble proteins can be used to distinguish between historical archives of bioagents and those produced from recent media. PMID:24814329

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

    NASA Technical Reports Server (NTRS)

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

    1974-01-01

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

  18. Processing, Assembly and Localization of a Bacillus anthracis Spore Protein

    DTIC Science & Technology

    2010-01-01

    anthracis (but not, for example, Bacillus megaterium ), a series of fine hair-like projections, also called a nap, extends from the exosporium (Aronson...3373–3378. Vary, P. S. (1994). Prime time for Bacillus megaterium . Microbiology 140, 1001–1013. Welkos, S. L., Cote, C. K., Rea, K. M. & Gibbs, P. H...Processing, assembly and localization of a Bacillus anthracis spore protein K. L. Moody,13 A. Driks,2 G. L. Rother,1 C. K. Cote,1 E. E. Brueggemann,3

  19. Encapsulation of Bacterial Spores in Nanoorganized Polyelectrolyte Shells (Postprint)

    DTIC Science & Technology

    2009-05-27

    features using in vitro self-assembly methods. Shell formation was based on layer-by-layer electrostatic assembly via the alternate adsorption of...small-angle neutron scattering. LbL assembly of natural polyelectrolytes ( chitosan , alginate, and hyaluronic acid) also allowed the encapsulation of...the spore dispersion and held for 15 min for adsorption completion. Polycations and poly- anions were adsorbed sequentially and washed with 1 mL of DI

  20. Long-term exposure of bacterial spores to space

    NASA Technical Reports Server (NTRS)

    Horneck, G.; Buecker, H.; Reitz, G.

    1992-01-01

    With the NASA mission of the Long Duration Exposure Facility (LDEF), the authors have obtained the opportunity to expose Bacillus subtilis spores for nearly six years to the space environment and to analyze their responses after retrieval. The experiment was mounted onto a side tray of LDEF facing space. Data shows that the chances of microorganisms surviving in free space will be greatly increased by adequate shielding against solar ultraviolet light.

  1. Optical Constant Determination of Bacterial Spores in the MIR

    DTIC Science & Technology

    2005-12-05

    reported by Gurton et al for Bacillus globigii20 (reclassified as a substrain of Bacillus atrophaeus ).21 Obtaining the optical constant spectra with...Constants from Particulates by Infrared Reflectance Microscopy Demonstrated with Polystyrene Microspheres and Bacillus subtilis Spores The potential...system containing Bacillus anthracis, resulting in 17 confirmed cases of inhalation or cutaneous anthrax. Of these cases, 5 proved fatal.1 Due to the

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

    PubMed

    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.

  3. Decontamination after a release of B. anthracis spores.

    PubMed

    Campbell, Chris G; Kirvel, Robert D; Love, Adam H; Bailey, Christopher G; Miles, Robin; Schweickert, Jerry; Sutton, Mark; Raber, Ellen

    2012-03-01

    Decontaminating civilian facilities or large urban areas following an attack with Bacillus anthracis poses daunting challenges because of the lack of resources and proven technologies. Nevertheless, lessons learned from the 2001 cleanups together with advances derived from recent research have improved our understanding of what is required for effective decontamination. This article reviews current decontamination technologies appropriate for use in outdoor environments, on material surfaces, within large enclosed spaces, in water, and on waste contaminated with aerosolized B. anthracis spores.

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

    DTIC Science & Technology

    2012-11-01

    Kingdom. 35. Setlow B, et al. 2009. Characterization of Bacillus megaterium spores lacking enzymes that degrade the spore cortex. J. Appl. Microbiol...REPORT Analysis of the effects of a gerP mutation on the germination of spores of Bacillus subtilis 14. ABSTRACT 16. SECURITY CLASSIFICATION OF... Bacillus subtilis spores with a gerP mutation triggered spore germination via nutrient germinant receptors (GRs) slowly, although this defect was

  5. Response of Bacillus Spores to Combinations of Germinative Compounds

    PubMed Central

    Foerster, Harold F.; Foster, J. W.

    1966-01-01

    Foerster, Harold F. (University of Texas, Austin), and J. W. Foster. Response of Bacillus spores to combinations of germinative compounds. J. Bacteriol. 91:1168–1177. 1966.—Spores of 21 strains of Bacillus megaterium and 25 other strains representing 13 species of Bacillus were produced under standardized conditions. The germination of a washed spore suspension of each strain was measured as a response to various combinations of 30 different germinative compounds. The strains were first typed with respect to their response to “primary” germination compounds, i.e., glucose, l-alanine, inosine, and l-alanine-inosine mixture, and also Na+ and K+. The second stage was the determination of the response to various organic and inorganic anions and cations, each strain being supplied with the “primary” compounds best for it. Marked differences in germination patterns were observed among species and strains of the same species. No relation to established taxonomic lines was evident. A nonspecific requirement for ions was found for all strains, but not all ions were effective. A striking degree of interchangeability of germinative chemicals was found. “Fractional germination” was very common. A mixture of l-alanine and inosine and various ions was the best germinative solution for most strains. Some anomalous germination patterns were encountered. Those studied included a strain whose cells lysed spontaneously upon germination and other strains for which l-leucine had striking germinative powers. PMID:4956331

  6. Uplift and Outflow of Bacterial Spores via Electric Field

    NASA Astrophysics Data System (ADS)

    Dehel, T.

    The questions of how did life arise and is there life on other planets are some of the most profound questions that humanity asks Although there has been controversial signs of past bacterial life in meteorites which originated on Mars and there are current claims of bacterial life high in the atmosphere the issues of origin by chemical process or contamination make these types of results arguable and they will likely remain that way until a comprehensive theory is developed to explain why the claims might be true This paper proposes a complete theory for the spread of bacterial life throughout the galaxy by combining current knowledge from the fields of bacteriology stellar evolution and space weather Here we show the possibility that the forces of uplift on a charged bacteria particle are sufficient bring at least some lighter types of bacteria high into the ionosphere and subsequently move the charged spore onto magnetic field lines The bacteria spore is then driven down the magnetotail where during a solar storm a structure known as a plasmoid is propelled radially outward into space at velocities exceeding solar system escape velocity From that point the plasmoids are capable of reaching Mars the outer planets and even others systems eventually depositing the bacterial spores either via comets or direct interaction with the receiving planet The solid observational evidence for the strength of the electric fields and the speeds that the plasmoids leave the magnetotail during geomagnetic storms provide a firm

  7. Arrhenius reconsidered: astrophysical jets and the spread of spores

    NASA Astrophysics Data System (ADS)

    Sheldon, Malkah I.; Sheldon, Robert B.

    2015-09-01

    In 1871, Lord Kelvin suggested that the fossil record could be an account of bacterial arrivals on comets. In 1903, Svante Arrhenius suggested that spores could be transported on stellar winds without comets. In 1984, Sir Fred Hoyle claimed to see the infrared signature of vast clouds of dried bacteria and diatoms. In 2012, the Polonnaruwa carbonaceous chondrite revealed fossilized diatoms apparently living on a comet. However, Arrhenius' spores were thought to perish in the long transit between stars. Those calculations, however, assume that maximum velocities are limited by solar winds to ~5 km/s. Herbig-Haro objects and T-Tauri stars, however, are young stars with jets of several 100 km/s that might provide the necessary propulsion. The central engine of bipolar astrophysical jets is not presently understood, but we argue it is a kinetic plasma instability of a charged central magnetic body. We show how to make a bipolar jet in a belljar. The instability is non-linear, and thus very robust to scaling laws that map from microquasars to active galactic nuclei. We scale up to stellar sizes and recalculate the viability/transit-time for spores carried by supersonic jets, to show the viability of the Arrhenius mechanism.

  8. Resistance of Alicyclobacillus acidoterrestris spores and biofilm to industrial sanitizers.

    PubMed

    dos Anjos, Márcia Maria; Ruiz, Suelen Pereira; Nakamura, Celso Vataru; de Abreu Filho, Benício Alves

    2013-08-01

    This study evaluated the adhesion and biofilm formation of Alicyclobacillus acidoterrestris on industrial orange juice processing equipment and the bactericidal efficacy of peracetic acid, sodium hypochlorite, and quaternary ammonia after biofilm formation. The efficacy of these sanitizers against the spores of this microorganism was also evaluated. Stainless steel and nylon surfaces exhibited higher cell adhesion levels than did polyvinyl chloride surfaces. Peracetic acid was the most effective in removing biofilms from all surfaces (P < 0.05) and also reduced bacterial counts by 3 log CFU/cm² on the surface of polyvinyl chloride, but the other sanitizers also reduced the bacterial counts by 2 log CFU/cm². Quaternary ammonia exhibited the optimal minimum sporicidal concentration, preventing spore germination after only 15 s of contact at a concentration of 82 ppm. The flow cytometry results indicated that the spores and cells had low incidences of plasma membrane lysis after treatment with sanitizer, suggesting that lysis is not the principal mode of action for these sanitizers on A. acidoterrestris.

  9. Pollen and spores as biological recorders of past ultraviolet irradiance

    NASA Astrophysics Data System (ADS)

    Jardine, Phillip E.; Fraser, Wesley T.; Lomax, Barry H.; Sephton, Mark A.; Shanahan, Timothy M.; Miller, Charlotte S.; Gosling, William D.

    2016-12-01

    Solar ultraviolet (UV) irradiance is a key driver of climatic and biotic change. Ultraviolet irradiance modulates stratospheric warming and ozone production, and influences the biosphere from ecosystem-level processes through to the largest scale patterns of diversification and extinction. Yet our understanding of ultraviolet irradiance is limited because no method has been validated to reconstruct its flux over timescales relevant to climatic or biotic processes. Here, we show that a recently developed proxy for ultraviolet irradiance based on spore and pollen chemistry can be used over long (105 years) timescales. Firstly we demonstrate that spatial variations in spore and pollen chemistry correlate with known latitudinal solar irradiance gradients. Using this relationship we provide a reconstruction of past changes in solar irradiance based on the pollen record from Lake Bosumtwi in Ghana. As anticipated, variations in the chemistry of grass pollen from the Lake Bosumtwi record show a link to multiple orbital precessional cycles (19–21 thousand years). By providing a unique, local proxy for broad spectrum solar irradiance, the chemical analysis of spores and pollen offers unprecedented opportunities to decouple solar variability, climate and vegetation change through geologic time and a new proxy with which to probe the Earth system.

  10. Fast and Sensitive Detection of Bacillus anthracis Spores by Immunoassay

    PubMed Central

    Volland, Hervé; Dano, Julie; Lamourette, Patricia; Sylvestre, Patricia; Mock, Michèle; Créminon, Christophe

    2012-01-01

    Bacillus anthracis is one of the most dangerous potential biological weapons, and it is essential to develop a rapid and simple method to detect B. anthracis spores in environmental samples. The immunoassay is a rapid and easy-to-use method for the detection of B. anthracis by means of antibodies directed against surface spore antigens. With this objective in view, we have produced a panel of monoclonal antibodies against B. anthracis and developed colorimetric and electrochemiluminescence (ECL) immunoassays. Using Meso Scale Discovery ECL technology, which is based on electrochemiluminescence (ECL) detection utilizing a sulfo-Tag label that emits light upon electrochemical stimulation (using a dedicated ECL plate reader, an electrical current is placed across the microplate with electrodes integrated into the bottom of the plate, resulting in a series of electrically induced reactions leading to a luminescent signal), a detection limit ranging between 0.3 × 103 and 103 CFU/ml (i.e., 30 to 100 spores per test), depending on the B. anthracis strain assayed, was achieved. In complex matrices (5 mg/ml of soil or simulated powder), the detection level (without any sample purification or concentration) was never altered more than 3-fold compared with the results obtained in phosphate-buffered saline. PMID:22773632

  11. Pollen and spores as biological recorders of past ultraviolet irradiance

    PubMed Central

    Jardine, Phillip E.; Fraser, Wesley T.; Lomax, Barry H.; Sephton, Mark A.; Shanahan, Timothy M.; Miller, Charlotte S.; Gosling, William D.

    2016-01-01

    Solar ultraviolet (UV) irradiance is a key driver of climatic and biotic change. Ultraviolet irradiance modulates stratospheric warming and ozone production, and influences the biosphere from ecosystem-level processes through to the largest scale patterns of diversification and extinction. Yet our understanding of ultraviolet irradiance is limited because no method has been validated to reconstruct its flux over timescales relevant to climatic or biotic processes. Here, we show that a recently developed proxy for ultraviolet irradiance based on spore and pollen chemistry can be used over long (105 years) timescales. Firstly we demonstrate that spatial variations in spore and pollen chemistry correlate with known latitudinal solar irradiance gradients. Using this relationship we provide a reconstruction of past changes in solar irradiance based on the pollen record from Lake Bosumtwi in Ghana. As anticipated, variations in the chemistry of grass pollen from the Lake Bosumtwi record show a link to multiple orbital precessional cycles (19–21 thousand years). By providing a unique, local proxy for broad spectrum solar irradiance, the chemical analysis of spores and pollen offers unprecedented opportunities to decouple solar variability, climate and vegetation change through geologic time and a new proxy with which to probe the Earth system. PMID:27976735

  12. Airborne pollen and spores of León (Spain)

    NASA Astrophysics Data System (ADS)

    Fernández-González, Delia; Suarez-Cervera, María; Díaz-González, Tomás; Valencia-Barrera, Rosa María

    1993-06-01

    A qualitative and quantitative analysis of airborne pollen and spores was carried out over 2 years (from September 1987 to August 1989) in the city of León. Slides were prepared daily using a volumetric pollen trap, which was placed on the Faculty of Veterinary Science building (University of León) 12m above ground-level. Fifty-one pollen types were observed; the most important of these were: Cupressaceae during the winter, Pinus and Quercus in spring, and Poaceae, Leguminosae and Chenopodiaceae in the summer. The results also showed the existence of a rich mould spore assemblage in the atmosphere. The group of Amerospores ( Penicillium, Aspergillus and Cladosporium) as well as Dictyospores ( Alternaria) were the most abundant; Puccinia was common in the air in August. Fluctuations in the total pollen and spores m3 of air were compared with meteorological parameters (temperature, relative humidity and rainfall). From the daily sampling of the atmosphere of León, considering the maximum and minimum temperature and duration of rainfall, the start of the pollen grain season was observed generally to coincide with a rise in temperature in the absence of rain.

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

    PubMed

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

    2010-10-01

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

  14. Long-term survival of bacterial spores in space

    NASA Technical Reports Server (NTRS)

    Horneck, G.; Bucker, H.; Reitz, G.

    1994-01-01

    On board of the NASA Long Duration Exposure Facility (LDEF), spores of Bacillus subtilis in monolayers (10(exp 6)/sample) or multilayers (10(exp 8)/sample) were exposed to the space environment for nearly six years and their survival was analyzed after retrieval. The response to space parameters, such as vacuum (10(exp -6) Pa), solar electromagnetic radiation up to the highly energetic vacuum-ultraviolet range 10(exp 9) J/sq m) and/or cosmic radiation (4.8 Gy), was studied and compared to the results of a simultaneously running ground control experiment. If shielded against solar ultraviolet (UV)-radiation, up to 80% of spores in multilayers survive in space. Solar UV-radiation, being the most deleterious parameter of space, reduces survival by 4 orders of magnitude or more. However, up to 10(exp 4) viable spores were still recovered, even in completely unprotected samples. Substances, such as glucose or buffer salts serve as chemical protectants. With this 6 year study in space, experimental data are provided to the discussion on the likelihood of 'Panspermia'.

  15. Raman spectroscopic study of Lactarius spores (Russulales, Fungi)

    NASA Astrophysics Data System (ADS)

    De Gussem, Kris; Vandenabeele, Peter; Verbeken, Annemieke; Moens, Luc

    2005-10-01

    Fungi are important organisms in ecosystems, in industrial and pharmaceutical production and are valuable food sources as well. Classical identification is often time-consuming and specialistic. In this study, Raman spectroscopy is applied to the analysis of fungal spores of Lactarius, an economically and ecologically important genus of Basidiomycota. Raman spectra of spores of Lactarius controversus Pers.: Fr., Lactarius lacunarum (Romagn.) ex Hora, Lactarius quieticolor Romagn. and Lactarius quietus (Fr.: Fr.) Fr. are reported for the first time. The spectra of these species show large similarity. These spectra are studied and compared with the Raman spectra of reference substances known to occur in macrofungi, including saccharides, lipids and some minor compounds that may serve as specific biomarkers (adenine, ergosterol and glycine). Most Raman bands could be attributed to specific components. In agreement with the biological role of fungal spores, high amounts of lipids were observed, the main fatty acid being oleate. In addition to different types of lipids and phospholipids, the polysaccharides chitin and amylopectin could be detected as well. The presence of trehalose is not equivocally shown, due to overlapping bands. Raman band positions are reported for the observed bands of the different species and reference products.

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

    NASA Astrophysics Data System (ADS)

    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 < 0.05), resulting from the nocturnal sporulation under near-saturated relative humidity. On the contrary, BVOC oxidation products showed higher levels in daytime than nighttime, indicating substantial photochemical SOA formation. Using tracer-based methods, we estimated that fungal spores account for 45 % of organic carbon (OC) in nighttime and 22 % in daytime, whereas BVOC oxidation products account for 15 and 19 %, respectively. To our knowledge, we present for the first time highly time-resolved results that fungal spores overwhelmed BVOC oxidation products in contributing to OA especially in nighttime. This study emphasizes the importance of both PBAPs and SOAs in forming forest organic aerosols.

  17. Kinetics of Death of Bacterial Spores at Elevated Temperatures

    PubMed Central

    Wang, Daniel I-C.; Scharer, Jeno; Humphrey, Arthur E.

    1964-01-01

    The kinetics of death of Bacillus stearothermophilus spores (FS 7954) suspended in phosphate buffer (pH 7) were studied over a temperature range of 127.2 to 143.8 C and exposure times of 0.203 to 4.150 sec. These short exposure were achieved by use of a tubular flow reactor in which a suspension of spores was injected into a hot flowing stream at the entrance of the reactor. Thermal equilibria of the suspension with the hot stream was achieved within 0.0006 sec. After flow through a fixed length of reactor, the stream containing the spores was cooled by flash vaporization and then assayed for viable count. The death rate data were fitted by a logarithmic expression. However, logarithmic death rate was only approximated in the tail or high-kill regions of exposure. Death rate constants obtained from this portion of the data were found to correlate by Arrhenius as well as Absolute Reaction Rate Theory relationships. Thermal-death time curves were found to correlate the data rather poorly. The activation energy and frequency constant for an Arrhenius relationship fit of the data were found to be 83.6 kcal/gmole and 1047.2 min-1, respectively. The standard enthalpy and entropy changes for an Absolute Reaction Rate Theory relationship fit of the data were found to be 84.4 kcal/gmole and 157 cal/gmole-K, respectively. PMID:14215978

  18. Long-term survival of bacterial spores in space.

    PubMed

    Horneck, G; Bucker, H; Reitz, G

    1994-10-01

    On board of the NASA Long Duration Exposure Facility (LDEF), spores of Bacillus subtilis in monolayers (10(6)/sample) or multilayers (10(8)/sample) were exposed to the space environment for nearly six years and their survival was analyzed after retrieval. The response to space parameters, such as vacuum (10(-6) Pa), solar electromagnetic radiation up to the highly energetic vacuum-ultraviolet range (10(9) J/m2) and/or cosmic radiation (4.8 Gy), was studied and compared to the results of a simultaneously running ground control experiment. If shielded against solar ultraviolet (UV)-radiation, up to 80 % of spores in multilayers survive in space. Solar UV-radiation, being the most deleterious parameter of space, reduces survival by 4 orders of magnitude or more. However, up to 10(4) viable spores were still recovered, even in completely unprotected samples. Substances, such as glucose or buffer salts serve as chemical protectants. With this 6 year study in space, experimental data are provided to the discussion on the likelihood of "Panspermia".

  19. Limited period of graviresponsiveness in germinating spores of Ceratopteris richardii

    NASA Technical Reports Server (NTRS)

    Edwards, E. S.; Roux, S. J.

    1994-01-01

    Rhizoids of the fern Ceratopteris richardii Brogn. usually emerge 40 h after germination is initiated by light, and more than 90% of them emerge growing in a downward direction. However, when the spores are germinated on a clinostat, the emerging rhizoids show no preferential orientation. This indicates that under normal 1 g conditions the initial growth direction of rhizoids can be oriented by gravity. If the orientation of the spores is changed 3 h or less after the start of germination, the growth direction of most emerging rhizoids becomes downward relative to the new orientation. However, if the orientation of the spores is changed by 180 degrees 8 h or more after germination is initiated by light, most rhizoids emerge growing upward; i.e., the same direction as if there had been no orientation change. Emerged rhizoids also do not change their direction of growth if their orientation is changed. These results indicate that the growth direction of emerging rhizoids is set by gravity prior to actual emergence, and that the time of full orientation responsiveness is limited to a period ranging from the initiation of germination to about 3-4 h after the start of germination. There is a gravity-oriented nuclear movement beginning at about 13 h after germination, and this movement appears to predict the initial growth direction of rhizoids.

  20. Characterization of Clostridium difficile Spores Lacking Either SpoVAC or Dipicolinic Acid Synthetase

    PubMed Central

    Donnelly, M. Lauren; Fimlaid, Kelly A.

    2016-01-01

    ABSTRACT The spore-forming obligate anaerobe Clostridium difficile is a leading cause of antibiotic-associated diarrhea around the world. In order for C. difficile to cause infection, its metabolically dormant spores must germinate in the gastrointestinal tract. During germination, spores degrade their protective cortex peptidoglycan layers, release dipicolinic acid (DPA), and hydrate their cores. In C. difficile, cortex hydrolysis is necessary for DPA release, whereas in Bacillus subtilis, DPA release is necessary for cortex hydrolysis. Given this difference, we tested whether DPA synthesis and/or release was required for C. difficile spore germination by constructing mutations in either spoVAC or dpaAB, which encode an ion channel predicted to transport DPA into the forespore and the enzyme complex predicted to synthesize DPA, respectively. C. difficile spoVAC and dpaAB mutant spores lacked DPA but could be stably purified and were more hydrated than wild-type spores; in contrast, B. subtilis spoVAC and dpaAB mutant spores were unstable. Although C. difficile spoVAC and dpaAB mutant spores exhibited wild-type germination responses, they were more readily killed by wet heat. Cortex hydrolysis was not affected by this treatment, indicating that wet heat inhibits a stage downstream of this event. Interestingly, C. difficile spoVAC mutant spores were significantly more sensitive to heat treatment than dpaAB mutant spores, indicating that SpoVAC plays additional roles in conferring heat resistance. Taken together, our results demonstrate that SpoVAC and DPA synthetase control C. difficile spore resistance and reveal differential requirements for these proteins among the Firmicutes. IMPORTANCE Clostridium difficile is a spore-forming obligate anaerobe that causes ∼500,000 infections per year in the United States. Although spore germination is essential for C. difficile to cause disease, the factors required for this process have been only partially characterized

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

    PubMed

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

    2015-12-01

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

  2. A natural O-ring optimizes the dispersal of fungal spores

    PubMed Central

    Fritz, Joerg A.; Seminara, Agnese; Roper, Marcus; Pringle, Anne; Brenner, Michael P.

    2013-01-01

    The forcibly ejected spores of ascomycete fungi must penetrate several millimetres of nearly still air surrounding sporocarps to reach dispersive airflows, and escape is facilitated when a spore is launched with large velocity. To launch, the spores of thousands of species are ejected through an apical ring, a small elastic pore. The startling diversity of apical ring and spore shapes and dimensions make them favoured characters for both species descriptions and the subsequent inference of relationships among species. However, the physical constraints shaping this diversity and the adaptive benefits of specific morphologies are not understood. Here, we develop an elastohydrodynamic theory of the spore's ejection through the apical ring and demonstrate that to avoid enormous energy losses during spore ejection, the four principal morphological dimensions of spore and apical ring must cluster within a nonlinear one-dimensional subspace. We test this prediction using morphological data for 45 fungal species from two different classes and 18 families. Our sampling encompasses multiple loss and gain events and potentially independent origins of this spore ejection mechanism. Although the individual dimensions of the spore and apical ring are only weakly correlated with each other, they collapse into the predicted subspace with high accuracy. The launch velocity appears to be within 2 per cent of the optimum for over 90 per cent of all forcibly ejected species. Although the morphological diversity of apical rings and spores appears startlingly diverse, a simple principle can be used to organize it. PMID:23782534

  3. Trichoderma asperellum spore dose depended modulation of plant growth in vegetable crops.

    PubMed

    Singh, Vivek; Upadhyay, Ram Sanmukh; Sarma, Birinchi Kumar; Singh, Harikesh Bahadur

    2016-12-01

    In the present study we bioprimed seeds of six vegetable crops (tomato, brinjal, chilli, okra, ridge gourd and guar) with different spore doses of Trichoderma asperellum BHUT8 ranging from 10(2) to 10(8) spores ml(-1) and the effect of biopriming was seen on seed germination and development. The most effective spore dose for enhancement in seed germination and radicle length was found to be 10(3) spores ml(-1) in tomato and ridge gourd, 10(4) spores ml(-1) in brinjal and okra while 10(6) spores ml(-1) in chilli and guar. At the most effective spore dose, the increase in germination percentage was 5, 146.15, 112.5, 5.4, 28.13 and 0% while increase in radicle length was 73.17, 50.83, 171.6, 107.35, 247.19 and 90.79% in tomato, brinjal, chilli, okra, ridge gourd and guar, respectively, compared to their controls. Higher spore dose i.e. 10(7)-10(8) spores ml(-1) and 10(6)-10(8) spores ml(-1) in tomato and brinjal, respectively, reduced seed germination percentage and radicle growth compared to their controls. Biopriming with T. asperellum BHUT8 also triggered various defense like responses such as high phenylpropanoid activities and lignifications in bioprimed tomato seedlings compared to the non-bioprimed tomato seedlings demonstrating possible use of BHUT8 against phytopathogens.

  4. Effects of sawdust pollution on the germination of fungal spores in Lagos Lagoon.

    PubMed

    Akpata, T V

    1987-01-01

    Four fungi, Aspergillus flavus, Aspergillus giganteus, Cladosporium oxysporum a and Trichoderma aureoviride, isolated from Lagos Lagoon, were tested for spore germination in aqueous sawdust extract of different hardwood species (Khaya ivorensis, Mitragyna ciliata and Triplochiton scleroxylon). Extracts of M. ciliata were inhibitory to spore germination especially at higher concentrations. Germ tubes of spores decreased in length with increase in extract concentration for C. oxysporum and T. aureoviride, while Aspergillus species showed increase in length up to a peak at 3% sawdust extract concentration, and thereafter germ tube lengths decreased with increasing concentration. Extracts of T. scleroxylon stimulated the spores and percentage germination increased at higher extract concentrations with no significant difference in germ tube length. Similarly, K. ivorensis had a stimulatory effect on spore germination and length of germ tube, especially at higher extract concentrations. Addition of soluble exogenous carbon and nitrogen sources to sawdust extract enhanced spore germination. Apart from A. flavus, which had only 9% germination, all the spores failed to germinate in lagoon water having 21% salinity. The spores were also inhibited by in lagoon water having 21 per thousand salinity. The spores were also inhibited by various inorganic salts, i.e. CaCl(2), KH(2)PO(4), MgSO(4) and NaCl, present in Lagos Lagoon. The results suggest that sawdust pollution causes enrichment of the lagoon, thereby enhancing spore germination.

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

    PubMed

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

    2016-01-01

    Association between arbuscular mycorrhizal fungi (AMF) and bacteria has long been studied. However, the factors influencing their association in the natural environment is still unknown. This study aimed to isolate bacteria associated with spore walls of AMF and identify their potential characters for association. Spores collected from coastal reclamation land were differentiated based on their morphology and identified by 18S rDNA sequencing as Funneliformis caledonium, Racocetra alborosea and Funneliformis mosseae. Bacteria associated with AMF spore walls were isolated after treating them with disinfection solution at different time intervals. After 0, 10 and 20 min of spore disinfection, 86, 24 and 10 spore associated bacteria (SAB) were isolated, respectively. BOX-PCR fingerprinting analysis showed that diverse bacterial communities were associated to AMF spores. Bacteria belonging to the same genera could associate with different AMF spores. Gram positive bacteria were more closely associated with AMF spores. Isolated SAB were characterized and tested for spore association characters such as chitinase, protease, cellulase enzymes and exopolysaccharide production (EPS). Among the 120 SAB, 113 SAB were able to show one or more characters for association and seven SAB did not show any association characters. The 16S rDNA sequence of SAB revealed that bacteria belonging to the phyla Firmicutes, Proteobacteria, Actinobacteria and Bactereiodes were associated with AMF spore walls.

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

    PubMed Central

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

    2016-01-01

    Association between arbuscular mycorrhizal fungi (AMF) and bacteria has long been studied. However, the factors influencing their association in the natural environment is still unknown. This study aimed to isolate bacteria associated with spore walls of AMF and identify their potential characters for association. Spores collected from coastal reclamation land were differentiated based on their morphology and identified by 18S rDNA sequencing as Funneliformis caledonium, Racocetra alborosea and Funneliformis mosseae. Bacteria associated with AMF spore walls were isolated after treating them with disinfection solution at different time intervals. After 0, 10 and 20 min of spore disinfection, 86, 24 and 10 spore associated bacteria (SAB) were isolated, respectively. BOX-PCR fingerprinting analysis showed that diverse bacterial communities were associated to AMF spores. Bacteria belonging to the same genera could associate with different AMF spores. Gram positive bacteria were more closely associated with AMF spores. Isolated SAB were characterized and tested for spore association characters such as chitinase, protease, cellulase enzymes and exopolysaccharide production (EPS). Among the 120 SAB, 113 SAB were able to show one or more characters for association and seven SAB did not show any association characters. The 16S rDNA sequence of SAB revealed that bacteria belonging to the phyla Firmicutes, Proteobacteria, Actinobacteria and Bactereiodes were associated with AMF spore walls. PMID:27479250

  7. Aspergillus fumigatus spore proteomics and genetics reveal that VeA represses DefA-mediated DNA damage response.

    PubMed

    Shin, Kwang-Soo; Park, Hee-Soo; Kim, Young; Heo, In-Beom; Kim, Young Hwan; Yu, Jae-Hyuk

    2016-10-04

    Aspergillus fumigatus reproduces and infects host by forming a high number of small asexual spores (conidia). The velvet proteins are global transcriptional regulators governing the complex process of conidiogenesis in this fungus. Here, to further understand the velvet-mediated regulation, we carried out comparative proteomic analyses of conidia of wild type (WT) and three velvet mutants (ΔveA, ΔvelB and ΔvosA). Cluster analysis of 184 protein spots showing at least 1.5-fold differential accumulation between WT and mutants reveal the clustering of WT- ΔveA and ΔvelB-ΔvosA. Among 43 proteins identified by Nano-LC-ESI-MS/MS, 23 including several heat shock proteins showed more than two-fold reduction in both the ∆velB and ∆vosA conidia. On the contrary, three proteins exhibited more than five-fold increase in ∆veA only, including the putative RNA polymerase II degradation factor DefA. The deletion of defA resulted in a reduced number of conidia and restricted colony growth. In addition, the defA deletion mutant conidia showed hypersensitivity against the DNA damaging agents NQO and MMS, while the ΔveA mutant conidia were more resistant against to NQO. Taken together, we propose that VeA controls protein level of DefA in conidia, which are dormant and equipped with multiple layers of protection against environmental cues.

  8. Turbulent Dispersion of Pathogenic Spores Within and Above Plant Canopies: Field Experiments and Lagrangian Modeling

    NASA Astrophysics Data System (ADS)

    Gleicher, S.; Chamecki, M.; Isard, S.; Katul, G. G.

    2012-12-01

    Plant disease epidemics caused by pathogenic spores are a common and consequential threat to agricultural crops. In most cases, pathogenic spores are produced and released deep inside plant canopies and must be transported out of the canopy region in order to infect other fields and spread the disease. The fraction of spores that "escape" the canopy is crucial in determining how fast and far these plant diseases will spread. The goal of this work is to use a field experiment, coupled with a Lagrangian Stochastic Model (LSM), to investigate how properties of canopy turbulence impact the dispersion of spores inside the canopy and the fraction of spores that escape from the canopy. An extensive field experiment was conducted to study spore dispersion inside and outside a corn canopy. The spores were released from point sources located at various depths inside the canopy. Concentration measurements were obtained inside and above the canopy by a 3-dimensional grid of spore collectors. The experimental measurements of mean spore concentration are used to validate a LSM for spore dispersion. In the LSM, flow field statistics used to drive the particle dispersion are specified by a second-order closure model for turbulence within plant canopies. The dispersion model includes spore deposition on and rebound from canopy elements. The combination of experimental and numerical simulations is used to quantify the fraction of spores that escape the canopy. Effects of release height, friction velocity, and canopy architecture on the escape fraction of spores are explored using the LSM, and implications for disease propagation are discussed.

  9. Carvacrol suppresses high pressure high temperature inactivation of Bacillus cereus spores.

    PubMed

    Luu-Thi, Hue; Corthouts, Jorinde; Passaris, Ioannis; Grauwet, Tara; Aertsen, Abram; Hendrickx, Marc; Michiels, Chris W

    2015-03-16

    The inactivation of bacterial spores generally proceeds faster and at lower temperatures when heat treatments are conducted under high pressure, and high pressure high temperature (HPHT) processing is, therefore, receiving an increased interest from food processors. However, the mechanisms of spore inactivation by HPHT treatment are poorly understood, particularly at moderately elevated temperature. In the current work, we studied inactivation of the spores of Bacillus cereus F4430/73 by HPHT treatment for 5 min at 600MPa in the temperature range of 50-100°C, using temperature increments of 5°C. Additionally, we investigated the effect of the natural antimicrobial carvacrol on spore germination and inactivation under these conditions. Spore inactivation by HPHT was less than about 1 log unit at 50 to 70°C, but gradually increased at higher temperatures up to about 5 log units at 100°C. DPA release and loss of spore refractility in the spore population were higher at moderate (≤65°C) than at high (≥70°C) treatment temperatures, and we propose that moderate conditions induced the normal physiological pathway of spore germination resulting in fully hydrated spores, while at higher temperatures this pathway was suppressed and replaced by another mechanism of pressure-induced dipicolinic acid (DPA) release that results only in partial spore rehydration, probably because spore cortex hydrolysis is inhibited. Carvacrol strongly suppressed DPA release and spore rehydration during HPHT treatment at ≤65°C and also partly inhibited DPA release at ≥65°C. Concomitantly, HPHT spore inactivation was reduced by carvacrol at 65-90°C but unaffected at 95-100°C.

  10. Role of YpeB in Cortex Hydrolysis during Germination of Bacillus anthracis Spores

    PubMed Central

    Bernhards, Casey B.

    2014-01-01

    The infectious agent of the disease anthrax is the spore of Bacillus anthracis. Bacterial spores are extremely resistant to environmental stresses, which greatly hinders spore decontamination efforts. The spore cortex, a thick layer of modified peptidoglycan, contributes to spore dormancy and resistance by maintaining the low water content of the spore core. The cortex is degraded by germination-specific lytic enzymes (GSLEs) during spore germination, rendering the cells vulnerable to common disinfection techniques. This study investigates the relationship between SleB, a GSLE in B. anthracis, and YpeB, a protein necessary for SleB stability and function. The results indicate that ΔsleB and ΔypeB spores exhibit similar germination phenotypes and that the two proteins have a strict codependency for their incorporation into the dormant spore. In the absence of its partner protein, SleB or YpeB is proteolytically degraded soon after expression during sporulation, rather than escaping the developing spore. The three PepSY domains of YpeB were examined for their roles in the interaction with SleB. YpeB truncation mutants illustrate the necessity of a region beyond the first PepSY domain for SleB stability. Furthermore, site-directed mutagenesis of highly conserved residues within the PepSY domains resulted in germination defects corresponding to reduced levels of both SleB and YpeB in the mutant spores. These results identify residues involved in the stability of both proteins and reiterate their codependent relationship. It is hoped that the study of GSLEs and interacting proteins will lead to the use of GSLEs as targets for efficient activation of spore germination and facilitation of spore cleanup. PMID:25022853

  11. Role of YpeB in cortex hydrolysis during germination of Bacillus anthracis spores.

    PubMed

    Bernhards, Casey B; Popham, David L

    2014-10-01

    The infectious agent of the disease anthrax is the spore of Bacillus anthracis. Bacterial spores are extremely resistant to environmental stresses, which greatly hinders spore decontamination efforts. The spore cortex, a thick layer of modified peptidoglycan, contributes to spore dormancy and resistance by maintaining the low water content of the spore core. The cortex is degraded by germination-specific lytic enzymes (GSLEs) during spore germination, rendering the cells vulnerable to common disinfection techniques. This study investigates the relationship between SleB, a GSLE in B. anthracis, and YpeB, a protein necessary for SleB stability and function. The results indicate that ΔsleB and ΔypeB spores exhibit similar germination phenotypes and that the two proteins have a strict codependency for their incorporation into the dormant spore. In the absence of its partner protein, SleB or YpeB is proteolytically degraded soon after expression during sporulation, rather than escaping the developing spore. The three PepSY domains of YpeB were examined for their roles in the interaction with SleB. YpeB truncation mutants illustrate the necessity of a region beyond the first PepSY domain for SleB stability. Furthermore, site-directed mutagenesis of highly conserved residues within the PepSY domains resulted in germination defects corresponding to reduced levels of both SleB and YpeB in the mutant spores. These results identify residues involved in the stability of both proteins and reiterate their codependent relationship. It is hoped that the study of GSLEs and interacting proteins will lead to the use of GSLEs as targets for efficient activation of spore germination and facilitation of spore cleanup.

  12. Live Cell Imaging of Germination and Outgrowth of Individual Bacillus subtilis Spores; the Effect of Heat Stress Quantitatively Analyzed with SporeTracker

    PubMed Central

    Vischer, Norbert O. E.; Smelt, Jan P. P. M.; Brul, Stanley; Manders, Erik M. M.

    2013-01-01

    Spore-forming bacteria are a special problem for the food industry as some of them are able to survive preservation processes. Bacillus spp. spores can remain in a dormant, stress resistant state for a long period of time. Vegetative cells are formed by germination of spores followed by a more extended outgrowth phase. Spore germination and outgrowth progression are often very heterogeneous and therefore, predictions of microbial stability of food products are exceedingly difficult. Mechanistic details of the cause of this heterogeneity are necessary. In order to examine spore heterogeneity we made a novel closed air-containing chamber for live imaging. This chamber was used to analyze Bacillus subtilis spore germination, outgrowth, as well as subsequent vegetative growth. Typically, we examined around 90 starting spores/cells for ≥4 hours per experiment. Image analysis with the purposely built program “SporeTracker” allows for automated data processing from germination to outgrowth and vegetative doubling. In order to check the efficiency of the chamber, growth and division of B. subtilis vegetative cells were monitored. The observed generation times of vegetative cells were comparable to those obtained in well-aerated shake flask cultures. The influence of a heat stress of 85°C for 10 min on germination, outgrowth, and subsequent vegetative growth was investigated in detail. Compared to control samples fewer spores germinated (41.1% less) and fewer grew out (48.4% less) after the treatment. The heat treatment had a significant influence on the average time to the start of germination (increased) and the distribution and average of the duration of germination itself (increased). However, the distribution and the mean outgrowth time and the generation time of vegetative cells, emerging from untreated and thermally injured spores, were similar. PMID:23536843

  13. FLUORESCENT-SERIOLOGICAL INVESTIGATIONS OF A PATHOGENIC FUNGUS (SPOROTRICHUM SCHENCKII),

    DTIC Science & Technology

    coloration of numerous other species of fungus no cross reactions with Sporotrichum schenkii were found. The use of this fluorescent coloring method for the diagnosis of Sporotrichosis is suggested. (Author)

  14. An insect parasitoid carrying an ochratoxin producing fungus

    NASA Astrophysics Data System (ADS)

    Vega, Fernando E.; Posada, Francisco; Gianfagna, Thomas J.; Chaves, Fabio C.; Peterson, Stephen W.

    2006-06-01

    The insect parasitoid Prorops nasuta has been introduced from Africa to many coffee-producing countries in an attempt to control the coffee berry borer. In this paper, we report on the sequencing of the ITS LSU-rDNA and beta-tubulin loci used to identify a fungus isolated from the cuticle of a P. nasuta that emerged from coffee berries infected with the coffee berry borer. The sequences were compared with deposits in GenBank and the fungus was identified as Aspergillus westerdijkiae. The fungus tested positive for ochratoxin A production, with varying levels depending on the media in which it was grown. These results raise the possibility that an insect parasitoid might be disseminating an ochratoxin-producing fungus in coffee plantations.

  15. An insect parasitoid carrying an ochratoxin producing fungus.

    PubMed

    Vega, Fernando E; Posada, Francisco; Gianfagna, Thomas J; Chaves, Fabio C; Peterson, Stephen W

    2006-06-01

    The insect parasitoid Prorops nasuta has been introduced from Africa to many coffee-producing countries in an attempt to control the coffee berry borer. In this paper, we report on the sequencing of the ITS LSU-rDNA and beta-tubulin loci used to identify a fungus isolated from the cuticle of a P. nasuta that emerged from coffee berries infected with the coffee berry borer. The sequences were compared with deposits in GenBank and the fungus was identified as Aspergillus westerdijkiae. The fungus tested positive for ochratoxin A production, with varying levels depending on the media in which it was grown. These results raise the possibility that an insect parasitoid might be disseminating an ochratoxin-producing fungus in coffee plantations.

  16. Dipicolinic Acid Release by Germinating Clostridium difficile Spores Occurs through a Mechanosensing Mechanism

    PubMed Central

    Francis, Michael B.

    2016-01-01

    ABSTRACT Classically, dormant endospores are defined by their resistance properties, particularly their resistance to heat. Much of the heat resistance is due to the large amount of dipicolinic acid (DPA) stored within the spore core. During spore germination, DPA is released and allows for rehydration of the otherwise-dehydrated core. In Bacillus subtilis, 7 proteins are encoded by the spoVA operon and are important for DPA release. These proteins receive a signal from the activated germinant receptor and release DPA. This DPA activates the cortex lytic enzyme CwlJ, and cortex degradation begins. In Clostridium difficile, spore germination is initiated in response to certain bile acids and amino acids. These bile acids interact with the CspC germinant receptor, which then transfers the signal to the CspB protease. Activated CspB cleaves the cortex lytic enzyme, pro-SleC, to its active form. Subsequently, DPA is released from the core. C. difficile encodes orthologues of spoVAC, spoVAD, and spoVAE. Of these, the B. subtilis SpoVAC protein was shown to be capable of mechanosensing. Because cortex degradation precedes DPA release during C. difficile spore germination (opposite of what occurs in B. subtilis), we hypothesized that cortex degradation would relieve the osmotic constraints placed on the inner spore membrane and permit DPA release. Here, we assayed germination in the presence of osmolytes, and we found that they can delay DPA release from germinating C. difficile spores while still permitting cortex degradation. Together, our results suggest that DPA release during C. difficile spore germination occurs though a mechanosensing mechanism. IMPORTANCE Clostridium difficile is transmitted between hosts in the form of a dormant spore, and germination by C. difficile spores is required to initiate infection, because the toxins that are necessary for disease are not deposited on the spore form. Importantly, the C. difficile spore germination pathway

  17. Fungus Ball in Concha Bullosa: A Rare Case with Anosmia

    PubMed Central

    Özkırıs, Mahmut; Kapusuz, Zeliha; Seçkın, Selda; Saydam, Levent

    2013-01-01

    Concha bullosa is the pneumatization of the concha and is one of the most common variations of the sinonasal anatomy. The histopathological changes caused by the infections which arise from the impaired aeration of conchal cavity are frequently found. Fungus ball of the nasal cavity is an extremely rare, fungal infection with only three cases reported previously. In this paper, we present the fourth fungus ball case which developed within a concha bullosa and presented with anosmia. PMID:23936708

  18. Biological control of Ascaris suum eggs by Pochonia chlamydosporia fungus.

    PubMed

    Ferreira, Sebastião Rodrigo; de Araújo, Jackson Victor; Braga, Fábio Ribeiro; Araujo, Juliana Milani; Frassy, Luiza Neme; Ferreira, Aloízio Soares

    2011-12-01

    Ascaris suum is a gastrointestinal nematode parasite of swines. The aim of this study was to observe Pochonia chlamydosporia fungus on biological control of A. suum eggs after fungus passage through swines gastrointestinal tract. Eighteen pigs, previously dewormed, were randomly divided into three groups: group 1, treated with the fungus isolate VC4; group 2, treated with the fungus isolate VC1 and group 3 did not receive fungus (control). In the treated groups, each animal received a 9 g single dose of mycelium mass containing P. chlamydosporia (VC1 or VC4). Thereafter, animal fecal samples were collected at the following intervals: 8, 12, 24, 36, 48, 72 and 96 h after treatment beginning and these were poured in Petri dishes containing 2% water-agar culture medium. Then, 1,000 A. suum eggs were poured into each dish and kept in an incubator at 26 °C and in the dark for 30 days. After this period, approximately 100 eggs were removed from each Petri dish and morphologically analyzed under light microscopy following the ovicidal activity parameters. The higher percentage observed for isolated VC4 eggs destruction was 57.5% (36 h) after fungus administration and for isolate VC1 this percentage was 45.8% (24 h and 72 h) (p > 0.01). P. chlamydosporia remained viable after passing through the gastrointestinal tract of swines, maintaining its ability of destroying A. suum eggs.

  19. Effect of washing on identification of Bacillus spores by principal-component analysis of fluorescence data

    NASA Astrophysics Data System (ADS)

    Kunnil, Joseph; Sarasanandarajah, Sivananthan; Chacko, Easaw; Reinisch, Lou

    2006-05-01

    The fluorescence spectra of Bacillus spores are measured at excitation wavelengths of 280, 310, 340, 370, and 400 nm. When cluster analysis is used with the principal-component analysis, the Bacillus globigii spores can be distinguished from the other species of Bacillus spores (B. cereus, B. popilliae, and B. thuringiensis). To test how robust the identification process is with the fluorescence spectra, the B. globigii is obtained from three separate preparations in different laboratories. Furthermore the fluorescence is measured before and after washing and redrying the B. globigii spores. Using the cluster analysis of the first two or three principal components of the fluorescence spectra, one is able to distinguish B. globigii spores from the other species, independent of preparing or washing the spores.

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

    PubMed

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

    2015-01-01

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

  1. The effects of dimethylsulfoxide (DMSO) on the radiation sensitivity of bacterial spores. [X-ray

    SciTech Connect

    Ewing, D.

    1982-01-01

    Dimethylsufoxide (DMSO) is a potent sensitizer of irradiated bacterial spores (Bacillus megaterium). It is effective under either anoxic or well-oxygenated conditions; in both cases, DMSO increases the response by a factor of 3 to 4. In water, the oxygen enhancement ratio (OER) with 260-kVp X rays is 1.9, while in pure DMSO, which is not toxic to spores, the OER is reduced to 1.1. Spores exposed to DMSO and then washed and resuspended in water still show most of the DMSO-characteristic sensitization, even when the spores are soaked in water for 24 hr before irradiation. This effect is not attributable to DMSO retention by the spore; instead, we suggest that DMSO causes a long-lasting change in a critical spore component that changes the way the entire cell responds to radiation.

  2. Optimization of survival and spore formation of Paenibacillus polymyxa SQR-21 during bioorganic fertilizer storage.

    PubMed

    Huo, Zhenhua; Zhang, Nan; Xu, Zhihui; Li, Shuqing; Zhang, Qiuxia; Qiu, Meihua; Yong, Xiaoyu; Huang, Qiwei; Zhang, Ruifu; Shen, Qi-rong

    2012-03-01

    The effects of storage temperature (20, 30 and 40 °C), inoculum type (pure spores, an equal mix of spores and vegetative cells and pure vegetative cells) and water content (20%, 30% and 40%) on the survival and spore formation of the biocontrol agent, Paenibacillus polymyxa SQR-21, in a bioorganic fertilizer were modeled in a 3×3×3 factorial design. Bacterial and spore populations were monitored by plate count and fluorescence in situ hybridization (FISH). Temperature significantly affected survival of inoculants after storage for 60 days. Populations were 1.48 (plate counting) or 1.71 (FISH) times greater when stored at 20 °C compared to 40 °C. Inoculation of the fertilizer with pure spores led to the highest spore formation percentage (67.6% for plate counting, 94.2% for FISH). This study provides useful information for preservation of bioorganic fertilizer.

  3. Involvement of alanine racemase in germination of Bacillus cereus spores lacking an intact exosporium.

    PubMed

    Venir, Elena; Del Torre, Manuela; Cunsolo, Vincenzo; Saletti, Rosaria; Musetti, Rita; Stecchini, Mara Lucia

    2014-02-01

    The L-alanine mediated germination of food isolated Bacillus cereus DSA 1 spores, which lacked an intact exosporium, increased in the presence of D-cycloserine (DCS), which is an alanine racemase (Alr) inhibitor, reflecting the activity of the Alr enzyme, capable of converting L-alanine to the germination inhibitor D-alanine. Proteomic analysis of the alkaline extracts of the spore proteins, which include exosporium and coat proteins, confirmed that Alr was present in the B. cereus DSA 1 spores and matched to that encoded by B. cereus ATCC 14579, whose spore germination was strongly affected by the block of conversion of L- to D-alanine. Unlike ATCC 14579 spores, L-alanine germination of B. cereus DSA 1 spores was not affected by the preincubation with DCS, suggesting a lack of restriction in the reactant accessibility.

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

    PubMed

    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.

  5. Cryopreservation of spores of Dicksonia sellowiana: an endangered tree fern indigenous to South and Central America.

    PubMed

    Rogge, G D; Viana, A M; Randi, A M

    2000-01-01

    Spores of Dicksonia sellowiana (Presl.) Hook., an endangered tree fern, were stored in liquid nitrogen. Surface sterilized spores were placed in 1 ml sterile polypropylene cryotubes and were plunged into liquid nitrogen cryo-cans for 15 minutes, 15 days, 1 month and 3 months. In all, of the treatments the percentage of germination was higher than the control (fresh spores). Germination in Dyer and MS media supplement with 10 (-7) M and 5 x 10(-7) M BA was also promoted as comparing to control. There was no difference between the germination of spores thawed rapidly in a water bath at 45 degree C during 5 minutes or slowly at room temperature. Cryopreservation seems to promote germination of some dormant spores of D. sellowiana. The pre-treatment in cryoprotective solution of dimethyl sulphoxide 15%(v/v) in 1 M glycerol inhibited the germination of cryopreserved spores

  6. Responses of Bacillus subtilis spores to space environment: results from experiments in space.

    PubMed

    Horneck, G

    1993-02-01

    Onboard of several spacecrafts (Apollo 16, Spacelab 1, LDEF), spores of Bacillus subtilis were exposed to selected parameters of space, such as space vacuum, different spectral ranges of solar UV-radiation and cosmic rays, applied separately or in combination, and we have studied their survival and genetic changes after retrieval. The spores survive extended periods of time in space--up to several years--, if protected against the high influx of solar UV-radiation. Water desorption caused by the space vacuum leads to structural changes of the DNA; the consequences are an increased mutation frequency and altered photobiological properties of the spores. UV-effects, such as killing and mutagenesis, are augmented, if the spores are in space vacuum during irradiation. Vacuum-specific photoproducts which are different from the 'spore photoproduct' may cause the synergistic response of spores to the simultaneous action of UV and vacuum. The experiments provide an experimental test of certain steps of the panspermia hypothesis.

  7. Isolation and characterization of outermost layer deficient mutant spores of Bacillus megaterium.

    PubMed

    Takubo, Y; Atarashi, M; Nishihara, T; Kondo, M

    1988-01-01

    Outermost layer deficient mutant spores of Bacillus megaterium ATCC 12872 were isolated by Urografin density gradient centrifugation after mutagenesis with ethyl methanesulfonate. Although the composition of the cortex peptidoglycan was the same as that of the parent spores, three major proteins (48, 36, and 22 K daltons) were missing, suggesting that these proteins are components of the outermost layer. All mutant spores were also found to have very hydrophobic surface by 'salt aggregation test,' which would facilitate selection of such mutants.

  8. Environmental microbiology as related to planetary quarantine. [water activity and temperature effects on bacterial spore survival

    NASA Technical Reports Server (NTRS)

    Pflug, I. J.

    1972-01-01

    The survival of Bacillus subtilis var. niger spores suspended in solutions of sucrose and glycerol at calculated water activities and varying temperatures was studied. The overall results indicated that as the water activity of the liquid decreased from .99 to .85, the heat resistance of the spores increased. The nature of the substance controlling the water activity, and the history of the spores prior to treatment also had an affect on their heat resistance.

  9. Nondestructive Biophysical Probes of the Basis and Mechanism of Resistance in Microbial Spores.

    DTIC Science & Technology

    1983-05-10

    Characterization of Various Salt Forms of Bacillus megaterium Spores R. E. MARQUIS, E. L. CARSTENSEN, S. Z. CHILD, AND G. R. BENDER Departments of...Microbiology and Electrical Engineering, The University of Rochester, Rochester, New York 14642 Spores of Bacillus megaterium ATCC 19213 were subjected to an...of various salt forms of Bacillus megaterium spores to the germinating action of hydrostatic pressure GARY R. BENDER AND ROBERT E. MARQUIS Department

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

    DTIC Science & Technology

    2013-01-01

    properties of Bacillus megaterium spores, with elevated Mn levels associated with increased spore resistance to wet or dry heat, UVC radiation, and...RA (1974). The formation of Bacillus megaterium spores having increased heat and radiation resistance and variable heat shock requirements due to... Bacillus megaterium by wet heat. Lett. Appl. Microbiol. 50:507-514. Daly MJ (2012) Death by protein damage in irradiated cells. DNA Repair 11:12-21

  11. Water properties in fern spores: sorption characteristics relating to water affinity, glassy states, and storage stability.

    PubMed

    Ballesteros, Daniel; Walters, Christina

    2007-01-01

    Ex situ conservation of ferns may be accomplished by maintaining the viability of stored spores for many years. Storage conditions that maximize spore longevity can be inferred from an understanding of the behaviour of water within fern spores. Water sorption properties were measured in spores of five homosporeous species of ferns and compared with properties of pollen, seeds, and fern leaf tissue. Isotherms were constructed at 5, 25, and 45 degrees C and analysed using different physicochemical models in order to quantify chemical affinity and heat (enthalpy) of sorption of water in fern spores. Fern spores hydrate slowly but dry rapidly at ambient relative humidity. Low Brunauer-Emmet-Teller monolayer values, few water-binding sites according to the D'Arcy-Watt model, and limited solute-solvent compatibility according to the Flory-Huggins model suggest that fern spores have low affinity for water. Despite the low water affinity, fern spores demonstrate relatively high values of sorption enthalpy (DeltaH(sorp)). Parameters associated with binding sites and DeltaH(sorp) decrease with increasing temperature, suggesting temperature- and hydration-dependent changes in volume of spore macromolecules. Collectively, these data may relate to the degree to which cellular structures within fern spores are stabilized during drying and cooling. Water sorption properties within fern spores suggest that storage at subfreezing temperatures will give longevities comparable with those achieved with seeds. However, the window of optimum water contents for fern spores is very narrow and much lower than that measured in seeds, making precise manipulation of water content imperative for achieving maximum longevity.

  12. Studies on Sporulation Optimization and Chracterization of Bacillus subtilis Spore Quality

    DTIC Science & Technology

    2011-12-01

    W-A. ; Pinyupa, P. Film coating of seeds with Bacillus cereus RS87 spores for early plant growth enhancement. Can. J. Microbial. 2008, 54, 861 - 867...Optimization and Characterization of Bacillus subtilis Spore Quality Sb. GRANT NUMBER Sc. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) Sd. PROJECT NUMBER Smith...Environmental Protection Agency for the production of Bacillus subtilis spores. The cells require 12- 14 days to sporulate. Our goal was to conduct a study

  13. Novel Strategies for Enhanced Removal of Persistent Bacillus anthracis Surrogates and Clostridium difficile Spores from Skin

    PubMed Central

    Nerandzic, Michelle M.; Rackaityte, Elze; Jury, Lucy A.; Eckart, Kevin; Donskey, Curtis J.

    2013-01-01

    Background Removing spores of Clostridium difficile and Bacillus anthracis from skin is challenging because they are resistant to commonly used antimicrobials and soap and water washing provides only modest efficacy. We hypothesized that hygiene interventions incorporating a sporicidal electrochemically generated hypochlorous acid solution (Vashe®) would reduce the burden of spores on skin. Methods Hands of volunteers were inoculated with non-toxigenic C. difficile spores or B. anthracis spore surrogates to assess the effectiveness of Vashe solution for reducing spores on skin. Reduction in spores was compared for Vashe hygiene interventions versus soap and water (control). To determine the effectiveness of Vashe solution for removal of C. difficile spores from the skin of patients with C. difficile infection (CDI), reductions in levels of spores on skin were compared for soap and water versus Vashe bed baths. Results Spore removal from hands was enhanced with Vashe soak (>2.5 log10 reduction) versus soap and water wash or soak (~2.0 log10 reduction; P <0.05) and Vashe wipes versus alcohol wipes (P <0.01). A combined approach of soap and water wash followed by soaking in Vashe removed >3.5 log10 spores from hands (P <0.01 compared to washing or soaking alone). Bed baths using soap and water (N =26 patients) did not reduce the percentage of positive skin cultures for CDI patients (64% before versus 57% after bathing; P =0.5), whereas bathing with Vashe solution (N =21 patients) significantly reduced skin contamination (54% before versus 8% after bathing; P =0.0001). Vashe was well-tolerated with no evidence of adverse effects on skin. Conclusions Vashe was safe and effective for reducing the burden of B. anthracis surrogates and C. difficile spores on hands. Bed baths with Vashe were effective for reducing C. difficile on skin. These findings suggest a novel strategy to reduce the burden of spores on skin. PMID:23844234

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

    DTIC Science & Technology

    2014-09-18

    individual organisms sporulate/germinate simultaneously. Spores are shown as light orange and ovular. Cells are rod-shaped and form long chains. Figure... inhalational ) [1]. Of greatest interest to the military is inhalational exposure which causes the most lethal disease. Typically pulmonary disease requires... inhalation of approximately 1000 separate spores into the lungs of a healthy person. So dissemination of respirable, dry spores by aerosol methods is

  15. The development and structure of thick-walled, multicellular, aerial spores in Diheterospora chlamydosporia (=Verticillium chlamydosporium).

    PubMed

    Cambell, W P; Griffiths, D A

    1975-07-01

    The aerial, thick-walled spores in Diheterospara chlamydosporia arose as terminal swellings on erect hyphae. Repeated septation of the continuously swelling spore resulted in a multicellular structure. Immediately after the onset of septation secondary wall material was laid down between the two-layered primary wall and the plasmalemma. The presence of secondary wall material indicates that the multicellular spore is a dictyochlamydospore and not an aleuriospore. The relationship between chlamydospores and aleuriospores in other fungi is discussed.

  16. The agricultural pathology of ant fungus gardens

    PubMed Central

    Currie, Cameron R.; Mueller, Ulrich G.; Malloch, David

    1999-01-01

    Gardens of fungus-growing ants (Formicidae: Attini) traditionally have been thought to be free of microbial parasites, with the fungal mutualist maintained in nearly pure “monocultures.” We conducted extensive isolations of “alien” (nonmutualistic) fungi from ant gardens of a phylogenetically representative collection of attine ants. Contrary to the long-standing assumption that gardens are maintained free of microbial pathogens and parasites, they are in fact host to specialized parasites that are only known from attine gardens and that are found in most attine nests. These specialized garden parasites, belonging to the microfungus genus Escovopsis (Ascomycota: anamorphic Hypocreales), are horizontally transmitted between colonies. Consistent with theory of virulence evolution under this mode of pathogen transmission, Escovopsis is highly virulent and has the potential for rapid devastation of ant gardens, leading to colony mortality. The specialized parasite Escovopsis is more prevalent in gardens of the more derived ant lineages than in gardens of the more “primitive” (basal) ant lineages. Because fungal cultivars of derived attine lineages are asexual clones of apparently ancient origin whereas cultivars of primitive ant lineages were domesticated relatively recently from free-living sexual stocks, the increased virulence of pathogens associated with ancient asexual cultivars suggests an evolutionary cost to cultivar clonality, perhaps resulting from slower evolutionary rates of cultivars in the coevolutionary race with their pathogens. PMID:10393936

  17. Secondary Metabolites from the Fungus Emericella nidulans

    PubMed Central

    Tarawneh, Amer H.; León, Francisco; Radwan, Mohamed M.; Rosa, Luiz H.

    2014-01-01

    A new polyketide derivative koninginin H (1), has been isolated from the fungus Emericella nidulans, together with koninginin E (2), koninginin A (3), trichodermatide B (4), citrantifidiol (5), (4S,5R)-4-hydroxy-5-methylfuran-2-one (6), the glycerol derivatives gingerglycolipid B (7), (2S)-bis[9Z,12Z]-1-O, 2-O-dilinoleoyl-3-O-[α-d-galactopyranosyl-(1″→6′)β-d-galactopyranosyl]glycerol (8), (2S)-bis[9Z,12Z]-1-O, 2-O-dilinoleoyl-3-O-β-d-galactopyranosylglycerol (9), the cerebroside flavuside B (10), and the known sterols β-sitosterol glucoside and ergosta-5,7,22-trien-3-ol. Their structures were established by extensive NMR studies (1H NMR, 13C NMR, DEPT, 1H–1H COSY, HSQC, HMBC) and mass spectrometry. The antibacterial, antimalarial, antifungal and antileishmanial activities of compounds 1-10 were examined and the results indicated that compound 4 showed good antifungal activity against Cryptococcus neoformans with an IC50 value of 4.9 μg /mL. PMID:24273867

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

    PubMed

    Ehgartner, Daniela; Herwig, Christoph; Neutsch, Lukas

    2016-06-01

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

  19. The role of initial spore adhesion in pellet and biofilm formation in Aspergillus niger.

    PubMed

    Priegnitz, Bert-Ewald; Wargenau, Andreas; Brandt, Ulrike; Rohde, Manfred; Dietrich, Sylvia; Kwade, Arno; Krull, Rainer; Fleissner, André

    2012-01-01

    Fungi grow on a great variety of organic and inorganic materials. Colony establishment and growth on solid surfaces require adhesion of spores and hyphae to the substrate, while cell-to-cell interactions among spores and/or hyphae are a prerequisite for the development of three-dimensional mycelial structures such as pellets or biofilms. Surface adherence has been described as a two-step process, comprised of the initial attachment of ungerminated conidia followed by further adhesion of the forming germ tubes and growing hyphae. In the present study, we analyzed the contribution of adhesion of ungerminated spores to pellet and biofilm formation in Aspergillus niger. Mutants deficient in melanin biosynthesis were constructed by the deletion of the alb1 gene, encoding a polyketide synthase essential for pigment biosynthesis. Δalb1 conidia have an altered surface structure and changed physicochemical surface properties. Spore aggregation in liquid culture as well as spore surface attachment differ between the wild type and the mutant in a pH-dependent manner. In liquid culture further pellet formation is unaffected by altered spore-spore interactions, indicating that germ tube and hyphal adherence can compensate for deficiencies in the initial step of spore attachment. In contrast, under conditions promoting adhesion of Δalb1 conidia to polymer surfaces the mutant forms more stable biofilms than the wild type, suggesting that initial spore adhesion supports sessile growth.

  20. Unique aggregation of anthrax (Bacillus anthracis) spores by sugar-coated single-walled carbon nanotubes.

    PubMed

    Wang, Haifang; Gu, Lingrong; Lin, Yi; Lu, Fushen; Meziani, Mohammed J; Luo, Pengju G; Wang, Wei; Cao, Li; Sun, Ya-Ping

    2006-10-18

    There has been significant interest in the binding of anthrax spores by molecular species, but with only limited success. Proteins and more recently peptides were used. However, despite the known presence of carbohydrates on the spore surface, carbohydrate-carbohydrate interactions have hardly been explored likely because of the lack of required specific platform for synthetic carbohydrates. We report the successful use of single-walled carbon nanotubes as a truly unique scaffold for displaying multivalent monosaccharide ligands that bind effectively to anthrax spores with divalent cation mediation to cause significant spore aggregation. The work should have far-reaching implications in development of countermeasure technologies.

  1. Utilization of low-pressure plasma to inactivate bacterial spores on stainless steel screws.

    PubMed

    Stapelmann, Katharina; Fiebrandt, Marcel; Raguse, Marina; Awakowicz, Peter; Reitz, Günther; Moeller, Ralf

    2013-07-01

    A special focus area of planetary protection is the monitoring, control, and reduction of microbial contaminations that are detected on spacecraft components and hardware during and after assembly. In this study, wild-type spores of Bacillus pumilus SAFR-032 (a persistent spacecraft assembly facility isolate) and the laboratory model organism B. subtilis 168 were used to study the effects of low-pressure plasma, with hydrogen alone and in combination with oxygen and evaporated hydrogen peroxide as a process gas, on spore survival, which was determined by a colony formation assay. Spores of B. pumilus SAFR-032 and B. subtilis 168 were deposited with an aseptic technique onto the surface of stainless steel screws to simulate a spore-contaminated spacecraft hardware component, and were subsequently exposed to different plasmas and hydrogen peroxide conditions in a very high frequency capacitively coupled plasma reactor (VHF-CCP) to reduce the spore burden. Spores of the spacecraft isolate B. pumilus SAFR-032 were significantly more resistant to plasma treatment than spores of B. subtilis 168. The use of low-pressure plasma with an additional treatment of evaporated hydrogen peroxide also led to an enhanced spore inactivation that surpassed either single treatment when applied alone, which indicates the potential application of this method as a fast and suitable way to reduce spore-contaminated spacecraft hardware components for planetary protection purposes.

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

    PubMed

    Tanaka, Takashi; Ito, Akiko; Kamikado, Hideaki

    2012-01-01

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

  3. Effect of medium components and culture conditions in Bacillus subtilis EA-CB0575 spore production.

    PubMed

    Posada-Uribe, Luisa F; Romero-Tabarez, Magally; Villegas-Escobar, Valeska

    2015-10-01

    Bacillus subtilis spores have important biotechnological applications; however, achieving both, high spore cell densities and sporulation efficiencies in fermentation, is poorly reported. In this study, medium components and culture conditions were optimized with different statistical methods to increase spore production of the plant growth promoting rhizobacteria B. subtilis EA-CB0575. Key medium components were determined with Plackett-Burman (PB) design, and the optimum concentration levels of two components (glucose, MgSO4·7H2O) were optimized with a full factorial and central composite design, achieving 1.37 × 10(9) CFU/mL of spore cell density and 93.5 % of sporulation efficiency in shake flask. The optimized medium was used to determine the effect of culture conditions on spore production at bioreactor level, finding that maintaining pH control did not affect significantly spore production, while the interaction of agitation and aeration rates had a significant effect on spore cell density. The overall optimization generated a 17.2-fold increase in spore cell density (8.78 × 10(9) CFU/mL) and 1.9-fold increase in sporulation efficiency (94.2 %) compared to that of PB design. These results indicate the potential of B. subtilis EA-CB0575 to produce both, high spore cell densities and sporulation efficiencies, with very low nutrient requirements and short incubation period which can represent savings of process production.

  4. Methyl Anthranilate, an Inhibitor for the Germination of Spores of Aerobic Bacilli

    PubMed Central

    Prasad, Chandan; Srinivasan, V. R.

    1969-01-01

    Methylanthranilate inhibited the germination of spores of aerobic bacilli without affecting growth and sporulation. The inhibition of germination could not be reversed by removal of methylanthranilate. PMID:4979580

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

    PubMed Central

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

    2003-01-01

    Recent interest in anthrax is due to its potential use in bioterrorism and as a biowarfare agent against civilian populations. The development of rapid and sensitive techniques to detect anthrax spores in suspicious specimens is the most important aim for public health. With a view to preventing exposure of laboratory workers to viable Bacillus anthracis spores, this study evaluated the suitability of PCR assays for detecting anthrax spores previously inactivated at 121°C for 45 min. The results indicate that heat treatment ensures the complete inactivation of B. anthracis spores without significantly affecting the efficiency of PCR assays. PMID:12574311

  6. Method and apparatus for detecting and quantifying bacterial spores on a surface

    NASA Technical Reports Server (NTRS)

    Ponce, Adrian (Inventor)

    2009-01-01

    A method and an apparatus for detecting and quantifying bacterial spores on a surface. In accordance with the method: a matrix including lanthanide ions is provided on the surface containing the bacterial spores; functionalized aromatic molecules are released from the bacterial spores on the surface; a complex of the lanthanide ion and the aromatic molecule is formed on the surface; the complex of the lanthanide ion and the aromatic molecule is excited to generate a characteristic luminescence of the complex on the surface; and the bacterial spores exhibiting the luminescence of the complex on the surface are detected and quantified.

  7. Species-Specific Peptide Ligands for the Detection of Bacillus anthracis Spores

    PubMed Central

    Williams, David D.; Benedek, Orsolya; Turnbough, Charles L.

    2003-01-01

    Currently available detectors for spores of Bacillus anthracis, the causative agent of anthrax, are inadequate for frontline use and general monitoring. There is a critical need for simple, rugged, and inexpensive detectors capable of accurate and direct identification of B. anthracis spores. Necessary components in such detectors are stable ligands that bind tightly and specifically to target spores. By screening a phage display peptide library, we identified a family of peptides, with the consensus sequence TYPXPXR, that bind selectively to B. anthracis spores. We extended this work by identifying a peptide variant, ATYPLPIR, with enhanced ability to bind to B. anthracis spores and an additional peptide, SLLPGLP, that preferentially binds to spores of species phylogenetically similar to, but distinct from, B. anthracis. These two peptides were used in tandem in simple assays to rapidly and unambiguously identify B. anthracis spores. We envision that these peptides can be used as sensors in economical and portable B. anthracis spore detectors that are essentially free of false-positive signals due to other environmental Bacillus spores. PMID:14532093

  8. Evaluation of germination, distribution, and persistence of Bacillus subtilis spores through the gastrointestinal tract of chickens.

    PubMed

    Latorre, J D; Hernandez-Velasco, X; Kallapura, G; Menconi, A; Pumford, N R; Morgan, M J; Layton, S L; Bielke, L R; Hargis, B M; Téllez, G

    2014-07-01

    Spores are popular as direct-fed microbials, though little is known about their mode of action. Hence, the first objective of the present study was to evaluate the in vitro germination and growth rate of Bacillus subtilis spores. Approximately 90% of B. subtilis spores germinate within 60 min in the presence of feed in vitro. The second objective was to determine the distribution of these spores throughout different anatomical segments of the gastrointestinal tract (GIT) in a chicken model. For in vivo evaluation of persistence and dissemination, spores were administered to day-of-hatch broiler chicks either as a single gavage dose or constantly in the feed. During 2 independent experiments, chicks were housed in isolation chambers and fed sterile corn-soy-based diets. In these experiments one group of chickens was supplemented with 10(6) spores/g of feed, whereas a second group was gavaged with a single dose of 10(6) spores per chick on day of hatch. In both experiments, crop, ileum, and cecae were sampled from 5 chicks at 24, 48, 72, 96, and 120 h. Viable B. subtilis spores were determined by plate count method after heat treatment (75°C for 10 min). The number of recovered spores was constant through 120 h in each of the enteric regions from chickens receiving spores supplemented in the feed. However, the number of recovered B. subtilis spores was consistently about 10(5) spores per gram of digesta, which is about a 1-log10 reduction of the feed inclusion rate, suggesting approximately a 90% germination rate in the GIT when fed. On the other hand, recovered B. subtilis spores from chicks that received a single gavage dose decreased with time, with only approximately 10(2) spores per gram of sample by 120 h. This confirms that B. subtilis spores are transiently present in the GIT of chickens, but the persistence of vegetative cells is presently unknown. For persistent benefit, continuous administration of effective B. subtilis direct-fed microbials as vegetative

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

    PubMed Central

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

    2011-01-01

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

  10. Live/Dead Bacterial Spore Assay Using DPA-Triggered Tb Luminescence

    NASA Technical Reports Server (NTRS)

    Ponce, Adrian

    2003-01-01

    A method of measuring the fraction of bacterial spores in a sample that remain viable exploits DPA-triggered luminescence of Tb(3+) and is based partly on the same principles as those described earlier. Unlike prior methods for performing such live/dead assays of bacterial spores, this method does not involve counting colonies formed by cultivation (which can take days), or counting of spores under a microscope, and works whether or not bacterial spores are attached to other small particles (i.e., dust), and can be implemented on a time scale of about 20 minutes.

  11. Two distinct groups within the Bacillus subtilis group display significantly different spore heat resistance properties.

    PubMed

    Berendsen, Erwin M; Zwietering, Marcel H; Kuipers, Oscar P; Wells-Bennik, Marjon H J

    2015-02-01

    The survival of bacterial spores after heat treatment and the subsequent germination and outgrowth in a food product can lead to spoilage of the food product and economical losses. Prediction of time-temperature conditions that lead to sufficient inactivation requires access to detailed spore thermal inactivation kinetics of relevant model strains. In this study, the thermal inactivation kinetics of spores of fourteen strains belonging to the Bacillus subtilis group were determined in detail, using both batch heating in capillary tubes and continuous flow heating in a micro heater. The inactivation data were fitted using a log linear model. Based on the spore heat resistance data, two distinct groups (p < 0.001) within the B. subtilis group could be identified. One group of strains had spores with an average D120 °C of 0.33 s, while the spores of the other group displayed significantly higher heat resistances, with an average D120 °C of 45.7 s. When comparing spore inactivation data obtained using batch- and continuous flow heating, the z-values were significantly different, hence extrapolation from one system to the other was not justified. This study clearly shows that heat resistances of spores from different strains in the B. subtilis group can vary greatly. Strains can be separated into two groups, to which different spore heat inactivation kinetics apply.

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

    PubMed

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

    2013-08-16

    A survey of dried mushrooms (Lentinula edodes (Shiitake) and Auricularia auricula (Wood Ear)) sourced from China was carried out to determine the natural contamination of these mushrooms with spores of proteolytic Clostridium botulinum and non-proteolytic C. botulinum. The mushrooms were collected from supermarkets and retailers in 21 cities in China during October 2008. Spore loads of C. botulinum in mushrooms have a degree of uncertainty and variability and this study contributes valuable data for determining prevalence of spores of C. botulinum in mushrooms. An optimized detection protocol that combined selective enrichment culture with multiplex PCR was used to test for spores of proteolytic and non-proteolytic C. botulinum. Detection limits were calculated, using a maximum likelihood protocol, from mushroom samples inoculated with defined numbers of spores of proteolytic C. botulinum or non-proteolytic C. botulinum. Based on the maximum likelihood detection limit, it is estimated that dried mushroom A. auricula contained <550spores/kg of proteolytic C. botulinum, and <350spores/kg of non-proteolytic C. botulinum. Dried L. edodes contained <1500spores/kg of proteolytic C. botulinum and it was not possible to determine reliable detection limits for spores of non-proteolytic C. botulinum using the current detection protocol.

  13. Proteomic and genomic characterization of highly infectious Clostridium difficile 630 spores.

    PubMed

    Lawley, Trevor D; Croucher, Nicholas J; Yu, Lu; Clare, Simon; Sebaihia, Mohammed; Goulding, David; Pickard, Derek J; Parkhill, Julian; Choudhary, Jyoti; Dougan, Gordon

    2009-09-01

    Clostridium difficile, a major cause of antibiotic-associated diarrhea, produces highly resistant spores that contaminate hospital environments and facilitate efficient disease transmission. We purified C. difficile spores using a novel method and show that they exhibit significant resistance to harsh physical or chemical treatments and are also highly infectious, with <7 environmental spores per cm(2) reproducibly establishing a persistent infection in exposed mice. Mass spectrometric analysis identified approximately 336 spore-associated polypeptides, with a significant proportion linked to translation, sporulation/germination, and protein stabilization/degradation. In addition, proteins from several distinct metabolic pathways associated with energy production were identified. Comparison of the C. difficile spore proteome to those of other clostridial species defined 88 proteins as the clostridial spore "core" and 29 proteins as C. difficile spore specific, including proteins that could contribute to spore-host interactions. Thus, our results provide the first molecular definition of C. difficile spores, opening up new opportunities for the development of diagnostic and therapeutic approaches.

  14. Development of a novel spectroscopic methodology for the unique determination of bacterial spores

    NASA Astrophysics Data System (ADS)

    Alexander, Troy A.

    2003-08-01

    A novel methodology has been developed for the determination (i.e., identification and quantification) of bacterial spores that may be useful in many applications; most notably, development of detection schemes toward potentially harmful biological agents such as Bacillus anthracis. In addition, this method would be useful as an environmental warning system where sterility is of importance (i.e., food preparation areas as well as invasive and minimally-invasive medical applications). This method is based on the infrared (1500 to 4000-nm) absorption of fatty acids and peptides extracted from the spore. The absorption spectra of several bacterial spore extracts in carbon disulfide solution have been measured. Further, the groups of absorption bands in this region are unique for each spore, which implies it may be possible to use this technique for their determination. The Bacillus spores studied were chosen because they are taxonomically close to each other as well as to Bacillus anthracis. Expectedly, the measured absorption bands are heavily overlapped since the extracted analytes are similar in structure for each Bacillus spore. Additionally, this makes it impossible to use a single wavelength for the determination of any bacterial spore species. However, it may be possible to use the infrared absorption technique in conjunction with the Partial Least Squares (PLS) regression method to develop statistical models for the determination of bacterial spores. Results will be presented concerning sampling, data treatment, and development of PLS models as well as application of these models in the determination of unknown Bacillus bacterial spores.

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

    SciTech Connect

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

    2010-04-12

    A promising, environmentally safe method for inactivating fungal spores of Penicillium digitatum, a difficult-to-inactivate food spoilage microorganism, was developed using a high-density nonequilibrium atmospheric pressure plasma (NEAPP). The NEAPP employing Ar gas had a high electron density on the order of 10{sup 15} cm{sup -3}. The spores were successfully and rapidly inactivated using the NEAPP, with a decimal reduction time in spores (D value) of 1.7 min. The contributions of ozone and UV radiation on the inactivation of the spores were evaluated and concluded to be not dominant, which was fundamentally different from the conventional sterilizations.

  16. Stable isotope labeling of entire Bacillus atrophaeus spores and vegetative cells using bioaerosol mass spectrometry.

    PubMed

    Czerwieniec, Gregg A; Russell, Scott C; Tobias, Herbert J; Pitesky, Maurice E; Fergenson, David P; Steele, Paul; Srivastava, Abneesh; Horn, Joanne M; Frank, Matthias; Gard, Eric E; Lebrilla, Carlito B

    2005-02-15

    Single vegetative cells and spores of Bacillus atrophaeus, formerly Bacillus subtilis var. niger, were analyzed using bioaerosol mass spectrometry. Key biomarkers were identified from organisms grown in 13C and 15N isotopically enriched media. Spore spectra contain peaks from dicipolinate and amino acids. The results indicate that compounds observed in the spectra correspond to material from the spore's core and not the exosporium. Standard compounds and mixtures were analyzed for comparison. The biomarkers for vegetative cells were clearly different from those of the spores, consisting mainly of phosphate clusters and amino acid fragments.

  17. The sps Gene Products Affect the Germination, Hydrophobicity, and Protein Adsorption of Bacillus subtilis Spores

    PubMed Central

    Cangiano, Giuseppina; Sirec, Teja; Panarella, Cristina; Isticato, Rachele; Baccigalupi, Loredana; De Felice, Maurilio

    2014-01-01

    The multilayered surface of the Bacillus subtilis spore is composed of proteins and glycans. While over 70 different proteins have been identified as surface components, carbohydrates associated with the spore surface have not been characterized in detail yet. Bioinformatic data suggest that the 11 products of the sps operon are involved in the synthesis of polysaccharides present on the spore surface, but an experimental validation is available only for the four distal genes of the operon. Here, we report a transcriptional analysis of the sps operon and a functional study performed by constructing and analyzing two null mutants lacking either all or only the promoter-proximal gene of the operon. Our results show that both sps mutant spores apparently have normal coat and crust but have a small germination defect and are more hydrophobic than wild-type spores. We also show that spores lacking all Sps proteins are highly adhesive and form extensive clumps. In addition, sps mutant spores have an increased efficiency in adsorbing a heterologous enzyme, suggesting that hydrophobic force is a major determinant of spore adsorption and indicating that a deep understanding of the surface properties of the spore is essential for its full development as a surface display platform. PMID:25239894

  18. Utilization of Low-Pressure Plasma to Inactivate Bacterial Spores on Stainless Steel Screws

    PubMed Central

    Stapelmann, Katharina; Fiebrandt, Marcel; Raguse, Marina; Awakowicz, Peter; Reitz, Günther

    2013-01-01

    Abstract A special focus area of planetary protection is the monitoring, control, and reduction of microbial contaminations that are detected on spacecraft co