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Sample records for aerobic spore count

  1. 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

  2. Use of aerobic spores as a surrogate for cryptosporidium oocysts in drinking water supplies.

    PubMed

    Headd, Brendan; Bradford, Scott A

    2016-03-01

    Waterborne illnesses are a growing concern among health and regulatory agencies worldwide. The United States Environmental Protection Agency has established several rules to combat the contamination of water supplies by cryptosporidium oocysts, however, the detection and study of cryptosporidium oocysts is hampered by methodological and financial constraints. As a result, numerous surrogates for cryptosporidium oocysts have been proposed by the scientific community and efforts are underway to evaluate many of the proposed surrogates. The purpose of this review is to evaluate the suitability of aerobic bacterial spores to serve as a surrogate for cryptosporidium oocysts in identifying contaminated drinking waters. To accomplish this we present a comparison of the biology and life cycles of aerobic spores and oocysts and compare their physical properties. An analysis of their surface properties is presented along with a review of the literature in regards to the transport, survival, and prevalence of aerobic spores and oocysts in the saturated subsurface environment. Aerobic spores and oocysts share many commonalities with regard to biology and survivability, and the environmental prevalence and ease of detection make aerobic spores a promising surrogate for cryptosporidium oocysts in surface and groundwater. However, the long-term transport and release of aerobic spores still needs to be further studied, and compared with available oocyst information. In addition, the surface properties and environmental interactions of spores are known to be highly dependent on the spore taxa and purification procedures, and additional research is needed to address these issues in the context of transport.

  3. Improved Aerobic Colony Count Technique for Hydrophobic Grid Membrane Filters

    PubMed Central

    Parrington, Lorna J.; Sharpe, Anthony N.; Peterkin, Pearl I.

    1993-01-01

    The AOAC International official action procedure for performing aerobic colony counts on hydrophobic grid membrane filters (HGMFs) uses Trypticase soy-fast green FCF agar (FGA) incubated for 48 h. Microbial growths are various shades of green on a pale green background, which can cause problems for automated as well as manual counting. HGMFs which had been incubated 24 or 48 h at 35°C on Trypticase soy agar were flooded underneath with 1 to 2 ml of 0.1% triphenyltetrazolium chloride (TTC) solution by simply lifting one corner of the filter while it was still on the agar and adding the reagent. Microbial growths on HGMFs were counted after color had been allowed to develop for 15 min at room temperature. With representative foods, virtually all colonies stained pink to red. Automated electronic counts made by using the MI-100 HGMF Interpreter were easier and more reliable than control HGMF counts made by the AOAC International official action procedure. Manual counting was easier as well because of increased visibility of the microbial growths. Except in the case of dairy products, 24-h TTC counts did not differ significantly from 48-h FGA counts, whereas the FGA counts at 24 h were always significantly lower, indicating that for many food products the HGMF TTC flooding method permits aerobic colony counts to be made after 24 h. PMID:16349033

  4. Improved aerobic colony count technique for hydrophobic grid membrane filters.

    PubMed

    Parrington, L J; Sharpe, A N; Peterkin, P I

    1993-09-01

    The AOAC International official action procedure for performing aerobic colony counts on hydrophobic grid membrane filters (HGMFs) uses Trypticase soy-fast green FCF agar (FGA) incubated for 48 h. Microbial growths are various shades of green on a pale green background, which can cause problems for automated as well as manual counting. HGMFs which had been incubated 24 or 48 h at 35 degrees C on Trypticase soy agar were flooded underneath with 1 to 2 ml of 0.1% triphenyltetrazolium chloride (TTC) solution by simply lifting one corner of the filter while it was still on the agar and adding the reagent. Microbial growths on HGMFs were counted after color had been allowed to develop for 15 min at room temperature. With representative foods, virtually all colonies stained pink to red. Automated electronic counts made by using the MI-100 HGMF Interpreter were easier and more reliable than control HGMF counts made by the AOAC International official action procedure. Manual counting was easier as well because of increased visibility of the microbial growths. Except in the case of dairy products, 24-h TTC counts did not differ significantly from 48-h FGA counts, whereas the FGA counts at 24 h were always significantly lower, indicating that for many food products the HGMF TTC flooding method permits aerobic colony counts to be made after 24 h.

  5. 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.

  6. Characterisation of aerobically grown non-spore-forming bacteria from paper mill pulps containing recycled fibres.

    PubMed

    Suihko, Maija-Liisa; Skyttä, Eija

    2009-01-01

    A total of 179 non-spore-forming bacteria aerobically growing on Nutrient Agar, Plate Count Agar or in specific enrichment conditions for salmonella, campylobacteria, listeria, yersinia or staphylococci, were isolated from 16 untreated paper mill pulps. After phenotypical screening the isolates were characterised by automated ribotyping and partial sequencing of the 16S rRNA gene. They could be divided into seven taxonomical classes representing 63 taxa (species): actinobacteria (11 species), bacilli (7), flavobacteria (3) alphaproteobacteria (10), betaproteobacteria (5), gammaproteobacteria (25) and sphingobacteria (2). Most of the gammaproteobacteria were enterobacteria, mainly species of the genera Enterobacter (7 species, 7 samples/3 mills) and Klebsiella (5 species, 6 samples/3 mills). Other commonly occurring bacteria were most closely related to Microbacterium barkeri (7 samples/3 mills), Cloacibacterium normanense (6 samples/2 mills), Pseudoxanthomonas taiwanensis (5 samples/2 mills) and Sphingobacterium composti (5 samples/1 mill). Sporadic isolates of Listeria innocua, L. monocytogenes, Enterococcus casseliflavus and Staphylococcus warneri were detected, from which only L. monocytogenes is considered to be a food pathogen. No isolates of the genera Campylobacter, Salmonella or Yersinia were detected. The detected bacteria may be harmful in process control, but the load of food pathogens with recycled fibres to paper machines is insignificant. Faecal contamination of the pulp samples was not indicated.

  7. 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

  8. Aerobic deterioration stimulates outgrowth of spore-forming Paenibacillus in corn silage stored under oxygen-barrier or polyethylene films.

    PubMed

    Borreani, Giorgio; Dolci, Paola; Tabacco, Ernesto; Cocolin, Luca

    2013-08-01

    The occurrence of Bacillus and Paenibacillus spores in silage is of great concern to dairy producers because their spores can survive pasteurization and some strains are capable of subsequently germinating and growing under refrigerated conditions in pasteurized milk. The objectives of this study were to verify the role of aerobic deterioration of corn silage on the proliferation of Paenibacillus spores and to evaluate the efficacy of oxygen-barrier films used to cover silage during fermentation and storage to mitigate these undesirable bacterial outbreaks. The trial was carried out on whole-crop maize (Zea mays L.) inoculated with a mixture of Lactobacillus buchneri, Lactobacillus plantarum, and Enterococcus faecium. A standard polyethylene film and a polyethylene-polyamide film with an enhanced oxygen barrier were used to produce the silage bags for this experiment. The silos were stored indoors at ambient temperature (18 to 22°C) and opened after 110 d. The silage was sampled after 0, 2, 5, 7, 9, and 14 d of aerobic exposure to quantify the growth of endospore-forming bacteria during the exposure of silages to air. Paenibacillus macerans (gram-positive, facultatively anaerobic bacteria) was able to develop during the aerobic exposure of corn silage. This species was present in the herbage at harvesting, together with clostridial spores, and survived ensiling fermentation; it constituted more than 60% of the anaerobic spore formers at silage opening. During silage spoilage, the spore concentration of P. macerans increased to values greater than 7.0 log10 cfu/g of silage. The use of different plastic films to seal silages affected the growth of P. macerans and the number of spores during aerobic exposure of silages. These results indicate that the number of Paenibacillus spores could greatly increase in silage after exposure to air, and that oxygen-barrier films could help to reduce the potential for silage contamination of this important group of milk spoilage

  9. Toxinogenic and spoilage potential of aerobic spore-formers isolated from raw milk.

    PubMed

    De Jonghe, Valerie; Coorevits, An; De Block, Jan; Van Coillie, Els; Grijspeerdt, Koen; Herman, Lieve; De Vos, Paul; Heyndrickx, Marc

    2010-01-01

    The harmful effects on the quality and safety of dairy products caused by aerobic spore-forming isolates obtained from raw milk were characterized. Quantitative assessment showed strains of Bacillus subtilis, the Bacillus cereus group, Paenibacillus polymyxa and Bacillus amyloliquefaciens to be strongly proteolytic, along with Bacillus licheniformis, Bacillus pumilus and Lysinibacillus fusiformis to a lesser extent. Lipolytic activity could be demonstrated in strains of B. subtilis, B. pumilus and B. amyloliquefaciens. Qualitative screening for lecithinase activity also revealed that P. polymyxa strains produce this enzyme besides the B. cereus group that is well-known for causing a 'bitty cream' defect in pasteurized milk due to lecithinase activity. We found a strain of P. polymyxa to be capable of gas production during lactose fermentation. Strains belonging to the species B. amyloliquefaciens, Bacillus clausii, Lysinibacillus sphaericus, B. subtilis and P. polymyxa were able to reduce nitrate. A heat-stable cytotoxic component other than the emetic toxin was produced by strains of B. amyloliquefaciens and B. subtilis. Heat-labile cytotoxic substances were produced by strains identified as B. amyloliquefaciens, B. subtilis, B. pumilus and the B. cereus group. Variations in expression levels between strains from the same species were noticed for all tests. This study emphasizes the importance of aerobic spore-forming bacteria in raw milk as the species that are able to produce toxins and/or spoilage enzymes are all abundantly present in raw milk. Moreover, we demonstrated that some strains are capable of growing at room temperature and staying stable at refrigeration temperatures.

  10. Hydrophobic grid membrane filter method for aerobic plate count in foods: collaborative study.

    PubMed

    Entis, P

    1986-01-01

    Twenty-one laboratories participated in a collaborative study to validate a hydrophobic grid membrane filter (HGMF) method for aerobic plate count by comparing its performance against the AOAC/APHA pour plate method. Raw milk, raw poultry, whole egg powder, flours, and spices were included in the study. Counts obtained by the HGMF and pour plate methods did not differ significantly, except in the case of whole egg powder, for which the HGMF method produced significantly higher counts. The hydrophobic grid membrane filter method for aerobic plate count in foods has been adopted official first action.

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

    PubMed

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

    2016-07-01

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

  12. Does Spore Count Matter in Fungal Allergy?: The Role of Allergenic Fungal Species

    PubMed Central

    Lin, Wan-Rou; Lee, Mey-Fann; Hsu, Ling-Yi; Tien, Chih-Jen; Shih, Feng-Ming; Hsiao, Shih-Ching

    2016-01-01

    Purpose Fungi have been known to be important aeroallergens for hundreds of years. Most studies have focused on total fungal concentration; however, the concentration of specific allergenic fungi may be more important on an individual basis. Methods Ten fungal allergic patients and 2 non-fungal allergic patients were enrolled. The patients with a decrease in physician or patient global assessment by more than 50% of their personal best were considered to have an exacerbation of allergic symptoms and to be in the active stage. Those who maintained their physician and patient global assessment scores at their personal best for more than 3 months were considered to be in the inactive stage. The concentrations of dominant fungi in the patients' houses and outdoors were measured by direct and viable counts at active and inactive stages. Results The exacerbation of allergic symptoms was not correlated with total fungal spore concentration or the indoor/outdoor ratio (I/O). Specific fungi, such as Cladosporium oxysporum (C. oxyspurum), C. cladosporioides, and Aspergillus niger (A. niger), were found to be significantly higher concentrations in the active stage than in the inactive stage. Presumed allergenic spore concentration threshold levels were 100 CFU/m3 for C. oxysporum, and 10 CFU/m3 for A. niger, Penicillium brevicompactum and Penicillium oxalicum. Conclusions The major factor causing exacerbation of allergic symptoms in established fungal allergic patients may be the spore concentration of specific allergenic fungi rather than the total fungal concentration. These results may be useful in making recommendations as regards environmental control for fungal allergic patients. PMID:27334778

  13. Aerobic spore-forming bacteria for assessing quality of drinking water produced from surface water.

    PubMed

    Mazoua, Stephane; Chauveheid, Eric

    2005-12-01

    Cryptosporidium and Giardia represent a major microbiological issue for drinking water production from surface water. As their monitoring through a treatment process is rather tedious and as low-concentration goals should be reached for drinking water, aerobic spore-forming bacteria (ASFB) have been studied as an indicator microorganism for a drinking water treatment plant using surface water. The results reveal that monitoring naturally occurring ASFB better highlights daily achievable performances and identifies unusual process events for global disinfection, for both physical and chemical treatment steps in a multi-barrier drinking water treatment plant. Advantages of ASFB over usual process parameters are that these microorganisms are more sensitive to process fluctuations. The use of ASFB also showed that the efficiency of ozone disinfection is not as significantly influenced by the water temperature as reported, despite similar or higher CT values applied during warmer periods. Thus, the disinfection of resistant microorganisms with ozone can also be an efficient process at lower water temperature. ASFB have been shown to be a conservative indicator for Cryptosporidium and Giardia up to a 1st stage filtration and the ASFB Log removals can be used to estimate Log removals for Cryptosporidium and Giardia: compared to ASFB, the Log removals for Cryptosporidium or Giardia are at least equal or 50% higher, respectively. Thus, the monitoring of ASFB along a drinking water treatment process could be a useful tool for performing risk analysis for parasites such as Cryptosporidium and Giardia, and would further allow integration of daily variability into a risk analysis.

  14. Lower white blood cell counts in elite athletes training for highly aerobic sports.

    PubMed

    Horn, P L; Pyne, D B; Hopkins, W G; Barnes, C J

    2010-11-01

    White cell counts at rest might be lower in athletes participating in selected endurance-type sports. Here, we analysed blood tests of elite athletes collected over a 10-year period. Reference ranges were established for 14 female and 14 male sports involving 3,679 samples from 937 females and 4,654 samples from 1,310 males. Total white blood cell counts and counts of neutrophils, lymphocytes and monocytes were quantified. Each sport was scaled (1-5) for its perceived metabolic stress (aerobic-anaerobic) and mechanical stress (concentric-eccentric) by 13 sports physiologists. Substantially lower total white cell and neutrophil counts were observed in aerobic sports of cycling and triathlon (~16% of test results below the normal reference range) compared with team or skill-based sports such as water polo, cricket and volleyball. Mechanical stress of sports had less effect on the distribution of cell counts. The lower white cell counts in athletes in aerobic sports probably represent an adaptive response, not underlying pathology.

  15. Evaluation of the petrifilm aerobic count plate for enumeration of aerobic marine bacteria from seawater and Caulerpa lentillifera.

    PubMed

    Kudaka, Jun; Horii, Toru; Tamanaha, Koji; Itokazu, Kiyomasa; Nakamura, Masaji; Taira, Katsuya; Nidaira, Minoru; Okano, Sho; Kitahara, Akio

    2010-08-01

    The enumeration and evaluation of the activity of marine bacteria are important in the food industry. However, detection of marine bacteria in seawater or seafood has not been easy. The Petrifilm aerobic count plate (ACP) is a ready-to-use alternative to the traditional enumeration media used for bacteria associated with food. The purpose of this study was to evaluate the usefulness of a simple detection and enumeration method utilizing the Petrifilm ACP for enumeration of aerobic marine bacteria from seawater and an edible seaweed, Caulerpa lentillifera. The efficiency of enumeration of total aerobic marine bacteria on Petrifilm ACP was compared with that using the spread plate method on marine agar with 80 seawater and 64 C. lentillifera samples. With sterile seawater as the diluent, a close correlation was observed between the method utilizing Petrifilm ACP and that utilizing the conventional marine agar (r=0.98 for seawater and 0.91 for C. lentillifera). The Petrifilm ACP method was simpler and less time-consuming than the conventional method. These results indicate that Petrifilm ACP is a suitable alternative to conventional marine agar for enumeration of marine microorganisms in seawater and C. lentillifera samples.

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

    PubMed

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

    2008-06-01

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

  17. Evaluation of the 3M™ Petrifilm™ Rapid Aerobic Count Plate for the Enumeration of Aerobic Bacteria: Collaborative Study, First Action 2015.13.

    PubMed

    Bird, Patrick; Flannery, Jonathan; Crowley, Erin; Agin, James; Goins, David; Jechorek, Robert

    2016-05-01

    The 3M™ Petrifilm™ Rapid Aerobic Count (RAC) Plate is a sample-ready culture medium system containing dual-sensor indicator technology for the rapid quantification of aerobic bacteria in food products. The 3M Petrifilm RAC Plate was compared to the U.S. Food and Drug Administration Bacteriological Analytical Manual (FDA BAM) Chapter 3 (Aerobic Plate Count) for the enumeration of aerobic bacteria in raw easy-peel shrimp and the Standard Methods for the Examination of Dairy Products (SMEDP) Chapter 6 (Standard Plate Count Method) for the enumeration of aerobic bacteria in pasteurized skim milk and instant nonfat dry milk (instant NFDM). The 3M Petrifilm RAC Plate was evaluated using a paired study design in a multilaboratory collaborative study following current AOAC validation guidelines. Three target contamination levels (low, 10-100 CFU/g; medium, 100-1000 CFU/g; and high 1000-10 000 CFU/g) were evaluated for naturally occurring aerobic microflora for each matrix. For raw easy-peel shrimp, duplicate 3M Petrifilm RAC Plates were enumerated after 24 ± 2 h incubation at both 32 and 35°C. Pasteurized skim milk 3M Petrifilm RAC Plates were enumerated after 24 ± 2 h incubation at 32°C, and instant NFDM 3M Petrifilm RAC Plates were enumerated after 48 ± 3 h incubation at 32°C. No statistical difference was observed between 3M Petrifilm RAC Plate and FDA BAM or SMEDP reference methods for each contamination level.

  18. The Influence of Time Spent in Outdoor Play on Daily and Aerobic Step Count in Costa Rican Children

    ERIC Educational Resources Information Center

    Morera Castro, Maria del Rocio

    2011-01-01

    The purpose of this study is to examine the influence of time spent in outdoor play (i.e., on weekday and weekend days) on daily (i.e., average step count) and aerobic step count (i.e., average moderate to vigorous physical activity [MVPA] during the weekdays and weekend days) in fifth grade Costa Rican children. It was hypothesized that: (a)…

  19. Sensitivity to lytic agents and DNA base composition of several aerobic spore-bearing bacilli.

    PubMed

    Candeli, A; Mastrandrea, V; Cenci, G; De Bartolomeo, A

    1978-01-01

    The authors studied the possible relationship between a genetic characteristic, like DNA base composition, and certain phenotypic characteristics, i.e., sensitivity to lytic agents, morphology of colonies, and biochemical reactions in 34 strains of spore-bearing bacilli. From the results obtained two groups of bacilli have been identified. The first group includes the species B. subtilis, B. pumilus, B. licheniformis, and B. firmus and one strain of B. megaterium. The mean value of the GC% of the DNA is 44.22 +/- 1.76. All the strains examined are highly sensitive to lysozyme and resistant to sodium lauryl sulphate (S.L.S.); the surface colonies have a "rhizoid" appearance and the microcolonies on slide microculture are star-shaped. The second group includes the species B. cereus, B. cereus var. mycoides, B. anthracis, and B. thuringiensis. The mean value of the GC% of the DNA is 33.65 +/- 0.59. All the strains belonging to this group are resistant to both lysozyme and S.L.S., and the surface macro-colonies and the microcolonies have a "medusae head" appearance. The two groups also have certain different biochemical reactions; e.g., anaerobic growth and the egg yolk reaction, with few exception, are negative for the first group and positive for the second; furthermore, the strains in the first group (with rare exceptions) cause fermentation in the three carbohydrates, glucose, arabinose, and xylose, while glucose only is fermented by all strains with one exception in the second group. The position of B. megaterium is not yet clear, although one strain may certainly be included in the first group. Lysis by lipase is extremely variable and does not correlate with any of the other characteristics studied. The other species studied in relation to the characteristics, considered in our research (B. coagulans, B. macerans, B. polymyxa, B. laterosporus, B. alvei, B. circulans, B. stearothermophilus, and B. brevis), are not susceptible to grouping, either in the first, or

  20. Comparison of 3M Petrifilm Aerobic Count Plates to standard plating methodology for use with AOAC antimicrobial efficacy methods 955.14, 955.15, 964.02, and 966.04 as an alternative enumeration procedure: collaborative study.

    PubMed

    Nelson, Maria T; LaBudde, Robert A; Tomasino, Stephen F; Pines, Rebecca M

    2013-01-01

    A multilaboratory study was conducted to determine the equivalence of the 3M Petrifilm Aerobic Count Plate and standard plating methodology for measuring viable bacteria and spores recovered from hard-surface carriers (stainless steel and porcelain), also known as "control carrier counts," used in AOAC antimicrobial efficacy test methods. Six laboratories participated in the study in which carriers inoculated with Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enterica, and spores of Bacillus subtilis were evaluated using 3M Petrifilm Aerobic Count (AC) plates and standard plating side-by-side. The data were analyzed using a matched-pair t-test to determine the between-method effect with confidence intervals. For all test organisms pooled across all laboratories, the mean difference in log10 concentration between the standard plate count method and 3M Petrifilm AC Plates was -0.012, with a 95% confidence interval of (-0.090, +0.066), which was well within the -0.5, +0.5 interval established as the acceptance criterion. The between-carrier SD averaged 0.139; the between-replicate SD was 0.050. The carrier reproducibility, given that a single replicate per carrier is done, was estimated to be 0.148. Although differences were seen in the final concentrations of the test organisms among laboratories, there were no statistical differences between the enumeration methods. Based on the results from this study, 3M Petrifilm AC Plates are equivalent to standard plating methodology and can be used as an alternative procedure for the enumeration of test organisms used in AOAC Methods 955.14, 955.15, 964.02, and 966.04.

  1. Evaluation of a Novel Dry Sheet Culture Method for Rapid Enumeration of Total Aerobic Count in Foods.

    PubMed

    Teramura, Hajime; Iwasaki, Mihoko; Ushiyama, Masashi; Ogihara, Hirokazu

    2015-10-01

    A novel dry sheet culture method (Sanita-kun ACplus; SkACp) for rapid enumeration of total viable count has been developed. This rehydrated plate system comprises an adhesive sheet, nonwoven fabric coated with nutrients, and two types of water absorption polymers. In addition, SkACp facilitates methods for both rapid count (rapid mode: 24-h incubation) and accurate enumeration (standard mode: 48-h incubation) because it not only contains conventional 2,3,5-triphenyltetrazolium chloride but also contains two kinds of new tetrazolium salts for rapid and accurate enumeration of total aerobic count. When SkACp was assessed with 91 microorganisms, 87 strains (95.6%), excluding lactic acid and psychrotrophic bacteria, formed red-colored colonies within 24 h, whereas all microorganisms tested formed colonies within 48 h. The SkACp method, with both 24 and 48 h of incubation, was compared with plate count agar (PCA) and 3M Petrifilm AC (PAC) by using 107 naturally contaminated foods. For all foods tested (n = 107), the linear correlation coefficients of 48-h counts on SkACp compared with PCA and PAC were 0.98 and 0.75, respectively, while the 24-h counts on SkACp compared with PCA and PAC were 0.77 and 0.96, respectively. For foods tested, excluding yogurt and lactic beverages ( n = 101), the linear correlation coefficients of 48-h counts on SkACp compared with PCA and PAC were 0.98 and 0.96, respectively, while the 24-h counts on SkACp compared with PCA and PAC were 0.96 and 0.95, respectively. These results demonstrated that SkACp (48 h) is a useful alternative for the enumeration of the total aerobic count for all foods, whereas SkACp (24 h) was also an effective method for rapid enumeration in foods, excluding yogurt and lactic beverages.

  2. Effect of 12-week-long aerobic training programme on body composition, aerobic capacity, complete blood count and blood lipid profile among young women

    PubMed Central

    Nowak, Robert; Jastrzębski, Zbigniew; Zarębska, Aleksandra; Bichowska, Marta; Drobnik-Kozakiewicz, Izabela; Radzimiński, Łukasz; Leońska-Duniec, Agata; Ficek, Krzysztof; Cięszczyk, Paweł

    2015-01-01

    Background Numerous data suggest that aerobic-type exercise improves lipoprotein-lipid profiles, cardiorespiratory fitness and body composition in young women. The aim of this study was to evaluate the biological response to high-low impact aerobic fitness among young women. Materials and methods Thirty-four young women aged 22 (19-24) years were divided into three groups: underweight (N = 10), normal weight (N = 12) and overweight (N = 12). Aerobic capacity, anthropometry and body composition together with complete blood count and lipid profile were determined before and after completion of a 12-week-long training period. Results The training programme caused a significant decrease in weight (by 4.3 kg, P = 0.003), body mass index (by 1.3 kg/m2, P = 0.003), free fat mass (by 2.1 kg, P = 0.002), total body water (by 0.4 kg, P = 0.036), percentage of fat (by 3 percent points, P = 0.002), all analyzed skinfolds thicknesses, as well as the lipid profile in overweight group, and no changes in normal weight group. Significant changes in weight (by 4.2 kg, P = 0.005), body mass index (by 0.9 kg/m2, P = 0.005), crus skinfold thickness (by 3.3 mm, P = 0.028), and in maximum oxygen uptake (by 2.49 mL/kg/min; P = 0.047) were observed among underweight women. No change in total blood count was observed in all groups. Conclusion Twelve-week-long fitness training programme of two alternating styles (low and high impact) has a beneficial effect on overweight young women. PMID:25672474

  3. 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.

  4. Effects of high-energy electron irradiation of chicken meat on Salmonella and aerobic plate count

    SciTech Connect

    Heath, J.L.; Owens, S.L.; Tesch, S.; Hannah, K.W. )

    1990-01-01

    Four experiments were used to determine the effects of high-energy irradiation on the number of aerobic microorganisms and Salmonella on broiler breasts and thighs. Irradiation ranging from 100 to 700 kilorads (krads) was provided by a commercial-scale, electron-beam accelerator. Irradiation of broiler breast and thigh pieces with electron beams at levels of 100, 200, 300, 400, 500, and 600 krads showed that levels as low as 100 krads would eliminate Salmonella. When 33 thighs were tested after irradiation at 200 krads, only one thigh tested presumptive positive. The total number of aerobic organisms was reduced by 2 to 3 log10 cycles at irradiation levels of 100, 200, 300, 400, 500, 600, and 700 krads. Increasing the dose above 100 krads gave little if any additional benefit.

  5. Sensitization to Alternaria and Cladosporium in patients with respiratory allergy and outdoor counts of mold spores in Ankara atmosphere, Turkey.

    PubMed

    Bavbek, Sevim; Erkekol, Ferda Oner; Ceter, Talip; Mungan, Dilşad; Ozer, Faruk; Pinar, Münevver; Misirligil, Zeynep

    2006-08-01

    Sensitization to Alternaria and Cladosporium has been reported to be 3% to 30% in European countries. However, in Turkey, there is limited data about the prevalence of sensitization to these molds and the intensity of the two mold spores in Ankara atmosphere. This study was designed to evaluate the sensitization to Alternaria and Cladosporium in patients with respiratory allergy in Ankara and also the concentration of the two molds in Ankara atmosphere. Allergic rhinitis and asthma patients living in Ankara were included in the study. Demographic and diagnostic data of the patients were recorded. A skin prick test with extracts supplied by three different laboratories was used to evaluate the sensitization to Alternaria and Cladosporium. Mold spores were measured using a Burkard 7-day recording volumetric spore trap in Ankara atmosphere during a year. Overall sensitization to the two molds was found to be 14.8%, and isolated Alternaria or Cladosporiumsensitization was 3%. Considering the positive reaction to at least one of the three suppliers, the sensitization rate was 11.9% and 8.1% for Alternaria and Cladosporium, respectively. Cochran's Q homogenization test demonstrated that the positive and negative reaction were not homogeneous among three laboratories. The total number of mold spores in Ankara atmosphere was 429,264 spores/m3 of which 75.5% and 6% were constituted by Cladosporium and Alternaria, respectively. The prevalence of Cladosporium and Alternaria sensitization in respiratory allergy patients is quite similar to European countries; however, our data indicate that commercial mold extracts should be standardized to establish the real sensitization rates. Additionally, considering the great numbers of these mold spores in Ankara atmosphere, long-term follow-up studies are needed to evaluate the relationship between the mold load and sensitization patterns.

  6. Inhibition of the growth of Paenibacillus larvae, the causal agent of American foulbrood of honeybees, by selected strains of aerobic spore-forming bacteria isolated from apiarian sources.

    PubMed

    Alippi, Adriana M; Reynaldi, Francisco J

    2006-03-01

    The bacterium Paenibacillus larvae, the causative agent of American foulbrood disease of honeybee larvae, occurs throughout the world and is found in many beekeeping areas of Argentina. The potential as biocontrol agents of antagonic aerobic spore-forming bacteria isolated from honey samples and other apiarian sources were evaluated. Each isolate was screened against one strain of Paenibacillus larvae (ATCC 9545) by using a perpendicular streak technique. Ten randomly selected bacterial strains from the group that showed the best antagonistic effect to P. larvae ATCC 9545 were selected for further study. These were identified as Bacillus subtilis (m351), B. pumilus (m350), B. licheniformis (m347), B. cereus (mv33), B. cereus (m387), B. cereus (m6c), B. megaterium (m404), Brevibacillus laterosporus (BLAT169), B. laterosporus (BLAT170), and B. laterosporus (BLAT171). The antagonistic strains were tested against 17 P. larvae strains from different geographical origins by means of a spot test in wells. The analysis of variance and posterior comparison of means by Tukey method (P < 0.01) showed that the best antagonists were B. megaterium (m404), B. licheniformis (m347), B. cereus (m6c), B. cereus (mv33), and B. cereus (m387).

  7. Intradialytic aerobic cycling exercise alleviates inflammation and improves endothelial progenitor cell count and bone density in hemodialysis patients

    PubMed Central

    Liao, Min-Tser; Liu, Wen-Chih; Lin, Fu-Huang; Huang, Ching-Feng; Chen, Shao-Yuan; Liu, Chuan-Chieh; Lin, Shih-Hua; Lu, Kuo-Cheng; Wu, Chia-Chao

    2016-01-01

    Abstract Inflammation, endothelial dysfunction, and mineral bone disease are critical factors contributing to morbidity and mortality in hemodialysis (HD) patients. Physical exercise alleviates inflammation and increases bone density. Here, we investigated the effects of intradialytic aerobic cycling exercise on HD patients. Forty end-stage renal disease patients undergoing HD were randomly assigned to either an exercise or control group. The patients in the exercise group performed a cycling program consisting of a 5-minute warm-up, 20 minutes of cycling at the desired workload, and a 5-minute cool down during 3 HD sessions per week for 3 months. Biochemical markers, inflammatory cytokines, nutritional status, the serum endothelial progenitor cell (EPC) count, bone mineral density, and functional capacity were analyzed. After 3 months of exercise, the patients in the exercise group showed significant improvements in serum albumin levels, the body mass index, inflammatory cytokine levels, and the number of cells positive for CD133, CD34, and kinase insert domain-conjugating receptor. Compared with the exercise group, the patients in the control group showed a loss of bone density at the femoral neck and no increases in EPCs. The patients in the exercise group also had a significantly greater 6-minute walk distance after completing the exercise program. Furthermore, the number of EPCs significantly correlated with the 6-minute walk distance both before and after the 3-month program. Intradialytic aerobic cycling exercise programs can effectively alleviate inflammation and improve nutrition, bone mineral density, and exercise tolerance in HD patients. PMID:27399127

  8. Tumebacillus permanentifrigoris gen. nov., sp. nov., an aerobic, spore-forming bacterium isolated from Canadian high Arctic permafrost.

    PubMed

    Steven, Blaire; Chen, Min Qun; Greer, Charles W; Whyte, Lyle G; Niederberger, Thomas D

    2008-06-01

    A Gram-positive, aerobic, rod-shaped bacterium (strain Eur1 9.5(T)) was isolated from a 9-m-deep permafrost sample from the Canadian high Arctic. Strain Eur1 9.5(T) could not be cultivated in liquid medium and grew over the temperature range 5-37 degrees C; no growth was observed at 42 degrees C and only slow growth was observed at 5 degrees C following 1 month of incubation. Eur1 9.5(T) grew over the pH range 5.5-8.9 and tolerated NaCl concentrations of 0-0.5 % (w/v). Eur1 9.5(T) grew heterotrophically on complex carbon substrates and chemolithoautotrophically on inorganic sulfur compounds, as demonstrated by growth on sodium thiosulfate and sulfite as sole electron donors. Eur1 9.5(T) contained iso-C(15 : 0) as the major cellular fatty acid and menaquinone 7 (MK-7) as the major respiratory quinone. The cell-wall peptidoglycan was of type A1gamma. The DNA G+C content was 53.1 mol%. The 16S rRNA gene sequence of strain Eur1 9.5(T) was only distantly related (

  9. Analytical performance issues: comparison of ATP bioluminescence and aerobic bacterial count for evaluating surface cleanliness in an Italian hospital.

    PubMed

    Amodio, Emanuele; Cannova, Lucia; Villafrate, Maria Rosaria; Merendino, Anna Maria; Aprea, Luigi; Calamusa, Giuseppe

    2014-01-01

    Contaminated hospital surfaces have been demonstrated to be an important environmental reservoir of microorganisms that can increase the risk of nosocomial infection in exposed patients. As a consequence, cleaning and disinfecting hospital environments play an important role among strategies for preventing healthcare-associated colonization and infections. The aim of the present study was to evaluate whether adenosine triphosphate (ATP) presence, measured by bioluminescence methods, can predict microbiological contamination of hospital surfaces. The study was carried out between September and December 2012 at the University Hospital "P. Giaccone" of Palermo. A total of 193 randomly selected surfaces (tables, lockers, furnishings) were sampled and analyzed in order to assess ATP levels (expressed as relative light units or RLU) and aerobic colony count (ACC) or presence of S. aureus. ACC had median values of 1.85 cfu/cm(2)(interquartile range = 4.16) whereas ATP median was 44.6 RLU/cm(2)(interquartile range = 92.3). Overall, 85 (44.0%) surfaces exceeded the established microbial benchmark: 73 (37.8%) exceeded the 2.5 cfu/cm(2)ACC standard, 5 (2.6%) surfaces were positive for S. aureus and 7 (3.6%) showed both the presence of S. aureus and an ACC of more than 2.5 cfu/cm(2). ACC and bioluminescence showed significant differences in the different surface sites (p < 0.001). A significant correlation was found between ACC and RLU values (p-value < 0.001; R(2)= 0.29) and increasing RLU values were significantly associated with a higher risk of failing the benchmark (p < 0.001). Our data suggest that bioluminescence could help in measuring hygienic quality of hospital surfaces using a quick and sensitive test that can be an useful proxy of microbial contamination; however, further analysis will be necessary to assess the cost-efficacy of this methodology before requiring incorporation in hospital procedures.

  10. Combined steam-ultrasound treatment of 2 seconds achieves significant high aerobic count and Enterobacteriaceae reduction on naturally contaminated food boxes, crates, conveyor belts, and meat knives.

    PubMed

    Musavian, Hanieh S; Butt, Tariq M; Larsen, Annette Baltzer; Krebs, Niels

    2015-02-01

    Food contact surfaces require rigorous sanitation procedures for decontamination, although these methods very often fail to efficiently clean and disinfect surfaces that are visibly contaminated with food residues and possible biofilms. In this study, the results of a short treatment (1 to 2 s) of combined steam (95°C) and ultrasound (SonoSteam) of industrial fish and meat transportation boxes and live-chicken transportation crates naturally contaminated with food and fecal residues were investigated. Aerobic counts of 5.0 to 6.0 log CFU/24 cm(2) and an Enterobacteriaceae spp. level of 2.0 CFU/24 cm(2) were found on the surfaces prior to the treatment. After 1 s of treatment, the aerobic counts were significantly (P < 0.0001) reduced, and within 2 s, reductions below the detection limit (<10 CFU) were reached. Enterobacteriaceae spp. were reduced to a level below the detection limit with only 1 s of treatment. Two seconds of steam-ultrasound treatment was also applied on two different types of plastic modular conveyor belts with hinge pins and one type of flat flexible rubber belt, all visibly contaminated with food residues. The aerobic counts of 3.0 to 5.0 CFU/50 cm(2) were significantly (P < 0.05) reduced, while Enterobacteriaceae spp. were reduced to a level below the detection limit. Industrial meat knives were contaminated with aerobic counts of 6.0 log CFU/5 cm(2) on the handle and 5.2 log CFU/14 cm(2) on the steel. The level of Enterobacteriaceae spp. contamination was approximately 2.5 log CFU on the handle and steel. Two seconds of steam-ultrasound treatment reduced the aerobic counts and Enterobacteriaceae spp. to levels below the detection limit on both handle and steel. This study shows that the steam-ultrasound treatment may be an effective replacement for disinfection processes and that it can be used for continuous disinfection at fast process lines. However, the treatment may not be able to replace efficient cleaning processes used to remove high

  11. Enumeration of total aerobic microorganisms in foods by SimPlate Total Plate Count-Color Indicator methods and conventional culture methods: collaborative study.

    PubMed

    Feldsine, Philip T; Leung, Stephanie C; Lienau, Andrew H; Mui, Linda A; Townsend, David E

    2003-01-01

    The relative efficacy of the SimPlate Total Plate Count-Color Indicator (TPC-CI) method (SimPlate 35 degrees C) was compared with the AOAC Official Method 966.23 (AOAC 35 degrees C) for enumeration of total aerobic microorganisms in foods. The SimPlate TPC-CI method, incubated at 30 degrees C (SimPlate 30 degrees C), was also compared with the International Organization for Standardization (ISO) 4833 method (ISO 30 degrees C). Six food types were analyzed: ground black pepper, flour, nut meats, frozen hamburger patties, frozen fruits, and fresh vegetables. All foods tested were naturally contaminated. Nineteen laboratories throughout North America and Europe participated in the study. Three method comparisons were conducted. In general, there was <0.3 mean log count difference in recovery among the SimPlate methods and their corresponding reference methods. Mean log counts between the 2 reference methods were also very similar. Repeatability (Sr) and reproducibility (SR) standard deviations were similar among the 3 method comparisons. The SimPlate method (35 degrees C) and the AOAC method were comparable for enumerating total aerobic microorganisms in foods. Similarly, the SimPlate method (30 degrees C) was comparable to the ISO method when samples were prepared and incubated according to the ISO method.

  12. Application of Microbiological Method Direct Epifluorescence Filter Techique/Aerobic Plate Count Agar in the Identification of Irradiated Herbs and Spices

    PubMed Central

    Di Schiavi, Maria Teresa; Foti, Marina; Mosconi, Maria Cristina; Mattiolo, Giuseppina; Cavallina, Roberta

    2014-01-01

    Irradiation is a preservation technology used to improve the safety and hygienic quality of food. Aim of this study was to assess the applicability and validity of the microbiological screening method direct epifluorescence filter technique (DEFT)/aerobic plate count (APC) (EN 13783:2001) for the identification of irradiated herbs and spices. Tests on non-irradiated and irradiated samples of dried herbs and spices were performed. The method was based on the comparison of APC and count obtained using DEFT. In accordance with the standard reference, this method is not applicable to samples with APC<103 colony forming units (CFU)/g and this is its main limit. The results obtained in our laboratories showed that in 50% of cases of non-irradiated samples and in 96% of the samples treated with ionising radiation, the method was not applicable due to a value of CFU/g <103. PMID:27800348

  13. Incidence and Diversity of Potentially Highly Heat-Resistant Spores Isolated at Dairy Farms

    PubMed Central

    Scheldeman, Patsy; Pil, Annelies; Herman, Lieve; De Vos, Paul; Heyndrickx, Marc

    2005-01-01

    The presence of highly heat-resistant spores of Bacillus sporothermodurans in ultrahigh-temperature or sterilized consumer milk has emerged as an important item in the dairy industry. Their presence is considered undesirable since they hamper the achievement of commercial sterility requirements. By using a selective 30-min heat treatment at 100°C, 17 Belgian dairy farms were screened to evaluate the presence, sources, and nature of potentially highly heat-resistant spores in raw milk. High numbers of these spores were detected in the filter cloth of the milking equipment and in green crop and fodder samples. About 700 strains were isolated after the selective heating, of which 635 could be screened by fatty acid methyl ester analysis. Representative strains were subjected to amplified ribosomal DNA restriction analysis, 16S rRNA gene sequencing, percent G+C content, and DNA-DNA reassociations for further identification. The strain collection showed a remarkable diversity, with representatives of seven aerobic spore-forming genera. Bacillus licheniformis and Bacillus pallidus were the most predominant species overall. Twenty-three percent of the 603 spore-forming isolates proved to belong to 18 separate novel species. These findings suggest that the selective heating revealed a pool of unknown organisms with a higher heat-resistant character. This study showed that high spore counts can occur at the dairy farm and that feed and milking equipment can act as reservoirs or entry points for potentially highly heat-resistant spores into raw milk. Lowering this spore load by good hygienic measures could probably further reduce the contamination level of raw milk, in this way minimizing the aerobic spore-forming bacteria that could lead to spoilage of milk and dairy products. Assessment and characterization of this particular flora are of great importance to allow the dairy or food industry to adequately deal with newly arising microbiological problems. PMID:15746351

  14. Effects of air exposure, temperature and additives on fermentation characteristics, yeast count, aerobic stability and volatile organic compounds in corn silage.

    PubMed

    Weiss, K; Kroschewski, B; Auerbach, H

    2016-10-01

    Ensiling conditions strongly influence fermentation characteristics, yeast count, and aerobic stability. Numerous volatile organic compounds including esters are produced, which may negatively affect feed intake and animal performance and air quality. In addition to a farm survey, 3 laboratory experiments were carried out to study the effects of air (by delayed sealing or by air infiltration during anaerobic storage), temperature (20 and 35°C), and various types of additives [blends of either sodium benzoate and sodium propionate (SBSP) or of sodium benzoate and potassium sorbate (SBPS); buffered mixture of formic and propionic acids (FAPA); homofermentative inoculant (LAB)]. After additive treatment, chopped whole corn plants were packed into 1.5-L glass jars and stored for several months. For treatments with air infiltration, glass jars with holes in the lid and body were used. The farm survey in 2009 revealed large variation in lactate, acetate, ethanol, n-propanol, and 1,2-propanediol concentrations. Whereas ethyl esters were detected in all silages, the mean ethyl lactate concentrations were higher than those for ethyl acetate (474 vs. 38mg/kg of dry matter). In the ensiling experiments, few unequivocal effects of the tested factors on the analyzed parameters were observed due to many interactions. Delayed ensiling without additives decreased lactic acid production but, in one trial, increased acetic acid and had no effect on ethanol. The effect of delayed sealing on yeast counts and aerobic stability differed widely among experiments. Air infiltration during fermentation tested in one trial did not alter lactic acid production, but resulted in more acetic acid in delayed and more ethanol than in promptly sealed untreated silages. Greater ethanol production was associated with increased yeast numbers. Storage at high temperature resulted in lower lactic acid and n-propanol, and a trend toward reduced ethanol production was observed. The additive FAPA

  15. Adequacy of Petrifilm™ Aerobic Count plates supplemented with de Man, Rogosa & Sharpe broth and chlorophenol red for enumeration of lactic acid bacteria in salami.

    PubMed

    de Castilho, Natália Parma Augusto; Okamura, Vivian Tiemi; Camargo, Anderson Carlos; Pieri, Fábio Alessandro; Nero, Luís Augusto

    2015-12-01

    The present study aimed to assess the performance of alternative protocols to enumerate lactic acid bacteria (LAB) in salami. Fourteen cultures and two mixed starter cultures were plated using six protocols: 1) Petrifilm™ Aerobic Count (AC) with MRS broth and chlorophenol red (CR), incubated under aerobiosis or 2) under anaerobiosis, 3) MRS agar with CR, 4) MRS agar with bromocresol purple, 5) MRS agar at pH5.7, and 6) All Purpose Tween agar. Samples of salami were obtained and the LAB microbiota was enumerated by plating according protocols 1, 2, 3 and 5. Regression analysis showed a significant correlation between the tested protocols, based on culture counts (p<0.05). Similar results were observed for salami, and no significant differences of mean LAB counts between selected protocols (ANOVA, p>0.05). Colonies were confirmed as LAB, indicating proper selectivity of the protocols. The results showed the adequacy of Petrifilm™ AC supplemented with CR for the enumeration of LAB in salami.

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

    PubMed

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

    2016-06-01

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

  17. 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.

  18. 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.

  19. Spore prevalence and toxigenicity of Bacillus cereus and Bacillus thuringiensis isolates from U.S. retail spices.

    PubMed

    Hariram, Upasana; Labbé, Ronald

    2015-03-01

    Recent incidents of foodborne illness associated with spices as the vehicle of transmission prompted this examination of U.S. retail spices with regard to Bacillus cereus. This study focused on the levels of aerobic-mesophilic spore-forming bacteria and B cereus spores associated with 247 retail spices purchased from five states in the United States. Samples contained a wide range of aerobic-mesophilic bacterial spore counts (< 200 to 8.3 × 10(7) CFU/g), with 19.1% of samples at levels above 10(5) CFU/g. For examples, paprika, allspice, peppercorns, and mixed spices had high levels of aerobic spores (> 10(7) CFU/g). Using a novel chromogenic agar, B. cereus and B. thuringiensis spores were isolated from 77 (31%) and 11 (4%) samples, respectively. Levels of B. cereus were <3 to 1,600 MPN/g. Eighty-eight percent of B. cereus isolates and 91% of B. thuringiensis isolates possessed at least one type of enterotoxin gene: HBL (hemolysin BL) or nonhemolytic enterotoxin (NHE). None of the 88 isolates obtained in this study possessed the emetic toxin gene (ces). Using commercially available immunological toxin detection kits, the toxigenicity of the isolates was confirmed. The NHE enterotoxin was expressed in 98% of B. cereus and 91% of B. thuringiensis isolates that possessed the responsible gene. HBL enterotoxin was detected in 87% of B. cereus and 100% of B. thuringiensis PCR-positive isolates. Fifty-two percent of B. cereus and 54% of B. thuringiensis isolates produced both enterotoxins. Ninety-seven percent of B. cereus isolates grew at 12°C, although only two isolates grew well at 9°C. The ability of these spice isolates to form spores, produce diarrheal toxins, and grow at moderately abusive temperatures makes retail spices an important potential vehicle for foodborne illness caused by B. cereus strains, in particular those that produce diarrheal toxins.

  20. Changes in Aerobic Plate and Escherichia coli-Coliform Counts and in Populations of Inoculated Foodborne Pathogens on Inshell Walnuts during Storage.

    PubMed

    Frelka, John C; Davidson, Gordon R; Harris, Linda J

    2016-07-01

    After harvest, inshell walnuts are dried using low-temperature forced air and are then stored in bins or silos for up to 1 year. To better understand the survival of bacteria on inshell walnuts, aerobic plate counts (APCs) and Escherichia coli?coliform counts (ECCs) were evaluated during commercial storage (10 to 12°C and 63 to 65% relative humidity) over 9 months. APCs decreased by 1.4 to 2.0 log CFU per nut during the first 5 months of storage, and ECCs decreased by 1.3 to 2.2 log CFU per nut in the first month of storage. Through the remaining 4 to 8 months of storage, APCs and ECCs remained unchanged (P > 0.05) or decreased by <0.15 log CFU per nut per month. Similar trends were observed on kernels extracted from the inshell walnuts. APCs and ECCs were consistently and often significantly higher on kernels extracted from visibly broken inshell walnuts than on kernels extracted from visibly intact inshell walnuts. Parameters measured in this study were used to determine the survival of five-strain cocktails of E. coli O157:H7, Listeria monocytogenes, and Salmonella inoculated onto freshly hulled inshell walnuts (∼8 log CFU/g) after simulated commercial drying (10 to 12 h; 40°C) and simulated commercial storage (12 months at 10°C and 65% relative humidity). Populations declined by 2.86, 5.01, and 4.40 log CFU per nut for E. coli O157:H7, L. monocytogenes, and Salmonella, respectively, after drying and during the first 8 days of storage. Salmonella populations changed at a rate of -0.33 log CFU per nut per month between days 8 and 360, to final levels of 2.83 ± 0.79 log CFU per nut. E. coli and L. monocytogenes populations changed by -0.17 log CFU per nut per month and -0.26 log CFU per nut per month between days 8 and 360, respectively. For some samples, E. coli or L. monocytogenes populations were below the limit of detection by plating (0.60 log CFU per nut) by day 183 or 148, respectively; at least one of the six samples was positive at each subsequent

  1. 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

  2. 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.

  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. 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.

  5. 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)

  6. 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.

  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. 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.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  10. 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

  11. 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.

  12. 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.

  13. A procedure for estimating Bacillus cereus spores in soil and stream-sediment samples - A potential exploration technique

    USGS Publications Warehouse

    Watterson, J.R.

    1985-01-01

    The presence of bacterial spores of the Bacillus cereus group in soils and stream sediments appears to be a sensitive indicator of several types of concealed mineral deposits, including vein-type gold deposits. The B. cereus assay is rapid, inexpensive, and inherently reproducible. The test, currently under investigation for its potential in mineral exploration, is recommended for use on a research basis. Among the aerobic spore-forming bacilli, only B. cereus and closely related strains produce an opaque zone in egg-yolk emulsion agar. This characteristic, also known as the Nagler of lecitho-vitellin reaction, has long been used to rapidly indentify and estimate presumptive B. cereus. The test is here adapted to permit rapid estimation of B. cereus spores in soil and stream-sediment samples. Relative standard deviation was 10.3% on counts obtained from two 40-replicate pour-plate determinations. As many as 40 samples per day can be processed. Enough procedural detail is included to permit investigation of the test in conventional geochemical laboratories using standard microbiological safety precautions. ?? 1985.

  14. 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 ...

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. Aerobic Tennis.

    ERIC Educational Resources Information Center

    Stewart, Michael J.; Ahlschwede, Robert

    1989-01-01

    Increasing the aerobic nature of tennis drills in the physical education class may be necessary if tennis is to remain a part of the public school curriculum. This article gives two examples of drills that can be modified by teachers to increase activity level. (IAH)

  20. 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

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

    PubMed

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

    1995-06-01

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

  3. 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

  4. 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.

  5. 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.

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

    PubMed Central

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

    2015-01-01

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

  7. 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.

  8. 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

  9. 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

  10. 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.

  11. 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.

  12. WBC count

    MedlinePlus

    Leukocyte count; White blood cell count ... blood is 4,500 to 11,000 white blood cells per microliter (mcL) or 4.5 to 11. ... LOW WHITE BLOOD CELL (WBC) COUNT A low number of WBCs is called leukopenia. A WBC count below 4500 is below normal ...

  13. 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.

  14. 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.

  15. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2013-03-01

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

  17. Evaluating the Transport of Bacillus subtilis Spores as a Potential Surrogate for Cryptosporidium parvum Oocysts.

    PubMed

    Bradford, Scott A; Kim, Hyunjung; Headd, Brendan; Torkzaban, Saeed

    2016-02-02

    The U.S. Environmental Protection Agency has recommended the use of aerobic spores as an indicator for Cryptosporidium oocysts when determining groundwater under the direct influence of surface water. Surface properties, interaction energies, transport, retention, and release behavior of B. subtilis spores were measured over a range of physicochemical conditions, and compared with reported information for C. parvum oocysts. Interaction energy calculations predicted a much larger energy barrier and a shallower secondary minimum for spores than oocysts when the solution ionic strength (IS) equaled 0.1, 1, and 10 mM, and no energy barrier when the IS = 100 mM. Spores and oocysts exhibited similar trends of increasing retention with IS and decreasing Darcy water velocity (qw), and the predicted setback distance to achieve a six log removal was always larger for spores than oocysts. However, low levels of observed spore and oocyst release significantly influenced the predicted setback distance, especially when the fraction of reversibly retained microbes (Frev) was high. An estimate for Frev was obtained from large release pulses of spore and oocyst when the IS was reduced to deionized water. The value of Frev always increased with qw, whereas an opposition trend for Frev with IS was observed for spores (decreasing) and oocysts (increasing).

  18. "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…

  19. 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

  20. 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.

  1. Combined effects of carbonation with heating and fatty acid esters on inactivation and growth inhibition of various bacillus spores.

    PubMed

    Klangpetch, Wannaporn; Nakai, Tomoe; Noma, Seiji; Igura, Noriyuki; Shimoda, Mitsuya

    2013-09-01

    The effects of carbonation treatment (1 to 5 MPa, 30 min) plus heat treatment (30 to 80°C, 30 min) in the presence of various fatty acid esters (FAEs; 0.05 and 0.1%, wt/vol) on counts of viable Bacillus subtilis spores were investigated. FAEs or carbonation alone had no inactivation or growth inhibition effects on B. subtilis spores. However, carbonation plus heat (CH; 80°C, 5 MPa, 30 min) in the presence of mono- and diglycerol fatty acid esters markedly decreased counts of viable spores, and the spore counts did not change during storage for 30 days. The greatest decrease in viable spore counts occurred in the presence of monoglycerol fatty acid esters. Under CH conditions, inactivation and/or growth inhibition occurred at only 80°C and increased with increasing pressure. The greatest decrease in spore counts (more than 4 log units) occurred with CH (80°C, 5 MPa, 30 min) in the presence of monoglycerol fatty acid esters. However, this treatment was less effective against Bacillus coagulans and Geobacillus stearothermophilus spores.

  2. 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.

  3. 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.

  4. Walking dead: Permeabilization of heat-treated Geobacillus stearothermophilus ATCC 12980 spores under growth-preventing conditions.

    PubMed

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

    2017-06-01

    Although heat treatment is probably the oldest and the most common method used to inactivate spores in food processes, the specific mechanism of heat killing of spores is still not fully understood. The purpose of this study is to investigate the evolution of the permeabilization and the viability of heat-treated spores during storage under growth-preventing conditions. Geobacillus stearothermophilus spores were heat-treated under various conditions of temperature and pH, and then stored under conditions of temperature and pH that prevent growth. Spore survival was evaluated by count plating immediately after heat treatment, and then during storage over a period of months. Flow cytometry analyses were performed to investigate the Syto 9 permeability of heat-treated spores. Sub-lethally heat-treated spores of G. stearothermophilus were physically committed to permeabilization after heat treatment. However, prolonged heat treatment may abolish the spore permeabilization and block heat-treated spores in the refractive state. However, viability loss and permeabilization during heat treatment seem to be two different mechanisms that occur independently, and the loss of permeabilization properties takes place at a much slower rate than spore killing. Under growth-preventing conditions, viable heat-treated spores presumably lose their viability due to the permeabilization phenomena, which makes them more susceptible to the action of adverse conditions precluding growth.

  5. Multiplicity Counting

    SciTech Connect

    Geist, William H.

    2015-12-01

    This set of slides begins by giving background and a review of neutron counting; three attributes of a verification item are discussed: 240Pueff mass; α, the ratio of (α,n) neutrons to spontaneous fission neutrons; and leakage multiplication. It then takes up neutron detector systems – theory & concepts (coincidence counting, moderation, die-away time); detector systems – some important details (deadtime, corrections); introduction to multiplicity counting; multiplicity electronics and example distributions; singles, doubles, and triples from measured multiplicity distributions; and the point model: multiplicity mathematics.

  6. Investigating the thermodynamic stability of Bacillus subtilis spore-uranium(VI) adsorption though surface complexation modeling

    NASA Astrophysics Data System (ADS)

    Harrold, Z.; Hertel, M.; Gorman-Lewis, D.

    2012-12-01

    Dissolved uranium speciation, mobility, and remediation are increasingly important topics given continued and potential uranium (U) release from mining operations and nuclear waste. Vegetative bacterial cell surfaces are known to adsorb uranium and may influence uranium speciation in the environment. Previous investigations regarding U(VI) adsorption to bacterial spores, a differentiated and dormant cell type with a tough proteinaceous coat, include U adsorption affinity and XAFS data. We investigated the thermodynamic stability of aerobic, pH dependent uranium adsorption to bacterial spore surfaces using purified Bacillus subtilis spores in solution with 5ppm uranium. Adsorption reversibility and kinetic experiments indicate that uranium does not precipitate over the duration of the experiments and equilibrium is reached within 20 minutes. Uranium-spore adsorption edges exhibited adsorption at all pH measured between 2 and 10. Maximum adsorption was achieved around pH 7 and decreased as pH increased above 7. We used surface complexation modeling (SCM) to quantify uranium adsorption based on balanced chemical equations and derive thermodynamic stability constants for discrete uranium-spore adsorption reactions. Site specific thermodynamic stability constants provide insight on interactions occurring between aqueous uranium species and spore surface ligands. The uranium adsorption data and SCM parameters described herein, also provide a basis for predicting the influence of bacterial spores on uranium speciation in natural systems and investigating their potential as biosorption agents in engineered systems.

  7. 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.

  8. Teaching Aerobic Fitness Concepts.

    ERIC Educational Resources Information Center

    Sander, Allan N.; Ratliffe, Tom

    2002-01-01

    Discusses how to teach aerobic fitness concepts to elementary students. Some of the K-2 activities include location, size, and purpose of the heart and lungs; the exercise pulse; respiration rate; and activities to measure aerobic endurance. Some of the 3-6 activities include: definition of aerobic endurance; heart disease risk factors;…

  9. RBC count

    MedlinePlus

    ... marrow disease that causes abnormal increase in RBCs ( polycythemia vera ) Your RBC count will increase for several ... Multiple myeloma Myelofibrosis Paroxysmal nocturnal hemoglobinuria (PNH) Peripheral Polycythemia vera Renal cell carcinoma Splenomegaly Transfusion reaction - hemolytic ...

  10. Reticulocyte count

    MedlinePlus

    Anemia - reticulocyte ... A higher than normal reticulocytes count may indicate: Anemia due to red blood cells being destroyed earlier than normal ( hemolytic anemia ) Bleeding Blood disorder in a fetus or newborn ( ...

  11. Tower counts

    USGS Publications Warehouse

    Woody, Carol Ann; Johnson, D.H.; Shrier, Brianna M.; O'Neal, Jennifer S.; Knutzen, John A.; Augerot, Xanthippe; O'Neal, Thomas A.; Pearsons, Todd N.

    2007-01-01

    Counting towers provide an accurate, low-cost, low-maintenance, low-technology, and easily mobilized escapement estimation program compared to other methods (e.g., weirs, hydroacoustics, mark-recapture, and aerial surveys) (Thompson 1962; Siebel 1967; Cousens et al. 1982; Symons and Waldichuk 1984; Anderson 2000; Alaska Department of Fish and Game 2003). Counting tower data has been found to be consistent with that of digital video counts (Edwards 2005). Counting towers do not interfere with natural fish migration patterns, nor are fish handled or stressed; however, their use is generally limited to clear rivers that meet specific site selection criteria. The data provided by counting tower sampling allow fishery managers to determine reproductive population size, estimate total return (escapement + catch) and its uncertainty, evaluate population productivity and trends, set harvest rates, determine spawning escapement goals, and forecast future returns (Alaska Department of Fish and Game 1974-2000 and 1975-2004). The number of spawning fish is determined by subtracting subsistence, sport-caught fish, and prespawn mortality from the total estimated escapement. The methods outlined in this protocol for tower counts can be used to provide reasonable estimates ( plus or minus 6%-10%) of reproductive salmon population size and run timing in clear rivers. 

  12. 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.

  13. 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.

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

    NASA Technical Reports Server (NTRS)

    Schanche, Bradley

    2012-01-01

    from the coupon to recover the spores. One hundred µl of sterile 10% PVA was applied to the surface of the coupon and allowed to dry for 1 hour at 37 C. The films were then removed using sterile scalpel and forceps and placed into a glass test tube containing 2 milliliters of sterile deionized water. The PVA film process was then repeated on each coupon one additional time to ensure recovery of the majority of spores. The second PVA film was added in the same glass tube as in the previous round. If the spores remained 100% viable, the test tubes should now contain between 5 X 10(exp 6) and 5 X 10(exp 7) spores per millimeter; however, it is expected that some loss of viability has occurred. In order to assess this loss, the number of colony forming, viable spores was counted. To count the colony forming units (CFUs), the spore containing solution was diluted in a process of 10-fold serial dilution by mixing successive solutions in a 100 microliter spore suspension to 900 microliter deionized H2O ratio. A sample dilution series revealed that 10(exp -3) and 10(exp -4) concentrations would be necessary for an accurate CFU count to be taken. For those two concentrations, a spread on a TSA plate was prepared and incubated at 32 C. For the samples exposed to UV radiation, the cell survivability was too low to establish a count from 100 microliter spread plating. Instead, no dilutions were performed and the entire 2 milliliter spore suspension was plated and incubated at 32 C. The plate's CFU counts were taken at 24 hours and 48 hours from the time of plating. At the end of the CFU counting the total surviving spores in each sample were calculated based on the number of CFUs that were observed per 100 microliters, or per 2 milliliters for the UV irradiated samples. The results of these calculations are shown in Figures 1 and 2.

  15. Assessment of Bacterial Spores in Solid Materials: Curriculum Improvements Partnership Award for the Integration of Research (CIPAIR)

    NASA Technical Reports Server (NTRS)

    Lavallee, Richard J.

    2012-01-01

    This summer, we quantified the release, by cryogenic grinding at liquid nitrogen temperatures, of microbes present in 4 different spacecraft solids: epoxy 9309, epoxy 9394, epoxy 9396, and a silicone coating. Three different samples of each material were prepared: aseptically prepared solid material, powdered material inoculated with a known spore count of Bacillus atrophaeus, and solid material artificially embedded with a known spore count of Bacillus atrophaeus. Samples were cryogenically ground as needed, and the powders were directly cultured to determine the number of microbial survivors per gram of material. Recovery rates were found to be highly material-dependent, varying from 0.2 to 50% for inoculated material surfaces and 0.002 to 0.5% for embedded spores. A study of the spore survival rate versus total grinding time was also performed, with results indicating that longer grinding time decreases recovery rates of viable spores.

  16. 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

  17. 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

  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. 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.

  1. Comparison of dry medium culture plates for mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products.

    PubMed

    Park, Junghyun; Kim, Myunghee

    2013-12-01

    This study was performed to compare the performance of Sanita-Kun dry medium culture plate with those of traditional culture medium and Petrifilm dry medium culture plate for the enumeration of the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet. Mesophilic aerobic bacteria were comparatively evaluated in milk, ice cream, ham, and codfish fillet using Sanita-Kun aerobic count (SAC), Petrifilm aerobic count (PAC), and traditional plate count agar (PCA) media. According to the results, all methods showed high correlations of 0.989~1.000 and no significant differences were observed for enumerating the mesophilic aerobic bacteria in the tested food products. SAC method was easier to perform and count colonies efficiently as compared to the PCA and PAC methods. Therefore, we concluded that the SAC method offers an acceptable alternative to the PCA and PAC methods for counting the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products.

  2. 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.

  3. White Blood Cell Count

    MedlinePlus

    ... limited. Home Visit Global Sites Search Help? White Blood Cell Count Share this page: Was this page helpful? ... Count; Leukocyte Count; White Count Formal name: White Blood Cell Count Related tests: Complete Blood Count , Blood Smear , ...

  4. Counting Populations

    ERIC Educational Resources Information Center

    Damonte, Kathleen

    2004-01-01

    Scientists use sampling to get an estimate of things they cannot easily count. A population is made up of all the organisms of one species living together in one place at the same time. All of the people living together in one town are considered a population. All of the grasshoppers living in a field are a population. Scientists keep track of the…

  5. Counting Penguins.

    ERIC Educational Resources Information Center

    Perry, Mike; Kader, Gary

    1998-01-01

    Presents an activity on the simplification of penguin counting by employing the basic ideas and principles of sampling to teach students to understand and recognize its role in statistical claims. Emphasizes estimation, data analysis and interpretation, and central limit theorem. Includes a list of items for classroom discussion. (ASK)

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

    NASA Astrophysics Data System (ADS)

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

    2008-06-01

    Temporal and spatial variations in airborne spore concentrations of selected allergenic and pathogenic fungi were examined in Dublin, Ireland, in 2005. Air samples were taken at four outdoor locations in the city every 2 weeks, coupled with measurements of meteorological conditions. Total culturable airborne fungal spore concentrations in Dublin ranged from 30-6800 colony forming units per cubic metre of air (CFU m-3) over the 12-month period. Cladosporium, Penicillium, Aspergillus and Alternaria spores were constantly present in the Dublin atmosphere, representing >20% of the total culturable spore count. Concentrations of Cladosporium increased significantly in summer and reached allergenic threshold levels, peaking at over 3200 CFU m-3 in August. Penicillium spore concentrations never reached allergenic threshold levels, with average concentrations of <150 CFU m-3. Alternaria conidia formed only 0.3% of the total culturable fungal spore count and concentrations never exceeded 50 CFU m-3, attributable to the coastal position of Dublin and its low levels of arable production. The opportunistic human pathogen Aspergillus fumigatus was present throughout the year in nominal concentrations (<10 CFU m-3), but sporadic high counts were also recorded (300-400 CFU m-3), the potential health implications of which give cause for concern. Spores of neither Cryptococcus neoformans nor Stachybotrys chartarum were detected, but airborne basidiospores of Schizophyllum commune were evidenced by the dikaryotization of monokaryon tester strains following exposure to the air. The relationships between airborne fungal spore concentrations and meteorological factors were analysed by redundancy analysis and revealed positive correlations between temperature and Cladosporium and relative humidity and Penicillium and Aspergillus.

  7. 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.

  8. A THUMBNAIL HISTORY OF HETEROTROPHIC PLATE COUNT (HPC) METHODOLOGY IN THE UNITED STATES

    EPA Science Inventory

    Over the past 100 years, the method of determining the number of bacteria in water, foods or other materials has been termed variously as: bacterial plate count, total plate count, total viable plate count, aerobic plate count, standard plate cound and more recently, heterotrophi...

  9. 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.

  10. 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

  11. Enumeration of Clostridium perfringens spores in groundwater samples: comparison of six culture media.

    PubMed

    Araujo, M; Sueiro, R A; Gómez, M J; Garrido, M J

    2004-05-01

    In order to investigate the ability of Fluorocult-supplemented TSC agar (TSCF (Fluorocult supplemented TSC-agar): prepared from Tryptose Sulfite Cycloserine Agar Base (Merck), D-cycloserine (Fluka Chemika, USA), and fluorocult TSC-Agar supplement (Merck)) for detecting spores of Clostridium perfringens in water, we analyzed groundwater samples, pretreated by heating to 80 degrees C/5 min, using this fluorogenic medium together with five other media: mCP agar (Panreac; Cultimed), TSC agar (Merck, Germany), TSN agar (Merck), and SPS agar (BBL, USA) by the membrane filtration technique, and Wilson-Blair agar (WB) following the still-in-force Spanish official method. Variance analysis of the data obtained shows statistically significant differences in the counts obtained between media employed in this work. The C. perfringens spore counts on mCP agar were significantly lower (P<0.05) than the corresponding values of TSC, TSCF, SPS, and WB media. No statistically significant differences were found between C. perfringens spore counts on TSCF compared with those of other methods used. On the other hand, the identification of typical and atypical colonies isolated from all media demonstrated that fluorogenic TSC agar was the most specific medium for C. perfringens spore recovery in groundwater samples. Additionally, the results obtained indicate that mCP agar, which is the reference method in the European Union, is not suitable medium for recovering C. perfringens spores from groundwater samples.

  12. Evolutionary clade affects resistance of Clostridium difficile spores to Cold Atmospheric Plasma

    PubMed Central

    Connor, Mairéad; Flynn, Padrig B.; Fairley, Derek J.; Marks, Nikki; Manesiotis, Panagiotis; Graham, William G.; Gilmore, Brendan F.; McGrath, John W.

    2017-01-01

    Clostridium difficile is a spore forming bacterium and the leading cause of colitis and antibiotic associated diarrhoea in the developed world. Spores produced by C. difficile are robust and can remain viable for months, leading to prolonged healthcare-associated outbreaks with high mortality. Exposure of C. difficile spores to a novel, non-thermal atmospheric pressure gas plasma was assessed. Factors affecting sporicidal efficacy, including percentage of oxygen in the helium carrier gas admixture, and the effect on spores from different strains representing the five evolutionary C. difficile clades was investigated. Strains from different clades displayed varying resistance to cold plasma. Strain R20291, representing the globally epidemic ribotype 027 type, was the most resistant. However all tested strains displayed a ~3 log reduction in viable spore counts after plasma treatment for 5 minutes. Inactivation of a ribotype 078 strain, the most prevalent clinical type seen in Northern Ireland, was further assessed with respect to surface decontamination, pH, and hydrogen peroxide concentration. Environmental factors affected plasma activity, with dry spores without the presence of organic matter being most susceptible. This study demonstrates that cold atmospheric plasma can effectively inactivate C. difficile spores, and highlights factors that can affect sporicidal activity. PMID:28155914

  13. Evolutionary clade affects resistance of Clostridium difficile spores to Cold Atmospheric Plasma

    NASA Astrophysics Data System (ADS)

    Connor, Mairéad; Flynn, Padrig B.; Fairley, Derek J.; Marks, Nikki; Manesiotis, Panagiotis; Graham, William G.; Gilmore, Brendan F.; McGrath, John W.

    2017-02-01

    Clostridium difficile is a spore forming bacterium and the leading cause of colitis and antibiotic associated diarrhoea in the developed world. Spores produced by C. difficile are robust and can remain viable for months, leading to prolonged healthcare-associated outbreaks with high mortality. Exposure of C. difficile spores to a novel, non-thermal atmospheric pressure gas plasma was assessed. Factors affecting sporicidal efficacy, including percentage of oxygen in the helium carrier gas admixture, and the effect on spores from different strains representing the five evolutionary C. difficile clades was investigated. Strains from different clades displayed varying resistance to cold plasma. Strain R20291, representing the globally epidemic ribotype 027 type, was the most resistant. However all tested strains displayed a ~3 log reduction in viable spore counts after plasma treatment for 5 minutes. Inactivation of a ribotype 078 strain, the most prevalent clinical type seen in Northern Ireland, was further assessed with respect to surface decontamination, pH, and hydrogen peroxide concentration. Environmental factors affected plasma activity, with dry spores without the presence of organic matter being most susceptible. This study demonstrates that cold atmospheric plasma can effectively inactivate C. difficile spores, and highlights factors that can affect sporicidal activity.

  14. Airborne pollen and fungal spores in Garki, Abuja (North-Central Nigeria).

    PubMed

    Ezike, Dimphna Nneka; Nnamani, Catherine V; Ogundipe, Oluwatoyin T; Adekanmbi, Olushola H

    2016-01-01

    The ambient atmosphere is dominated with pollen and spores, which trigger allergic reactions and diseases and impact negatively on human health. A survey of pollen and fungal spores constituents of the atmosphere of Garki, Abuja (North-Central Nigeria) was carried out for 1 year (June 1, 2011-May 31, 2012). The aim of the study was to determine the prevalence and abundance of pollen and fungal spores in the atmosphere and their relationship with meteorological parameters. Airborne samples were trapped using modified Tauber-like pollen trap, and the recipient solutions were subjected to acetolysis. Results revealed the abundance of fungal spores, pollen, fern spores, algal cysts and diatoms in decreasing order of dominance. The atmosphere was qualitatively and quantitatively dominated by pollen during the period of late rainy/harmattan season than the rainy season. Numerous fungal spores were trapped throughout the sampling periods among which Alternaria spp., Fusarium spp., Cladosporium spp. and Curvularia spp. dominated. These fungi have been implicated in allergic diseases and are dermatophytic, causing diverse skin diseases. Other pathogenic fungi found in the studied aeroflora were Dreschlera spp., Helminthosporium spp., Torula spp., Pithomyces spp., Tetraploa spp., Nigrospora ssp., Spadicoides spp., Puccinia spp. and Erysiphe graminis. Total pollen and fungal spores counts do not show significant correlation with meteorological parameters.

  15. Reticulocyte Count Test

    MedlinePlus

    ... Reticulocyte Count Related tests: Red Blood Cell Count ; Hemoglobin ; Hematocrit ; Complete Blood Count ; Blood Smear ; Erythropoietin ; Vitamin ... on a complete blood count (CBC) , RBC count , hemoglobin or hematocrit , to help determine the cause To ...

  16. 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

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

    NASA Astrophysics Data System (ADS)

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

    2012-08-01

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

  18. Petrifilm plates for enumeration of bacteria counts in goat milk

    Technology Transfer Automated Retrieval System (TEKTRAN)

    PetrifilmTM Aerobic Count (AC) and Coliform Count (CC) plates were validated against standard methods for enumeration of coliforms, total bacteria, and psychrotrophic bacteria in raw (n = 39) and pasteurized goat milk (n = 17) samples. All microbiological data were transformed into log form and sta...

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

    NASA Astrophysics Data System (ADS)

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

    2010-05-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  2. In-hospital source of airborne Penicillium species spores.

    PubMed Central

    Streifel, A J; Stevens, P P; Rhame, F S

    1987-01-01

    Between 16 July and 1 October 1984, prospectively monitored corridor air samples from a bone marrow transplant station revealed a marked increase in airborne thermotolerant Penicillium spores. Simultaneous cultures of outside air showed lower spore counts, which were unchanged before, during, and after the corridor outburst, establishing that the source was within the hospital. Although the corridor was equipped with recirculating high-efficiency particulate air filtration units which provided 16 air changes per h, the mean corridor air count rose to 64.4 thermotolerant Penicillium CFU/m3 during the outburst period. The in-hospital source was ultimately traced to rotting cabinet wood enclosing a sink with leaking pipes in the medication room. It produced approximately 5.5 X 10(5) thermotolerant Penicillium CFU/h. In a patient room supplied by corridor air, an in-room recirculating high-efficiency particulate air filter reduced the mean thermotolerant Penicillium count to 2.2 CFU/m3. No patient illness or colonization occurred as a result of this event, although the cabinet wood, after sterilization, was shown to sustain abundant growth of Aspergillus fumigatus and Aspergillus flavus. Wet organic substrates should be avoided in hospital areas with immunosuppressed patients. Images PMID:3539981

  3. Influence of temperature and organic load on chemical disinfection of Geobacillus steareothermophilus spores, a surrogate for Bacillus anthracis.

    PubMed

    Guan, Jiewen; Chan, Maria; Brooks, Brian W; Rohonczy, Liz

    2013-04-01

    This study evaluated the influence of temperature and organic load on the effectiveness of domestic bleach (DB), Surface Decontamination Foam (SDF), and Virkon in inactivating Geobacillus stearothermophilus spores, which are a surrogate for Bacillus anthracis spores. The spores were suspended in light or heavy organic preparations and the suspension was applied to stainless steel carrier disks. The dried spore inoculum was covered with the disinfectants and the disks were then incubated at various temperatures. At -20°C, the 3 disinfectants caused less than a 2.0 log10 reduction of spores in both organic preparations during a 24-h test period. At 4°C, the DB caused a 4.4 log10 reduction of spores in light organic preparations within 2 h, which was about 3 log10 higher than what was achieved with SDF or Virkon. In heavy organic preparations, after 24 h at 4°C the SDF had reduced the spore count by 4.5 log10, which was about 2 log10 higher than for DB or Virkon. In general, the disinfectants were most effective at 23°C but a 24-h contact time was required for SDF and Virkon to reduce spore counts in both organic preparations by at least 5.5 log10. Comparable disinfecting activity with DB only occurred with the light organic load. In summary, at temperatures as low as 4°C, DB was the most effective disinfectant, inactivating spores within 2 h on surfaces with a light organic load, whereas SDF produced the greatest reduction of spores within 24 h on surfaces with a heavy organic load.

  4. Differences in concentrations of allergenic pollens and spores at different heights on an agricultural farm in West Bengal, India.

    PubMed

    Chakraborty, P; Gupta-Bhattacharya, S; Chowdhury, I; Majumdar, M R; Chanda, S

    2001-01-01

    The aim of the study was to assess the vertical profile of the major airborne pollen and spore concentration in the lower heights (up to six meters) and to check their allergenic potential causing respiratory allergy in agricultural workers. The study was conducted using rotorod samplers mounted at different heights at weekly intervals for two consecutive years (November 1997-October 1999). The major pollen grains and fungal spores (from mass culture) were collected in bulk and studied by skin-prick tests to detect allergenicity. Of the recorded pollen, 10 major and perennial types (e.g., Poaceae, Cheno-Amaranthaceae, Cyperaceae, Areca, etc.) were considered for comparative analyses. The tree pollen count showed more or less good correlation with increasing heights, whereas herb/shrub members are dominant at lower heights during all the three seasons (winter, summer and rains). The 10 major and perennial fungal spore types included Aspergilli group, Cladosporium, Nigrospora, etc. The smaller spores were dominant at greater heights and larger spores and conidia were more prevalent at lower levels. The total spore count was higher just after the rainy season during winter. In terms of allergenicity, Saccharum officinarum (sugar cane) of Poaceae, showed highest reactivity (70.58%) in skin test carried out in 189 adult agricultural field workers with respiratory disorders living inside the study area. Among fungal spores, Aspergillus japonicus was the strongest allergen, evoking 74.07% positive reactions. Drechslera oryzae, the pathogen causing brown spot of rice was also found to be a potent allergen.

  5. Bacillus odysseyi sp. nov., a round-spore-forming bacillus isolated from the Mars Odyssey spacecraft

    NASA Technical Reports Server (NTRS)

    La Duc, Myron T.; Satomi, Masataka; Venkateswaran, Kasthuri

    2004-01-01

    A round-spore-forming Bacillus species that produces an exosporium was isolated from the surface of the Mars Odyssey spacecraft. This novel species has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA-DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus and is a Gram-positive, aerobic, rod-shaped, endospore-forming eubacterium. Ultrathin sections of the spores showed the presence of an exosporium, spore coat, cortex and core. 16S rDNA sequence similarities between this strain, Bacillus fusiformis and Bacillus silvestris were approximately 96% and DNA-DNA reassociation values with these two bacilli were 23 and 17%, respectively. Spores of the novel species were resistant to desiccation, H2O2 and UV and gamma radiation. Of all strains tested, the spores of this strain were the most consistently resistant and survived all of the challenges posed, i.e. exposure to conditions of desiccation (100% survival), H2O2 (26% survival), UV radiation (10% survival at 660 J m(-2)) and gamma radiation (0.4% survival). The name proposed for this novel bacterium is Bacillus odysseyi sp. nov.; the type strain is 34hs-1T (=ATCC PTA-4993T=NRRL B-30641T=NBRC 100172T).

  6. Bacillus odysseyi sp. nov., a round-spore-forming bacillus isolated from the Mars Odyssey spacecraft.

    PubMed

    La Duc, Myron T; Satomi, Masataka; Venkateswaran, Kasthuri

    2004-01-01

    A round-spore-forming Bacillus species that produces an exosporium was isolated from the surface of the Mars Odyssey spacecraft. This novel species has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA-DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus and is a Gram-positive, aerobic, rod-shaped, endospore-forming eubacterium. Ultrathin sections of the spores showed the presence of an exosporium, spore coat, cortex and core. 16S rDNA sequence similarities between this strain, Bacillus fusiformis and Bacillus silvestris were approximately 96% and DNA-DNA reassociation values with these two bacilli were 23 and 17%, respectively. Spores of the novel species were resistant to desiccation, H2O2 and UV and gamma radiation. Of all strains tested, the spores of this strain were the most consistently resistant and survived all of the challenges posed, i.e. exposure to conditions of desiccation (100% survival), H2O2 (26% survival), UV radiation (10% survival at 660 J m(-2)) and gamma radiation (0.4% survival). The name proposed for this novel bacterium is Bacillus odysseyi sp. nov.; the type strain is 34hs-1T (=ATCC PTA-4993T=NRRL B-30641T=NBRC 100172T).

  7. What Is Aerobic Dancing?

    MedlinePlus

    ... aerobics can reach up to six times the force of gravity, which is transmitted to each of the 26 bones in the foot. Because of the many side-to-side motions, shoes need an arch design that will compensate ...

  8. 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.

  9. 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

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

    SciTech Connect

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

    2011-04-01

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

  11. Comparison of Innovative Molecular Approaches and Standard Spore Assays for Assessment of Surface Cleanliness ▿

    PubMed Central

    Cooper, Moogega; La Duc, Myron T.; Probst, Alexander; Vaishampayan, Parag; Stam, Christina; Benardini, James N.; Piceno, Yvette M.; Andersen, Gary L.; Venkateswaran, Kasthuri

    2011-01-01

    A bacterial spore assay and a molecular DNA microarray method were compared for their ability to assess relative cleanliness in the context of bacterial abundance and diversity on spacecraft surfaces. Colony counts derived from the NASA standard spore assay were extremely low for spacecraft surfaces. However, the PhyloChip generation 3 (G3) DNA microarray resolved the genetic signatures of a highly diverse suite of microorganisms in the very same sample set. Samples completely devoid of cultivable spores were shown to harbor the DNA of more than 100 distinct microbial phylotypes. Furthermore, samples with higher numbers of cultivable spores did not necessarily give rise to a greater microbial diversity upon analysis with the DNA microarray. The findings of this study clearly demonstrated that there is not a statistically significant correlation between the cultivable spore counts obtained from a sample and the degree of bacterial diversity present. Based on these results, it can be stated that validated state-of-the-art molecular techniques, such as DNA microarrays, can be utilized in parallel with classical culture-based methods to further describe the cleanliness of spacecraft surfaces. PMID:21652744

  12. Comparison of innovative molecular approaches and standard spore assays for assessment of surface cleanliness.

    PubMed

    Cooper, Moogega; La Duc, Myron T; Probst, Alexander; Vaishampayan, Parag; Stam, Christina; Benardini, James N; Piceno, Yvette M; Andersen, Gary L; Venkateswaran, Kasthuri

    2011-08-01

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

  13. 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

  14. Fluorescence-based methods for the detection of pressure-induced spore germination and inactivation

    NASA Astrophysics Data System (ADS)

    Baier, Daniel; Reineke, Kai; Doehner, Isabel; Mathys, Alexander; Knorr, Dietrich

    2011-03-01

    The application of high pressure (HP) provides an opportunity for the non-thermal preservation of high-quality foods, whereas highly resistant bacterial endospores play an important role. It is known that the germination of spores can be initiated by the application of HP. Moreover, the resistance properties of spores are highly dependent on their physiological states, which are passed through during the germination. To distinguish between different physiological states and to detect the amount of germinated spores after HP treatments, two fluorescence-based methods were applied. A flow cytometric method using a double staining with SYTO 16 as an indicator for germination and propidium iodide as an indicator for membrane damage was used to detect different physiological states of the spores. During the first step of germination, the spore-specific dipicolinic acid (DPA) is released [P. Setlow, Spore germination, Curr. Opin. Microbiol. 6 (2003), pp. 550-556]. DPA reacts with added terbium to form a distinctive fluorescent complex. After measuring the fluorescence intensity at 270 nm excitation wavelength in a fluorescence spectrophotometer, the amount of germinated spores can be determined. Spores of Bacillus subtilis were treated at pressures from 150 to 600 MPa and temperatures from 37 °C to 60 °C in 0.05 M ACES buffer solution (pH 7) for dwell times of up to 2 h. During the HP treatments, inactivation up to 2log 10 cycles and thermal sensitive populations up to 4log 10 cycles could be detected by plate counts. With an increasing number of thermal sensitive spores, an increased proportion of spores in germinated states was detected by flow cytometry. Also the released amount of DPA increased during the dwell times. Moreover, a clear pressure-temperature-time-dependency was shown by screening different conditions. The fluorescence-based measurement of the released DPA can provide the opportunity of an online monitoring of the germination of spores under HP inside

  15. Determination of spore concentration in Bacillus thuringiensis through the analysis of dipicolinate by capillary zone electrophoresis.

    PubMed

    He, Jin; Luo, Xiaofeng; Chen, Shouwen; Cao, Lili; Sun, Ming; Yu, Ziniu

    2003-04-25

    A new capillary zone electrophoresis (CZE) method for the analysis of dipicolinic acid, a specific component found in spores but not in vegetative cells, was used to determine spore concentration in Bacillus thuringiensis according to the relationship between the spore concentration and the content of dipicolinate. The quantitative relationship was established by using purified spores. Electrolyte conditions that affected the separation efficiency of dipicolinate and the reproducibility were investigated. With 10 mM phosphate, 10 mM ethylenediaminetetraacetic acid and 0.25 mM tetradecyltrimethylammonium bromide at pH 6.2 as the carrier electrolyte, dipicolinate can be determined within 8 min at an applied voltage of -25 kV (anode at detector) and a capillary temperature of 25 degrees C. The method has a high separation efficiency with which the number of theoretical plates is above 300,000 plates m(-1). The relative standard deviations for migration time and peak area are less than 0.5% and 2.0%, respectively. The detection limit for dipicolinate was 10 ng ml(-1), which corresponds to 7.2 x 10(5) spores ml(-1). The method was used to determine spores in fermentation broths, and the results obtained agreed well with the values obtained by plate counting.

  16. 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.

  17. 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

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

    PubMed

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

    1999-02-01

    The effects of nisin and monolaurin, alone and in combination, were investigated on Bacillus licheniformis spores in milk at 37 degrees C. In the absence of inhibitors, germinated spores developed into growing vegetative cells and started sporulation at the end of the exponential phase. In the presence of nisin (25 IU ml-1), spore outgrowth was inhibited (4 log10 reduction at 10 h). Regrowth appeared between 10 and 24 h and reached a high population level (1.25 x 10(8) cfu ml-1) after 7 d. Monolaurin (250 micrograms ml-1) had a bacteriostatic effect during the first 10 h but thereafter, regrowth occurred slowly with a population level after 7 d (4 x 10(5) cfu ml-1) lower than that of nisin. Different combined effects of nisin (between 0 and 42 IU ml-1), monolaurin (ranging from 0 to 300 micrograms ml-1), pH values (between 5.0 and 7.0) and spore loads (10(3), 10(4), 10(5) spores ml-1) were investigated using a Doehlert matrix in order to study the main effects of these factors and the different interactions. Results were analysed using the Response Surface Methodology (RSM) and indicated that nisin and monolaurin had no action on spores before germination; only pH values had a significant effect (P < or = 0.001), i.e. spore count decreased as the pH value increased in relation to germination. Sublethal concentrations of nisin (30 IU ml-1) and monolaurin (100 micrograms ml-1) in combination acted synergistically on outgrown spores and vegetative cells, showing total inhibition at pH 6.0, without regrowth, within 7 d at 37 degrees C.

  19. 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.

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. Promoting Bacillus cereus spore germination for subsequent inactivation by mild heat treatment.

    PubMed

    Løvdal, Irene Stranden; Hovda, Maria Befring; Granum, Per Einar; Rosnes, Jan Thomas

    2011-12-01

    Sublethal heat treatment may activate dormant spores and thereby potentiate the conversion of spores to vegetative cells. As the germinated spore is known to possess lower heat resistance than its dormant counterpart, it has been postulated that double heat treatment, i.e., spore heat activation followed by germination and then by heat inactivation, can be used to control spores in foods. Production of refrigerated processed foods of extended durability often includes more than one heat treatment of the food components. This work simulates conventional heat treatment procedures and evaluates double heat treatment as a method to improve spore control in model food matrixes of meat broth and cream sauce. Bacillus cereus NVH 1230-88 spores were supplemented in food model matrixes and heat activated at 70°C and then heat inactivated at 80 or 90°C. The samples were held at 29 to 30°C for 1 h between primary and secondary heat treatments, to allow spore germination. Nutrients naturally present in the food matrixes, e.g., amino acids and inosine, could act as germinants that induce germination. The levels of germinants could be too low to produce effective germination within 1 h. Following primary heat treatment, some samples were therefore supplemented with a combination of L-alanine and inosine, a germinant mixture known to be effective for B. cereus spores. In both matrixes, a combination of double heat treatment (heat activation, germination, and inactivation) and addition of germinants gave a reduction in spore counts equivalent to or greater than that obtained with a single heat treatment for 12 min at 90°C. Addition of germinants was essential to induce effective germination in cream sauce during 1 h at 29 to 30°C, and germinants were therefore a crucial supplement to obtain an effect of double heat treatment in this matrix. These data will be valuable when setting up temperature-time-germinant combinations for an optimized spore reduction in mild

  6. 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, ...

  7. Dance--Aerobic and Anaerobic.

    ERIC Educational Resources Information Center

    Cohen, Arlette

    1984-01-01

    This article defines and explains aerobic exercise and its effects on the cardiovascular system. Various studies on dancers are cited indicating that dance is an anaerobic activity with some small degree of aerobic benefit. (DF)

  8. 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.

  9. 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.

  10. Implementation of Aerobic Programs.

    ERIC Educational Resources Information Center

    American Alliance for Health, Physical Education, Recreation and Dance (AAHPERD).

    This information is intended for health professionals interested in implementing aerobic exercise programs in public schools, institutions of higher learning, and business and industry workplaces. The papers are divided into three general sections. The introductory section presents a basis for adhering to a health fitness lifestyle, using…

  11. Aerobic Anoxygenic Phototrophic Bacteria

    PubMed Central

    Yurkov, Vladimir V.; Beatty, J. Thomas

    1998-01-01

    The aerobic anoxygenic phototrophic bacteria are a relatively recently discovered bacterial group. Although taxonomically and phylogenetically heterogeneous, these bacteria share the following distinguishing features: the presence of bacteriochlorophyll a incorporated into reaction center and light-harvesting complexes, low levels of the photosynthetic unit in cells, an abundance of carotenoids, a strong inhibition by light of bacteriochlorophyll synthesis, and the inability to grow photosynthetically under anaerobic conditions. Aerobic anoxygenic phototrophic bacteria are classified in two marine (Erythrobacter and Roseobacter) and six freshwater (Acidiphilium, Erythromicrobium, Erythromonas, Porphyrobacter, Roseococcus, and Sandaracinobacter) genera, which phylogenetically belong to the α-1, α-3, and α-4 subclasses of the class Proteobacteria. Despite this phylogenetic information, the evolution and ancestry of their photosynthetic properties are unclear. We discuss several current proposals for the evolutionary origin of aerobic phototrophic bacteria. The closest phylogenetic relatives of aerobic phototrophic bacteria include facultatively anaerobic purple nonsulfur phototrophic bacteria. Since these two bacterial groups share many properties, yet have significant differences, we compare and contrast their physiology, with an emphasis on morphology and photosynthetic and other metabolic processes. PMID:9729607

  12. Aerobic Dance in Public Schools.

    ERIC Educational Resources Information Center

    Chiles, Barbara Ann; Moore, Suzanne

    1981-01-01

    Aerobic dance offers a challenging workout in a social atmosphere. Though some physical education instructors tend to exclude dance units from the curriculum, most could teach aerobic dance if they had a basic knowledge of aerobic routines. The outline for a unit to be used in the class is presented. (JN)

  13. 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.

  14. [Saprophytic and opportunistic non spore-forming anaerobic microflora of the vagina (author's transl)].

    PubMed

    Cavazzini, G; Folegatti, M R; Segala, V; Cenci, P

    1980-01-01

    A microbiological survey has been carried out on 179 healthy, child-bearing aged, non-pregnant women, with the aim to evaluate the incidence of anaerobic non-spore forming bacteria in the normal vaginal flora. This group of bacteria has been isolated in 50.3% of women, with a clear prevalence of "anaerobic Streptococci " and Bacteroides, followed by Fusobacterium and Veillonella. No Propionibacterium, Eubacterium or Bifidobacterium have been isolated. According to many Authors the non-spore forming anaerobes must be considered opportunistic bacteria, responsible of many infections of the female genital tract, especially when associated with other aerobic or facultative bacteria. Antibiograms have demonstrated a wide spectrum of activity of chloramphenicol and clindamycin; although not widely distributed, antibacterial activity have also shown metronidazole, penicillins, cephalosporins and lincomycin.

  15. Online monitoring of Escherichia coli and Bacillus thuringiensis spore inactivation after advanced oxidation treatment.

    PubMed

    Sherchan, Samendra P; Snyder, Shane A; Gerba, Charles P; Pepper, Ian L

    2014-01-01

    Various studies have shown that advanced oxidation processes (AOPs) such as UV light in combination with hydrogen peroxide is an efficient process for the removal of a large variety of emerging contaminants including microorganisms. The mechanism of destruction in the presence of hydrogen peroxide (H2O2) is the enhanced formation of hydroxyl (·OH) radicals, which have a high oxidation potential. The goal of this study was to utilize in-line advanced oxidation to inactivate microbes, and document the inactivation via an in-line, real-time sensor. Escherichia coli cells and Bacillus thuringiensis spores were exposed to UV/H2O2 treatment in DI water, and the online sensor BioSentry(®) was evaluated for its potential to monitor inactivation in real-time. B. thuringiensis was selected as a non-pathogenic surrogate for B. anthracis, the causative agent of anthrax and a proven biological weapon. UV radiation and UV/H2O2 exposure resulted in a >6 log10 reduction of the viable culturable counts of E. coli vegetative cells, and a 3 log10 reduction of B. thuringiensis spores. Scanning electron microscopy of the treated samples revealed severe damage on the surface of most E. coli cells, yet there was no significant change observed in the morphology of the B. thuringiensis spores. Following AOP exposure, the BioSentry sensor showed an increase in the categories of unknown, rod and spores counts, but overall, did not correspond well with viable count assays. Data from this study show that advanced oxidation processes effectively inactivate E. coli vegetative cells, but not B. thuringiensis spores, which were more resistant to AOP. Further, the BioSentry in-line sensor was not successful in documenting destruction of the microbial cells in real-time.

  16. Quantifying spore viability of the honey bee pathogen Nosema apis using flow cytometry.

    PubMed

    Peng, Yan; Lee-Pullen, Tracey F; Heel, Kathy; Millar, A Harvey; Baer, Boris

    2014-05-01

    Honey bees are hosts to more than 80 different parasites, some of them being highly virulent and responsible for substantial losses in managed honey bee populations. The study of honey bee pathogens and their interactions with the bees' immune system has therefore become a research area of major interest. Here we developed a fast, accurate and reliable method to quantify the viability of spores of the honey bee gut parasite Nosema apis. To verify this method, a dilution series with 0, 25, 50, 75, and 100% live N. apis was made and SYTO 16 and Propidium Iodide (n = 35) were used to distinguish dead from live spores. The viability of spores in each sample was determined by flow cytometry and compared with the current method based on fluorescence microscopy. Results show that N. apis viability counts using flow cytometry produced very similar results when compared with fluorescence microscopy. However, we found that fluorescence microscopy underestimates N. apis viability in samples with higher percentages of viable spores, the latter typically being what is found in biological samples. A series of experiments were conducted to confirm that flow cytometry allows the use of additional fluorescent dyes such as SYBR 14 and SYTOX Red (used in combination with SYTO 16 or Propidium Iodide) to distinguish dead from live spores. We also show that spore viability quantification with flow cytometry can be undertaken using substantially lower dye concentrations than fluorescence microscopy. In conclusion, our data show flow cytometry to be a fast, reliable method to quantify N. apis spore viabilities, which has a number of advantages compared with existing methods.

  17. Examination of B. subtilis var. niger Spore Killing by Dry Heat Methods

    NASA Technical Reports Server (NTRS)

    Kempf, Michael J.; Kirschner, Larry E.

    2004-01-01

    Dry heat microbial reduction is the only NASA approved sterilization method to reduce the microbial bioburden on space-flight hardware prior to launch. Reduction of the microbial bioburden on spacecraft is necessary to meet planetary protection requirements specific for the mission. Microbial bioburden reduction also occurs if a spacecraft enters a planetary atmosphere (e.g., Mars) and is heated due to frictional forces. Temperatures reached during atmospheric entry events (>200 C) are sufficient to damage or destroy flight hardware and also kill microbial spores that reside on the in-bound spacecraft. The goal of this research is to determine the survival rates of bacterial spores when they are subjected to conditions similar to those the spacecraft would encounter (i.e., temperature, pressure, etc.). B. subtilis var. niger spore coupons were exposed to a range of temperatures from 125 C to 200 C in a vacuum oven (at <1 Torr). After the exposures, the spores were removed by sonication, dilutions were made, and the spores were plated using the pour plate method with tryptic soy agar. After 3 days incubation at 32 C, the number of colony-forming units was counted. Lethality rate constants and D-values were calculated at each temperature. The calculated D-values were: 27 minutes (at 125 C), 13 minutes (at 135 C), and <0.1 minutes (at 150 C). The 125 C and 135 C survivor curves appeared as concavedownward curves. The 150 C survivor curve appeared as a straight-line. Due to the prolonged ramp-up time to the exposure conditions, spore killing during the ramp-up resulted in insufficient data to draw curves for exposures at 160 C, 175 C, and 200 C. Exploratory experiments using novel techniques, with short ramp times, for performing high temperature exposures were also examined. Several of these techniques, such as vacuum furnaces, thermal spore exposure vessels, and laser heating of the coupons, will be discussed.

  18. Bacterial counts associated with poultry processing at different sampling times.

    PubMed

    Geornaras, I; von Holy, A

    2000-01-01

    Aerobic plate counts, Enterobacteriaceae counts and Pseudomonas counts were performed on neck skin samples from six processing steps in a poultry abattoir at three different sampling times. Sampling time 1 was shortly after start-up of processing operations, time 2 after a tea break which was preceded by a cold water rinse-down of equipment surfaces, and time 3 before shut-down. No significant differences (P > 0.05) in microbial numbers of neck skin samples were observed between the three sampling times at the six sampling sites. At this particular processing plant, therefore, sampling at any time of the processing shift would thus not lead to significantly different bacterial counts of neck skins. The lowest aerobic plate counts, over all three sampling times, were obtained for neck skins sampled after spray washing, and the highest for neck skins sampled after packaging. This indicated the efficacy of the washing step in reducing microbial contamination but subsequent re-contamination of carcasses. Despite the Pseudomonas counts of neck skins being lower than the Enterobacteriaceae counts at the beginning of processing, packaging of carcasses resulted in Pseudomonas counts that were higher than the Enterobacteriaceae counts.

  19. 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

  20. 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.

  1. 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

  2. All about Carbohydrate Counting

    MedlinePlus

    Toolkit No. 14 All About Carbohydrate Counting What is carbohydrate counting? Carbohydrate counting is a way to plan your meals. It can help ... Diabetes Association, Inc. 2/14 Toolkit No. 14: All About Carbohydrate Counting continued The chart at the ...

  3. Efficacy of gaseous chlorine dioxide in inactivating Bacillus cereus spores attached to and in a biofilm on stainless steel.

    PubMed

    Nam, Hyegyeong; Seo, Hyun-Sun; Bang, Jihyun; Kim, Hoikyung; Beuchat, Larry R; Ryu, Jee-Hoon

    2014-10-01

    We evaluated the lethal activity of gaseous chlorine dioxide (ClO2) against Bacillus cereus spores attached to and in biofilm formed on a stainless steel surface. Aqueous ClO2 was prepared by mixing sulfuric acid (5% w/v) with sodium chlorite (10mg/mL), and gaseous ClO2 was produced by vaporization of aqueous ClO2 in an air-tight container. The concentration of gaseous ClO2 in the air within the container increased rapidly at first but gradually decreased over time. The lethality of gaseous ClO2 against B. cereus spores attached to stainless steel coupons (SSCs) and in biofilm formed by the pathogen on SSCs was determined. The B. cereus spores attached to SSCs (5.3±0.1logCFU/coupon) were completely inactivated within 1h at 25°C when treated with gaseous ClO2 (peak concentration: 115.3±5.0 parts per million [ppm]). The total number of vegetative cells and spores in biofilm formed by B. cereus on SSCs was 5.9±0.3logCFU/coupon; the spore count was 5.3±0.1logCFU/coupon. The vegetative cells and spores in biofilm were completely inactivated within 6h (peak concentration: 115.3±5.0ppm). Results show that B. cereus spores in biofilms are more resistant to gaseous ClO2 than are attached spores not in biofilms. Gaseous ClO2 was, nevertheless, very effective in killing B. cereus spores in biofilm on the surface of stainless steel. Results show promise for application of gaseous ClO2 to enhance the microbiological safety of foods that may come in contact with stainless steel and possibly other hard surfaces on which B. cereus biofilms have formed.

  4. 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.

  5. 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

  6. Total airborne mold particle sampling: evaluation of sample collection, preparation and counting procedures, and collection devices.

    PubMed

    Godish, Diana; Godish, Thad

    2008-02-01

    This study was conducted to evaluate (i) procedures used to collect, prepare, and count total airborne mold spore/particle concentrations, and (ii) the relative field performance of three commercially available total airborne mold spore/particle sampling devices. Differences between factory and laboratory airflow calibration values of axial fan-driven sampling instruments (used in the study) indicated a need for laboratory calibration using a mass flow meter to ensure that sample results were accurately calculated. An aniline blue-amended Calberla's solution adjusted to a pH of 4.2-4.4 provided good sample mounting/counting results using Dow Corning high vacuum grease, Dow Corning 280A adhesive, and Dow Corning 316 silicone release spray for samples collected using mini-Burkard and Allergenco samplers. Count variability among analysts was most pronounced in 5% counts of relatively low mold particle deposition density samples and trended downward with increased count percentage and particle deposition density. No significant differences were observed among means of 5, 10, and 20% counts and among analysts; a significant interaction effect was observed between analysts' counts and particle deposition densities. Significantly higher mini-Burkard and Air-O-Cell total mold spore/particle counts for 600x vs. 400x (1.9 and 2.3 x higher, respectively), 1000x vs. 600x (1.9 and 2.2 x higher, respectively) and 1000x vs. 400x (3.6 and 4.6 x higher, respectively) comparisons indicated that 1000x magnification counts best quantified total airborne mold spore/particles using light microscopy, and that lower magnification counts may result in unacceptable underreporting of airborne mold spore/particle concentrations. Modest but significantly higher (1.2x) total mold spore concentrations were observed with Allergenco vs. mini-Burkard samples collected in co-located, concurrently operated sampler studies; moderate but significantly higher mini-Burkard count values (1.4x) were

  7. 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.

  8. 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

  9. 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.

  10. [Microbiological research methods of drinking water regulation in West Germany from 1986. Suitability of the specifications of DIN 38411, Part 7, for the detection of sulfite-reducing, spore-forming anaerobes (Clostridia)].

    PubMed

    Schneider, J; Edenharder, R; Borneff, J

    1988-01-01

    The drinking-water regulations of the Federal Republic of Germany, from 22.05.1986, contains in paragraph 1 the instructions: "Drinking-water must be free of pathogens", and further in paragraph 11, "Responsibilities of the employer or other owner of a water supplying facility", include that: "The official authority may direct, that the employer...of a water supplying facility has to extend or has to cause to extend the microbiological examinations in order to determine, that...sulfite-reducing, spore-forming anaerobes (Clostridia) can not be detected in 20 ml of water..." The drinking-water regulations do not prescribe a bacteriological examination method in detail. Appendix 1 rules only that the examination for sulfite-reducing, spore-forming anaerobes (Clostridia) has to be performed after heating the sample to 75 degrees C (+/- 5 degrees C) for 10 min, by either the multiple-tube or membrane filtration method and cultivation in DRCM1-medium. If growth occurs, the presence of Clostridia must be confirmed by anaerobic and aerobic subcultivation. Furthermore, a DIN-instruction (DIN 38411, part 7) exists, which prescribes a detailed procedure for multiple-tube and membrane filtration methods, but does not provide for strict anaerobiosis. We were, however, unable to detect Clostridia in a multitude of water samples with the methods of the DIN-regulation. In order to examine if neglect of strict anaerobiosis was the reason for these failures, we checked the suitability of the DIN-regulation for the isolation of Clostridia from drinking water. In preliminary tests we examined up to four strains of the species C. botulinum, C. cadaveris, C. cochlearium, C. difficile, C. innocuum, C. perfringens and C. tertium for their ability to form heat-resistent spores in four sporulation media. It was, however, not possible to find a medium, in which all strains could sporulate within one week. In order to characterize the detection of these anaerobes in water, one particularly

  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. Bacillus thermoamylovorans Spores with Very-High-Level Heat Resistance Germinate Poorly in Rich Medium despite the Presence of ger Clusters but Efficiently upon Exposure to Calcium-Dipicolinic Acid

    PubMed Central

    Berendsen, Erwin M.; Krawczyk, Antonina O.; Klaus, Verena; de Jong, Anne; Boekhorst, Jos; Eijlander, Robyn T.

    2015-01-01

    High-level heat resistance of spores of Bacillus thermoamylovorans poses challenges to the food industry, as industrial sterilization processes may not inactivate such spores, resulting in food spoilage upon germination and outgrowth. In this study, the germination and heat resistance properties of spores of four food-spoiling isolates were determined. Flow cytometry counts of spores were much higher than their counts on rich medium (maximum, 5%). Microscopic analysis revealed inefficient nutrient-induced germination of spores of all four isolates despite the presence of most known germination-related genes, including two operons encoding nutrient germinant receptors (GRs), in their genomes. In contrast, exposure to nonnutrient germinant calcium-dipicolinic acid (Ca-DPA) resulted in efficient (50 to 98%) spore germination. All four strains harbored cwlJ and gerQ genes, which are known to be essential for Ca-DPA-induced germination in Bacillus subtilis. When determining spore survival upon heating, low viable counts can be due to spore inactivation and an inability to germinate. To dissect these two phenomena, the recoveries of spores upon heat treatment were determined on plates with and without preexposure to Ca-DPA. The high-level heat resistance of spores as observed in this study (D120°C, 1.9 ± 0.2 and 1.3 ± 0.1 min; z value, 12.2 ± 1.8°C) is in line with survival of sterilization processes in the food industry. The recovery of B. thermoamylovorans spores can be improved via nonnutrient germination, thereby avoiding gross underestimation of their levels in food ingredients. PMID:26341201

  14. Aerobic landfill bioreactor

    SciTech Connect

    Hudgins, Mark P; Bessette, Bernard J; March, John; McComb, Scott T.

    2000-01-01

    The present invention includes a method of decomposing municipal solid waste (MSW) within a landfill by converting the landfill to aerobic degradation in the following manner: (1) injecting air via the landfill leachate collection system (2) injecting air via vertical air injection wells installed within the waste mass; (3) applying leachate to the waste mass using a pressurized drip irrigation system; (4) allowing landfill gases to vent; and (5) adjusting air injection and recirculated leachate to achieve a 40% to 60% moisture level and a temperature between 120.degree. F. and 140.degree. F. in steady state.

  15. Aerobic landfill bioreactor

    SciTech Connect

    Hudgins, Mark P; Bessette, Bernard J; March, John C; McComb, Scott T.

    2002-01-01

    The present invention includes a system of decomposing municipal solid waste (MSW) within a landfill by converting the landfill to aerobic degradation in the following manner: (1) injecting air via the landfill leachate collection system (2) injecting air via vertical air injection wells installed within the waste mass; (3) applying leachate to the waste mass using a pressurized drip irrigation system; (4) allowing landfill gases to vent; and (5) adjusting air injection and recirculated leachate to achieve a 40% to 60% moisture level and a temperature between 120.degree. F. and 140.degree. F. in steady state.

  16. Aerobic landfill bioreactor

    SciTech Connect

    Hudgins, M.P.; Bessette, B.J.; March, J.; McComb, S.T.

    2000-02-15

    The present invention includes a method of decomposing municipal solid waste (MSW) within a landfill by converting the landfill to aerobic degradation in the following manner: (1) injecting air via the landfill leachate collection system (2) injecting air via vertical air injection wells installed within the waste mass; (3) applying leachate to the waste mass using a pressurized drip irrigation system; (4) allowing landfill gases to vent; and (5) adjusting air injection and recirculated leachate to achieve a 40% to 60% moisture level and a temperature between 120 F and 140 F in steady state.

  17. Effect of media, additives, and incubation conditions on the recovery of high pressure and heat-injured Clostridium botulinum spores.

    PubMed

    Reddy, N R; Tetzloff, R C; Skinner, G E

    2010-08-01

    The effect of additives and post-treatment incubation conditions on the recovery of high pressure and heat-injured (i.e., processed at 620 MPa and 95 and 100 degrees C for 5 min) spores of Clostridium botulinum strains, 62-A (proteolytic type A) and 17-B (nonproteolytic type B) was studied. High pressure and heat-injured spores were inoculated into TPGY (Trypticase-Peptone-Glucose-Yeast extract) anaerobic broth media containing additives (lysozyme, L-alanine, L-aspartic acid, dipicolonic acid, sodium bicarbonate, and sodium lactate) at various concentrations (0-10 microg/ml) individually or in combination. The spore counts of high pressure and heat-injured 62-A and 17-B recovered from TPGY broth containing lysozyme (10 microg/ml) incubated for 4 months versus that recovered from peptone-yeast extract-glucose-starch (PYGS) plating agar containing lysozyme (10 microg/ml) incubated under anaerobic conditions for 5 days were also compared. None of the additives either individually or in combination in TPGY broth improved recovery of injured spore enumeration compared to processed controls without additives. Addition of lysozyme at concentrations of 5 and 10 microg/ml in TPGY broth improved initial recovery of injured spores of 17-B during the first 4 days of incubation but did not result in additional recovery at the end of the 4 month incubation compared to the processed control without lysozyme. Adding lysozyme at a concentration of 10 microg/ml to PYGS plating agar resulted in no effect on the recovery of high pressure and heat-injured 62-A and 17-B spores. The recovery counts of high pressure and heat-injured spores of 62-A and 17-B were lower (i.e., <1.0 log units) with PYGS plating agar compared to the MPN method using TPGY broth as the growth medium.

  18. 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

  19. The exosporium of B. cereus contains a binding site for gC1qR/p33: implication in spore attachment and/or entry.

    PubMed

    Ghebrehiwet, Berhane; Tantral, Lee; Titmus, Mathew A; Panessa-Warren, Barbara J; Tortora, George T; Wong, Stanislaus S; Warren, John B

    2007-01-01

    B. cereus, is a member of a genus of aerobic, gram-positive, spore-forming rod-like bacilli, which includes the deadly, B. anthracis. Preliminary experiments have shown that gC1qR binds to B. cereus spores that have been attached to microtiter plates. The present studies were therefore undertaken, to examine if cell surface gC1qR plays a role in B. cereus spore attachment and/or entry. Monolayers of human colon carcinoma (Caco-2) and lung cells were grown to confluency on 6 mm coverslips in shell vials with gentle swirling in a shaker incubator. Then, 2 microl of a suspension of strain SB460 B. cereus spores (3x10(8)/ml, in sterile water), were added and incubated (1-4 h; 36 degrees C) in the presence or absence of anti-gC1qR mAb-carbon nanoloops. Examination of these cells by EM revealed that: (1) When B. cereus endospores contacted the apical Caco-2 cell surface, or lung cells, gC1qR was simultaneously detectable, indicating upregulation of the molecule. (2) In areas showing spore contact with the cell surface, gC1qR expression was often adjacent to the spores in association with microvilli (Caco-2 cells) or cytoskeletal projections (lung cells). (3) Furthermore, the exosporia of the activated and germinating spores were often decorated with mAb-nanoloops. These observations were further corroborated by experiments in which B.cereus spores were readily taken up by monocytes and neutrophils, and this uptake was partially inhibited by mAb 60.11, which recognizes the C1q binding site on gC1qR. Taken together, the data suggest a role, for gC1qR at least in the initial stages of spore attachment and/or entry.

  20. The Exosporium of B.cereus Contains a Binding Site for gC1qR/p33: Implication in Spore Attachment and/or Entry.

    SciTech Connect

    GHEBREHIWET,B.; TANTRAL, L.; TITMUS, M.A.; PANESSA-WARREN, B.J.; TORTORA, G.T.; WONG, S.S.; WARREN, J.B.

    2008-01-01

    B. cereus, is a member of a genus of aerobic, gram-positive, spore-forming rod-like bacilli, which includes the deadly, B. anthracis. Preliminary experiments have shown that gC1qR binds to B.cereus spores that have been attached to microtiter plates. The present studies were therefore undertaken, to examine if cell surface gC1qR plays a role in B.cereus spore attachment and/or entry. Monolayers of human colon carcinoma (Caco-2) and lung cells were grown to confluency on 6 mm coverslips in shell vials with gentle swirling in a shaker incubator. Then, 2 {micro}l of a suspension of strain SB460 B.cereus spores (3x10{sup 8}/ml, in sterile water), were added and incubated (1-4 h; 36{sup 0} C) in the presence or absence of anti-gC1qR mAb-carbon nanoloops. Examination of these cells by EM revealed that: (1) When B. cereus endospores contacted the apical Caco-2 cell surface, or lung cells, gClqR was simultaneously detectable, indicating upregulation of the molecule. (2) In areas showing spore contact with the cell surface, gClqR expression was often adjacent to the spores in association with microvilli (Caco-2 cells) or cytoskeletal projections (lung cells). (3) Furthermore, the exosporia of the activated and germinating spores were often decorated with mAb-nanoloops. These observations were further corroborated by experiments in which B.cereus spores were readily taken up by monocytes and neutrophils, and this uptake was partially inhibited by mAb 60.11, which recognizes the C1q binding site on gC1qR. Taken together, the data suggest a role, for gC1qR at least in the initial stages of spore attachment and/or entry.

  1. The Big Pumpkin Count.

    ERIC Educational Resources Information Center

    Coplestone-Loomis, Lenny

    1981-01-01

    Pumpkin seeds are counted after students convert pumpkins to jack-o-lanterns. Among the activities involved, pupils learn to count by 10s, make estimates, and to construct a visual representation of 1,000. (MP)

  2. 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.

  3. 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.

  4. 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

  5. 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

  6. 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

  7. Exercise, Animal Aerobics, and Interpretation?

    ERIC Educational Resources Information Center

    Oliver, Valerie

    1996-01-01

    Describes an aerobic activity set to music for children that mimics animal movements. Example exercises include walking like a penguin or jumping like a cricket. Stresses basic aerobic principles and designing the program at the level of children's motor skills. Benefits include reaching people who normally don't visit nature centers, and bridging…

  8. Seasonal occurrence and molecular diversity of clostridia species spores along cheesemaking streams of 5 commercial dairy plants.

    PubMed

    Bermúdez, Jorge; González, Marcela J; Olivera, Jorge A; Burgueño, Juan A; Juliano, Pablo; Fox, Edward M; Reginensi, Stella M

    2016-05-01

    Five commercial dairy plants were monitored over a 17-mo period to determine the seasonal occurrence of Clostridium spores in streams from the cheesemaking process. Every 2 mo, samples of raw milk (RM), separated cream (SC), pasteurized and standardized vat milk (PSVM), PSVM + lysozyme (PSVM+L), and manufactured cheese aged for 60 to 90 d were processed for analysis. Molecular diversity of the main species identified was determined using repetitive element palindromic PCR. The mean anaerobic spore counts (μ ± SE) were 3.16±0.054, 3.00±0.054, 2.89±0.059, and 2.03±0.054 log10 most probable number/L for RM, PSVM, PSVM+L, and SC, respectively. Although spore counts did not differ between dairy plants, seasonal variation was observed; spore counts of RM, PSVM, and PSVM+L were higher during winter (June to August) and summer (December to February) months, but no seasonal variation was seen in SC counts. The most frequently isolated species was Clostridium tyrobutyricum, ranging from 50 to 58.3% of isolates from milk and cream samples. Clostridium sporogenes was the second most common species identified (16.7-21.1%); Clostridium beijerinckii and Clostridium butyricum were also found, although at lower prevalence (7.9-13.2%). Analysis of the C. tyrobutyricum and C. sporogenes population structure through repetitive element palindromic PCR indicated a high diversity, with unique isolates found in each positive sample. The occurrence of Clostridia spores in incoming streams to cheesemaking was most prominent in the winter and summer seasons, with higher prevalence of C. tyrobutyricum in the months of June and August.

  9. 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.

  10. 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.

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

    PubMed

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

    2015-01-01

    Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger maximizes the diameter of the vesicle and the spore chain length. The length and width of the conidiophore stalk and vesicle were reduced nearly twofold in a ΔveA strain. The latter implies a fourfold reduced surface area to develop chains of spores. Over and above this, the conidial chain length was approximately fivefold reduced. The calculated 20-fold reduction in formation of conidia by ΔveA fits the 8- to 17-fold decrease in counted spore numbers. Notably, morphology of the ΔveA conidiophores of A. niger was very similar to that of wild-type Aspergillus sydowii. This suggests that VeA is key in conidiophore architecture diversity in the fungal kingdom. The finding that biomass formation of the A. niger ΔveA strain was reduced twofold shows that VeA not only impacts dispersion capacity but also colonization capacity of A. niger.

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

    PubMed

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

    2015-06-01

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

  13. Regional-scale simulations of fungal spore aerosols using an emission parameterization adapted to local measurements of fluorescent biological aerosol particles

    NASA Astrophysics Data System (ADS)

    Hummel, M.; Hoose, C.; Gallagher, M.; Healy, D. A.; Huffman, J. A.; O'Connor, D.; Pöschl, U.; Pöhlker, C.; Robinson, N. H.; Schnaiter, M.; Sodeau, J. R.; Toprak, E.; Vogel, H.

    2014-04-01

    Fungal spores as a prominent type of primary biological aerosol particles (PBAP) have been incorporated into the COSMO-ART regional atmospheric model, using and comparing three different emission parameterizations. Two literature-based emission rates derived from fungal spore colony counts and chemical tracer measurements were used as a parameterization baseline for this study. A third, new emission parameterization was adapted to field measurements of fluorescent biological aerosol particles (FBAP) from four locations across Northern Europe. FBAP concentrations can be regarded as a lower estimate of total PBAP concentrations. Size distributions of FBAP often show a distinct mode at approx. 3 μm, corresponding to a diameter range characteristic for many fungal spores. Previous studies have suggested the majority of FBAP in several locations are dominated by fungal spores. Thus, we suggest that simulated fungal spore concentrations obtained from the emission parameterizations can be compared to the sum of total FBAP concentrations. A comparison reveals that parameterized estimates of fungal spore concentrations based on literature numbers underestimate measured FBAP concentrations. In agreement with measurement data, the model results show a diurnal cycle in simulated fungal spore concentrations, which may develop partially as a consequence of a varying boundary layer height between day and night. Measured FBAP and simulated fungal spore concentrations also correlate similarly with simulated temperature and humidity. These meteorological variables, together with leaf area index, were chosen to drive the new emission parameterization discussed here. Using the new emission parameterization on a model domain covering Western Europe, fungal spores in the lowest model layer comprise a fraction of 15% of the total aerosol mass over land and reach average number concentrations of 26 L-1. The results confirm that fungal spores and biological particles may account for a

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. 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

  19. 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.

  20. Effects of muscle strengthening versus aerobic exercise program in fibromyalgia.

    PubMed

    Bircan, Ciğdem; Karasel, Seide Alev; Akgün, Berrin; El, Ozlem; Alper, Serap

    2008-04-01

    The purpose of this study was to compare the effects of aerobic training with a muscle-strengthening program in patients with fibromyalgia. Thirty women with fibromyalgia were randomized to either an aerobic exercise (AE) program or a strengthening exercise (SE) program for 8 weeks. Outcome measures included the intensity of fibromyalgia-related symptoms, tender point count, fitness (6-min walk distance), hospital anxiety and depression (HAD) scale, and short-form health survey (SF-36). There were significant improvements in both groups regarding pain, sleep, fatigue, tender point count, and fitness after treatment. HAD-depression scores improved significantly in both groups while no significant change occurred in HAD-anxiety scores. Bodily pain subscale of SF-36 and physical component summary improved significantly in the AE group, whereas seven subscales of SF-36, physical component summary, and mental component summary improved significantly in the SE group. When the groups were compared after treatment, there were no significant differences in pain, sleep, fatigue, tender point count, fitness, HAD scores, and SF-36 scores. AE and SE are similarly effective at improving symptoms, tender point count, fitness, depression, and quality of life in fibromyalgia.

  1. Occurrence of Cladosporium spp. and Alternaria spp. spores in Western, Northern and Central-Eastern Poland in 2004-2006 and relation to some meteorological factors

    NASA Astrophysics Data System (ADS)

    Grinn-Gofroń, Agnieszka; Rapiejko, Piotr

    2009-08-01

    The concentration of airborne spores of Cladosporium spp. and Alternaria spp. has been investigated at three monitoring stations situated along the west-north and central-east transect in Poland (Szczecin, Olsztyn, Warszawa,) i.e. from a height of 100 m to 149 m above sea level. The aerobiological monitoring of fungal spores was performed by means of three Lanzoni volumetric spore traps. Cladosporium spp. spores were dominant at all the stations. The highest Cladosporium spp. and Alternaria spp. numbers of spores were observed at all the cities in July and August. Statistically significant correlations have been found between the Cladosporium spp. and Alternaria spp. concentration in the air and the mean air temperature, amount of precipitation, air pressure and relative air humidity. The spore count of Cladosporium spp. and Alternaria spp. is determined by the diversity of local flora and weather conditions, especially by the air temperature. The identification of factors, which influence and shape spore concentrations, may significantly improve the current methods of allergy prevention.

  2. 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

  3. 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).

  4. 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

  5. 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.

  6. Inactivation effects of neutral reactive-oxygen species on Penicillium digitatum spores using non-equilibrium atmospheric-pressure oxygen radical source

    NASA Astrophysics Data System (ADS)

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

    2013-10-01

    The effectiveness of atomic and excited molecular oxygen species at inactivating Penicillium digitatum spores was quantitatively investigated by measuring these species and evaluating the spore inactivation rate. To avoid the effects of ultraviolet light and charged species, a non-equilibrium atmospheric-pressure radical source, which supplies only neutral radicals, was employed. Ground-state atomic oxygen (O(3Pj)) and excited molecular oxygen (O2(1Δg)) species were measured using vacuum ultraviolet absorption spectroscopy. The inactivation rate of spores was evaluated using the colony count method. The lifetimes of O(3Pj) and O2(1Δg) in an argon gas ambient at atmospheric pressure were found to be about 0.5 ms and much more than tens of ms, and their spore inactivation rates were about 10-17 cm3 s-1 and much lower than 10-21 cm3 s-1, respectively.

  7. Spore population, colonization, species diversity and factors influencing the association of arbuscular mycorrhizal fungi with litchi trees in India.

    PubMed

    Kumar, Vinod; Kumar, Rajesh; Kumar, Ajit; Anal, Dubedi

    2016-01-01

    Abundance and diversity of arbuscular mycorrhizal fungi (AMF) in association with litchi (Litchi chinensis Sonn.) trees were studied during 2012-2013, where orchard soil had high pH (7.42-9.53) and salinity (0.07- 0.39 dSm(-1)). A total of 105 rhizospheric soil and root samples were collected considering variables like location, age of tree, cultivar and production management. Results showed that spore count was in the range of 1-22 g(-1) soil. All the examined root segments had colonization of AMF, which ranged between 3.3 to 90.0%. AMF community comprised of Glomus mosseae, G. intaradices, G. constricta, G. coronatum, G. fasciculatum, G. albidum, G. hoi, G. multicauli, Acaulospora scrobiculata, A. laevis, Rhizophagus litchi and Entrophosphora infrequens. Higher spore density and AMF colonization were observed at medium level (13-28 kg ha(-1)) of available phosphorus that decreased ('r' = -0.21 for spore density, -0.48 for root colonization) with increasing soil phosphorus. While nitrogen did not influence the AMF association, a weak negative linear relationship with AMF colonization ('r' = -0.30) was apparent in the medium level (112-200 kg ha(-1)) of potash. Micronutrients (Zn, Fe, Cu, Mn and B) did not affect spore density (zero or a very weak linear correlation) but influenced root colonization ('r' = -0.53 to -0.44), the effect being more prominent above critical limits. Nutritionally sufficient, irrigated litchi orchards had greater spore count (46% samples having 5-22 spores g(-1) soil) and colonization (> 50% in 37.4% roots examined) than nutrient deficient, non-irrigated orchards, indicating essentiality of a threshold nutrients and moisture regime for the association. AMF symbiosis was influenced by cultivar (greater in 'China'), but tree age was not correlated to mycorrhizal association. A consortium of native species coupled with the understanding of nutrient effects on AMF would be useful for field application in litchi.

  8. Rapid counting of liquid-borne microorganisms by light scattering spectrometry.

    PubMed

    Mainelis, Gediminas; Górny, Rafał; Willeke, Klaus; Reponen, Tiina

    2005-01-01

    Fast and sensitive techniques are needed to determine microorganism presence in liquid samples. In this research, the feasibility of using light scattering spectrometry for enumerating the biological particles in liquid samples was investigated. A particle size spectrometer was used to count six commonly found microbial species suspended in liquid with and without microbiological stains applied: Pseudomonas fluorescens, Micrococcus spp. vegetative cells and Bacillus subtilis var. niger endospores were stained with Acridine Orange and Crystal Violet, while Cladosporium cladosporioides, Penicillium melinii and Aspergillus versicolor fungi were stained with Acridine Orange and Lactophenol Cotton Blue. The counts obtained with the spectrometer were compared with those obtained with a phase-contrast microscope. It was found that the spectrometer counted about 32 % of non-stained B. subtilis endospores and this percentage increased to almost 90 % for stained endospores. Among the investigated species of fungi, the counting efficiency of P. melinii was the only one significantly affected by the application of the stain Lactophenol Cotton Blue: the fraction of counted fungal spores increased from 64 % (non-stained spores) to about 100 % (stained spores). The observed difference in counting efficiency may serve as a basis for differentiating biological from non-biological particles in liquid samples.

  9. Mid-infrared versus far-infrared (THz) relative intensities of room-temperature Bacillus spores

    NASA Astrophysics Data System (ADS)

    Johnson, Timothy J.; Valentine, Nancy B.; Sharpe, Steven W.

    2005-02-01

    We have simultaneously recorded the mid-IR and far-IR (a.k.a. terahertz, THz) spectra of the sporulated form of five Bacillus bacteria: Bacillus subtilis ATCC 49760, B. subtilis ATCC 6051, Bacillus thuringiensis subsp. kurstaki ATCC 35866, Bacillus globigii 01, and Bacillus atrophaeus 49337. The 295 K spectra were recorded from ˜8 to 6000 cm -1 using spore counts on the order of 10 9 deposited onto windows transparent in both the mid- and far-infrared. The results indicate that any room-temperature THz absorption features due to the spores are at least 28 times weaker (based on p-p noise) than the corresponding mid-IR amide I band.

  10. Use of aerobic spores as a surrogate for cryptosporidium oocysts in drinking water and supplies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Waterborne illnesses are a growing concern among health agencies worldwide and regulatory efforts to prevent microbial contamination of water supplies are constantly evolving to stay ahead of the threat. The United States Environmental Protection Agency has established several rules to combat the co...

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. Potential for growth of Clostridium perfringens from spores in pork scrapple during cooling.

    PubMed

    Juneja, Vijay K; Porto-Fett, Anna C S; Gartner, Kelly; Tufft, Linda; Luchansky, John B

    2010-02-01

    We conducted stabilization studies to determine the ability of Clostridium perfringens spores to germinate and grow during exponential cooling of a commercial formulation of pork scrapple. Scrapple was inoculated with a mixture of three strains of C. perfringens spores (NTCC 8238, NCTC 8239, and ATCC 10288), vacuum packaged, and reheated (20 min/93.3 degrees C) in a circulating water bath. The cooked samples were cooled (30 s) in an ice bath before being transferred to a programmable water bath to cool through the temperature range of 54.4 degrees C to 7.2 degrees C in 12, 14, or 21 h to simulate deviations from the required cooling time of 6.5 h. After cooling, the samples, in duplicate, were analyzed to determine if growth from spores had occurred. The samples were plated onto tryptose-sulfite-cycloserine agar and incubated anaerobically at 37 degrees C for 48 h before counting the colonies. Minimal growth (less than 1.0 log) was observed during a 12- or 14 h cooling period. However, when the time to achieve 7.2 degrees C was extended to 21 h, C. perfringens spores germinated and grew from an inoculum of approximately 3.0 log(10) to approximately 7.8 log(10) CFU/g. Thus, scrapple must be cooled after cooking to 7.2 degrees C within 6.5 h, but for no more than 14 h, to prevent a food safety hazard from outgrowth of C. perfringens spores during cooling.

  2. Survival and germination of Clostridium perfringens spores during heating and cooling of ground pork.

    PubMed

    Márquez-González, M; Cabrera-Díaz, E; Hardin, M D; Harris, K B; Lucia, L M; Castillo, A

    2012-04-01

    The effect of heating rate on the heat resistance, germination, and outgrowth of Clostridium perfringens spores during cooking of cured ground pork was investigated. Inoculated cured ground pork portions were heated from 20 to 75°C at a rate of 4, 8, or 12°C/h and then held at 75°C for 48 h. No significant differences (P > 0.05) in the heat resistance of C. perfringens spores were observed in cured ground pork heated at 4, 8, or 12°C/h. At heating rates of 8 and 12°C/h, no significant differences in the germination and outgrowth of spores were observed (P > 0.05). However, when pork was heated at 4°C/h, growth of C. perfringens occurred when the temperature of the product was between 44 and 56°C. In another set of experiments, the behavior of C. perfringens spores under temperature abuse conditions was studied in cured and noncured ground pork heated at 4°C/h and then cooled from 54.4 to 7.2°C within 20 h. Temperature abuse during cooling of noncured ground pork resulted in a 2.8-log CFU/g increase in C. perfringens. In cured ground pork, C. perfringens decreased by 1.1 log CFU/g during cooling from 54.4 to 36.3°C and then increased by 0.9 log CFU/g until the product reached 7.2°C. Even when the initial level of C. perfringens spores in cured ground pork was 5 log CFU/g, the final counts after abusive cooling did not exceed 3.4 log CFU/g. These results suggest that there is no risk associated with C. perfringens in cured pork products under the tested conditions.

  3. Warm and dry weather accelerates and elongates Cladosporium spore seasons in Poland.

    PubMed

    Kasprzyk, Idalia; Kaszewski, Boguslaw Michal; Weryszko-Chmielewska, Elzbieta; Nowak, Malgorzata; Sulborska, Aneta; Kaczmarek, Joanna; Szymanska, Agata; Haratym, Weronika; Jedryczka, Malgorzata

    Temperature is the environmental factor that systematically changes for decades and, as in plants and animals, can significantly affect the growth and development of fungi, including the abundance of their sporulation. During the time of study (2010-2012), a rapid increase in air temperature was observed in Poland, which coincided with the substantial decrease in rainfall. The increase in annual mean temperatures at three monitoring sites of this study was 0.9 °C in Lublin and Rzeszow (east Poland) and 2.0 °C in Poznan (west Poland). Such warming of air masses was comparable to the average global air temperature rise in the period of 1880-2012 accounting for 0.85 °C, as reported by the Intergovernmental Panel on Climate Change. Moreover, there was a substantial decrease in rainfall, ranging from 32.7 % (Poznan) to 43.0 % (Rzeszow). We have demonstrated that under such conditions the mean and median values of total Cladosporium spore counts significantly increased and the spore seasons were greatly accelerated. Moreover, earlier start and later end of the season caused its extension, lasting from over 20 days in Rzeszow to around 60 days in Lublin and Poznan, when the cumulative amount of 5-95 % of spores was considered. The time of reaching the cumulative amount of 50 % of spores was up to 25 days earlier (difference in Poznan between 2010 and 2012). There was also a striking acceleration of the date of the maximal Cladosporium spore concentration per cubic metre of air (26 days for Lublin, 43 for Poznan and 56 for Rzeszow).

  4. 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

  5. Outgrowth inhibition of Clostridium beijerinckii spores by a bacteriocin-producing lactic culture in ovine milk cheese.

    PubMed

    Garde, Sonia; Avila, Marta; Arias, Ramón; Gaya, Pilar; Nuñez, Manuel

    2011-10-17

    In the manufacture of model cheeses, ovine milk was deliberately contaminated with spores of Clostridium beijerinckii INIA 63, a wild isolate from Manchego cheese with late blowing defect, and inoculated with nisin- and lacticin 481-producing Lactococcus lactis subsp. lactis INIA 415 as starter, to test its potential to prevent the late blowing defect, or with L. lactis subsp. lactis INIA 415-2, a spontaneous mutant not producing bacteriocins. Cheeses made individually with the lactococcal strains, without clostridial spores, served as controls. Cheese made with clostridial spores and L. lactis subsp. lactis INIA 415-2 showed late blowing defect after 120days of ripening. Spoilt cheese also showed lower concentrations of lactic acid, and higher levels of acetic, propionic and butyric acids, and of other volatile compounds such as 2-propanol and 1-butanol, than control cheese. In addition, cheese made with the bacteriocin producer did not show any late blowing symptoms, despite its spore counts similar to those of blown cheese, pointing to outgrowth inhibition of C. beijerinckii spores by bacteriocins. Besides, cheese made with the bacteriocin producer showed similar concentrations of lactic acid and volatile compounds than control cheese. Inclusion of L. lactis subsp. lactis INIA 415 in starter cultures seems a feasible method to prevent late blowing defect in cheese without altering its sensory characteristics.

  6. 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

  7. Averaging of TNTC counts.

    PubMed Central

    Haas, C N; Heller, B

    1988-01-01

    When plate count methods are used for microbial enumeration, if too-numerous-to-count results occur, they are commonly discarded. In this paper, a method for consideration of such results in computation of an average microbial density is developed, and its use is illustrated by example. PMID:3178211

  8. Anarthria impairs subvocal counting.

    PubMed

    Cubelli, R; Nichelli, P; Pentore, R

    1993-12-01

    We studied subvocal counting in two pure anarthric patients. Analysis showed that they performed definitively worse than normal subjects free to articulate subvocally and their scores were in the lower bounds of the performances of subjects suppressing articulation. These results suggest that subvocal counting is impaired after anarthria.

  9. Complexities of Counting.

    ERIC Educational Resources Information Center

    Stake, Bernadine Evans

    This document focuses on one child's skip counting methods. The pupil, a second grade student at Steuben School, in Kankakee, Illinois, was interviewed as she made several attempts at counting twenty-five poker chips on a circular piece of paper. The interview was part of a larger study of "Children's Conceptions of Number and Numeral,"…

  10. Counting Sheep in Basque

    ERIC Educational Resources Information Center

    Araujo, Frank P.

    1975-01-01

    Demonstrates the interplay of a cognitive system, the Basque numerative system, and a behavioral one, counting sheep. The significant features of the Basque numerative system are analyzed; then it is shown how use of these features facilitates the counting of sheep on open ranges by Basque sheep farmers in California. (Author/RM)

  11. 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

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. 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

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. AUTOMATIC COUNTING APPARATUS

    DOEpatents

    Howell, W.D.

    1957-08-20

    An apparatus for automatically recording the results of counting operations on trains of electrical pulses is described. The disadvantages of prior devices utilizing the two common methods of obtaining the count rate are overcome by this apparatus; in the case of time controlled operation, the disclosed system automatically records amy information stored by the scaler but not transferred to the printer at the end of the predetermined time controlled operations and, in the case of count controlled operation, provision is made to prevent a weak sample from occupying the apparatus for an excessively long period of time.

  2. 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

  3. 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.

  4. Evaluation of Petrifilm method for enumerating aerobic bacteria in Crottin goat cheese.

    PubMed

    de Sousa, G B; Tamagnini, L M; González, R D; Budde, C E

    2005-01-01

    The Petrifilm Aerobic Count Plate (ACP) developed by 3M laboratories, is a ready-to-use culture medium system, useful for the enumeration of aerobic bacteria in food. Petrifilm was compared with a standard method in several different food products with satisfactory results. However, many studies showed that bacterial counts in Petrifilm were significantly lower than those obtained with conventional methods in fermented food. The purpose of this study was to compare the Petrifilm method for enumerating aerobic bacteria with a conventional method (PCA) in Crottin goat's cheese. Thirty samples were used for the colony count. The mean count and standard deviation were 7.18 +/- 1.17 log CFU g(-1) on PCA and 7.11 +/- 1.05 log CFU g(-1) on Petrifilm. Analysis of variance revealed no significant differences between both methods (t = 1.33, P = 0.193). The Pearson correlation coefficient (0.971, P = 0.0001) indicated a strong linear relationship between the Petrifilm and the standard method. The results showed that Petrifilm is suitable and a convenient alternative to this standard method for the enumeration of aerobic flora in goat soft cheese.

  5. Understanding Blood Counts

    MedlinePlus

    ... Heart, Kidneys, Liver and Lung Function Infections Iron Overload Low Blood Counts Pain Disease- and Treatment-Related ... cancer research around the world and provides free information and support services. Privacy Policy Security Copyright Link ...

  6. Blood Count Tests

    MedlinePlus

    ... white blood cells (WBC), and platelets. Blood count tests measure the number and types of cells in ... helps doctors check on your overall health. The tests can also help to diagnose diseases and conditions ...

  7. Calorie count - Fast food

    MedlinePlus

    ... count - fast food FOOD ITEM SERVING SIZE CALORIES Breakfast Foods Dunkin Donuts Egg White Veggie Wrap 1 ... Cheese Biscuit Sandwich 1 sandwich 510 BK Ultimate Breakfast Platter 1 platter 1190 McDonalds Fruit 'n Yogurt ...

  8. Carbohydrate Counting and Diabetes

    MedlinePlus

    ... are the other main nutrients. Carbohydrates include sugars, starches, and fiber. Carbohydrate counting can help you control ... called starchy vegetables because they are high in starch. These vegetables have more carbohydrates per serving than ...

  9. Inventory count strategies.

    PubMed

    Springer, W H

    1996-02-01

    An important principle of accounting is that asset inventory needs to be correctly valued to ensure that the financial statements of the institution are accurate. Errors is recording the value of ending inventory in one fiscal year result in errors to published financial statements for that year as well as the subsequent fiscal year. Therefore, it is important that accurate physical counts be periodically taken. It is equally important that any system being used to generate inventory valuation, reordering or management reports be based on consistently accurate on-hand balances. At the foundation of conducting an accurate physical count of an inventory is a comprehensive understanding of the process coupled with a written plan. This article presents a guideline of the physical count processes involved in a traditional double-count approach.

  10. 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.

  11. 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

  12. Developmental intestinal aerobic microflora in the kori bustard (Ardeotis kori).

    PubMed

    Naldo, J L; Silvanose, C D; Samour, J H; Bailey, T A

    1998-01-01

    A study was carried out to investigate the normal aerobic bacterial flora of developing kori bustard (Ardeotis kori) chicks, captive bred at the National Avian Research Center, Abu Dhabi, United Arab Emirates. Faecal samples were collected from 14 birds at different ages from the first day of hatching until 99 days old and were cultured for aerobic bacteria. Several bacterial species were isolated from the cultures, they included Escherichia coli, Streptococcus viridians, Enterococcus faecalis, Klebsiella oxytoca, Proteus spp., Enterobacter, spp. and Serratia marcescens. Gram-negative bacilli were isolated from all but one of the faecal samples collected. They were also the predominant bacteria, accounting for between 55.6 and 73.4% of the mean colony count of faecal cultures from all age groups. E. coli was the most frequently isolated bacteria, the frequency and mean colony count increased as the birds grew older. Gram-positive cocci were isolated from between 50 and 100% of the faecal samples from all age groups, and they accounted for between 26.6 and 44.4% of the mean colony count. Results from this study indicated that Gram-negative bacilli and Gram-positive cocci can be isolated frequently from the faeces of developing, clinically normal, captive bred kori bustard chicks.

  13. 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

  14. 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

  15. 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.

  16. Neutron counting with cameras

    SciTech Connect

    Van Esch, Patrick; Crisanti, Marta; Mutti, Paolo

    2015-07-01

    A research project is presented in which we aim at counting individual neutrons with CCD-like cameras. We explore theoretically a technique that allows us to use imaging detectors as counting detectors at lower counting rates, and transits smoothly to continuous imaging at higher counting rates. As such, the hope is to combine the good background rejection properties of standard neutron counting detectors with the absence of dead time of integrating neutron imaging cameras as well as their very good spatial resolution. Compared to Xray detection, the essence of thermal neutron detection is the nuclear conversion reaction. The released energies involved are of the order of a few MeV, while X-ray detection releases energies of the order of the photon energy, which is in the 10 KeV range. Thanks to advances in camera technology which have resulted in increased quantum efficiency, lower noise, as well as increased frame rate up to 100 fps for CMOS-type cameras, this more than 100-fold higher available detection energy implies that the individual neutron detection light signal can be significantly above the noise level, as such allowing for discrimination and individual counting, which is hard to achieve with X-rays. The time scale of CMOS-type cameras doesn't allow one to consider time-of-flight measurements, but kinetic experiments in the 10 ms range are possible. The theory is next confronted to the first experimental results. (authors)

  17. 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.

  18. 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.

  19. Die aerobe Glykolyse der Tumorzelle

    NASA Astrophysics Data System (ADS)

    Schneider, Friedhelm

    1981-01-01

    A high aerobic glycolysis (aerobic lactate production) is the most significant feature of the energy metabolism of rapidly growing tumor cells. Several mechanisms, which may be different in different cell lines, seem to be involved in this characteristic of energy metabolism of the tumor cell. Changes in the cell membrane leading to increased uptake and utilization of glucose, a high level of fetal types of isoenzymes, a decreased number of mitochondria and a reduced capacity to metabolize pyruvate are some factors which must be taken into consideration. It is not possible to favour one of them at the present time.

  20. [Evaluation of normal aerobic skin flora (author's transl)].

    PubMed

    Crémieux, A; Cazac, J L

    1980-01-01

    This work attempts the quantitative and qualitative evaluation of the bacterial population from two different areas: elbow and groin. Bacteria are recovered using the method of Williamson and Kligman modified by Fleurette and Transy. Aerobic flora is determined from bacterial counts on various media. Results show a density of 475 to 630 bacteria/cm2 for elbow, and 1.9 to 2.4 X 10(5) bacteria/cm2 for groin (geometric and arithmetic mean, respectively). Percentages of different species and types are calculated, and skin population is represented by a circular diagram.

  1. The Transition from Aerobic to Anaerobic Metabolism.

    ERIC Educational Resources Information Center

    Skinner, James S.; McLellan, Thomas H.

    1980-01-01

    The transition from aerobic to anaerobic metabolism is discussed. More research is needed on different kinds of athletes and athletic activities and how they may affect aerobic and anaerobic metabolisms. (CJ)

  2. Culturability of Bacillus spores on aerosol collection filters exposed to airborne combustion products of Al, Mg, and B·Ti.

    PubMed

    Adhikari, Atin; Yermakov, Michael; Indugula, Reshmi; Reponen, Tiina; Driks, Adam; Grinshpun, Sergey A

    2016-05-01

    Destruction of bioweapon facilities due to explosion or fire could aerosolize highly pathogenic microorganisms. The post-event air quality assessment is conducted through air sampling. A bioaerosol sample (often collected on a filter for further culture-based analysis) also contains combustion products, which may influence the microbial culturability and, thus, impact the outcome. We have examined the interaction between spores deposited on collection filters using two simulants of Bacillus anthracis [B. thuringiensis (Bt) and B. atrophaeus (referred to as BG)] and incoming combustion products of Al as well as Mg and B·Ti (common ingredient of metalized explosives). Spores extracted from Teflon, polycarbonate, mixed cellulose ester (MCE), and gelatin filters (most common filter media for bioaerosol sampling), which were exposed to combustion products during a short-term sampling, were analyzed by cultivation. Surprisingly, we observed that aluminum combustion products enhanced the culturability of Bt (but not BG) spores on Teflon filters increasing the culturable count by more than an order of magnitude. Testing polycarbonate and MCE filter materials also revealed a moderate increase of culturability although gelatin did not. No effect was observed with either of the two species interacting on either filter media with products originated by combustion of Mg and B·Ti. Sample contamination, spore agglomeration, effect of a filter material on the spore survival, changes in the spore wall ultrastructure and germination, as well as other factors were explored to interpret the findings. The study raises a question about the reliability of certain filter materials for collecting airborne bio-threat agents in combustion environments.

  3. Fast counting electronics for neutron coincidence counting

    DOEpatents

    Swansen, James E.

    1987-01-01

    An amplifier-discriminator is tailored to output a very short pulse upon an above-threshold input from a detector which may be a .sup.3 He detector. The short pulse output is stretched and energizes a light emitting diode (LED) to provide a visual output of operation and pulse detection. The short pulse is further fed to a digital section for processing and possible ORing with other like generated pulses. Finally, the output (or ORed output ) is fed to a derandomizing buffer which converts the rapidly and randomly occurring pulses into synchronized and periodically spaced-apart pulses for the accurate counting thereof. Provision is also made for the internal and external disabling of each individual channel of amplifier-discriminators in an ORed plurality of same.

  4. Fast counting electronics for neutron coincidence counting

    DOEpatents

    Swansen, J.E.

    1985-03-05

    An amplifier-discriminator is tailored to output a very short pulse upon an above-threshold input from a detector which may be a /sup 3/He detector. The short pulse output is stretched and energizes a light emitting diode (LED) to provide a visual output of operation and pulse detection. The short pulse is further fed to a digital section for processing and possible ORing with other like generated pulses. Finally, the output (or ORed output) is fed to a derandomizing buffer which converts the rapidly and randomly occurring pulses into synchronized and periodically spaced-apart pulses for the accurate counting thereof. Provision is also made for the internal and external disabling of each individual channel of amplifier-discriminators in an ORed plurality of same.

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

    PubMed

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

    2010-05-01

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

  6. Feeding horses with industrially manufactured pellets with fungal spores to promote nematode integrated control.

    PubMed

    Hernández, José Ángel; Arroyo, Fabián Leonardo; Suárez, José; Cazapal-Monteiro, Cristiana Filipa; Romasanta, Ángel; López-Arellano, María Eugenia; Pedreira, José; de Carvalho, Luis Manuel Madeira; Sánchez-Andrade, Rita; Arias, María Sol; de Gives, Pedro Mendoza; Paz-Silva, Adolfo

    2016-10-15

    The usefulness of pellets industrially manufactured with spores of parasiticide fungi as a contribution to integrated nematode control was assessed in grazing horses throughout sixteen months. Two groups of 7 Pura Raza Galega autochthonous horses (G-T and G-P) were dewormed pour-on (1mg Ivermectin/kg bw) at the beginning of the trial, and other group (G-C) remained untreated. The G-P was provided daily with commercial pellets to which was added a mixture of fungal spores during the industrial manufacturing (2×10(6) spores of Mucor circinelloides and same dose of Duddingtonia flagrans/kg), and G-T and G-C received pellets without spores. The efficacy of the parasiticidal strategy was assessed by estimating the reduction in the faecal egg counts (FECR) and in the number of horses shedding eggs in the faeces (PHR), and also the egg reappearance periods (ERP). Blood analyses were performed to identify the changes in the red and white cell patterns. To ascertain if horses developed an IgG humoral response against the fungi, antigenic products collected from M. circinelloides and D. flagrans were exposed to the horse sera by using an ELISA. The faecal elimination of eggs of Parascaris equorum and strongyles ceased 2 weeks after treatment in G-T and G-P, thus the values of FECR and PHR were 100%. No P. equorum-eggs were detected later, and the strongyle egg reappearance period was 28 weeks in G-P, and 8 weeks in G-T. Strongyle egg-output values remained lower than 300 eggs per gram of faeces in the G-P, whereas numbers between 330 and 772 in G-C and G-T were recorded. Normal values for the erythrocytes, haemoglobin and haematocrit in horses consuming pellets with spores were recorded, and lower than normal in the other groups. Sensitization of horses to the fungal species was disproven. It is concluded that feeding horses with pellets industrially manufactured with fungal spores represents a very useful tool to implement an integrated control of helminths affecting

  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. Chlorine Dioxide Inactivation of Cryptosporidium parvum Oocysts and Bacterial Spore Indicators

    PubMed Central

    Chauret, Christian P.; Radziminski, Chris Z.; Lepuil, Michael; Creason, Robin; Andrews, Robert C.

    2001-01-01

    Cryptosporidium parvum, which is resistant to chlorine concentrations typically used in water treatment, is recognized as a significant waterborne pathogen. Recent studies have demonstrated that chlorine dioxide is a more efficient disinfectant than free chlorine against Cryptosporidium oocysts. It is not known, however, if oocysts from different suppliers are equally sensitive to chlorine dioxide. This study used both a most-probable-number–cell culture infectivity assay and in vitro excystation to evaluate chlorine dioxide inactivation kinetics in laboratory water at pH 8 and 21°C. The two viability methods produced significantly different results (P < 0.05). Products of disinfectant concentration and contact time (Ct values) of 1,000 mg · min/liter were needed to inactivate approximately 0.5 log10 and 2.0 log10 units (99% inactivation) of C. parvum as measured by in vitro excystation and cell infectivity, respectively, suggesting that excystation is not an adequate viability assay. Purified oocysts originating from three different suppliers were evaluated and showed marked differences with respect to their resistance to inactivation when using chlorine dioxide. Ct values of 75, 550, and 1,000 mg · min/liter were required to achieve approximately 2.0 log10 units of inactivation with oocysts from different sources. Finally, the study compared the relationship between easily measured indicators, including Bacillus subtilis (aerobic) spores and Clostridium sporogenes (anaerobic) spores, and C. parvum oocysts. The bacterial spores were found to be more sensitive to chlorine dioxide than C. parvum oocysts and therefore could not be used as direct indicators of C. parvum inactivation for this disinfectant. In conclusion, it is suggested that future studies address issues such as oocyst purification protocols and the genetic diversity of C. parvum, since these factors might affect oocyst disinfection sensitivity. PMID:11425712

  9. Arthritis and Aerobic Exercise: A Review.

    ERIC Educational Resources Information Center

    Ike, Robert W.; And Others

    1989-01-01

    Arthritic patients who regularly do aerobic exercise make significant gains in aerobic and functional status, and in subjective areas like pain tolerance and mood. Still, they are often advised to curtail physical activity. Guidelines are presented for physicians prescribing aerobic exercise. An exercise tolerance test is recommended. (SM)

  10. Interpretation of galaxy counts

    SciTech Connect

    Tinsely, B.M.

    1980-10-01

    New models are presented for the interpretation of recent counts of galaxies to 24th magnitude, and predictions are shown to 28th magnitude for future comparison with data from the Space Telescope. The results supersede earlier, more schematic models by the author. Tyson and Jarvis found in their counts a ''local'' density enhancement at 17th magnitude, on comparison with the earlier models; the excess is no longer significant when a more realistic mixture of galaxy colors is used. Bruzual and Kron's conclusion that Kron's counts show evidence for evolution at faint magnitudes is confirmed, and it is predicted that some 23d magnitude galaxies have redshifts greater than unity. These may include spheroidal systems, elliptical galaxies, and the bulges of early-type spirals and S0's, seen during their primeval rapid star formation.

  11. Whose interests count?

    PubMed

    Brudney, Daniel; Lantos, John D

    2014-10-01

    Whose interests should count and how should various interests be balanced at the pediatric patient's bedside? The interests of the child patient clearly count. Recently, however, many authors have argued that the family's interests also count. But how should we think about the interests of others? What does it mean to talk about "the family" in this context? Does it really just mean the interests of each individual family member? Or is the family itself a moral entity that has interests of its own independent of the interests of each of its members? Are such interests important only as they affect the patient's interest or also for their own sake? In this special supplement to Pediatrics, a group of pediatricians, philosophers, and lawyers grapple with these questions. They examine these issues from different angles and reach different conclusions. Jointly, they demonstrate the ethical importance and, above all, the ethical complexity of the family's role at the bedside.

  12. 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

  13. 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

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. 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

  19. 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

  20. Enumeration of Clostridium perfringens spores in human feces: comparison of four culture media.

    PubMed

    Harmon, S M; Kautter, D A

    1987-01-01

    Enumeration of Clostridium perfringens spores was compared using 4 culture media. Duplicate 1 g portions of 35 stools (25 from C. perfringens food poisoning outbreaks and 10 from normal stools) were heat treated 20 min at 75 degrees C and tested on tryptose-sulfite-cycloserine (TSC) agar, trypticase-soy-blood (TSB) agar, lactose-sulfite (LS) medium, and iron milk (IM) medium. Dilutions were plated directly onto TSB and TSC, and a 3-tube most probable number determination was made with each specimen in LS and IM incubated at 45 degrees C. TSB was easiest to use and nonhemolytic food poisoning strains were readily differentiated from the normal hemolytic biotype on this medium. Confirmed counts on TSC and TSB were similar for all specimens, but counts of 8 of 25 outbreak specimens were 2-4 log units lower in LS and IM than on plating media; spores in specimens associated with 2 of 5 outbreaks were intolerant of the elevated temperatures. Results showed that elevated temperature MPN methods in LS and IM are inappropriate for the examination of outbreak stools.

  1. Spore Germination and Carbon Metabolism in Fusarium solani V. Changes in Anaerobic Metabolism and Related Enzyme Activities during Development 1

    PubMed Central

    Cochrane, Vincent W.; Cochrane, Jean C.

    1966-01-01

    Macroconidia of Fusarium solani f. phascoli have no detectable capacity to respire glucose anaerobically; germinated spores and mycelium, on the other hand, ferment glucose, although slowly. Extracts of ungerminated spores contain hexokinase, phosphohexoisomerase, phosphofructokinase, aldolase, triose phosphate dehydrogenase, triose phosphate isomerase, phosphoglyceric kinase, enolase, phosphoglyceric mutase, pyruvate kinase, and pyruvate decarboxylase. It follows, therefore, that the appearance of fermentative capacity during spore germination cannot be ascribed to the de novo synthesis of any of these enzymes. During germination and mycelial development the specific activity of all of the enzymes named except phosphohexoisomerase and aldolase increases 2- to 8-fold. Specific activity of all of the enzymes is substantially higher than the fermentative capacity of intact cells, i.e., none is limiting to anaerobic respiration. The enzymatic assay data are consistent with a conclusion reached earlier on the basis of studies of aerobic glucose metabolism, that the process of germination involves an acceleration of pre-existing metabolic systems rather than an appearance of new pathways. PMID:16656324

  2. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2012-03-01

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

  4. Administration of Bacillus coagulans in calves: recovery from faecal samples and evaluation of functional aspects of spores.

    PubMed

    Ripamonti, Barbara; Agazzi, Alessandro; Baldi, Antonella; Balzaretti, Claudia; Bersani, Carla; Pirani, Silvia; Rebucci, Raffaella; Savoini, Giovanni; Stella, Simone; Stenico, Alberta; Domeneghini, Cinzia

    2009-12-01

    An investigation was carried out into the recovery from calf faeces of Bacillus coagulans spores added to the feed as probiotic. For this purpose, Bacillus coagulans spores (9 log₁₀ CFU g⁻¹) were given daily to 10 calves during the whole farming periods; another 10 calves acted as controls. Throughout the trial the faecal spore counts were significantly (P < 0.01) higher in the treated group than in the controls (averaging 2.1 x 10⁵ vs 3.7 x 10⁴ CFU g⁻¹). Bacterial cells were recovered from faecal samples and ribotyping matched the strain isolated from faecal sample to the clone administered to the animals. In addition, the recovered cells were found to maintain their functionality aspects of acid production, survival in artificial gastric juice and in the presence of bile, and attachment to human intestinal epithelial cells. The results further elucidate the fate of spore formers administered to calves, and this will help in the development of new species-specific nutritional strategies.

  5. LOW ENERGY COUNTING CHAMBERS

    DOEpatents

    Hayes, P.M.

    1960-02-16

    A beta particle counter adapted to use an end window made of polyethylene terephthalate was designed. The extreme thinness of the film results in a correspondingly high transmission of incident low-energy beta particles by the window. As a consequence, the counting efficiency of the present counter is over 40% greater than counters using conventional mica end windows.

  6. WY Kids Count.

    ERIC Educational Resources Information Center

    Wyoming Kids Count, Cheyenne.

    This WY Kids Count brochure uses the metaphor of children's building blocks to present information on the current well-being of Wyoming children and to advocate enhancing the lives of young children. Each block (i.e., each develop the brochure) presents concerns in a separate area: (1) poverty, highlighting the number of children living in…

  7. What Counts as Evidence?

    ERIC Educational Resources Information Center

    Dougherty Stahl, Katherine A.

    2014-01-01

    Each disciplinary community has its own criteria for determining what counts as evidence of knowledge in their academic field. The criteria influence the ways that a community's knowledge is created, communicated, and evaluated. Situating reading, writing, and language instruction within the content areas enables teachers to explicitly…

  8. 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.

  9. Correlation of pollen counts and number of hospital visits of asthmatic and allergic rhinitis patients

    PubMed Central

    Singh, Nishtha; Singh, Udaiveer; Singh, Dimple; Daya, Mangal; Singh, Virendra

    2017-01-01

    Aims and Objectives: Environmental pollens are known to cause exacerbation of symptoms of patients with allergic rhinitis (AR) and asthma. During pollen months, number of patients visiting hospital has been shown to increase in some studies. However, in India, such studies are lacking. Therefore, we aimed to study pollen counts and to find its correlation with number of new patients attending Asthma Bhawan for 2 years. Materials and Methods: Aerobiological sampling was done using Burkard 24 h spore trap system. The site selected for the entrapment of the air spore was the building of Asthma Bhawan situated at Vidhyadhar Nagar, Jaipur. New patients coming with problems of respiratory allergy such as AR or asthma were recruited in the study. Skin prick tests (SPTs) were carried out after obtaining consent in these patients. Monthly pollen counts of trees, weeds and grasses were correlated with the number of new patients. Pollen calendar was prepared for 2 years. Results: Average annual pollen count during 2011 and 2012 were 14,460.5. In the analysis, 37 types of species or families were identified. Pollen count showed two seasonal peaks during March–April and from August to October. January and June showed the lowest pollen counts in 2 years. Average monthly count of grass pollens showed significant correlation with number of new patients (r = 0.59). However, monthly pollen count of trees and weeds did not correlate. The correlation of the pollen count of individual pollen with the SPT positivity to that pollen showed significant correlation with Chenopodium album only. Conclusions: It can be concluded that there were two peaks of pollen count in a year during March–April and August–October. Average monthly pollen counts of grass were significantly correlated with the number of hospital visits of new patients. PMID:28360459

  10. Calcium precipitate induced aerobic granulation.

    PubMed

    Wan, Chunli; Lee, Duu-Jong; Yang, Xue; Wang, Yayi; Wang, Xingzu; Liu, Xiang

    2015-01-01

    Aerobic granulation is a novel biotechnology for wastewater treatment. This study refined existing aerobic granulation mechanisms as a sequencing process including formation of calcium precipitate under alkaline pH to form inorganic cores, followed by bacterial attachment and growth on these cores to form the exopolysaccharide matrix. Mature granules comprised an inner core and a matrix layer and a rim layer with enriched microbial strains. The inorganic core was a mix of different crystals of calcium and phosphates. Functional strains including Sphingomonas sp., Paracoccus sp. Sinorhizobium americanum strain and Flavobacterium sp. attached onto the cores. These functional strains promote c-di-GMP production and the expression by Psl and Alg genes for exopolysaccharide production to enhance formation of mature granules.

  11. Aerobic microbial enhanced oil recovery

    SciTech Connect

    Torsvik, T.; Gilje, E.; Sunde, E.

    1995-12-31

    In aerobic MEOR, the ability of oil-degrading bacteria to mobilize oil is used to increase oil recovery. In this process, oxygen and mineral nutrients are injected into the oil reservoir in order to stimulate growth of aerobic oil-degrading bacteria in the reservoir. Experiments carried out in a model sandstone with stock tank oil and bacteria isolated from offshore wells showed that residual oil saturation was lowered from 27% to 3%. The process was time dependent, not pore volume dependent. During MEOR flooding, the relative permeability of water was lowered. Oxygen and active bacteria were needed for the process to take place. Maximum efficiency was reached at low oxygen concentrations, approximately 1 mg O{sub 2}/liter.

  12. WWOX loss activates aerobic glycolysis.

    PubMed

    Abu-Remaileh, Muhannad; Seewaldt, Victoria L; Aqeilan, Rami I

    2015-01-01

    Cancer cells undergo reprogramming of glucose metabolism to limit energy production to glycolysis-a state known as "aerobic glycolysis." Hypoxia-inducible factor 1 (HIF1α) is a transcription factor that regulates many genes responsible for this switch. As discussed here, new data suggest that the tumor suppressor WW domain-containing oxidoreductase (WWOX) modulates HIF1α, thereby regulating this metabolic state.

  13. WWOX loss activates aerobic glycolysis

    PubMed Central

    Abu-Remaileh, Muhannad; Seewaldt, Victoria L; Aqeilan, Rami I

    2015-01-01

    Cancer cells undergo reprogramming of glucose metabolism to limit energy production to glycolysis—a state known as “aerobic glycolysis.” Hypoxia-inducible factor 1 (HIF1α) is a transcription factor that regulates many genes responsible for this switch. As discussed here, new data suggest that the tumor suppressor WW domain-containing oxidoreductase (WWOX) modulates HIF1α, thereby regulating this metabolic state. PMID:27308416

  14. 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.

  15. 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

  16. 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.

  17. 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

  18. 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.

  19. Aerobic Metabolism of Streptococcus agalactiae

    PubMed Central

    Mickelson, M. N.

    1967-01-01

    Streptococcus agalactiae cultures possess an aerobic pathway for glucose oxidation that is strongly inhibited by cyanide. The products of glucose oxidation by aerobically grown cells of S. agalactiae 50 are lactic and acetic acids, acetylmethylcarbinol, and carbon dioxide. Glucose degradation products by aerobically grown cells, as percentage of glucose carbon, were 52 to 61% lactic acid, 20 to 23% acetic acid, 5.5 to 6.5% acetylmethylcarbinol, and 14 to 16% carbon dioxide. There was no evidence for a pentose cycle or a tricarboxylic acid cycle. Crude cell-free extracts of S. agalactiae 50 possessed a strong reduced nicotinamide adenine dinucleotide (NADH2) oxidase that is also cyanide-sensitive. Dialysis or ultrafiltration of the crude, cell-free extract resulted in loss of NADH2 oxidase activity. Oxidase activity was restored to the inactive extract by addition of the ultrafiltrate or by addition of menadione or K3Fe(CN)6. Noncytochrome iron-containing pigments were present in cell-free extracts of S. agalactiae. The possible participation of these pigments in the respiration of S. agalactiae is presently being studied. PMID:4291090

  20. The Watt Count System

    NASA Technical Reports Server (NTRS)

    1981-01-01

    The Watt Count System combines aerospace and computer technology, an engineering systems design approach and the environmental control expertise of a group of engineers who worked on Apollo. First step of the system is a computerized energy consumption analysis based on plans for a new home. Company calculates heating and cooling loads and customizes the most energy efficient system for the particular structure analyzed. A quality control engineer assures that insulation and ducting are installed to specifications. Analysis and design techniques are so accurate that company is able to give homeowner a two-year guarantee that the home's energy consumption will not exceed the kilowatt hour level determined by the computer in step one. In an annual energy performance comparison, a Watt Count home used 45 percent less energy than a conventionally equipped home.

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

    PubMed Central

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

    2017-01-01

    Premature spoilage and varying product quality due to microbial contamination still constitute major problems in the production of microfiltered and pasteurized extended shelf life (ESL) milk. Spoilage-associated bacteria may enter the product either as part of the raw milk microbiota or as recontaminants in the dairy plant. To identify spoilage-inducing bacteria and their routes of entry, we analyzed end products for their predominant microbiota as well as the prevalence and biodiversity of psychrotolerant spores in bulk tank milk. Process analyses were performed to determine the removal of psychrotolerant spores at each production step. To detect transmission and recontamination events, strain typing was conducted with isolates obtained from all process stages. Microbial counts in 287 ESL milk packages at the end of shelf life were highly diverse ranging from <1 to 7.9 log cfu/mL. In total, 15% of samples were spoiled. High G+C Gram-positive bacteria were the most abundant taxonomic group, but were responsible for only 31% of spoilage. In contrast, psychrotolerant spores were isolated from 55% of spoiled packages. In 90% of samples with pure cultures of Bacillus cereus sensu lato and Paenibacillus spp., counts exceeded 6 log cfu/mL. In bulk tank milk, the concentration of psychrotolerant spores was low, accounting for merely 0.5 ± 0.8 MPN/mL. Paenibacillus amylolyticus/xylanexedens was by far the most dominant species in bulk tank milk (48% of all isolates), but was never detected in ESL milk, pointing to efficient removal during manufacturing. Six large-scale process analyses confirmed a high removal rate for psychrotolerant spores (reduction by nearly 4 log-units). B. cereus sensu lato, on the contrary, was frequently found in spoiled end products, but was rarely detected in bulk tank milk. Due to low counts in bulk tank samples and efficient spore removal during production, we suggest that shelf life is influenced only to a minor extent by raw

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. Genome-Wide Transcriptional Profiling of Clostridium perfringens SM101 during Sporulation Extends the Core of Putative Sporulation Genes and Genes Determining Spore Properties and Germination Characteristics

    PubMed Central

    Xiao, Yinghua; van Hijum, Sacha A. F. T.; Abee, Tjakko; Wells-Bennik, Marjon H. J.

    2015-01-01

    The formation of bacterial spores is a highly regulated process and the ultimate properties of the spores are determined during sporulation and subsequent maturation. A wide variety of genes that are expressed during sporulation determine spore properties such as resistance to heat and other adverse environmental conditions, dormancy and germination responses. In this study we characterized the sporulation phases of C. perfringens enterotoxic strain SM101 based on morphological characteristics, biomass accumulation (OD600), the total viable counts of cells plus spores, the viable count of heat resistant spores alone, the pH of the supernatant, enterotoxin production and dipicolinic acid accumulation. Subsequently, whole-genome expression profiling during key phases of the sporulation process was performed using DNA microarrays, and genes were clustered based on their time-course expression profiles during sporulation. The majority of previously characterized C. perfringens germination genes showed upregulated expression profiles in time during sporulation and belonged to two main clusters of genes. These clusters with up-regulated genes contained a large number of C. perfringens genes which are homologs of Bacillus genes with roles in sporulation and germination; this study therefore suggests that those homologs are functional in C. perfringens. A comprehensive homology search revealed that approximately half of the upregulated genes in the two clusters are conserved within a broad range of sporeforming Firmicutes. Another 30% of upregulated genes in the two clusters were found only in Clostridium species, while the remaining 20% appeared to be specific for C. perfringens. These newly identified genes may add to the repertoire of genes with roles in sporulation and determining spore properties including germination behavior. Their exact roles remain to be elucidated in future studies. PMID:25978838

  9. Detection of the dipicolinic acid biomarker in Bacillus spores using Curie-point pyrolysis mass spectrometry and Fourier transform infrared spectroscopy.

    PubMed

    Goodacre, R; Shann, B; Gilbert, R J; Timmins, E M; McGovern, A C; Alsberg, B K; Kell, D B; Logan, N A

    2000-01-01

    Thirty-six strains of aerobic endospore-forming bacteria confirmed by polyphasic taxonomic methods to belong to Bacillus amyloliquefaciens, Bacillus cereus, Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis (including Bacillus niger and Bacillus globigii), Bacillus sphaericus, and Brevi laterosporus were grown axenically on nutrient agar, and vegetative and sporulated biomasses were analyzed by Curie-point pyrolysis mass spectrometry (PyMS) and diffuse reflectance-absorbance Fourier-transform infrared spectroscopy (FT-IR). Chemometric methods based on rule induction and genetic programming were used to determine the physiological state (vegetative cells or spores) correctly, and these methods produced mathematical rules which could be simply interpreted in biochemical terms. For PyMS it was found that m/z 105 was characteristic and is a pyridine ketonium ion (C6H3ON+) obtained from the pyrolysis of dipicolinic acid (pyridine-2,6-dicarboxylic acid; DPA), a substance found in spores but not in vegetative cells; this was confirmed using pyrolysis-gas chromatography/mass spectrometry. In addition, a pyridine ring vibration at 1447-1439 cm-1 from DPA was found to be highly characteristic of spores in FT-IR analysis. Thus, although the original data sets recorded hundreds of spectral variables from whole cells simultaneously, a simple biomarker can be used for the rapid and unequivocal detection of spores of these organisms.

  10. A genetic algorithm-Bayesian network approach for the analysis of metabolomics and spectroscopic data: application to the rapid identification of Bacillus spores and classification of Bacillus species

    PubMed Central

    2011-01-01

    Background The rapid identification of Bacillus spores and bacterial identification are paramount because of their implications in food poisoning, pathogenesis and their use as potential biowarfare agents. Many automated analytical techniques such as Curie-point pyrolysis mass spectrometry (Py-MS) have been used to identify bacterial spores giving use to large amounts of analytical data. This high number of features makes interpretation of the data extremely difficult We analysed Py-MS data from 36 different strains of aerobic endospore-forming bacteria encompassing seven different species. These bacteria were grown axenically on nutrient agar and vegetative biomass and spores were analyzed by Curie-point Py-MS. Results We develop a novel genetic algorithm-Bayesian network algorithm that accurately identifies sand selects a small subset of key relevant mass spectra (biomarkers) to be further analysed. Once identified, this subset of relevant biomarkers was then used to identify Bacillus spores successfully and to identify Bacillus species via a Bayesian network model specifically built for this reduced set of features. Conclusions This final compact Bayesian network classification model is parsimonious, computationally fast to run and its graphical visualization allows easy interpretation of the probabilistic relationships among selected biomarkers. In addition, we compare the features selected by the genetic algorithm-Bayesian network approach with the features selected by partial least squares-discriminant analysis (PLS-DA). The classification accuracy results show that the set of features selected by the GA-BN is far superior to PLS-DA. PMID:21269434

  11. 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

  12. 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.

  13. 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.

  14. Toxigenic potential and heat survival of spore-forming bacteria isolated from bread and ingredients.

    PubMed

    De Bellis, Palmira; Minervini, Fiorenza; Di Biase, Mariaelena; Valerio, Francesca; Lavermicocca, Paola; Sisto, Angelo

    2015-03-16

    Fifty-four spore-forming bacterial strains isolated from bread ingredients and bread, mainly belonging to the genus Bacillus (including Bacillus cereus), together with 11 reference strains were investigated to evaluate their cytotoxic potential and heat survival in order to ascertain if they could represent a risk for consumer health. Therefore, we performed a screening test of cytotoxic activity on HT-29 cells using bacterial culture filtrates after growing bacterial cells in Brain Heart Infusion medium and in the bread-based medium Bread Extract Broth (BEB). Moreover, immunoassays and PCR analyses, specifically targeting already known toxins and related genes of B. cereus, as well as a heat spore inactivation assay were carried out. Despite of strain variability, the results clearly demonstrated a high cytotoxic activity of B. cereus strains, even if for most of them it was significantly lower in BEB medium. Cytotoxic activity was also detected in 30% of strains belonging to species different from B. cereus, although, with a few exceptions (e.g. Bacillus simplex N58.2), it was low or very low. PCR analyses detected the presence of genes involved in the production of NHE, HBL or CytK toxins in B. cereus strains, while genes responsible for cereulide production were not detected. Production of NHE and HBL toxins was also confirmed by specific immunoassays only for B. cereus strains even if PCR analyses revealed the presence of related toxin genes also in some strains of other species. Viable spore count was ascertained after a heat treatment simulating the bread cooking process. Results indicated that B. amyloliquefaciens strains almost completely survived the heat treatment showing less than 2 log-cycle reductions similarly to two strains of B. cereus group III and single strains belonging to Bacillus subtilis, Bacillus mojavensis and Paenibacillus spp. Importantly, spores from strains of the B. cereus group IV exhibited a thermal resistance markedly lower than B

  15. 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

  16. 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.

  17. Monitoring airborne fungal spores in an experimental indoor environment to evaluate sampling methods and the effects of human activity on air sampling.

    PubMed Central

    Buttner, M P; Stetzenbach, L D

    1993-01-01

    Aerobiological monitoring was conducted in an experimental room to aid in the development of standardized sampling protocols for airborne microorganisms in the indoor environment. The objectives of this research were to evaluate the relative efficiencies of selected sampling methods for the retrieval of airborne fungal spores and to determine the effect of human activity on air sampling. Dry aerosols containing known concentrations of Penicillium chrysogenum spores were generated, and air samples were taken by using Andersen six-stage, Surface Air System, Burkard, and depositional samplers. The Andersen and Burkard samplers retrieved the highest numbers of spores compared with the measurement standard, an aerodynamic particle sizer located inside the room. Data from paired samplers demonstrated that the Andersen sampler had the highest levels of sensitivity and repeatability. With a carpet as the source of P. chrysogenum spores, the effects of human activity (walking or vacuuming near the sampling site) on air sampling were also examined. Air samples were taken under undisturbed conditions and after human activity in the room. Human activity resulted in retrieval of significantly higher concentrations of airborne spores. Surface sampling of the carpet revealed moderate to heavy contamination despite relatively low airborne counts. Therefore, in certain situations, air sampling without concomitant surface sampling may not adequately reflect the level of microbial contamination in indoor environments. PMID:8439150

  18. The effect of Perasafe and sodium dichloroisocyanurate (NaDCC) against spores of Clostridium difficile and Bacillus atrophaeus on stainless steel and polyvinyl chloride surfaces.

    PubMed

    Block, C

    2004-06-01

    Clostridium difficile is an important cause of nosocomial diarrhoea. The aim of this study was to evaluate the potential for Perasafe, a recently introduced biocide, to contribute to control of C. difficile spores in the patient environment, in comparison with the chlorine-releasing agent sodium dichloroisocyanurate (NaDCC). These agents were evaluated against a water control, in a surface test on stainless steel and polyvinyl chloride (PVC) floor covering, materials commonly found in the hospital environment. The organisms studied were a toxigenic clinical isolate of C. difficile, and Bacillus atrophaeus (formerly B. subtilis var niger). The data indicate that in our in vitro system, Perasafe was significantly more active than NaDCC (1000 ppm available chlorine) against C. difficile spores dried on stainless steel surfaces, and against B. atrophaeus on PVC floor covering material, achieving mean log10 reduction factors in viable counts of 6 and 5.5, respectively, at 10 min exposures. Perasafe appeared to be less lethal in 10 min exposures to C. difficile spores fixed on PVC floor covering material. In general, 1000 ppm chlorine generated from NaDCC showed lower log10 reduction factors in viable counts at 10 min, ranging from 0.7 to 1.5, than Perasafe which ranged from 2.7 to 6.0. The potential efficacy of Perasafe in reducing the density of C. difficile spores in the patient environment in hospitals, nursing homes or other long-stay facilities should be evaluated in field studies.

  19. Regional-scale simulations of fungal spore aerosols using an emission parameterization adapted to local measurements of fluorescent biological aerosol particles

    NASA Astrophysics Data System (ADS)

    Hummel, M.; Hoose, C.; Gallagher, M.; Healy, D. A.; Huffman, J. A.; O'Connor, D.; Pöschl, U.; Pöhlker, C.; Robinson, N. H.; Schnaiter, M.; Sodeau, J. R.; Stengel, M.; Toprak, E.; Vogel, H.

    2015-06-01

    Fungal spores as a prominent type of primary biological aerosol particles (PBAP) have been incorporated into the COSMO-ART (Consortium for Small-scale Modelling-Aerosols and Reactive Trace gases) regional atmospheric model. Two literature-based emission rates for fungal spores derived from fungal spore colony counts and chemical tracer measurements were used as a parameterization baseline for this study. A third, new emission parameterization for fluorescent biological aerosol particles (FBAP) was adapted to field measurements from four locations across Europe. FBAP concentrations can be regarded as a lower estimate of total PBAP concentrations. Size distributions of FBAP often show a distinct mode at approx. 3 μm, corresponding to a diameter range characteristic for many fungal spores. Previous studies for several locations have suggested that FBAP are in many cases dominated by fungal spores. Thus, we suggest that simulated FBAP and fungal spore concentrations obtained from the three different emission parameterizations can be compared to FBAP measurements. The comparison reveals that simulated fungal spore concentrations based on literature emission parameterizations are lower than measured FBAP concentrations. In agreement with the measurements, the model results show a diurnal cycle in simulated fungal spore concentrations, which may develop partially as a consequence of a varying boundary layer height between day and night. Temperature and specific humidity, together with leaf area index (LAI), were chosen to drive the new emission parameterization which is fitted to the FBAP observations. The new parameterization results in similar root mean square errors (RMSEs) and correlation coefficients compared to the FBAP observations as the previously existing fungal spore emission parameterizations, with some improvements in the bias. Using the new emission parameterization on a model domain covering western Europe, FBAP in the lowest model layer comprise a

  20. Dynamics Associated with Prolonged Ensiling and Aerobic Deterioration of Total Mixed Ration Silage Containing Whole Crop Corn

    PubMed Central

    Wang, Huili; Ning, Tingting; Hao, Wei; Zheng, Mingli; Xu, Chuncheng

    2016-01-01

    This study investigated the dynamics associated with prolonged ensiling and aerobic deterioration of whole crop corn (WCC) silages and total mixed ration (TMR) silages containing WCC (C-TMR silages) to clarify the differences that account for the enhanced aerobic stability of TMR silages. Laboratory-scale barrel silos were randomly opened after 7, 14, 28, and 56 d of ensiling and were subjected to analyses of fermentation quality, microbial and temperature dynamics during aerobic exposure. WCC and C-TMR silages were both well preserved and microorganisms were inhibited with prolonged ensiling, including lactic acid bacteria. Yeast were inhibited to below the detection limit of 500 cfu/g fresh matter within 28 d of ensiling. Aerobic stability of both silages was enhanced with prolonged ensiling, whereas C-TMR silages were more aerobically stable than WCC silages for the same ensiling period. Besides the high moisture content, the weak aerobic stability of WCC silage is likely attributable to the higher lactic acid content and yeast count, which result from the high water-soluble carbohydrates content in WCC. After silo opening, yeast were the first to propagate and the increase in yeast levels is greater than that of other microorganisms in silages before deterioration. Besides, increased levels of aerobic bacteria were also detected before heating of WCC silages. The temperature dynamics also indicated that yeast are closely associated with the onset of the aerobic deterioration of C-TMR silage, whereas for WCC silages, besides yeast, aerobic bacteria also function in the aerobic deterioration. Therefore, the inclusion of WCC might contribute to the survival of yeast during ensiling but not influence the role of yeast in deterioration of C-TMR silages. PMID:26732329

  1. High Red Blood Cell Count

    MedlinePlus

    Symptoms High red blood cell count By Mayo Clinic Staff A high red blood cell count is an increase in oxygen-carrying cells in your bloodstream. Red blood cells transport oxygen from your lungs to tissues throughout ...

  2. 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

  3. 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.

  4. 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.

  5. 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

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. Lower limb loading in step aerobic dance.

    PubMed

    Wu, H-W; Hsieh, H-M; Chang, Y-W; Wang, L-H

    2012-11-01

    Participation in aerobic dance is associated with a number of lower extremity injuries, and abnormal joint loading seems to be a factor in these. However, information on joint loading is limited. The purpose of this study was to investigate the kinetics of the lower extremity in step aerobic dance and to compare the differences of high-impact and low-impact step aerobic dance in 4 aerobic movements (mambo, kick, L step and leg curl). 18 subjects were recruited for this study. High-impact aerobic dance requires a significantly greater range of motion, joint force and joint moment than low-impact step aerobic dance. The peak joint forces and moments in high-impact step aerobic dance were found to be 1.4 times higher than in low-impact step aerobic dance. Understanding the nature of joint loading may help choreographers develop dance combinations that are less injury-prone. Furthermore, increased knowledge about joint loading may be helpful in lowering the risk of injuries in aerobic dance instructors and students.

  11. Reduced bacterial colony count of anaerobic bacteria is associated with a worsening in lung clearance index and inflammation in cystic fibrosis.

    PubMed

    O'Neill, Katherine; Bradley, Judy M; Johnston, Elinor; McGrath, Stephanie; McIlreavey, Leanne; Rowan, Stephen; Reid, Alastair; Bradbury, Ian; Einarsson, Gisli; Elborn, J Stuart; Tunney, Michael M

    2015-01-01

    Anaerobic bacteria have been identified in abundance in the airways of cystic fibrosis (CF) subjects. The impact their presence and abundance has on lung function and inflammation is unclear. The aim of this study was to investigate the relationship between the colony count of aerobic and anaerobic bacteria, lung clearance index (LCI), spirometry and C-Reactive Protein (CRP) in patients with CF. Sputum and blood were collected from CF patients at a single cross-sectional visit when clinically stable. Community composition and bacterial colony counts were analysed using extended aerobic and anaerobic culture. Patients completed spirometry and a multiple breath washout (MBW) test to obtain LCI. An inverse correlation between colony count of aerobic bacteria (n = 41, r = -0.35; p = 0.02), anaerobic bacteria (n = 41, r = -0.44, p = 0.004) and LCI was observed. There was an inverse correlation between colony count of anaerobic bacteria and CRP (n = 25, r = -0.44, p = 0.03) only. The results of this study demonstrate that a lower colony count of aerobic and anaerobic bacteria correlated with a worse LCI. A lower colony count of anaerobic bacteria also correlated with higher CRP levels. These results indicate that lower abundance of aerobic and anaerobic bacteria may reflect microbiota disruption and disease progression in the CF lung.

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

    PubMed

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

    2017-02-01

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

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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

  18. Survival of anaerobic and aerobic bacteria in a nonsupportive gassed transport system.

    PubMed Central

    Chow, A W; Cunningham, P J; Guze, L B

    1976-01-01

    Survival of anaerobic and aerobic bacteria in a commercially available, non-supportive, gassed (oxygen-free) transport container (Anaport) was evaluated quantitatively. Saline-suspended obligate anaerobes survived significantly better in the gassed container in aerobic control tubes (P less than 0.025, t test), and counts were virtually unchanged after 8 h of holding. Similarly, initial counts and relative proportions of a mixture of Bacteroides fragilis and Staphylococcus aureus were maintained for 72 h. The value of the gassed transport system was less apparent when microorganisms were suspended in nutrient broth. The major advantage of the gassed transport system appears to be for holding of specimens collected by saline irrigation. PMID:1254710

  19. Counting RG flows

    SciTech Connect

    Gukov, Sergei

    2016-01-05

    Here, interpreting renormalization group flows as solitons interpolating between different fixed points, we ask various questions that are normally asked in soliton physics but not in renormalization theory. Can one count RG flows? Are there different "topological sectors" for RG flows? What is the moduli space of an RG flow, and how does it compare to familiar moduli spaces of (supersymmetric) dowain walls? Analyzing these questions in a wide variety of contexts -- from counting RG walls to AdS/CFT correspondence -- will not only provide favorable answers, but will also lead us to a unified general framework that is powerful enough to account for peculiar RG flows and predict new physical phenomena. Namely, using Bott's version of Morse theory we relate the topology of conformal manifolds to certain properties of RG flows that can be used as precise diagnostics and "topological obstructions" for the strong form of the C-theorem in any dimension. Moreover, this framework suggests a precise mechanism for how the violation of the strong C-theorem happens and predicts "phase transitions" along the RG flow when the topological obstruction is non-trivial. Along the way, we also find new conformal manifolds in well-known 4d CFT's and point out connections with the superconformal index and classifying spaces of global symmetry groups.

  20. Counting RG flows

    DOE PAGES

    Gukov, Sergei

    2016-01-05

    Here, interpreting renormalization group flows as solitons interpolating between different fixed points, we ask various questions that are normally asked in soliton physics but not in renormalization theory. Can one count RG flows? Are there different "topological sectors" for RG flows? What is the moduli space of an RG flow, and how does it compare to familiar moduli spaces of (supersymmetric) dowain walls? Analyzing these questions in a wide variety of contexts -- from counting RG walls to AdS/CFT correspondence -- will not only provide favorable answers, but will also lead us to a unified general framework that is powerfulmore » enough to account for peculiar RG flows and predict new physical phenomena. Namely, using Bott's version of Morse theory we relate the topology of conformal manifolds to certain properties of RG flows that can be used as precise diagnostics and "topological obstructions" for the strong form of the C-theorem in any dimension. Moreover, this framework suggests a precise mechanism for how the violation of the strong C-theorem happens and predicts "phase transitions" along the RG flow when the topological obstruction is non-trivial. Along the way, we also find new conformal manifolds in well-known 4d CFT's and point out connections with the superconformal index and classifying spaces of global symmetry groups.« less

  1. Neural networks counting chimes.

    PubMed Central

    Amit, D J

    1988-01-01

    It is shown that the ideas that led to neural networks capable of recalling associatively and asynchronously temporal sequences of patterns can be extended to produce a neural network that automatically counts the cardinal number in a sequence of identical external stimuli. The network is explicitly constructed, analyzed, and simulated. Such a network may account for the cognitive effect of the automatic counting of chimes to tell the hour. A more general implication is that different electrophysiological responses to identical stimuli, at certain stages of cortical processing, do not necessarily imply synaptic modification, a la Hebb. Such differences may arise from the fact that consecutive identical inputs find the network in different stages of an active temporal sequence of cognitive states. These types of networks are then situated within a program for the study of cognition, which assigns the detection of meaning as the primary role of attractor neural networks rather than computation, in contrast to the parallel distributed processing attitude to the connectionist project. This interpretation is free of homunculus, as well as from the criticism raised against the cognitive model of symbol manipulation. Computation is then identified as the syntax of temporal sequences of quasi-attractors. PMID:3353371

  2. 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

  3. 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

  4. 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

  5. Standardization of Spore Inactivation Method for PMA-PhyloChip Analysis

    NASA Technical Reports Server (NTRS)

    Schrader, Michael

    2011-01-01

    In compliance with the Committee on Space Research (COSPAR) planetary protection policy, National Aeronautics and Space Administration (NASA) monitors the total microbial burden of spacecraft as a means for minimizing the inadvertent transfer of viable contaminant microorganisms to extraterrestrial environments (forward contamination). NASA standard assay-based counts are used both as a proxy for relative surface cleanliness and to estimate overall microbial burden as well as to assess whether forward planetary protection risk criteria are met for a given mission, which vary by the planetary body to be explored and whether or not life detection missions are present. Despite efforts to reduce presence of microorganisms from spacecraft prior to launch, microbes have been isolated from spacecraft and associated surfaces within the extreme conditions of clean room facilities using state of the art molecular technologies. Development of a more sensitive method that will better enumerate all viable microorganisms from spacecraft and associated surfaces could support future life detection missions. Current culture-based (NASA standard spore assay) and nucleic-acid-based polymerase chain reaction (PCR) methods have significant shortcomings in this type of analysis. The overall goal of this project is to evaluate and validate a new molecular method based on the use of a deoxyribonucleic acid (DNA) intercalating agent propidium monoazide (PMA). This is used in combination with DNA microarray (PhyloChip) which has been shown to identify very low levels of organisms on spacecraft associated surfaces. PMA can only penetrate the membrane of dead cells. Once penetrated, it intercalates the DNA and, upon photolysis using visible light it produces stable DNA monoadducts. This allows DNA to be unavailable for further PCR analysis. The specific aim of this study is to standardize the spore inactivation method for PMA-PhyloChip analysis. We have used the bacterial spores Bacillus

  6. 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.

  7. 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

  8. 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...

  9. Validation of the Peel Plate™ AC for Detection of Total Aerobic Bacteria in Dairy and Nondairy Products.

    PubMed

    Salter, Robert S; Durbin, Gregory W; Bird, Patrick; Fisher, Kiel; Crowley, Erin; Hammack, Thomas; Chen, Yi; Clark, Dorn; Ziemer, Wayne

    2016-01-01

    Peel Plate™ AC (aerobic count) is a low-profile plastic 47 mm culture dish with adhesive top that contains a dried standard plate count medium with oxidation/reduction indicator triphenyl tetrazolium chloride (TTC) that turns red with dehydrogenase enzyme activity of growing aerobic bacteria. The method provides a conventional quantitative count with simple rehydration and incubation for 48 ± 3 h at 35 ± 1°C for most food matrixes and 32 ± 1°C for 48 ± 3 h for dairy products. Dairy matrixes claimed and supported with total aerobic count data are whole milk, skim milk, chocolate milk (2% fat), light cream (20% fat), pasteurized whole goat milk, ultra-high temperature pasteurized milk, nonfat dried milk, lactose-reduced milk, strawberry milk, raw cow milk, raw goat milk, raw sheep milk, condensed skim milk, and vanilla ice cream. Food matrixes claimed for aerobic count detection are raw ground beef, environmental sponge of stainless steel, raw ground turkey, dry dog food, liquid whole pasteurized eggs, milk chocolate, poultry carcass rinse, and large animal carcass sponge. The method has been independently evaluated for aerobic count in dairy products: whole milk, skim milk, chocolate milk, and light cream. The method was also independently evaluated for aerobic count in food matrixes: ground beef and sponge rinse from stainless steel surfaces. In the matrix study, each matrix was assessed separately at each contamination level in comparison to an appropriate reference method. Colony counts were determined for each level and then log10-transformed. The transformed data were evaluated for repeatability, mean comparison between methods with 95% confidence interval (CI), and r(2). A CI range of (-0.5, 0.5) on the mean difference was used as the acceptance criterion to establish significant statistical differences between methods. The evaluations demonstrate that the Peel Plate AC provides no statistical differences across most of the matrixes with r(2) > 0

  10. Skeletal Muscle Hypertrophy after Aerobic Exercise Training

    PubMed Central

    Konopka, Adam R.; Harber, Matthew P.

    2014-01-01

    Current dogma suggests aerobic exercise training has minimal effect on skeletal muscle size. We and others have demonstrated that aerobic exercise acutely and chronically alters protein metabolism and induces skeletal muscle hypertrophy. These findings promote an antithesis to the status quo by providing novel perspective on skeletal muscle mass regulation and insight into exercise-countermeasures for populations prone to muscle loss. PMID:24508740

  11. Aerobic rice mechanization: techniques for crop establishment

    NASA Astrophysics Data System (ADS)

    Khusairy, K. M.; Ayob, H.; Chan, C. S.; Fauzi, M. I. Mohamed; Mohamad Fakhrul, Z. O.; Shahril Shah, G. S. M.; Azlan, O.; Rasad, M. A.; Hashim, A. M.; Arshad, Z.; E, E. Ibrahim; Saifulizan, M. N.

    2015-12-01

    Rice being the staple food crops, hundreds of land races in it makes the diversity of rice crops. Aerobic rice production was introduced which requires much less water input to safeguard and sustain the rice production and conserve water due to decreasing water resources, climatic changes and competition from urban and industrial users. Mechanization system plays an important role for the success of aerobic rice cultivation. All farming activities for aerobic rice production are run on aerobic soil conditions. Row seeder mechanization system is developed to replace conventional seeding technique on the aerobic rice field. It is targeted for small and the large scale aerobic rice farmers. The aero - seeder machine is used for the small scale aerobic rice field, while the accord - seeder is used for the large scale aerobic rice field. The use of this mechanization machine can eliminate the tedious and inaccurate seeding operations reduce labour costs and increases work rate. The machine is easy to operate and it can increase crop establishment rate. It reduce missing hill, increasing planting and crop with high yield can be produce. This machine is designed for low costs maintenance and it is easy to dismantle and assemble during maintenance and it is safe to be used.

  12. Aerobic Fitness for the Moderately Retarded.

    ERIC Educational Resources Information Center

    Bauer, Dan

    1981-01-01

    Intended for physical education teachers, the booklet offers ideas for incorporating aerobic conditioning into programs for moderately mentally retarded students. An explanation of aerobic fitness and its benefits is followed by information on initiating a fitness program with evaluation of height, weight, body fat, resting heart rate, and…

  13. Aerobic Dancing--A Rhythmic Sport.

    ERIC Educational Resources Information Center

    Sorensen, Jacki

    Fitness programs now and in the future must offer built-in cardiovascular conditioning, variety, novelty, and change to meet the physical, mental, and emotional needs of our society. Aerobic dancing (dancing designed to train and strengthen the heart, lungs, and vascular system) is one of the first indoor group Aerobic exercise programs designed…

  14. Mixed Production of Filamentous Fungal Spores for Preventing Soil-Transmitted Helminth Zoonoses: A Preliminary Analysis

    PubMed Central

    Arias, M. S.; Cazapal-Monteiro, C. F.; Suárez, J.; Miguélez, S.; Francisco, I.; Arroyo, F. L.; Suárez, J. L.; Paz-Silva, A.; Sánchez-Andrade, R.; Mendoza de Gives, P.

    2013-01-01

    Helminth zoonoses are parasitic infections shared by humans and animals, being the soil-transmitted helminths (STHs) mainly caused by roundworms (ascarids) and hookworms. This study was aimed to assess the individual and/or mixed production of two helminth-antagonistic fungi, one ovicide (Mucor circinelloides) and other predator (Duddingtonia flagrans). Fungi were grown both in Petri plates and in a submerged culture (composed by water, NaCl, Na2HPO4· 12 H2O, and wheat (Triticum aestivum)). A Fasciola hepatica recombinant protein (FhrAPS) was incorporated to the cultures to improve fungal production. All the cultured plates showed fungal growth, without difference in the development of the fungi when grown alone or mixed. High counts of Mucor spores were produced in liquid media cultures, and no significant differences were achieved regarding single or mixed cultures, or the incorporation of the FhrAPS. A significantly higher production of Duddingtonia spores after the incorporation of the FhrAPS was observed. When analyzing the parasiticide efficacy of the fungal mixture, viability of T. canis eggs reduced to 51%, and the numbers of third stage cyathostomin larvae reduced to 4%. It is concluded, the capability of a fungal mixture containing an ovicide (Mucor) and a predator species (Duddingtonia) for growing together in a submerged medium containing the FhrAPS offers a very interesting tool for preventing STHs. PMID:23710451

  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. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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.

  11. Evaluation of counting error due to colony masking in bioaerosol sampling.

    PubMed

    Chang, C W; Hwang, Y H; Grinshpun, S A; Macher, J M; Willeke, K

    1994-10-01

    Colony counting error due to indistinguishable colony overlap (i.e., masking) was evaluated theoretically and experimentally. A theoretical model to predict colony masking was used to determine colony counting efficiency by Monte Carlo computer simulation of microorganism collection and development into CFU. The computer simulation was verified experimentally by collecting aerosolized Bacillus subtilis spores and examining micro- and macroscopic colonies. Colony counting efficiency decreased (i) with increasing density of collected culturable microorganisms, (ii) with increasing colony size, and (iii) with decreasing ability of an observation system to distinguish adjacent colonies as separate units. Counting efficiency for 2-mm colonies, at optimal resolution, decreased from 98 to 85% when colony density increased from 1 to 10 microorganisms cm-2, in contrast to an efficiency decrease from 90 to 45% for 5-mm colonies. No statistically significant difference (alpha = 0.05) between experimental and theoretical results was found when colony shape was used to estimate the number of individual colonies in a CFU. Experimental colony counts were 1.2 times simulation estimates when colony shape was not considered, because of nonuniformity of actual colony size and the better discrimination ability of the human eye relative to the model. Colony surface densities associated with high counting accuracy were compared with recommended upper plate count limits and found to depend on colony size and an observation system's ability to identify overlapped colonies. Correction factors were developed to estimate the actual number of collected microorganisms from observed colony counts.(ABSTRACT TRUNCATED AT 250 WORDS)

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. [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.

  19. 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.

  20. 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.

  1. Combined effects of heat, nisin and acidification on the inactivation of Clostridium sporogenes spores in carrot-alginate particles: from kinetics to process validation.

    PubMed

    Naim, F; Zareifard, M R; Zhu, S; Huizing, R H; Grabowski, S; Marcotte, M

    2008-10-01

    Combined effects of mild temperatures, acidification and nisin on the thermal resistance of Clostridium sporogenes ATCC 11437 spores were assessed. Inoculated carrot-alginate particles were used as a solid-food model for the validation of the spore inactivation during the flow of a solid-liquid food system through the holding tube of an aseptic processing unit. Inactivation kinetics was studied in a water bath with the spores inoculated into carrot-alginate particles and in Sorensen's phosphate buffer. For temperatures of 70-90 degrees C, D-values in the buffer were 24.9-5.7 min, much lower than those evaluated for the particles (115.1-22.2 min). Statistical analyses showed significant synergistic effects of temperature and pH on spore inactivation for both media. Acidification reduced the heat resistance of the spores by reducing the D-values. Nisin was not significantly effective at the lower concentrations (up to 750 IU/g). The combination of 90 degrees C, pH: 4.5 and 500IU/g nisin resulted in a ten-fold decrease of the D-value for spores inoculated in the particles (from 111.1 to 10.6 min). Microbial validation tests were conducted using a pilot-scale aseptic processing unit with a mixture of carrot cubes (10%) and carrier liquid of 2%-carboxymethylcellulose solution (90%). Spore-inoculated carrot-alginate particles (initial counts of 106 CFU/g, obtained after come-up-time pre-heat) with pH 3.5 and 2000 IU/g nisin were processed at 90 degrees C in the aseptic processing unit. Microbial analysis showed no spore survivors in the particles after passing through the holding tube (5.2-6.0 min of residence time). The proposed combination of these hurdles significantly enhanced the spore inactivation rate (D(90)=1.17 min) as compared to that for thermal treatment only (D(90)=19.6 min).

  2. 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 ...

  3. ATP bioluminescence rapid detection of total viable count in soy sauce.

    PubMed

    Yan, Shou-Lei; Miao, Su-Na; Deng, Shao-Ya; Zou, Min-Juan; Zhong, Fo-Sheng; Huang, Wen-Biao; Pan, Si-Yi; Wang, Qing-Zhang

    2012-01-01

    The adenosine triphosphate (ATP) bioluminescence rapid determination method may be useful for enumerating the total viable count (TVC) in soy sauce, as it has been previously used in food and beverages for sanitation with good precision. However, many factors interfere with the correlation between total aerobic plate counts and ATP bioluminescence. This study investigated these interfering factors, including ingredients of soy sauce and bacteria at different physiological stages. Using the ATP bioluminescence method, TVC was obtained within 4 h, compared to 48 h required for the conventional aerobic plate count (APC) method. Our results also indicated a high correlation coefficient (r = 0.90) between total aerobic plate counts and ATP bioluminescence after filtration and resuscitation with special medium. The limit of quantification of the novel detection method is 100 CFU/mL; there is a good linear correlation between the bioluminescence intensity and TVC in soy sauce in the range 1 × 10(2) -3 × 10(4) CFU/mL and even wider. The method employed a luminescence recorder (Tristar LB-941) and 96-well plates and could analyse 50-100 samples simultaneously at low cost. In this study, we evaluated and eliminated the interfering factors and made the ATP bioluminescence rapid method available for enumerating TVC in soy sauce.

  4. 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

  5. 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

  6. Snow in the city as a spore bank of potentially pathogenic fungi.

    PubMed

    Ejdys, Elżbieta; Biedunkiewicz, Anna; Dynowska, Maria; Sucharzewska, Ewa

    2014-02-01

    This study evaluates the role of snow as a specific ecological niche and a vector in fungal spreading with particular emphasis on potential pathogens in seasonally and daily changing conditions. The experimental material was fungi isolated from the atmospheric air, snow cover, and fragments of ice and soil from underneath the snow cover. The total count of microfungi in the air before snowfall, i.e. in the autumn, reached 1756.1 CFU/m(3) on average. After the first snowfalls, it dropped to 85.2 CFU/m(3). The analyzed samples of snow cover contained from 101.6 to 8500.0 CFU/m(3) of fungi. Furthermore, 26 species of yeast and yeast-like fungi were isolated from the experimental material. Amongst the analyzed species, 13 were potential anthropopathogens. Though another three species were isolated from organ ontocenoses, i.e. Candida intermedia, Saccharomyces bayanus and Zygosaccharomyces rouxii, their pathogenic potential has not yet been explicitly confirmed. The results of the presented study may be applied in predicting concentrations of fungal spores responsible for mycoses. The first snowfalls significantly reduced the number of colony-forming units of fungi in the air. Under conditions of temperate climate, snow becomes a temporary bank of yeast-like fungi spores and while it melts cells of deposited microfungi migrate to the atmosphere. Hence, individuals with impaired immunity or in the course of immunosuppression or recovery should avoid long walks during periods of snow melting. The count of fungi in urban bioaerosol during the melt may be reduced through systematic removal of snow cover, which is a significant reservoir of potential pathogens. In addition, it should be noted that even a typical psychrophilic strain, capable of surviving at a temperature of 37°C, may bear a significant pathogenic potential.

  7. Aerobic fitness testing: an update.

    PubMed

    Stevens, N; Sykes, K

    1996-12-01

    This study confirms that all three tests are reliable tools for the assessment of cardiorespiratory fitness and the prediction of aerobic capacity. While this particular study consisted of active, youthful subjects, subsequent studies at University College Chester have found similar findings with larger databases and a wider cross-section of subjects. The Astrand cycle test and Chester step test are submaximal tests with error margins of 5-15 per cent and therefore, not as precise as maximal testing. However, they still give a reasonably accurate reflection of an individual's fitness without the cost, time, effort and risk on the part of the subject. The bleep test is a low-cost maximal test designed for well-motivated, active individuals who are used to running to physical exhaustion. Used on other groups, results will not accurately reflect cardiorespiratory fitness values. While all three tests have inherent advantages and disadvantages, perhaps the most important factors are the knowledge and skills of the tester. Without a sound understanding of the physiological principles underlying these tests, and the ability to conduct an accurate assessment and evaluation of results in a knowledgeable and meaningful way, then the credibility of the tests and the results become suspect. However, used correctly, aerobic capacity tests can provide valuable baseline data about the fitness levels of individuals and data from which exercise programmes may be developed. The tests also enable fitness improvements to be monitored, help to motivate participants by establishing reasonable and achievable goals, assist in risk stratification and facilitate participants' education about the importance of physical fitness for work and for life. Since this study was completed, further tests have been repeated on 140 subjects of a wider age and ability range. This large database confirms the results found in this study.

  8. Aerobic glycolysis and lymphocyte transformation

    PubMed Central

    Hume, David A.; Radik, Judith L.; Ferber, Ernst; Weidemann, Maurice J.

    1978-01-01

    1. The role of enhanced aerobic glycolysis in the transformation of rat thymocytes by concanavalin A has been investigated. Concanavalin A addition doubled [U-14C]glucose uptake by rat thymocytes over 3h and caused an equivalent increased incorporation into protein, lipids and RNA. A disproportionately large percentage of the extra glucose taken up was converted into lactate, but concanavalin A also caused a specific increase in pyruvate oxidation, leading to an increase in the percentage contribution of glucose to the respiratory fuel. 2. Acetoacetate metabolism, which was not affected by concanavalin A, strongly suppressed pyruvate oxidation in the presence of [U-14C]glucose, but did not prevent the concanavalin A-induced stimulation of this process. Glucose uptake was not affected by acetoacetate in the presence or absence of concanavalin A, but in each case acetoacetate increased the percentage of glucose uptake accounted for by lactate production. 3. [3H]Thymidine incorporation into DNA in concanavalin A-treated thymocyte cultures was sensitive to the glucose concentration in the medium in a biphasic manner. Very low concentrations of glucose (25μm) stimulated DNA synthesis half-maximally, but maximum [3H]thymidine incorporation was observed only when the glucose concentration was raised to 1mm. Lactate addition did not alter the sensitivity of [3H]-thymidine uptake to glucose, but inosine blocked the effect of added glucose and strongly inhibited DNA synthesis. 4. It is suggested that the major function of enhanced aerobic glycolysis in transforming lymphocytes is to maintain higher steady-state amounts of glycolytic intermediates to act as precursors for macromolecule synthesis. PMID:310305

  9. Possible overestimation of surface disinfection efficiency by assessment methods based on liquid sampling procedures as demonstrated by in situ quantification of spore viability.

    PubMed

    Grand, I; Bellon-Fontaine, M-N; Herry, J-M; Hilaire, D; Moriconi, F-X; Naïtali, M

    2011-09-01

    The standard test methods used to assess the efficiency of a disinfectant applied to surfaces are often based on counting the microbial survivors sampled in a liquid, but total cell removal from surfaces is seldom achieved. One might therefore wonder whether evaluations of microbial survivors in liquid-sampled cells are representative of the levels of survivors in whole populations. The present study was thus designed to determine the "damaged/undamaged" status induced by a peracetic acid disinfection for Bacillus atrophaeus spores deposited on glass coupons directly on this substrate and to compare it to the status of spores collected in liquid by a sampling procedure. The method utilized to assess the viability of both surface-associated and liquid-sampled spores included fluorescence labeling with a combination of Syto 61 and Chemchrome V6 dyes and quantifications by analyzing the images acquired by confocal laser scanning microscopy. The principal result of the study was that the viability of spores sampled in the liquid was found to be poorer than that of surface-associated spores. For example, after 2 min of peracetic acid disinfection, less than 17% ± 5% of viable cells were detected among liquid-sampled cells compared to 79% ± 5% or 47% ± 4%, respectively, when the viability was evaluated on the surface after or without the sampling procedure. Moreover, assessments of the survivors collected in the liquid phase, evaluated using the microscopic method and standard plate counts, were well correlated. Evaluations based on the determination of survivors among the liquid-sampled cells can thus overestimate the efficiency of surface disinfection procedures.

  10. 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

  11. 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

  12. 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

  13. Fit women are not able to use the whole aerobic capacity during aerobic dance.

    PubMed

    Edvardsen, Elisabeth; Ingjer, Frank; Bø, Kari

    2011-12-01

    Edvardsen, E, Ingjer, F, and Bø, K. Fit women are not able to use the whole aerobic capacity during aerobic dance. J Strength Cond Res 25(12): 3479-3485, 2011-This study compared the aerobic capacity during maximal aerobic dance and treadmill running in fit women. Thirteen well-trained female aerobic dance instructors aged 30 ± 8.17 years (mean ± SD) exercised to exhaustion by running on a treadmill for measurement of maximal oxygen uptake (VO(2)max) and peak heart rate (HRpeak). Additionally, all subjects performed aerobic dancing until exhaustion after a choreographed videotaped routine trying to reach the same HRpeak as during maximal running. The p value for statistical significance between running and aerobic dance was set to ≤0.05. The results (mean ± SD) showed a lower VO(2)max in aerobic dance (52.2 ± 4.02 ml·kg·min) compared with treadmill running (55.9 ± 5.03 ml·kg·min) (p = 0.0003). Further, the mean ± SD HRpeak was 182 ± 9.15 b·min in aerobic dance and 192 ± 9.62 b·min in treadmill running, giving no difference in oxygen pulse between the 2 exercise forms (p = 0.32). There was no difference in peak ventilation (aerobic dance: 108 ± 10.81 L·min vs. running: 113 ± 11.49 L·min). In conclusion, aerobic dance does not seem to be able to use the whole aerobic capacity as in running. For well endurance-trained women, this may result in a lower total workload at maximal intensities. Aerobic dance may therefore not be as suitable as running during maximal intensities in well-trained females.

  14. Compton suppression gamma-counting: The effect of count rate

    USGS Publications Warehouse

    Millard, H.T.

    1984-01-01

    Past research has shown that anti-coincidence shielded Ge(Li) spectrometers enhanced the signal-to-background ratios for gamma-photopeaks, which are situated on high Compton backgrounds. Ordinarily, an anti- or non-coincidence spectrum (A) and a coincidence spectrum (C) are collected simultaneously with these systems. To be useful in neutron activation analysis (NAA), the fractions of the photopeak counts routed to the two spectra must be constant from sample to sample to variations must be corrected quantitatively. Most Compton suppression counting has been done at low count rate, but in NAA applications, count rates may be much higher. To operate over the wider dynamic range, the effect of count rate on the ratio of the photopeak counts in the two spectra (A/C) was studied. It was found that as the count rate increases, A/C decreases for gammas not coincident with other gammas from the same decay. For gammas coincident with other gammas, A/C increases to a maximum and then decreases. These results suggest that calibration curves are required to correct photopeak areas so quantitative data can be obtained at higher count rates. ?? 1984.

  15. Kids Count [and] Families Count in Delaware: Fact Book, 1998.

    ERIC Educational Resources Information Center

    Nelson, Carl, Ed.; Wilson, Nancy, Ed.

    This Kids Count report is combined with Families Count, and provides information on statewide trends affecting children and families in Delaware. The first statistical profile is based on 10 main indicators of child well-being: (1) births to teens; (2) low birth weight babies; (3) infant mortality; (4) child deaths; (5) teen deaths; (6) juvenile…

  16. "Just" Counting: Young Children's Oral Counting and Enumeration

    ERIC Educational Resources Information Center

    Threlfall, John; Bruce, Bob

    2005-01-01

    This article focuses on the specific skills and abilities of young children in oral counting and enumeration. Responses to an oral counting task and an enumeration task by a sample (n=93) of 3- and 4-year old children attending a range of pre-five establishments in an urban district of northern England are described. The findings, whilst providing…

  17. Impact of different water activities (aw) adjusted by solutes on high pressure high temperature inactivation of Bacillus amyloliquefaciens spores

    PubMed Central

    Sevenich, Robert; Reineke, Kai; Hecht, Philipp; Fröhling, Antje; Rauh, Cornelia; Schlüter, Oliver; Knorr, Dietrich

    2015-01-01

    Much research has been conducted to comprehend the mechanisms of high pressure (HP) inactivation of spores in aqueous systems but for food model systems these information are scarce. In these systems spores can interact with ingredients which then could possibly lead to retarded or reduced inactivation, which can cause a problem for the sterilization process. The protective mechanism of a reduced aw-value is still unclear. HP processing might prove valuable to overcome protective effects of solutes and achieve shorter process times for sterilization under HP. To gain insight into the underlying mechanisms five aw-values (0.9, 0.92, 0.94, 0.96, 1) were adjusted with two different solutes (NaCl, sucrose). Solutions were inoculated with spores of Bacillus amyloliquefaciens and treated at 105, 110, and 115°C at 600 MPa. Further a thermal inactivation was conducted at the same temperatures for a comparison with the HP data. Afterward, the influence of HP high temperature treatment on the inactivation, the dipicolinic acid (DPA)-release and membrane constitution was assessed by plate count, HPLC and flow cytometry (FCM). The results show that during HP treatments sucrose and salt both have a protective effect, in which the influence of sucrose on the retarded inactivation is higher. The threshold water activities (aw), which is 0.94, here salt and sucrose have a significant influence on the inactivation. The comparison of thermal (105–115°C) and HP and high temperature (600 MPa, 105–115°C) treated samples showed that the time needed to achieve a 4–5 log10 inactivation is reduced from 45 (aw = 1) to 75 (aw = 0.9) min at 105°C to 3 (aw = 1) to 15 (aw = 0.9) minutes at 600 MPa and 105°C. The release of DPA is the rate limiting step of the inactivation and therefore monitoring the release is of great interest. The DPA-release is slowed down in high concentrated solutions (e.g., sucrose, salt) in comparison to aw 1. Since there is a difference in the way the

  18. 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.

  19. 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.

  20. 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

  1. Effects of an esterase-producing inoculant on fermentation, aerobic stability, and neutral detergent fiber digestibility of corn silage.

    PubMed

    Kang, T W; Adesogan, A T; Kim, S C; Lee, S S

    2009-02-01

    This experiment evaluated effects of an inoculant containing esterase-producing bacteria on fermentation, aerobic stability, in situ dry matter digestibility (DMD), and neutral detergent fiber (NDF) digestibility (NDFD) of corn silage. Two corn hybrids grown on adjacent fields [Croplan Genetics 851RR2 (CS1) and Vigoro 61R36 (CS2)] were harvested at approximately 39% dry matter. Each forage was conserved in quadruplicate in 20-L mini silos with or without application of an inoculant at a level to achieve 1.0 x 10(4) cfu/g of Lactobacillus casei and 1.0 x 10(5) cfu/g of Lactobacillus buchneri. After 110 d of ensiling, silos were opened and silages were analyzed for chemical composition, fermentation indices, microbial counts, and aerobic stability. In situ DMD, 24-h and 48-h DMD, and NDFD were measured by incubating ground (6-mm) samples in triplicate in each of 2 lactating, fistulated dairy cows fed a corn silage-based diet. Inoculation decreased concentrations of total fermentation acids and lactate, as well as lactate to acetate ratio, and increased propionate concentration compared with the uninoculated control in CS1 but not CS2. Inoculation tended to decrease yeast counts of CS1 but increased yeast counts and tended to increase the mold counts of CS2. Consequently, inoculation improved the aerobic stability of CS1 by 57.3 h (98%) but decreased that of CS2 by 20.5 h (20%). Inoculation also increased the potentially degradable fraction of CS1 and the total degradable fraction, 24-h and 48-h DMD, and 48-h NDFD of CS2. Inoculation of CS1 modified the fermentation, improved the aerobic stability, and increased the potentially degradable DM fraction. Inoculation of CS2 did not affect fermentation, but decreased the aerobic stability and increased the total degradable DM fraction, 24-h and 48-h DMD, and 48-h NDFD.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. Low White Blood Cell Count

    MedlinePlus

    Symptoms Low white blood cell count By Mayo Clinic Staff A low white blood cell count (leukopenia) is a decrease in disease-fighting cells ( ... a decrease in a certain type of white blood cell (neutrophil). The definition of low white blood cell ...

  7. Young Children Counting at Home

    ERIC Educational Resources Information Center

    Griffiths, Rose

    2007-01-01

    Learning to count is something that most children start to do by the time they are about two, and parents know from first-hand experience that family members play a big part in helping with this complex process. In this article, the author describes a project involving families sharing effective counting activities. The project called…

  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. The effects of aerobic training on children's creativity, self-perception, and aerobic power.

    PubMed

    Herman-Tofler, L R; Tuckman, B W

    1998-10-01

    The article examines whether participation in an aerobic exercise program (AE), as compared with a traditional physical education class (PE), significantly increased children's perceived athletic competence, physical appearance, social acceptance, behavioral conduct, and global self-worth; increased their figural creativity; and improved aerobic power as measured by an 800-meter run around a track. Further research on the effects of different types of AE is discussed, as well as the need for aerobic conditioning in the elementary school.

  11. 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

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. 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.

  17. Conditioning and Aerobics for Older Americans.

    ERIC Educational Resources Information Center

    Hansen, Joyce

    1980-01-01

    A class designed for the maintenance and gradual improvement of senior citizens' physical fitness includes relaxation training, flexibility and stretching exercises, interval training activities (designed as a link between less strenuous exercise and more strenuous activities), and aerobic exercises. (CJ)

  18. The rise of oxygen and aerobic biochemistry.

    PubMed

    Saito, Mak A

    2012-01-11

    Analysis of conserved protein folding domains across extant genomes by Kim et al. in this issue of Structure provides insights into the timing of some of the earliest aerobic metabolisms to arise on Earth.

  19. Neuromodulation of Aerobic Exercise—A Review

    PubMed Central

    Heijnen, Saskia; Hommel, Bernhard; Kibele, Armin; Colzato, Lorenza S.

    2016-01-01

    Running, and aerobic exercise in general, is a physical activity that increasingly many people engage in but that also has become popular as a topic for scientific research. Here we review the available studies investigating whether and to which degree aerobic exercise modulates hormones, amino acids, and neurotransmitters levels. In general, it seems that factors such as genes, gender, training status, and hormonal status need to be taken into account to gain a better understanding of the neuromodular underpinnings of aerobic exercise. More research using longitudinal studies and considering individual differences is necessary to determine actual benefits. We suggest that, in order to succeed, aerobic exercise programs should include optimal periodization, prevent overtraining and be tailored to interindividual differences, including neuro-developmental and genetically-based factors. PMID:26779053

  20. The Energetics of Aerobic versus Anaerobic Respiration.

    ERIC Educational Resources Information Center

    Champion, Timothy D.; Schwenz, Richard W.

    1990-01-01

    Background information, laboratory procedures, and a discussion of the results of an experiment designed to investigate the difference in energy gained from the aerobic and anaerobic oxidation of glucose are presented. Sample experimental and calculated data are included. (CW)

  1. 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.

  2. Surface Structure of Aerobically Oxidized Diamond Nanocrystals

    DTIC Science & Technology

    2014-10-27

    distribution is unlimited. Surface Structure of Aerobically Oxidized Diamond Nanocrystals The views, opinions and/or findings contained in this report...2211 diamond nanocrystals, REPORT DOCUMENTATION PAGE 11. SPONSOR/MONITOR’S REPORT NUMBER(S) 10. SPONSOR/MONITOR’S ACRONYM(S) ARO 8. PERFORMING...Room 254, Mail Code 8725 New York, NY 10027 -7922 ABSTRACT Surface Structure of Aerobically Oxidized Diamond Nanocrystals Report Title We investigate

  3. Aerobic biodegradation of selected monoterpenes.

    PubMed

    Misra, G; Pavlostathis, S G; Perdue, E M; Araujo, R

    1996-07-01

    Batch experiments were conducted to assess the biotransformation potential of four hydrocarbon monoterpenes (d-limonene, alpha-pinene, gamma-terpinene, and terpinolene) and four alcohols (arbanol, linalool, plinol, and alpha-terpineol) under aerobic conditions at 23 degrees C. Both forest-soil extract and enriched cultures were used as inocula for the biodegradation experiments conducted first without, then with prior microbial acclimation to the monoterpenes tested. All four hydrocarbons and two alcohols were readily degraded. The increase in biomass and headspace CO2 concentrations paralleled the depletion of monoterpenes, thus confirming that terpene disappearance was the result of biodegradation accompanied by microbial growth and mineralization. Plinol resisted degradation in assays using inocula from diverse sources, while arbanol degraded very slowly. A significant fraction of d-limonene-derived carbon was accounted for as non-extractable, dissolved organic carbon, whereas terpineol exhibited a much higher degree of utilization. The rate and extent of monoterpene biodegradation were not significantly affected by the presence of dissolved natural organic matter.

  4. Aerobic catabolism of bile acids.

    PubMed Central

    Leppik, R A; Park, R J; Smith, M G

    1982-01-01

    Seventy-eight stable cultures obtained by enrichment on media containing ox bile or a single bile acid were able to utilize one or more bile acids, as well as components of ox bile, as primary carbon sources for growth. All isolates were obligate aerobes, and most (70) were typical (48) or atypical (22) Pseudomonas strains, the remainder (8) being gram-positive actinomycetes. Of six Pseudomonas isolates selected for further study, five produced predominantly acidic catabolites after growth on glycocholic acid, but the sixth, Pseudomonas sp. ATCC 31752, accumulated as the principal product a neutral steroid catabolite. Optimum growth of Pseudomonas sp. ATCC 31752 on ox bile occurred at pH 7 to 8 and from 25 to 30 degrees C. No additional nutrients were required to sustain good growth, but growth was stimulated by the addition of ammonium sulfate and yeast extract. Good growth was obtained with a bile solids content of 40 g/liter in shaken flasks. A near-theoretical yield of neutral steroid catabolites, comprising a major (greater than 50%) and three minor products, was obtained from fermentor growth of ATCC 31752 in 6.7 g of ox bile solids per liter. The possible commercial exploitation of these findings to produce steroid drug intermediates for the pharmaceutical industry is discussed. PMID:7149711

  5. Mineralization and kinetics of Reactive Brilliant Red X-3B by a combined anaerobic-aerobic bioprocess inoculated with the coculture of fungus and bacterium.

    PubMed

    Shi, Shengnan; Ma, Fang; Sun, Tieheng; Li, Ang; Zhou, Jiti; Qu, Yuanyuan

    2014-01-01

    Mineralization of Reactive Brilliant Red X-3B by a combined anaerobic-aerobic process which was inoculated with the co-culture of Penicillium sp. QQ and Exiguobacterium sp. TL was studied. The optimal conditions of decolorization were investigated by response surface methodology as follows: 132.67 g/L of strain QQ wet spores, 1.09 g/L of strain TL wet cells, 2.25 g/L of glucose, 2.10 g/L of yeast extract, the initial dye concentration of 235.14 mg/L, pH 6.5, and 33 °C. The maximal decolorization rate was about 96 % within 12 h under the above conditions. According to the Haldane kinetic equation, the maximal specific decolorization rate was 89.629 mg/g˙h. It was suggested that in the anaerobic-aerobic combined process, decolorization occurred in the anaerobic unit and chemical oxygen demand (COD) was mainly removed in the aerobic one. Inoculation of fungus QQ in the anaerobic unit was important for mineralization of X-3B. Besides, the divided anaerobic-aerobic process showed better performance of COD removal than the integrated one. It was suggested that the combined anaerobic-aerobic process which was inoculated with co-culture was potentially useful for the field application.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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

  16. 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.

  17. Aerobic Exercise Protects Retinal Function and Structure from Light-Induced Retinal Degeneration

    PubMed Central

    Lawson, Eric C.; Han, Moon K.; Sellers, Jana T.; Chrenek, Micah A.; Hanif, Adam; Gogniat, Marissa A.

    2014-01-01

    Aerobic exercise is a common intervention for rehabilitation of motor, and more recently, cognitive function (Intlekofer and Cotman, 2013; Wood et al., 2012). While the underlying mechanisms are complex, BDNF may mediate much of the beneficial effects of exercise to these neurons (Ploughman et al., 2007; Griffin et al., 2011; Real et al., 2013). We studied the effects of aerobic exercise on retinal neurons undergoing degeneration. We exercised wild-type BALB/c mice on a treadmill (10 m/min for 1 h) for 5 d/week or placed control mice on static treadmills. After 2 weeks of exercise, mice were exposed to either toxic bright light (10,000 lux) for 4 h to induce photoreceptor degeneration or maintenance dim light (25 lux). Bright light caused 75% loss of both retinal function and photoreceptor numbers. However, exercised mice exposed to bright light had 2 times greater retinal function and photoreceptor nuclei than inactive mice exposed to bright light. In addition, exercise increased retinal BDNF protein levels by 20% compared with inactive mice. Systemic injections of a BDNF tropomyosin-receptor-kinase (TrkB) receptor antagonist reduced retinal function and photoreceptor nuclei counts in exercised mice to inactive levels, effectively blocking the protective effects seen with aerobic exercise. The data suggest that aerobic exercise is neuroprotective for retinal degeneration and that this effect is mediated by BDNF signaling. PMID:24523530

  18. Aerobic exercise protects retinal function and structure from light-induced retinal degeneration.

    PubMed

    Lawson, Eric C; Han, Moon K; Sellers, Jana T; Chrenek, Micah A; Hanif, Adam; Gogniat, Marissa A; Boatright, Jeffrey H; Pardue, Machelle T

    2014-02-12

    Aerobic exercise is a common intervention for rehabilitation of motor, and more recently, cognitive function (Intlekofer and Cotman, 2013; Wood et al., 2012). While the underlying mechanisms are complex, BDNF may mediate much of the beneficial effects of exercise to these neurons (Ploughman et al., 2007; Griffin et al., 2011; Real et al., 2013). We studied the effects of aerobic exercise on retinal neurons undergoing degeneration. We exercised wild-type BALB/c mice on a treadmill (10 m/min for 1 h) for 5 d/week or placed control mice on static treadmills. After 2 weeks of exercise, mice were exposed to either toxic bright light (10,000 lux) for 4 h to induce photoreceptor degeneration or maintenance dim light (25 lux). Bright light caused 75% loss of both retinal function and photoreceptor numbers. However, exercised mice exposed to bright light had 2 times greater retinal function and photoreceptor nuclei than inactive mice exposed to bright light. In addition, exercise increased retinal BDNF protein levels by 20% compared with inactive mice. Systemic injections of a BDNF tropomyosin-receptor-kinase (TrkB) receptor antagonist reduced retinal function and photoreceptor nuclei counts in exercised mice to inactive levels, effectively blocking the protective effects seen with aerobic exercise. The data suggest that aerobic exercise is neuroprotective for retinal degeneration and that this effect is mediated by BDNF signaling.

  19. White blood cell counting system

    NASA Technical Reports Server (NTRS)

    1972-01-01

    The design, fabrication, and tests of a prototype white blood cell counting system for use in the Skylab IMSS are presented. The counting system consists of a sample collection subsystem, sample dilution and fluid containment subsystem, and a cell counter. Preliminary test results show the sample collection and the dilution subsystems are functional and fulfill design goals. Results for the fluid containment subsystem show the handling bags cause counting errors due to: (1) adsorption of cells to the walls of the container, and (2) inadequate cleaning of the plastic bag material before fabrication. It was recommended that another bag material be selected.

  20. Testing for aerobic heterotrophic bacteria allows no prediction of contamination with potentially pathogenic bacteria in the output water of dental chair units

    PubMed Central

    Bristela, Margit; Skolka, Astrid; Schmid-Schwap, Martina; Piehslinger, Eva; Indra, Alexander; Wewalka, Günther; Stauffer, Fritz

    2012-01-01

    Background: Currently, to our knowledge, quality of output water of dental chair units is not covered by specific regulations in the European Union, and national recommendations are heterogeneous. In Germany, water used in dental chair units must follow drinking water quality. In the United States of America, testing for aerobic heterotrophic bacteria is recommended. The present study was performed to evaluate whether the counts of aerobic heterotrophic bacteria correlate with the presence of potentially pathogenic bacteria such as Legionella spp. or Pseudomonas aeruginosa. Methods: 71 samples were collected from 26 dental chair units with integrated disinfection device and 31 samples from 15 outlets of the water distribution pipework within the department were examined. Samples were tested for aerobic heterotrophic bacteria at 35°C and 22°C using different culture media and for Legionella spp. and for Pseudomonas aeruginosa. Additionally, strains of Legionella pneumophila serogroup 1 were typed with monoclonal antibodies and representative samples of Legionella pneumophila serogroup 1 were typed by sequence based typing. Results: Our results showed a correlation between different agars for aerobic heterotrophic bacteria but no correlation for the count of aerobic heterotrophic bacteria and the presence of Legionella spp. or Pseudomonas aeruginosa. Conclusion: Testing for aerobic heterotrophic bacteria in output water or water distribution pipework within the departments alone is without any value for predicting whether the water is contaminated with potentially pathogenic bacteria like Legionella spp. or Pseudomonas aeruginosa. PMID:22558046

  1. 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

  2. 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.

  3. 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

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. 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.

  13. 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

  14. Counting Triangles to Sum Squares

    ERIC Educational Resources Information Center

    DeMaio, Joe

    2012-01-01

    Counting complete subgraphs of three vertices in complete graphs, yields combinatorial arguments for identities for sums of squares of integers, odd integers, even integers and sums of the triangular numbers.

  15. 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.

  16. 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

  17. 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...

  18. Aerobic Excercise and Research Opportunities to Benefit Impaired Children. (Project AEROBIC). Final Report.

    ERIC Educational Resources Information Center

    Idaho Univ., Moscow.

    The final report summarizes accomplishments of Project AEROBIC (Aerobic Exercise and Research Opportunities to Benefit Impaired Children), which provided a physical education exercise program for severely, profoundly, and multiply handicapped children aged 10-21. Activities are outlined for the 3 year period and include modification of exercise…

  19. Effect of long term anaerobic and intermittent anaerobic/aerobic starvation on aerobic granules.

    PubMed

    Pijuan, Maite; Werner, Ursula; Yuan, Zhiguo

    2009-08-01

    The effect of long term anaerobic and intermittent anaerobic/aerobic starvation on the structure and activity of aerobic granules was studied. Aerobic granular sludge treating abattoir wastewater and achieving high levels of nutrient removal was subjected to 4-5 week starvation under anaerobic and intermittent anaerobic/aerobic conditions. Microscopic pictures of granules at the beginning of the starvation period presented a round and compact surface morphology with a much defined external perimeter. Under both starvation conditions, the morphology changed at the end of starvation with the external border of the granules surrounded by floppy materials. The loss of granular compactness was faster and more pronounced under anaerobic/aerobic starvation conditions. The release of Ca(2+) at the onset of anaerobic/aerobic starvation suggests a degradation of extracellular polymeric substances. The activity of ammonia oxidizing bacteria was reduced by 20 and 36% during anaerobic and intermittent anaerobic/aerobic starvation, respectively. When fresh wastewater was reintroduced, the granules recovered their initial morphology within 1 week of normal operation and the nutrient removal activity recovered fully in 3 weeks. The results show that both anaerobic and intermittent anaerobic/aerobic conditions are suitable for maintaining granule structure and activity during starvation.

  20. 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.

  1. Counting on Using a Number Game

    ERIC Educational Resources Information Center

    Betts, Paul

    2015-01-01

    Counting all and counting on are distinct counting strategies that can be used to compute such quantities as the total number of objects in two sets (Wright, Martland, and Stafford 2010). Given five objects and three more objects, for example, children who use counting all to determine quantity will count both collections; that is, they count…

  2. Use of alternative carrier materials in AOAC Official Method 2008.05, efficacy of liquid sporicides against spores of Bacillus subtilis on a hard, nonporous surface, quantitative three-step method.

    PubMed

    Tomasino, Stephen F; Rastogi, Vipin K; Wallace, Lalena; Smith, Lisa S; Hamilton, Martin A; Pines, Rebecca M

    2010-01-01

    The quantitative Three-Step Method (TSM) for testing the efficacy of liquid sporicides against spores of Bacillus subtilis on a hard, nonporous surface (glass) was adopted as AOAC Official Method 2008.05 in May 2008. The TSM uses 5 x 5 x 1 mm coupons (carriers) upon which spores have been inoculated and which are introduced into liquid sporicidal agent contained in a microcentrifuge tube. Following exposure of inoculated carriers and neutralization, spores are removed from carriers in three fractions (gentle washing, fraction A; sonication, fraction B; and gentle agitation, fraction C). Liquid from each fraction is serially diluted and plated on a recovery medium for spore enumeration. The counts are summed over the three fractions to provide the density (viable spores per carrier), which is log10-transformed to arrive at the log density. The log reduction is calculated by subtracting the mean log density for treated carriers from the mean log density for control carriers. This paper presents a single-laboratory investigation conducted to evaluate the applicability of using two porous carrier materials (ceramic tile and untreated pine wood) and one alternative nonporous material (stainless steel). Glass carriers were included in the study as the reference material. Inoculated carriers were evaluated against three commercially available liquid sporicides (sodium hypochlorite, a combination of peracetic acid and hydrogen peroxide, and glutaraldehyde), each at two levels of presumed efficacy (medium and high) to provide data for assessing the responsiveness of the TSM. Three coupons of each material were evaluated across three replications at each level; three replications of a control were required. Even though all carriers were inoculated with approximately the same number of spores, the observed counts of recovered spores were consistently higher for the nonporous carriers. For control carriers, the mean log densities for the four materials ranged from 6.63 for

  3. 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

  4. 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.

  5. 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

  6. 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

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

    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.

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. 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 ...

  13. 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.

  14. 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...

  15. Standardization of 241Am by digital coincidence counting, liquid scintillation counting and defined solid angle counting.

    PubMed

    Balpardo, C; Capoulat, M E; Rodrigues, D; Arenillas, P

    2010-01-01

    The nuclide (241)Am decays by alpha emission to (237)Np. Most of the decays (84.6%) populate the excited level of (237)Np with energy of 59.54 keV. Digital coincidence counting was applied to standardize a solution of (241)Am by alpha-gamma coincidence counting with efficiency extrapolation. Electronic discrimination was implemented with a pressurized proportional counter and the results were compared with two other independent techniques: Liquid scintillation counting using the logical sum of double coincidences in a TDCR array and defined solid angle counting taking into account activity inhomogeneity in the active deposit. The results show consistency between the three methods within a limit of a 0.3%. An ampoule of this solution will be sent to the International Reference System (SIR) during 2009. Uncertainties were analysed and compared in detail for the three applied methods.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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

  3. 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.

  4. 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...

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. Survival of foodborne pathogenic bacteria (Bacillus cereus, Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Listeria monocytogenes) and Bacillus cereus spores in fermented alcoholic beverages (beer and refined rice wine).

    PubMed

    Kim, S A; Kim, N H; Lee, S H; Hwang, I G; Rhee, M S

    2014-03-01

    Only limited information is available on the microbiological safety of fermented alcoholic beverages because it is still a common belief that such beverages do not provide a favorable environment for bacterial growth and survival. Thus, in this study, we examined the survival of major foodborne pathogens and spores in fermented alcoholic beverages. Foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Staphylococcus aureus) and B. cereus spores (initial population, 3 to 4 log CFU/ml) were inoculated separately into three types of beer and refined rice wine, which were then stored at 5 and 22°C. Bacterial counts were assayed periodically for up to 28 days. Vegetative B. cereus counts decreased rapidly, whereas B. cereus spore counts remained constant (P > 0.05) for a long period of time in all beverages. Vegetative B. cereus cells formed spores in beer at 5 and 22°C, and the spores survived for long periods. Among vegetative cells, E. coli O157:H7 had the highest survival (only 1.49 to 1.56 log reduction during 28 days in beer at 5°C). Beer and refined rice wine supported microbial survival from several days to several weeks. Our results appear to contradict the common belief that pathogens cannot survive in alcoholic beverages. Long-term survival of pathogens (especially B. cereus and E. coli O157:H7) in beer and refined rice wine should be taken into consideration by the manufacturers of these beverages. This study provides basic information that should help further research into microbial survival in alcoholic beverages and increase the microbiological safety regulation of fermented alcoholic beverages.

  10. 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

  11. 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

  12. 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.

  13. Evidence for a role of biosurfactants produced by Pseudomonas fluorescens in the spoilage of fresh aerobically stored chicken meat.

    PubMed

    Mellor, Glen E; Bentley, Jessica A; Dykes, Gary A

    2011-08-01

    Fresh chicken meat is a fat-rich environment and we therefore hypothesised that production of biosurfactants to increase bioavailability of fats may represent one way in which spoilage bacteria might enhance the availability of nutrients. Numbers of Pseudomonas were determined on a total of 20 fresh and 20 spoiled chicken thighs with skin. A total of 400 randomly isolated Pseudomonas colonies from fresh (200) and spoiled (200) chicken were screened for the presence of biosurfactant production. Biosurfactant producing strains represented 5% and 72% of the Pseudomonas spp. isolates from fresh (mean count 2.3 log(10) cfu g(-1)) and spoiled (mean count 7.4 log(10) cfu g(-1)) chicken skin, respectively. Partially-purified biosurfactants derived from a subgroup of four Pseudomonasfluorescens strains obtained through the screening process were subsequently used to investigate the role that the addition of these compounds plays in the spoilage of aerobically stored chicken. Emulsification potential of the four selected biosurfactants was measured against a range of hydrocarbons and oils. All four biosurfactants displayed a greater ability to emulsify rendered chicken fat than hydrocarbons (paraffin liquid, toluene and hexane) and oils (canola, olive, sunflower and vegetable). Storage trials (4 °C) of chicken meat treated with the four selected biosurfactants revealed a significantly greater (P < 0.05) total aerobic count in biosurfactant treated samples, as compared to untreated samples on each day (0, 1, 2, 3) of storage. For biosurfactant treated samples the greatest increase in total aerobic count (1.3-1.7 log(10) cfu g(-1)) occurred following one day of incubation. These results indicate that biosurfactants produced by Pseudomonas spp. may play an important role in the spoilage of aerobically stored chicken meat by making nutrients more freely available and providing strains producing them with a competitive advantage.

  14. Hanford whole body counting manual

    SciTech Connect

    Palmer, H.E.; Rieksts, G.A.; Lynch, T.P.

    1990-06-01

    This document describes the Hanford Whole Body Counting Program as it is administered by Pacific Northwest Laboratory (PNL) in support of the US Department of Energy--Richland Operations Office (DOE-RL) and its Hanford contractors. Program services include providing in vivo measurements of internally deposited radioactivity in Hanford employees (or visitors). Specific chapters of this manual deal with the following subjects: program operational charter, authority, administration, and practices, including interpreting applicable DOE Orders, regulations, and guidance into criteria for in vivo measurement frequency, etc., for the plant-wide whole body counting services; state-of-the-art facilities and equipment used to provide the best in vivo measurement results possible for the approximately 11,000 measurements made annually; procedures for performing the various in vivo measurements at the Whole Body Counter (WBC) and related facilities including whole body counts; operation and maintenance of counting equipment, quality assurance provisions of the program, WBC data processing functions, statistical aspects of in vivo measurements, and whole body counting records and associated guidance documents. 16 refs., 48 figs., 22 tabs.

  15. 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.

  16. 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

  17. 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

  18. Therapeutic aspects of aerobic dance participation.

    PubMed

    Estivill, M

    1995-01-01

    An ethnographic analysis of aerobic dance exercise culture was conducted to determine the impact of the culture on the mind-body connection. After a review of the predominant theories on the relationship between vigorous exercise and elevated mood, aerobic dance participants' experiences are reported to illustrate how cognitive experience and self-esteem may be influenced. Interviews revealed that some participants achieved a pleasantly altered state of consciousness and respite from depression and stress. The relationship of the work ethic to achievement of participant satisfaction is underscored.

  19. 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

  20. LINEAR COUNT-RATE METER

    DOEpatents

    Henry, J.J.

    1961-09-01

    A linear count-rate meter is designed to provide a highly linear output while receiving counting rates from one cycle per second to 100,000 cycles per second. Input pulses enter a linear discriminator and then are fed to a trigger circuit which produces positive pulses of uniform width and amplitude. The trigger circuit is connected to a one-shot multivibrator. The multivibrator output pulses have a selected width. Feedback means are provided for preventing transistor saturation in the multivibrator which improves the rise and decay times of the output pulses. The multivibrator is connected to a diode-switched, constant current metering circuit. A selected constant current is switched to an averaging circuit for each pulse received, and for a time determined by the received pulse width. The average output meter current is proportional to the product of the counting rate, the constant current, and the multivibrator output pulse width.