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1.
Biofilms enable foodborne pathogens to resist removal from surfaces, survive disinfection and elude detection. This study evaluated the use of Calcofluor, which binds to polysaccharides containing β-D-glucans, to detect biofilms produced by Salmonella enterica serovar Berta and Salmonella enterica serovar Typhimurium DT104 (St DT104), Escherichia coli, Aeromonas hydrophila, Vibrio cholerae O139 and Hyphomonas adhaerens. Biofilms produced by St DT104, S. berta and V. cholerae on five types of surfaces (glass, polypropylene, Teflon™, stainless steel and aluminum) were detected by Calcofluor. Results suggest the potential use of Calcofluor as probes of foodborne pathogens in biofilms.  相似文献   

2.
Chen H 《Food microbiology》2007,24(3):197-204
Survival curves of six foodborne pathogens suspended in ultra high-temperature (UHT) whole milk and exposed to high hydrostatic pressure at 21.5 degrees C were obtained. Vibrio parahaemolyticus was treated at 300 MPa and other pathogens, Listeria monocytogenes, Escherichia coli O157:H7, Salmonella enterica serovar Enteritidis, Salmonella enterica serovar Typhimurium, and Staphylococcus aureus were treated at 600 MPa. All the survival curves showed a rapid initial drop in bacterial counts followed by tailing caused by a diminishing inactivation rate. A linear model and two nonlinear models were fitted to these data and the performances of these models were compared using mean square error (MSE) values. The log-logistic and Weibull models consistently produced better fits to the inactivation data than the linear model. The mean MSE value of the linear model was 6.1, while the mean MSE values were 0.7 for the Weibull model and 0.3 for the log-logistic model. There was no correlation between pressure resistance and the taxonomic group the bacteria belong to. The order, most to least pressure-sensitive, of the single strains tested was: V. parahaemolyticus (gram negative)相似文献   

3.
In this study, identification of environmental reservoirs of Salmonella enterica subsp. enterica serovar Typhimurium (abbreviated as Salmonella Typhimurium) in sediments, water, and aquatic flora collected from the Ganges River (Ganges riverine material) was carried out by adopting a two-step strategy. Step 1 comprised a selective serovar-specific capture of Salmonella Typhimurium from potential reservoirs. Step 2 involved culture-free detection of selectively captured Salmonella Typhimurium by ttr gene-specific molecular beacon (MB) based quantitative polymerase chain reaction (q-PCR). The ttr gene-specific MB designed in this study could detect 1 colony-forming unit (cfu)/PCR captured by serovar-specific DNA aptamer. Sediments, water, and aquatic flora collected from the Ganges River were highly contaminated with Salmonella Typhimurium. The preanalytical step in the form of serovar-specific DNA aptamer-based biocapture of bacterial cells was found to enhance the sensitivity of the fluorescent probe in the presence of nonspecific DNA . Information about the presence of environmental reservoirs of Salmonella Typhimurium in the Ganges River region may pave the way for forecasting and management of nontyphoidal salmonellosis in south Asia.  相似文献   

4.
Recent events both here and abroad have focused attention on the need for ensuring a safe and secure food supply. Although much has been written about the potential of particular select agents in bioterrorism, we must consider seriously the more mundane pathogens, especially those that have been implicated previously in foodborne outbreaks of human disease, as possible agents of bioterrorism. Given their evolutionary history, the enteric pathogens are more diverse than agents such as Bacillus anthracis, Francisella tularensis, or Yersinia pestis. This greater diversity, however, is a double-edged sword; although diversity affords the opportunity for unequivocal identification of an organism without the need for whole-genome sequencing, the same diversity can confound definitive forensic identification if boundaries are not well defined. Here, we discuss molecular approaches used for the identification of Salmonella enterica, Escherichia coli, and Shigella spp. and viral pathogens and discuss the utility of these approaches to the field of microbial molecular forensics.  相似文献   

5.
Sensitivities of foodborne pathogens to pressure changes   总被引:1,自引:0,他引:1  
Eight foodborne pathogens were suspended in ultrahigh-temperature whole milk and treated at pressure levels of 0.1 to 690 MPa at 21.5 degrees C for 10 min. There was no clear trend in pressure resistance between gram-negative and gram-positive organisms. The order of the single strains tested, from most to least pressure sensitive, was Vibrio parahaemolyticus < Yersinia enterocolitica < Listeria monocytogenes < Salmonella enterica serovar Typhimurium < S. enterica serovar Enteritidis < Escherichia coli O157:H7 approximately equal to Staphylococcus aureus < Shigella flexneri. For each organism there existed a pressure range in which log(number of survivors) had a near linear relationship when plotted versus treatment pressure level. In this study, a decimal reduction pressure (Dp) value was defined and used to measure the sensitivity of these pathogens to pressure changes. L. monocytogenes and V. parahaemolyticus were most sensitive to pressure changes, and S. flexneri was most resistant. The D(P) values were 16.3 MPa for L. monocytogenes, 21.7 MPa for V. parahaemolyticus, and 127.0 MPa for S. flexneri. The most pressure-resistant gram-negative bacterium, S. flexneri, and most pressure-resistant gram-positive bacterium, S. aureus, were treated at 50 degrees C and pressures of 0.1 to 650 MPa for 10 min. High temperature considerably enhanced pressure inactivation of these two organisms and affected their sensitivities to pressure changes. The effect of treatment time on the D(P) values of L. monocytogenes and V. parahaemolyticus was also determined, and it was found that it did not significantly affect their D(P) values.  相似文献   

6.
Biopreservatives are being developed to inhibit the growth of foodborne pathogens and thus improve food safety. The lactoperoxidase system (LPS) is a naturally occurring system that has potential for use as an antimicrobial agent in foods. Growth of single strains of the pathogens Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium, Yersinia enterocolitica, Pseudomonas aeruginosa and beef microflora were assessed on LPS-treated meat surfaces in an experimental system. Beef cubes inoculated with approximately 10(4) cfu cm(-2) of bacteria were treated with the LPS and incubated at 37 degrees C for 24 h, 12 degrees C for 7 days or in a chilling regime: 12 to -1 degrees C over 1 week and held at -1 degrees C for 4 weeks. Treatment with LPS was more effective at storage temperatures non-permissive for rapid bacterial growth with strong inhibition of growth achieved on LPS-treated cubes at 12 degrees C and reduction in pathogen viable counts at chilling temperatures. At chilling temperatures, the LPS inhibited the growth of native pseudomonads but did not prevent the development of native lactic acid bacteria.  相似文献   

7.
The feasibility of using two primers internal to the stdA gene (which encodes the fimbrial major subunit of the std fimbrial gene cluster in Salmonella enterica serovar Typhi) to detect Salmonella by PCR was explored. The 518-bp stdA specific sequence was conserved among 268 strains from 45 serovars of S. enterica. One Salmonella bongori CCUG 30042 strain and 34 non-Salmonella strains did not possess this sequence. A sensitivity test revealed that the stdA-specific primer set detected 3.4 x 10(-1) pg of genomic DNA and 3.0 x 10(5) CFU/ml with serial dilutions of Salmonella Typhimurium cells. In vitro testing for specificity using pig carcass sponge samples contaminated with Salmonella Typhimurium also was performed. An initial Salmonella Typhimurium inoculum of 4.4 x 10(1) CFU/ml in pig carcass exudates reached the stdA primer detection level after preenrichment in buffered peptone water at 37 degrees C for 18 h in the presence of indigenous non-Salmonella flora at 4.0 X 10(7) CFU/ml, but the detection level decreased to 4.4 x 10(0) CFU/ml after selective enrichment in Rappaport-Vassiliadis R10 broth for 18 h at 42 degrees C. The PCR method with primers specific for stdA is a quick and sensitive tool for detecting S. enterica, which is an important cause of foodborne disease.  相似文献   

8.
Salmonella enterica serovar Typhimurium is a major foodborne pathogen throughout the world. Until now, the specific target genes for the detection and identification of serovar Typhimurium have not been developed. To determine the specific probes for serovar Typhimurium, the genes of serovar Typhimurium LT2 that were expected to be unique were selected with the BLAST (Basic Local Alignment Search Tool) program within GenBank. The selected genes were compared with 11 genomic sequences of various Salmonella serovars by BLAST. Of these selected genes, 10 were expected to be specific to serovar Typhimurium and were not related to virulence factor genes of Salmonella pathogenicity island or to genes of the O and H antigens of Salmonella. Primers for the 10 selected genes were constructed, and PCRs were evaluated with various genomic DNAs of Salmonella and non-Salmonella strains for the specific identification of Salmonella serovar Typhimurium. Among all the primer sets for the 10 genes, STM4497 showed the highest degree of specificity to serovar Typhimurium. In this study, a specific primer set for Salmonella serovar Typhimurium was developed on the basis of the comparison of genomic sequences between Salmonella serovars and was validated with PCR. This method of comparative genomics to select target genes or sequences can be applied to the specific detection of microorganisms.  相似文献   

9.
当今食源性疾病已成为一个全球关注的食品卫生问题,常见的细菌性食物中毒的病原菌有:致病性大肠杆菌(特别是出血性大肠杆菌O157:H7)、沙门氏菌、志贺氏菌、致病性弧菌(包括霍乱弧菌和副溶血弧菌)、金黄色葡萄球菌、单增李斯特菌、空肠弯曲菌以及阪崎克罗诺杆菌等。单独检测一种致病菌已无法满足现今社会对同时高效检测多种致病菌的要求,为建立一种同时对上述9种食品中常见致病菌的检测方法,本研究分别以上述9种菌的特异性基因为靶基因,创新性地结合一对通用引物,建立能同时检测多种食源性致病菌的多重HRM-real time PCR检测体系。结果表明该多重HRM-real time PCR检测体系通过两管PCR反应可对上述9种食品中常见致病菌进行有效的检测与区分,且具有良好的特异性和灵敏度。  相似文献   

10.
The antibacterial activity of a chitooligosaccharide mixture prepared by digestion of shrimp chitosan with cellulase at 50 degrees C for 14 h was evaluated. Sugars with 1 to 8 degrees of polymer (DP) were found in this chitooligosaccharide mixture, and the weight percentage of sugars with DP > or = 6 was 44.3%. Minimal lethal concentrations of this mixture against Aeromonas hydrophila, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella Typhimurium, Shigella dysenteriae, Staphylococcus aureus, Vibrio cholerae, and Vibrio parahaemolyticus in nutrient broth were 5 to 29 ppm, which were much lower than those of the chitosan reactant (50 to 1,000 ppm). The antibacterial activity of this mixture in the sterilized milk against E. coli O157, L. monocytogenes, Salmonella Typhimurium, and S. aureus was much stronger at 4 degrees C than at 37 degrees C. When raw milk was supplemented with either 0.24% or 0.48% (wt/vol) of this oligosaccharide mixture and stored at 4 degrees C for 12 days, its mesophilic and psychrotrophic counts were reduced by at least 3 log cycles, and there was very little change in pH. In addition, this mixture retarded the growth of Salmonella species and caused quicker reduction of Staphylococcus species in raw milk. Accordingly, the shelf life of raw milk at 4 degrees C was extended by at least 4 days.  相似文献   

11.
The adhesion properties of Bifidobacterium longum and Bifidobacterium catenulatum strains with an acquired resistance to acid and their ability to competitively exclude Salmonella enterica serovar Typhimurium, Escherichia coli, Listeria monocytogenes, Enterobacter sakazakii, and Clostridium difficile from adhering to human intestinal mucus were evaluated and compared with the results when the same experiments were run with the original acid-sensitive strains. In half of the four studied cases, the acid-resistant derivative showed a greater ability to adhere to human intestinal mucus than the original strain. The ability of bifidobacteria to inhibit pathogen adhesion to mucus was not generally improved by the acquisition of acid resistance. In contrast, three of the four acid-resistant strains showed a greater ability to displace preadhered pathogens than the original strains, especially preadhered Salmonella Typhimurium and C. difficile. Overall, the induction of acid resistance in bifidobacteria could be a strategy when selecting strains with enhanced stability and improved surface properties that favor their potential functionality as probiotics against specific pathogens.  相似文献   

12.
Seafood is a leading commodity implicated in foodborne disease outbreaks in the United States. Seafood importation rose dramatically in the past 3 decades and now contributes to more than 80% of the total U.S. seafood supply. However, limited data are available on the microbiological safety of imported seafood. In this study, we obtained a total of 171 salmon, shrimp, and tilapia samples imported from 12 countries in three retail stores in Baton Rouge, LA. The total microbial population and the prevalence and antimicrobial susceptibilities of six major foodborne-pathogen genera (Campylobacter, Escherichia coli, Listeria, Salmonella, Shigella, and Vibrio) were determined. The aerobic plate counts (APC) for the 171 samples averaged 4.96 log CFU/g, with samples from Chile carrying the highest mean APC of 6.53 log CFU/g and fresh samples having a significantly higher mean APC than frozen ones (P < 0.0001). There were 27 samples (15.8%) with unacceptable microbiological quality (APC > 7 log CFU/g). By culture, no sample tested positive for Campylobacter coli, Shigella, or Vibrio vulnificus. Campylobacter jejuni and Salmonella enterica serovar Typhimurium were each recovered once from farm-raised tilapia from China. By PCR, 17.5 and 32.2% of the samples were positive for Salmonella and Shigella, respectively. The overall prevalence rates of other target bacteria were low, ranging from 4.1% for Listeria monocytogenes to 9.4% for E. coli. All of the Vibrio parahaemolyticus isolates recovered were from shrimp, and 63.3% showed intermediate resistance to ampicillin. Both C. jejuni isolates possessed a rare resistance to gentamicin, while 75% of L. monocytogenes isolates were resistant to nitrofurantoin. Taken together, these findings suggest potential food safety hazards associated with imported seafood and warrant further large-scale studies.  相似文献   

13.
沙门氏菌是造成我国食源性疾病的常见细菌之一,也是肉类消费过程中密切监测的重点对象。在2018年~2021年期间,共计采集了南京市545份猪肉源食品样本,利用选择性培养法分离得到44株沙门氏菌,采用血清学方法和分子生物学方法(MLST)对其进行分型鉴定,并分析其耐药性。结果表明,市售样本中共计检出44株沙门氏菌,平均检出率为8.07%,其中内脏样本检出率相对最高(检出率为30.49%);血清型分析表明,检出的菌株中以鼠伤寒沙门氏菌(Salmonella typhimurium)、罗森氏沙门氏菌(Salmonella rissen)、德尔卑沙门氏菌(Salmonella derby)和伦敦沙门氏菌(Salmonella london)4种血清型沙门氏菌最为常见;基因分型表明,ST19型为猪肉源中优势沙门氏菌株,占比为20.45%;检出的沙门氏菌中,38株菌株对四环素具有明显耐药性,占全部检出菌株的86.36%,而对头孢他啶、头孢噻肟和头孢西丁的耐药不超过10%。此外,对检出的1株多重耐药性沙门氏菌耐药基因分析发现,共筛查出了包括7大类抗生素及与耐药相关的基因。该实验详细分析了南京市场猪肉源食品中沙门氏菌污染状况,也为后期沙门氏菌的综合防治提供了理论依据。  相似文献   

14.
With three pairs of primers, a multiplex PCR assay was established for the simultaneous detection of Escherichia coli 0157:H7, Salmonella, and Shigella. Under the optimized conditions, the assay yielded a 252-bp product from E. coli O157:H7, a 429-bp product from Salmonella Typhimurium, and a 620-bp product from Shigella flexneri, respectively. When the DNA extraction of multiple target organisms was included in the same reaction, two or three corresponding amplicons of different sizes were observed. In the specificity test, 10 E. coli O157:H7 strains and one E. coli O157:NM strain showed the expected 252-bp amplicon. Seven other E. coli strains yielded no signal. Additionally, the 429-bp amplicon was produced from 20 Salmonella strains covering 16 serotypes, whereas the 620-bp amplicon was generated from 11 Shigella strains covering 4 species. No nonspecific amplification was observed with DNA from 48 other bacterial strains. Following a 24-h enrichment, the developed assay could concurrently detect the three pathogens at initial inoculation levels of approximately 8 x 10(-1) CFU/g (or CFU/ml) in apple cider, cantaloupe, lettuce, tomato, and watermelon and 8 x 10(1) CFU/g in alfalfa sprouts. The whole procedure can be easily completed within 30 h. The multiplex PCR assay can potentially be a simple, rapid, and efficient tool for presumptive and simultaneous screening of apple cider and produce for contamination by E. coli O157:H7, Salmonella, and/or Shigella.  相似文献   

15.
Salmonella strains (n = 75) isolated from foods involved in foodborne outbreaks occurred in Rio Grande do Sul State, Brazil, during 1999 and 2000 were studied. Strains were serotyped and submitted to PCR analysis to verify the prevalence of Salmonella plasmid virulence (spvR) regulatory gene. Among the 75 isolates, 73 (97%) were classified as Salmonella enterica serovar Enteritidis. All of the Salmonella strains isolated in 1999 were classified as serotype Enteritidis, whereas in 2000 two isolates were serotyped as Salmonella Derby and Salmonella Typhimurium. Regarding the prevalence of spvR gene, 62 strains (82.7%) were PCR positive, and a positive correlation (P < 0.05) between the strains of Salmonella Enteritidis and the presence of spvR gene was demonstrated, which suggests that this gene is a characteristic of the Salmonella Enteritidis analyzed.  相似文献   

16.
A survey of the presence of Salmonella and Shigella in freshly squeezed orange juice and related samples was conducted in Guadalajara, Mexico. One hundred samples of freshly squeezed orange juice were collected from 49 street booths and 51 small food service establishments. In addition, 75 fresh orange samples, each consisting of five orange units, and 75 wiping cloths were collected from the same establishments from which juice had been collected. Salmonella was isolated from 14, 20, and 23% of samples of orange juice, orange surfaces, and wiping cloths collected from street vendors, while Shigella was isolated from 6, 17, and 5% of these samples. In general, the frequency of isolation of these pathogens in samples from juice serving establishments at public markets was significantly lower than that found among street vendors (P < 0.05). Salmonella enterica serotypes Agona, Typhimurium, and Anatum were found in orange juice, fresh oranges, and wiping cloth samples, while serotype Mexico was found on fresh oranges and in wiping cloths and serotypes Muenchen and Panama were found only in wiping cloth samples. Regarding Shigella species, Shigella sonnei was found in all three types of sample tested; Shigella dysenteriae was found in juice and orange samples, Shigella boydii in orange and wiping cloth samples, and Shigella flexneri on oranges only. Thirty-one percent and 39% of the juice samples showed aerobic plate counts of > or = 5.0 log CFU/ml and Escherichia coli counts of > 3.0 log CFU/ml, respectively. These high counts may indicate poor sanitation and potential exposure to fecal contamination either in the raw materials or during the orange-crushing and juice-serving process. These data may be useful for a further risk assessment of Salmonella or Shigella in unpasteurized, freshly squeezed juice.  相似文献   

17.
Salmonella enterica serovar Choleraesuis may cause swine salmonellosis and human infection. Because the conventional method for detection of this Salmonella serovar may take 3 to 5 days, a PCR method for detection was evaluated. By comparing the sequence of the phase 1 flagellin (fliC) gene of Salmonella Choleraesuis with that of other Salmonella serovars and of other bacteria species available in GenBank, two PCR primers (flinC-F and flinC-R) were designed. Using these primers, all 97 Salmonella Choleraesuis strains assayed generated the expected PCR product, with a molecular mass of 963 bp. Except for S. enterica Paratyphi C, Salmonella isolates other than Salmonella Choleraesuis and non-Salmonella isolates, including strains of Enterobacteriaceae, all generated negative PCR results. Salmonella Paratyphi C could be differentiated from Salmonella Choleraesuis through the use of primers designed from the viaB gene. When Salmonella Choleraesuis isolates from swine stool, pork, liver, feed, and human whole blood samples were assayed with a preenrichment step, as low as 1 CFU/g or ml of the original sample could be detected.  相似文献   

18.
The U.S. Food and Drug Administration recognizes that raw seed sprouts are an important cause of foodborne disease and is now recommending that either spent irrigation water or final product be screened for Salmonella and Escherichia coli O157:H7 as a means of assuring the safety of product intended for consumption. In an effort to streamline such testing efforts, a simple method to preconcentrate pathogens from sprouts and spent irrigation water was investigated to facilitate the direct (without prior cultural enrichment) detection of pathogens using the PCR technique. Alfalfa sprouts and spent irrigation water were seeded with Salmonella enterica serovar Typhimurium and E. coli O157:H7 at 10(-1) to 106 CFU/g or CFU/ml, respectively. Samples were blended (sprouts only) and then centrifuged at high speed to sediment the total bacterial population. The precipitate was processed for DNA isolation, PCR amplification, and amplicon confirmation by Southern hybridization. Mean pathogen recoveries after centrifugation ranged from 96 to 99% for both pathogens in both matrices. Using primers targeting the invA gene for Salmonella Typhimurium and the stx genes of E. coli O157:H7, it was possible to detect both pathogens in alfalfa sprouts at seeding concentrations as low as 10 CFU/g. PCR detection limits for both pathogens from spent irrigation water were 10(-1) CFU/ml, the equivalent of 100 CFU/liter of water. Because spent irrigation water is constitutionally simple, it is particularly well suited for bacterial concentration by simple centrifugation steps. In this study, progress was made toward development of a rapid, inexpensive, and sensitive method for the detection of sprout-associated pathogens that is relevant to current industrial practices and needs.  相似文献   

19.
In order to investigate contamination of chicken farms with Salmonella, feed and eggs were sampled from 16 commercial layer farms in eastern Japan between 1993 and 1998 and cultured for salmonellae. Salmonella enterica subsp. enterica isolates belonging to 19 serovars were obtained from the feed. Six of the 19 serotypes, including Salmonella serovar Enteritidis, were observed in isolates recovered from the eggs. Salmonella serovar Enteritidis strains obtained from a feed sample and egg contents in a layer farm showed pulsed-field gel electrophoresis patterns that were genetically related and belonged to a single phage type, suggesting that the contamination of the farms was linked to the occurrence of salmonellae in feed.  相似文献   

20.
A total of 4,581 Salmonella strains isolated from nonhuman sources, including foodstuffs associated with foodborne Salmonella outbreaks, from January 1996 through December 2000 were serotyped at the Enteropathogens Laboratory, Instituto Adolfo Lutz, S?o Paulo, Brazil. Among the 123 different serotypes identified, Salmonella enterica subsp. enterica serotype Enteritidis (Salmonella Enteritidis) was the most prevalent (32.7%), ranking first for almost every kind of source. The next most common serotypes were Salmonella Senftenberg (10.3%), Salmonella Hadar (6.8%), Salmonella Agona (5.1%), and Salmonella Typhimurium (2.4%). Rough strains belonging to the subspecies S. enterica subsp. enterica (4.8%), S. enterica subsp. arizonae (<1%), S. enterica subsp. diarizonae (<1%), and S. enterica subsp. houtenae (<1%) were also detected. Foodstuffs (including poultry meat for consumption) contained 38.1% of the studied Salmonella strains, poultry flocks (from several farms under salmonellosis control by the owners) contained 21.7%, the environment contained 10.6%, sewage contained 9.4%, water contained 6.6%, animal feed contained 4.4%, chill water from poultry-processing operations contained 2.2%, and other sources contained 7.0%. Foodstuffs extensively contaminated with Salmonella strains were poultry meat (40%), cow meat (11%), desserts (8%), mayonnaise (6%), sausage (5%), and unpasteurized shell eggs (4%), and there were several other food sources (26%). Homemade mayonnaise was the most common vehicle for Salmonella foodborne outbreaks, and Salmonella Enteritidis was the serotype most isolated (95%) from that source. According to these data and previously published data concerning Salmonella strains isolated in S?o Paulo State, almost the same serotypes have predominated among nonhuman sources for the last decade.  相似文献   

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