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1.
Jin JD Lee DS Shin EK Kim SJ Jung R Hahn TW 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(12):1321-1326
Salmonella enterica subspecies enterica serovar Gallinarum biovar Gallinarum is the causative agent of fowl typhoid in chickens, outbreaks of which have devastated poultry populations in Korea since 1992. In order to identify genetic differences among S. Gallinarum isolates, bacteria were examined using the random amplified polymorphic DNA (RAPD) method. Of 13 arbitrary primers screened initially, the primer designated as universal rice primer-6 (URP-6) was selected for subsequent typing assays because it produced a distinctive and reproducible DNA fingerprint for a S. Gallinarum reference strain. URP-6-based RAPD analysis assigned 30 S. Gallinarum isolates into 6 types, with 26 isolates (86.6%) belonging to 2 major RAPD types. The distribution of virulence genes in S. Gallinarum isolates was examined by Southern hybridization. All tested isolates had the invasion gene, invA, the virulence plasmid gene, spvB, and the S. Enteritidis fimbrial gene, sefC. The distribution of virulence genes among S. Gallinarum isolates did not correlate with any specific RAPD type. 相似文献
2.
Hybridization studies with a DNA probe derived from the virulence region of the 60 Mdal plasmid of Salmonella typhimurium. 总被引：8，自引：1，他引：7 
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下载免费PDF全文 C Poppe R Curtiss rd P A Gulig C L Gyles 《Canadian journal of veterinary research》1989,53(4):378-384
Plasmid DNA of 68 strains of Salmonella that belonged to 18 serovars and exhibited 48 different plasmid profiles was examined for hybridization with a 32P-labelled DNA probe which consisted of a 3750 base pairs (bp) HindIII-HindIII fragment derived from the virulence region of the 60 megadalton (Mdal) plasmid of Salmonella typhimurium. The 32 Mdal plasmid of S. cholerae-suis, the 50 Mdal plasmid of S. dublin, the 36 Mdal plasmid of S. enteritidis, the 60 Mdal plasmid of S. gallinarum, the 60 Mdal plasmid of S. pullorum, and the 60 Mdal plasmid of S. typhimurium, plasmids that have been associated with virulence, all hybridized with the probe. Digestion of plasmid DNA of these strains with PvuII and hybridization with the probe revealed that the plasmids of strains of all six serovars contained fragments of approximately 2520 and 1520 bp that hybridized with the probe. Similarly, hybridization with BglI digests of DNA of the virulence-associated plasmids of strains of these six serovars showed that all six plasmids contained a fragment of approximately 3690 bp that hybridized with the probe. No other plasmids of these strains nor any plasmids of 12 other Salmonella serovars hybridized with the probe. Chromosomal DNA did not hybridize with the probe. The 60 Mdal plasmids of S. gallinarum and S. pullorum showed similar digestion patterns with restriction endonucleases BglI, BglII and PvuII. 相似文献
3.
鸡沙门氏菌脉冲场凝胶电泳分型研究 总被引：1，自引：0，他引：1
为应用脉冲场凝胶电泳(PFGE)从分子水平上对禽源沙门氏菌之间的差别进行分析和研究,本研究采用自动微生物鉴定仪对12株疑似鼠伤寒沙门氏菌和15株疑似鸡白痢沙门氏菌进行了鉴定.以限制性核酸内切酶Xba I对其全基因组DNA进行酶切、PFGE分型,Info Quest FP聚类分析软件对电泳结果进行分析.结果显示,共鉴定出11株鼠伤寒沙门氏菌、1株圣保罗沙门氏菌、6株阿姆斯特丹沙门氏菌、5株亚拉巴马沙门氏菌、1株鸡白痢沙门氏菌、2株纽波特沙门氏菌和2株肠炎沙门氏菌,一共分为12个PFGE型.实验结果表明,江苏地区存在有不常见的沙门氏菌血清型,同种血清型之间的基因型差别较小(相似值为0.85～1),不同血清型之间差别较大(相似值为0.45～0.70).PFGE能从分子水平上对禽源沙门氏菌的基因组DNA进行分型. 相似文献
4.
Osek J 《Veterinary microbiology》2000,71(3-4):211-222
Forty-six Escherichia coli strains isolated from post-weaning diarrhea of pigs were analysed for their phenotypic and genotypic properties. The isolates were of serogroups O138, O139, and O141 and most of them possessed hemolytic activities. PCR analysis showed that 34 of the isolates harboured the genes for shiga toxin 2e and 32 strains possessed the genes for heat-stable enterotoxins I and II. Ten strains had the fedA gene of F18 fimbriae. The genetic relationships among all isolates were tested by random amplified polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus (ERIC) PCR analyses. Using the RAPD test with two different primers, six fingerprints were distinguished whereas the ERIC analysis revealed only three DNA patterns. Some strains possessing identical phenotypic and genotypic virulence determinants exhibited distinct RAPD profiles and some isolates with different pathogenic markers showed the same RAPD and ERIC pictures. Thus, RAPD, and to a less extent ERIC techniques, revealed intra- and interserogroup genotypic variations among the E. coli strains analyzed. 相似文献
5.
Zamperini K Soni V Waltman D Sanchez S Theriault EC Bray J Maurer JJ 《Avian diseases》2007,51(4):958-964
Although Salmonella remains one of the leading causes of foodborne illnesses in the United States, the Salmonella enterica serovars and genetic types associated with most infections appear to fluctuate over time. Recently, the Center for Disease Control and Prevention (CDC) has reported an increase in cases of salmonellosis caused by Salmonella 4,[5],12:i:-. Similarly, this unusual Salmonella serovar has been isolated from cattle and poultry in the state of Georgia. We examined the genetic relatedness of Salmonella 4,[5],12:i:-, isolated from several different poultry companies and dairy farms in Georgia, by pulsed-field gel electrophoresis (PFGE). Several Salmonella 4,[5],12:i:- isolates had PFGE patterns identical or similar to PFGE patterns of Salmonella Typhimurium isolated from numerous animal sources. We identified distinct PFGE patterns for Salmonella 4,[5],12:i:- and matching Salmonella Typhimurium PFGE patterns, identifying four "distinct" strains. We focused a more specific analysis on the poultry Salmonella 4,[5],12:i:- and Salmonella Typhimurium isolates and found that of these Salmonella 4,[5],12:i:- isolates, 32% lacked the entire phase 2 antigen gene, fljB; 61% contained partial deletion(s); and 4% had partial deletion(s) in fljB and an adjacent gene hin, 5' to fljB. Thirteen percent contained smaller deletions or point mutations not identified by our DNA probes. The Salmonella 4,[5],12:i:- isolates were positive for several genes present in the Salmonella Typhimurium, including lpfE (100%), sseI(96%), and spvC (93%). Genetic analysis indicates independent, spontaneous mutations in fljB in at least four distinct Salmonella Typhimurium strains of animal origin circulating in nature. 相似文献
6.
Genomic fingerprinting of pigeon Streptococcus gallolyticus strains of different virulence by randomly amplified polymorphic DNA (RAPD) analysis 总被引：2，自引：0，他引：2
Baele M Vanrobaeys M Vaneechoutte M De Herdt P Devriese LA Haesebrouck F 《Veterinary microbiology》2000,71(1-2):103-111
Randomly amplified polymorphic DNA (RAPD) analysis was performed on 95 pigeon S. gallolyticus strains of different virulence and belonging to different biotypes and different culture supernatant phenotypes as determined by SDS-PAGE. Four distinct RAPD patterns, designated A, B, C and D, were distinguished using primer OPM6 (5'CTGGGCAACT). All 76 strains generating RAPD pattern A or B were designated highly virulent on the basis of their SDS-PAGE pattern. Five of seven strains generating RAPD pattern C and 11 of 12 strains generating RAPD pattern D belonged to the moderately virulent and low virulent culture supernatant phenotype groups, respectively. Only one RAPD group C strain belonged to a highly virulent culture supernatant phenotype group. There was a correlation between biotype and RAPD patterns. These findings indicate that there is a high correlation between RAPD pattern and virulence for pigeons. Therefore, RAPD typing seems a rapid, reliable method to distinguish pigeon S. gallolyticus strains of high, moderate and low virulence. 相似文献
7.
沙门菌肠毒素基因克隆及序列分析 总被引：1，自引：1，他引：0
研究常见的不同血清型沙门菌肠毒素(stn)基因核苷酸序列之间的差异及其分布情况。根据沙门菌的stn核苷酸序列设计一对引物,应用PCR技术,分别对肠炎沙门菌、鼠伤寒沙门菌和鸡白痢沙门菌进行PCR扩增,对扩增产物进行克隆及序列分析,并用所设计的引物检测7种血清型沙门菌(42株)。结果显示,3种沙门菌经PCR均扩增出749 bp的特异条带,DNA序列分析证实,沙门菌的stn核苷酸序列比较保守,42株沙门菌stn的检出率为100%。本试验成功克隆出沙门菌的stn,调查其在不同血清型沙门菌中的分布及序列分析,为进一步研究stn致病机理及研制减毒沙门菌活菌疫苗奠定了基础。 相似文献
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Mycobacterium avium is an important veterinary pathogen causing avian tuberculosis in birds. The aim of the study was to evaluate the genetic relatedness in M. avium isolates from deep tissues of farmed lesser white-fronted geese with avian tuberculosis and in samples from the farm environment. The strains were analyzed by two PCR-based typing methods, inverted repeat (IR) typing and random amplified polymorphic DNA (RAPD) analysis. The primers for the inverted repeats of the insertion sequences IS1245 and IS1311 were used in IR typing, and the RAPD analysis was performed with six primers. Seven of the nine avian strains yielded an identical pattern in the IR typing, but they could be divided into two groups in the RAPD analysis. The remaining two bird isolates had an identical IR pattern (IR cluster II) which they shared with two environmental isolates. However, the RAPD analysis revealed that these environmental isolates had a RAPD pattern (RAPD cluster VI) distinct and different from either of the bird isolates (RAPD clusters II and IV). In all, four M. avium strains were verified as being inducers of avian tuberculosis in birds, and all were distinct from the three environmental strains identified. Thus, the results did not confirm the preliminary idea that a single strain had caused the epidemic. The polymorphism among M. avium strains highlighted the great biodiversity among an M. avium population even in a limited environmental setting during a short time span, and indicated the high susceptibility to avian tuberculosis of lesser white-fronted geese. 相似文献
10.
Lopes VC Velayudhan BT Halvorson DA Lauer DC Gast RK Nagaraja KV 《American journal of veterinary research》2004,65(5):538-543
OBJECTIVE: To compare molecular typing methods for the differentiation of Salmonella enterica serovar Enteritidis phage type (PT) 4 isolates that allowed for the determination of their genetic relatedness. SAMPLE POPULATION: 27 Salmonella Enteritidis PT 4 strains isolated in the United States and Europe. PROCEDURE: Several molecular typing methods were performed to assess their ability to genetically differentiate among Salmonella Enteritidis PT 4 isolates. Results of pulse-field gel electrophoresis (PFGE), repetitive polymerase chain reaction (PCR) assay, 16S rRNA gene sequencing, random amplification of polymorphic DNA (RAPD), PCR-restriction fragment length polymorphism of 16S rRNA, and antimicrobial susceptibility were evaluated. RESULTS: Compared with results for other techniques, results for the RAPD typing method with the RAPD1 primer reveal that it was the most discriminatory fingerprinting technique, and it allowed us to cluster Salmonella Enteritidis PT 4 isolates on the basis of their genetic similarity. CONCLUSIONS AND CLINICAL RELEVANCE: This study revealed the value of RAPD with the RAPD1 primer as a tool for epidemiologic investigations of Salmonella Enteritidis PT 4. It can be used in conjunction with PFGE and phage typing to determine the genetic relatedness of Salmonella Enteritidis isolates involved in outbreaks of disease. A reliable and highly discriminatory method for epidemiologic investigations is critical to allow investigators to identify the source of infections and consequently prevent the spread of Salmonella Enteritidis PT 4. 相似文献
11.
Molecular investigation of 16 strains, conventionally identified to be Malassezia pachydermatis, isolated from dogs in Japan was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD band patterns of 13 clinical isolates were identical to that of standard strain of M. pachydermatis (CBS-1879). The other three clinical isolates were different from the standard strain of M. pachydermatis in RAPD patterns, and two of the three isolates were identical. About 620 bp genomic DNA fragments of the CHS2 gene were amplified from the same 16 clinical isolates of M. pachydermatis by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of the 16 clinical isolates revealed that the 13 strains were genetically very close to the standard strain of M. pachydermatis and the other two isolates were genetically close to the standard strain of M. furfur rather than M. pachydermatis. The remaining one isolate was phylogenetically distinct from all the seven Malassezia species reported so far. 相似文献
12.
根据鸡白痢沙门菌与鸡伤寒沙门菌的rfbS基因在第237和第598位碱基的不同，设计和合成了等位基因特异性PCR引物，建立了快速检测鸡白痢沙门菌的PCR方法，并应用该方法对鸡白痢沙门菌临床分离样品进行了PCR鉴定。结果显示，该PCR方法能够特异性地鉴定鸡白痢沙门菌，检测灵敏度达100PgDNA。对35个经常规方法鉴定的鸡白痢沙门菌分离株应用等位基因特异性PCR方法进行鉴定，鉴定出33株鸡白痢沙门菌，符合率为94．3％。表明，建立的等位基因特异性PCR方法能够准确而快速地鉴定鸡白痢沙门菌。 相似文献
13.
J Osek 《Research in veterinary science》1999,67(2):197-198
The genetic relatedness among 72 Escherichia coli strains of serotype O149:K91 isolated from pigs with diarrhoea was investigated by randomly amplified polymorphic DNA (RAPD) analysis. Fimbrial and toxic virulence markers of the isolates were also tested. Amplification with primer 1254 resulted in three different RAPD types whereas primer 1290 generated one RAPD profile only. Based on the RAPD and fimbrial/toxin types the strains were classified into five distinct groups. 相似文献
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OBJECTIVE: To identify swimming motility in Salmonella pullorum isolates and to characterize the flagellar proteins produced by motile isolates. SAMPLE POPULATION: 30 S pullorum isolates and isolates of 7 other Salmonella sp. PROCEDURE: Salmonella pullorum isolates were inoculated into high motility medium to evaluate swimming motility. Putative flagellar proteins were purified from the organisms and analyzed by means of gel electrophoresis and western blotting procedures, using various antisera specific for flagellar proteins. Antisera shown to be reactive with putative flagellar proteins were incorporated into the growth medium to examine their effects on motility of the isolates. RESULTS: All S pullorum isolates had evidence of swimming motility. Two putative flagellar proteins were purified from 2 of the S pullorum isolates: a 60 to 62 kd protein shown to react with antiserum specific for type y flagellar protein, and a 58 to 59 kd protein shown to react with antiserum specific for type d flagellar protein and with antibody reactive to a highly conserved flagellar epitope found on various Enterobacteriaceae. Antiserum specific for type d flagellar protein inhibited swimming motility of S pullorum isolates, but antiserum specific for type y flagellar protein did not. CONCLUSIONS: Results suggest that S pullorum isolates can be induced to manifest swimming motility when grown on medium with a low agar concentration and possess a 58 to 59 kd protein of d serotype and a second protein of 60 to 62 kd that also may be a flagellar protein. 相似文献
16.
Ceelen LM Haesebrouck F Favoreel H Ducatelle R Decostere A 《Veterinary microbiology》2006,113(1-2):45-53
Helicobacter pullorum has been associated with diarrhoea, gastroenteritis and liver disease in humans and with hepatitis and enteritis in poultry. The purpose of the present study was to examine whether cytolethal distending toxin was present among 10 poultry and three human H. pullorum isolates and whether a different level of cytolethal distending toxin activity was noted. A PCR assay was performed to detect the cdtB gene. In addition, epithelial Hep-2 cells inoculated with sonicate from all strains were observed microscopically and DNA analysis of these cells was done by flow cytometry. All H. pullorum isolates harboured the cdtB gene, but functional cytolethal distending toxin activity was only demonstrated in the human H. pullorum strain CCUG 33839. A significant number of cells treated with sonicate from this strain were enlarged. The nuclei were distended proportionally. Giant cells and multinucleated cells were observed as well. In addition, stress fibers accumulated. DNA analysis by flow cytometry revealed 31.0% of these cells at the S/G2 stage of the cell cycle. The tested poultry and human H. pullorum isolates all possess the cdtB gene, but under the circumstances adopted in this study only the human strain CCUG 33839 seems to show biological activity typically for CDT in vitro. 相似文献
17.
鸡白痢沙门氏菌耐药性的监测研究 总被引：42，自引：0，他引：42
采用WHO推荐的Kirby-Bauer法对我国部分地区1962－1999年间346株鸡白痢沙门氏菌进行了药物敏感性测定。结果表明，在近40年时间里，鸡白痢沙门氏菌对氨苄青霉素、壮观霉素、复方磺胺、磺胺异恶性唑、甲氧苄胺嘧啶、羧苄青霉素、四环素、链霉素、青霉素的耐药率显著增强（P＜0.01)。菌株的多重耐药率明显增加。60年代以二耐菌株为主（37.0%)，70年代以四耐、五耐菌株居多（60.5%)，80年代五耐、六耐、七耐菌株占大多数（80.2%)，90年代仅七耐以上菌株就达83.7%。不同年代菌株显示出不尽相同的耐药谱，且呈现出不断增宽的趋势。通过流行病学调查发现，60年代至90年代，鸡白痢沙门氏菌对青霉素、链霉素、磺胺类药物、四环素、卡那霉素、氯霉素、庆大霉素等药物的耐药性大多呈现出显著增强的变化趋势，这充分说明细菌耐药性的形成和发展与抗菌药物长期反复使用有着极为密切的关系。研究对于指导临诊合理用药及控制疾病流行具有重要的现实意义。 相似文献
18.
A polymerase chain reaction assay, amplifying a 1027 base pair portion of the 23S rDNA gene, was evaluated for identification of the intestinal spirochaete Serpulina intermedia. A total of 34 strains of S. intermedia isolated from pigs and chickens and 195 strains of other related species were tested. The optimised assay correctly identified all the S. intermedia strains, but generated 11 false positive reactions, giving a test sensitivity of 100% and a test specificity of 94.3%. The false positive reactions were generated from strains of four different species of intestinal spirochaetes, and the product was of the original predicted size. This suggests that the primer sites selected on the 23S rRNA gene were not completely specific for S. intermedia. Pulsed-field gel electrophoresis was then developed to investigate diversity amongst the S. intermedia strains. All strains tested had distinct DNA banding patterns using Mlu1, although three isolates from chickens on the same farm appeared closely related. The collection exhibited considerable genetic diversity, and strains from pigs and chickens were distributed in clusters throughout the dendrogram produced. The most closely related porcine and avian strains shared only 62% similarity. 相似文献
19.
Ramasoota P Krovacek K Chansiripornchai N Mörner AP Svenson SB 《Acta veterinaria Scandinavica》2000,41(3):249-259
A standardized-reagents commercial kit for random amplified polymorphic DNA (RAPD) analysis was used for typing 58 Escherichia coli strains that were recovered from the milk of sows, having coliform mastitis, within a single swineherd in Sweden. Previously, the 58 E. coli strains were characterized serologically and profiled biochemically. They were also evaluated for their serum resistance and their ability to adhere to fibronectin and bovine fetal fibroblasts. The RAPD analysis was fast, easily performed, and required only a nanogram of DNA. The indistinguishable banding patterns obtained with repeated analyses of 2 isolates from each strain demonstrated that RAPD analysis using standardized beads is a technique that provides reproducible results for typing E. coli strains that cause mastitis in sows. The results of the RAPD analyses demonstrated that E. coli sow mastitis strains are highly variable in serotype, biochemical profiles, virulence factors, and RAPD type, and that all 58 strains can be differentiated by means of the RAPD technique. The strains grouped into 24 RAPD types by combining the results of 2 primers, and into 38 groups by combining the results of serotype and RAPD type. No relationship between serotypes, virulence factors and RAPD types was found. 相似文献
20.
Genotyping of Staphylococcus aureus isolated from various sites on farms with dairy sheep using pulsed-field gel electrophoresis 总被引：1，自引：0，他引：1
We investigated the genetic diversity of 179 Staphylococcus aureus isolates recovered from various sites in 10 farms producing cheeses manufactured with raw ewe's milk. Isolates were collected from handcrafted cheeses, bulk tank milk, milk from half-udders, skin abscesses on the udder if present, hands and anterior nares of farmers, and air of the milking area. The isolates were typed using pulsed-field gel electrophoresis (PFGE) of DNA SmaI digests and compared to other isolates of S. aureus isolated in different hosts or in different locations. The results showed that nine farms were contaminated by S. aureus isolates with identical banding patterns (named OV) or by genetically related isolates (named OV'). These dominant banding patterns were found in a variable proportion of the samples from each farm (range: 11-100%). Most of the strains isolated from nasal carriage or strains isolated from other regions or from other animal species had different PFGE patterns to OV or OV', except for three strains. These results show that a single clone of S. aureus is widely distributed both in infected mammary glands and in cheese produced from raw milk. This study confirms that infected mammary glands are the main source of the contamination of dairy products in sheep. 相似文献