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Detection of viable and dead Listeria monocytogenes on gouda-like cheeses by real-time PCR 总被引:4,自引:0,他引:4
AIMS: Surface contamination by Listeria monocytogenes of gouda-like cheeses during processing represents a potential public health problem. The aim of this work was to develop novel real-time PCR diagnostics to detect the presence of viable, dead or viable but not culturable (VBNC) cells on gouda-like cheeses. METHODS AND RESULTS: We used ethidium monoazide bromide (EMA)-PCR for direct quantification of viable and dead cells, while semiquantitative detection of culturable cells below the PCR detection limit (c. 100 CFU g(-1)) was obtained by combining growth and real-time PCR. We were able to quantify the fraction of >0.5% viable cells in a background of dead cells by EMA-PCR, given that the viable cell concentration was above the PCR detection limit. The combined growth and real-time PCR complemented the EMA-PCR, and enabled semiquantitative detection of low levels of culturable cells (10 and 100 CFU g(-1)). SIGNIFICANCE AND IMPACT OF THE STUDY: The significance of this work is that we have developed a novel concept for detection of viable and potentially infectious L. monocytogenes. 相似文献
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《African Journal of Aquatic Science》2013,38(2):199-203
The biodiversity of heterotrophic viable bacteria (209 isolates) in the hypersaline Bardawil Lagoon, Egypt, was studied. Composition and extracellular activities of viable culturable heterotrophic bacteria (VCHB) in the water and in non-colonised and seagrass-colonised sediments of Bardawil Lagoon were determined bimonthly during 1997 and 1998. The average ± SD total Kjeldhal nitrogen was 1.69 ± 0.44 mg l?1 in water, 335.95 ± 19.22 mg kg?1 in colonised sediments, and 215.5 ± 16.0 mg kg?1 in non-colonised sediments. Exoenzymatic bacterial activity (glycosidase) presented a seasonal trend with average values of 1.02 ±0.16 μM cm?3 min?1 in colonised sediment samples and was 0.36 ± 0.27 μM cm?3 min?1 in non-colonised sediments. Mean of VCHB was 4 017 ± 565 cfu g?1 and 1 195 ± 242 cfu g?1 for colonised and non-colonised sediments, respectively. Bacterial isolates from Bardawil Lagoon water and sediments yielded a wide diversity of VCHB: a total of 209 different species, belonging to 13 genera from the water and 12 genera from the sediments. 相似文献
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Epifluorescence microscope methods, namely BacLight, direct epifluorescence filter technique and Rhodamine 123, consistently underestimated plate bacterial counts in a 4-chlorophenol degrading consortium. Cells capable of passing through 0.2 microm filters, referred as 'ultramicrocells', were found. Although cell counts were higher when traditional methods were used, BacLight and direct epifluorescence filter technique were convenient techniques for the systematic monitoring of bacteria involved in biodegradation processes, as results were consistent and available within a short time. 相似文献
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AIMS: This study was designed to evaluate potential effects of sampling duration on observed concentrations of airborne culturable mould and bacteria on selected media. METHODS AND RESULTS: Airborne culturable mould and bacteria from lightly to moderately contaminated environments were collected on selected culture media using two co-located, concurrently operated, Andersen N-6 samplers for five sampling durations in the range of 1-10 min. Differences in mean concentrations, as well as linear relationships between sampling duration and both concentration and variability, were evaluated using nonparametric procedures. For the five sampling durations, there were no significant differences in mean concentrations of mould; for bacteria, there were significant differences, with a trend of decreasing concentrations as sampling duration increased. Data variability decreased with increasing sampling duration for both mould and bacteria. CONCLUSIONS: Airborne culturable mould concentrations were similar for sampling durations in the range of 1-10 min. Airborne bacteria concentrations tended to trend downwards with sampling durations exceeding 3 min. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has shown that sampling durations of 1-10 min are appropriate for collection of airborne culturable mould on malt extract agar (MEA) and dichloran glycerol agar (DG-18); based on the apparent trend of decreasing bacterial sample concentrations associated with increasing sampling duration, sampling durations of 相似文献
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冷箭竹根际土壤中可培养细菌的多样性 总被引:1,自引:0,他引:1
为了解冷箭竹(Bashania fangiana)根际土壤中可培养细菌的多样性,2006年5月从四川卧龙自然保护区冷箭竹根际土壤中共分离出50株具不同菌落形态的细菌.16S rDNA序列分析结果表明:50株菌分属于10个属26个种.27株属于变形菌门γ亚群(Gammaproteobaeteria)(42.3%)、9株属于厚壁菌门(Firmicutes)(26.9%)、4株属于放线菌门(Actinobacteria)(15.4%)、6株属于变形菌门β亚群(Betaproteobacteria)(7.7%)、3株属于变形菌门α亚群(Alphaproteobacteria)(3.9%),1株与土地杆菌属(Pedobacter)关系密切.假单胞菌属(Pseudomonas)和芽孢杆菌属(Bacillus)为优势菌属.2株菌为可能的新种或属.研究表明冷箭竹根际土壤中含有较为丰富的微生物多样性. 相似文献
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Aims: To determine whether nonspore‐forming biothreat agents can survive and persist in potable water that does not contain a disinfectant. Methods and Results: Autoclaved, de‐chlorinated Atlanta municipal water was inoculated with eight isolates of bacterial biothreat agents (106 CFU ml?1). The inoculated water samples were incubated at 5, 8 (Francisella tularensis only) or 25°C and assayed for viability by culture and by the presence of metabolic activity as measured by esterase activity (ScanRDI, AES Chemunex). Viability as determined by culture varied from 1 to 30 days, depending upon the organism and the temperature of the water. All organisms were determined viable as measured by esterase activity for the entire 30 days, regardless of the incubation temperature. Conclusion: Francisella tularensis was culturable for at least 21 days if held at 8°C. The remaining nonspore‐forming bacterial biothreat agents were found to be metabolically active for at least 30 days in water held at 5 or 25°C. Significance and Impact of the Study: The data can assist public health officials to determine the safety of drinking water after contamination with a biothreat agent. 相似文献
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Senoh M Ghosh-Banerjee J Ramamurthy T Colwell RR Miyoshi S Nair GB Takeda Y 《Microbiology and immunology》2012,56(5):342-345
Viable but nonculturable (VBNC) Vibrio cholerae non-O1/non-O139, V. parahaemolyticus, enterohemorrhagic Escherichia coli, enterotoxigenic E. coli, enteropathogenic E. coli, Shigella flexneri, and Salmonella enterica were converted to the culturable state by co-culture with selected eukaryotic cells, e.g., HT-29, Caco-2, T84, HeLa, Intestine 407, and CHO cells. 相似文献
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Changes in intrinsic fluorescence during the production of viable but nonculturable Escherichia coli
D C Roselle M Seaver J D Eversole 《Journal of industrial microbiology & biotechnology》1998,20(5):265-267
The potential of intrinsic fluorescence spectroscopy to detect and differentiate viable but nonculturable bacteria in the
presence of culturable bacteria was explored. Escherichia coli cells, starved for 210 days in nutrient-free normal saline, show new fluorescence emissions near 400 and 440 nm, and reduced
emission near 340 nm.
Received 7 July 1997/ Accepted in revised form 26 November 1997 相似文献
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北欧海海水可培养细菌多样性 总被引:1,自引:0,他引:1
【目的】为了解北欧海表层海水可培养细菌多样性与所在水文环境的关系。【方法】利用2216E、R2A和海水培养基对该海域暖流区、寒流区、海盆区及交汇区等多个区域不同站位的表层海水样品中的可培养细菌进行分离培养,通过16S r RNA基因测序对分离的菌株进行分类鉴定,并构建系统发育树进行系统发育分析。【结果】从北欧海表层海水中共分离到407株细菌,通过RFLP分析选取其中154株进行测序,结果表明此154株细菌分属于3个门,18个属,27个种。3个门包括变形菌门、厚壁菌门和放线菌门,优势属为假交替单胞菌属、嗜冷杆菌属等,优势种为食琼脂假交替单胞菌、海雪嗜冷杆菌等,并分离到闪烁交替单胞菌等多株嗜冷菌。比较不同区域的微生物多样性可以看出,γ-变形菌纲的细菌在各个区域均占较高比例。交汇区的细菌多样性最高,分离到了10个不同属的细菌,而海盆区细菌多样性最低,只分离到了4种。除了海盆区外,其他3个区域的样品中都分离到了特有的类群。【结论】从以上结果可以看出,北欧海域有较为丰富的微生物资源,且交汇区微生物多样性较其他区域高。 相似文献
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Copper amendment of agricultural soil selects for bacterial antibiotic resistance in the field 总被引:1,自引:0,他引:1
AIMS: The objective of this study was to determine whether Cu-amendment of field plots affects the frequency of Cu resistance, and antibiotic resistance patterns in indigenous soil bacteria. METHODS AND RESULTS: Soil bacteria were isolated from untreated and Cu-amended field plots. Cu-amendment significantly increased the frequency of Cu-resistant isolates. A panel of isolates were characterized by Gram-reaction, amplified ribosomal DNA restriction analysis and resistance profiling against seven antibiotics. More than 95% of the Cu-resistant isolates were Gram-negative. Cu-resistant Gram-negative isolates had significantly higher incidence of resistance to ampicillin, sulphanilamide and multiple (> or =3) antibiotics than Cu-sensitive Gram-negative isolates. Furthermore, Cu-resistant Gram-negative isolates from Cu-contaminated plots had significantly higher incidence of resistance to chloramphenicol and multiple (> or =2) antibiotics than corresponding isolates from control plots. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this field experiment show that introduction of Cu to agricultural soil selects for Cu resistance, but also indirectly selects for antibiotic resistance in the Cu-resistant bacteria. Hence, the widespread accumulation of Cu in agricultural soils worldwide could have a significant effect on the environmental selection of antibiotic resistance. 相似文献
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Arana I Orruño M Pérez-Pascual D Seco C Muela A Barcina I 《FEMS microbiology ecology》2007,62(1):1-11
After induction of the viable but nonculturable (VBNC) state in Escherichia coli populations, we analysed abiotic and biotic factors suggested to promote the resuscitation process. The response to the stressing conditions implied the formation of three subpopulations, culturable, VBNC and nonviable. In most adverse situations studied, the VBNC subpopulation did not represent the dominant fraction, decreasing with time. This suggests that, in most cases, the VBNC is not a successful phenotype. Combining methods of dilution and inhibition of remaining culturable cells, we designed a working protocol in order to distinguish unequivocally between regrowth and resuscitation. Reversion of abiotic factors inducing nonculturability as well as prevention of additional oxidative stress did not provoke resuscitation. Participation of biotic factors was studied by addition of supernatants from different origin without positive results. These results indicate that the E. coli strain used is not able to resuscitate from the VBNC state. VBNC cells release into the surrounding medium, and could thus aid in the survival of persisting culturable cells. The formation of a VBNC subpopulation could thus be considered as an adaptive process, designed for the benefit of the population as a whole. 相似文献
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Measurements of total and culturable bacteria in the alfresco atmosphere using a wet-cyclone sampler 总被引:2,自引:0,他引:2
Alfresco (def. clean, outdoor) airborne bacteria were collected with a commercially available wet-cyclone bioaerosol sampler
to demonstrate its use, sample processing and resultant observations of total and culturable bacteria in mid-summer in the
mid-Willamette River Valley, OR. Some critiques of the system are given. The maximum and minimum total and culturable airborne
bacterial concentrations in the samples were 5.9 × 105 and 8.8 × 102 cells m−3, and 1.3 × 104 and 3.1 CFU m−3, respectively. What is thought to be a diurnal cycle was also observed for both fractions with highest concentrations during
the day and lowest at dawn and dusk. The culturable bacteria as a percentage of the total, was maximal at mid-day (≈ 3%) and
minimal at early morning and late evening (≈ 0.5–2%). Contrarily, the total bacteria in the downwind dust plume of a grass
seed combine was 2.9 × 106 cells m−3 and of these approximately 73% were culturable, a much greater culturable percentage than found in the alfresco outdoor atmosphere. 相似文献
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Changes of bacterial communities in the rhizosphere of sugarcane under elevated concentration of atmospheric CO2 下载免费PDF全文
Diogo P. da Costa Armando C. F. Dias Simone R. Cotta Daniella Vilela Pedro A. M. de Andrade Vivian H. Pellizari Fernando Dini Andreote 《Global Change Biology Bioenergy》2018,10(2):137-145
It is believed that climate change will influence most of interactions that sustain life on Earth. Among these, the recruitment exerted by plants in their roots vicinity can change, leading to differential assemblages of microbiomes in the rhizosphere. We approached this issue analyzing the variations in the composition of bacterial communities in the rhizosphere of sugarcane cultivated under two concentrations of atmospheric CO2 (350 or 700 ppm). In addition to the analysis of bacterial community, the use of DNA‐SIP allowed the comparison of bacterial groups assimilating roots exudates (based on 13C‐labeled DNA) in both conditions, in a period of 8 days after the CO2 pulse. The separation of 13C‐DNA indicated the low but increasing frequency of labeling in the rhizosphere, as averages of 0.6, 2.4 and 5.0% of total DNA were labeled after 2, 4, and 8 days after the 13CO2 pulse, respectively. Based on large‐scale sequencing of the V6 region in the gene 16S rRNA, we found an increase in the bacterial diversity in the 13C‐DNA along the sampling period. We also describe the occurrence of distinct bacterial groups assimilating roots exudates from sugarcane cultivated under each CO2 concentration. Bacilli, Gammaproteobacteria, and Clostridia showed high affinity for the C‐sources released by sugarcane under 350 ppm of CO2, while under elevated concentration of CO2, the assimilation of roots exudates was prevalently made by members of Bacilli and Betaproteobacteria. The communities became more similar along time (4 and 8 days after CO2 pulse), in both concentrations of CO2, electing Actinobacteria, Sphingobacteriia, and Alphaproteobacteria as the major cross‐feeders on sugarcane exudates. In summary, we described the bacterial groups with higher affinity to assimilate roots exudates in the rhizosphere of sugarcane, and also demonstrated that the rhizosphere community can be differentially assembled in a future scenario with increased contents of CO2. 相似文献
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Tsutomu Yoshida Shoko Shinoda Tsuneya Matsumoto Satoru Watarai 《Bioscience, biotechnology, and biochemistry》2013,77(12):3093-3095
A strain of Alcaligenes isolated from soil was a good producer of β-glucuronidase, and the enzyme was purified from the cell-free extract by sequential column chromatography on DEAE-Toyopearl, Toyopearl HW-55F, and Phenyl-Sepharose CL-4B. By these procedures, two β-glucuronidases designated as β-glucuronidases I and II were purified 240- and 508-fold, respectively. β-Glucuronidase I, with a molecular weight of 75,000, had an optimum pH at 7.5 and the enzyme II, with a molecular weight of 300,000, had maximum activity at pH 6.0. Both enzymes were strongly inhibited by saccharo-1,4-lactone, glucaro-δ-lactam, p-chloromercuribenzoate, Hg2+, and N-bromosuccinimide. β-Glucuronidase I was active toward estrogen-3-β-glucuronides and inert toward β-glucuronide conjugates of menthol, estrogen-17β-, estrogen-16α-, androsterone-3α-, testosterone-17β-, cortisol-17α-. β-Glucuronidase II hydrolyzed all of these substrates. β-Glucuronidase I was inhibited by phenolphthalein and its glucuronide. 相似文献
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Enumeration and presumptive identification of some functional groups of bacteria in the rumen of dairy cows fed grass silage-based diets 总被引:5,自引:0,他引:5
Abstract Samples of rumen ingesta from two rumen-fistulated dairy cows fed grass silage-based diets were examined for numbers and types of bacteria that developed colonies on rumen fluid-agar media designated to support the growth of (a) a wide range of species, (b) cellulolytic bacteria, (c) lactate-fermenting bacteria, (d) non-fermentative bacteria. The most numerous species was Bacteroides ruminicola followed by Butyrivibrio fibrisolvens . The most abundant cellulolytic species were Eubacterium cellulosolvens and Ruminococcus flavefaciens. Megasphaera elsdenii and Selenomonas ruminantium were important lactate fermenters but an unidentified bacterium that grew poorly on maintenance medium was by far the most numerous among bacteria isolated from lactate-containing medium. One strain remained sufficiently viable to show that it fermented lactate to propionate and acetate. 相似文献
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Effect of green manure on Pythium spp. population and microbial communities in intensive cropping systems 总被引:4,自引:0,他引:4
Saprophytic soil-borne pathogens can be either actively suppressed by organic amendments or enhanced, depending on soil health conditions. This can be deleterious in the event of selection of a soil-borne population by previous soil management and short crop rotation. Trials were performed in the open field and in pots, using naturally infected soil from intensive crop systems, i.e., soil from fields with 8 years of strawberry cultivation. The aim was to study short-term response of Pythium and soil microbial populations to green manure. The use of green manure in these naturally infested soils, 3–10 weeks after fresh tissue incorporation, caused Pythium populations to increase concurrent with an increase in soil microbial populations, and did not result in the suppression of the pathogen. A more elaborate trial was performed under controlled conditions, amending soil with fresh wheat plant material, air-dried wheat plant material and an organic fertilizer with a high level of humic substances. Although compared to the original soil, all amendments caused a similar increase in organic matter content and small differences in soil respiration, incorporation of fresh, not decomposed, plant material strongly increased Pythium, while the organic fertilizer did not affect the original level of the pathogen population. The increase in total number of fungi and bacteria did not have any suppressive effect on the Pythium population in naturally infested soil used for this study. 相似文献