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1.
为了解某肉种鸡场沙门菌感染来源、类型及其对药物和噬菌体的敏感性,研究对种鸡场内外养殖环境和孵化场死胚进行了沙门菌采样检测。样品通过亚硒酸盐胱氨酸增菌液增菌培养后接种显色培养基进行沙门菌分离,并通过生化试验、PCR和血清型进一步鉴定;通过MIC测定了解沙门菌分离株对10种抗菌药物的药敏性;通过双层平板法检测了一株沙门菌噬菌体BS3对分离菌株的裂解谱。结果显示:从采集的1850份样品中分离到45株沙门菌,其中肠炎沙门菌为优势血清型(60%);沙门菌分离株对10种抗菌药物具有不同程度耐药并具多重耐药性;噬菌体BS3能够裂解75.5%的沙门菌分离株。提示应根据药敏结果选择适宜药物进行沙门菌感染防控,针对沙门菌的高度耐药性,噬菌体的筛选与应用将具有重要价值。  相似文献   

2.
为了解某蛋鸡场金黄色葡萄球菌和沙门菌对抗菌药物的耐药情况,采集重庆市某蛋鸡场48份羽毛样品、123份肛拭子样品,将羽毛样品经MH高盐培养液增菌后划线接种于MH高盐平板,将肛拭子样品经沙门菌增菌液增菌后划线接种于S.S.平板。将疑似菌落接种LB液体培养基培养后提取基因组,PCR扩增金黄色葡萄球菌特异性nuc基因和沙门菌特异性invA基因片段。对分离鉴定的金黄色葡萄球菌和沙门菌进行药物敏感性试验。最终共分离鉴定出9株金黄色葡萄球菌、6株沙门菌。药敏试验结果表明,分离的金黄色葡萄球菌和沙门菌耐药严重,且存在多重耐药。本研究基本摸清了该蛋鸡场金黄色葡萄球菌和沙门菌对常见抗菌药物的敏感性,为该蛋鸡场的临床用药提供了参考。  相似文献   

3.
为了解徐州地区鸭源沙门菌的感染情况,对徐州市场和超市300份样品(鸭肉及副产品)、6个养鸭场的380份样品(鸭泄殖腔肛拭子、粪便以及饲料和用具等)以及50例临床发病鸭的脏器组织样品(肝、脾、盲肠等)进行增菌培养,通过细菌分离培养特性、生化反应和血清型鉴定,成功分离到56株鸭源沙门菌。市场及超市产品共分离到6株沙门菌,分离率为2%,优势血清型为肠炎沙门菌;鸭场共分离到26株沙门菌,分离率为6.8%,优势血清型为肠炎沙门菌;临床发病鸭分离到24株,分离率为48%,其优势血清型为印第安纳沙门菌和肠炎沙门菌。对21株沙门菌进行ERIC-PCR分子分型,分成18类基因型,遗传相似性在59%~100%,结果显示出良好的多态性,获得了较好的指纹图谱数据。  相似文献   

4.
美国科宝公司的工作人员在2种初级增菌培养基中定量接种沙门菌样本后,确定复苏家禽环境基质中沙门菌最有效和快速的方法。样品随机接种2种培养基,即缓冲蛋白胨水(BPW)+酵母提取物与含碘的四磺酸盐煌绿增菌液(TT)。将冷冻沙门菌培养菌解冻后,连续10倍稀释,取100μL稀释液接种到增菌液中,分别在42℃和37℃条件下孵育24h后,接种到改良半固体RV培养基(MSRV)中42℃二次孵育。经过次级增  相似文献   

5.
为了了解重庆市荣昌区畜禽源沙门菌的流行特征和耐药情况,从荣昌区的一家规模化生猪养殖场和一家肉鹅养殖场分别采集动物粪便样品80份和30份,样品经过前增菌、选择性增菌、平板划线接种、挑取可疑菌落革兰氏染色并扩增沙门菌特异性基因invA,从而鉴定沙门菌分离株。对沙门菌分离株进行血清型分型和药物敏感性试验。试验结果显示,110份样品中共分离到8株沙门菌,其中猪源沙门菌6株,鹅源沙门菌2株。沙门菌血清型鉴定表明,8株沙门菌中有5株为鼠伤寒沙门菌,1株为乙型副伤寒沙门菌,1株为圣保罗沙门菌,1株沙门菌尚不能判断。药敏试验表明,沙门菌分离株对四环素类抗菌药物的耐药现象最为严重,其次为酰胺醇类、青霉素类和氨基糖苷类。8株沙门菌分离株菌均为多重耐药菌株,尤其需要注意的是,分别有1株鹅源和猪源沙门菌对16种和17种抗菌药物产生耐药,多重耐药现象严重。  相似文献   

6.
一沙门氏菌的检测 1常规细菌学分离方法(GB/T13091-2002) 第一步,前增菌,使样品在含有营养的非选择性培养基中增菌,使受损伤的沙门氏菌细胞恢复到稳定的生理状态。第二步,选择性增菌,在含选择性抑制剂的促生长培养基中间,样品进一步增菌。  相似文献   

7.
本文旨在研究中国华东地区沙门菌分布情况及其优势菌的血清型变异情况.收集2010年10月-2012年8月来源于江苏、安徽、上海等地区的1 730份禽源样品,进行增菌、纯化、PCR鉴定、血清学试验及生化试验,并对7株血清型有变异的疑似鸡白痢沙门菌进行进一步序列测定,对血清型进行分析.结果表明,本次调查中共分离沙门菌87株,包括鼠伤寒沙门菌、德尔卑沙门菌、圣保罗沙门菌、鸡白痢沙门菌及阿巴特图巴沙门菌,并且在鸡白痢沙门菌血清型H相存在Hd、Hg阳性的现象.结果提示,中国禽类沙门菌感染以鸡白痢沙门菌感染为主,且存在血清型变异情况,变异概率呈上升趋势.  相似文献   

8.
针对河南某大型肉牛场牛猝死的现象,采集饲料样品、牛血液进行微量元素硒检测;对病死牛无菌采取病料进行分离培养和细菌学检查,通过菌体形态、鉴别培养基培养、动物试验、细菌自动鉴定仪检测等方法对病原菌进行了鉴定。结果表明,饲料样品、牛血液中微量元素硒含量合格;病原菌为集团肠杆菌、沙门菌。牛猝死不是因硒缺乏引起的,而是因为牛只突然换料应激后免疫力降低,导致集团肠杆菌,肠炎沙门菌(溶血型)和肠炎沙门菌属沙门菌引起的急性死亡。  相似文献   

9.
噬菌蛭弧菌为一类杆形、弧形或螺旋形的革兰阴性菌,可以寄生并裂解其他细菌。本试验以禽源大肠杆菌、沙门菌以及环境水体中分离的1株大肠杆菌为宿主菌,采用双层琼脂培养法成功地从环境水体中分离到3株噬菌蛭弧菌,分别命名为BDD、BDE、BDH。电镜观察证明菌体大小约0.33μm×0.78μm,单个弧形,一端有一根长鞭毛。基因序列分析表明,分离株与已发表的噬菌蛭弧菌属其他菌株16S rRNA基因序列的同源范围分别为96.0%~99.9%。通过对不同细菌的裂解试验证明,3个分离株对大肠杆菌、沙门菌、摩根摩根氏菌、变形杆菌和阴沟肠杆菌等均具有裂解作用,但对铜绿假单胞菌、鸭疫里默氏菌和金黄色葡萄球菌不敏感。  相似文献   

10.
对2020年在宁夏和新疆部分动物养殖场和屠宰场分离的沙门菌(Salmonella)进行了相关抗菌药物的耐药性调查,以了解沙门菌的流行特点以及耐药性情况,一方面为国家动物源细菌耐药网提供数据,另一方面为相关养殖场临床抗菌药物的合理使用提供科学依据。首先采用沙门菌选择性培养基进行菌株的分离、纯化及鉴定,采用微量肉汤稀释法检测16种抗菌药物对分离菌株的最小抑菌浓度(MICs),采用玻片凝集法对分离菌株进行血清型鉴定。结果显示:试验共分离到60株沙门菌,其血清型有16种,主要包括鼠伤寒沙门菌(S. Typhimurium)、里森沙门菌(S. Rissen)、汤普森沙门菌(S. Thompson)、肠炎沙门菌(S. Entertidis)、德尔卑沙门菌(S. Derby)、明斯特沙门菌(S. Muenster),检出率分别为26.67%(16株)、15.00%(9株)、11.67%(7株)、10.00%(6株)、8.33%(5株)、6.67%(4株),其他9种血清型的检出率为1%~5%。分离菌株对磺胺异噁唑、四环素和氨苄西林耐药程度较高,耐药率分别达75.00%、70.00%和66.67%;对奥...  相似文献   

11.
OBJECTIVE: To compare 3 alternative culture techniques for the detection of Salmonella organisms in swine feces with a modification of the International Standard Organization (ISO) 6579 standard protocol. SAMPLE POPULATION: Fecal samples from swine herds suspected of having Salmonella infections. PROCEDURE: 4 experiments were performed to evaluate the following: 1) diagnostic sensitivity of the selective preenrichment and rapid isolation novel technology (SPRINT) protocol, compared with that of the modified ISO protocol; 2) detection limit of the SPRINT protocol for Salmonella organisms; 3) use of tetrathionate-novobiocin (TTN) broth, compared with selenite cysteine (SC) broth for selective enrichment; and 4) use of universal preenrichment (UPE) broth, compared with buffered peptone water (BPW) for preenrichment of samples prior to the use of modified semisolid Rappaport-Vassiliadis (MSRV) plates. RESULTS: Comparing the Salmonella culture results of 183 swine fecal samples, the diagnostic sensitivity of the SPRINT protocol (0.86) was not significantly different than the diagnostic sensitivity of the modified ISO protocol (0.80), although it was 24 hours faster. The SPRINT protocol could detect 5 of the 6 investigated Salmonella serotypes at inoculation concentrations of < 10 colony-forming units (CFU)/25 g of uncontaminated feces. The TTN broth performed significantly better than the SC broth for selective enrichment of Salmonella organisms. There was no significant difference in results of preenrichment of samples between the use of UPE broth or BPW. CONCLUSIONS AND CLINICAL RELEVANCE: The SPRINT protocol may provide a faster alternative for isolation of Salmonella organisms from swine fecal samples. Furthermore, the use of TTN broth instead of SC broth may increase the sensitivity of the modified ISO 6579 protocol.  相似文献   

12.
133 samples of food were investigated for comparison of a recent commercially available 1-2 test (Biocontrol) with a cultural standard method (non-selective pre-enrichment in buffered peptone water, selective enrichment in Rappaport-Vassiliadis-medium and selenite brilliant-green mannitol enrichment broth, inoculation on two selective agars) for presence of Salmonella. The 1-2 test showed in positive results an accuracy ("sensitivity") of 94.7% and in negative results an assurance ("specificity") of 97.9% and is therefore considered suitable for detection of Salmonella contaminated food.  相似文献   

13.
OBJECTIVES: To determine the prevalence of Salmonella infections in horses at necropsy. DESIGN: Cross-sectional prevalence survey. ANIMALS: 102 horses. PROCEDURE: Mesenteric lymph nodes were collected from horses that were necropsied. Horses had died or were euthanatized because of severe disease or at the request of the owner. Twenty-eight of the horses were racehorses euthantized following acute catastrophic injuries on the racetrack. Mesenteric lymph nodes were submitted for Salmonella culture via direct plating of tissue specimens on MacConkey agar and by use of 4 enrichment culture techniques that used tetrathionate and selenite enrichment broth and brilliant green and Salmonella-Shigella selective plating media. RESULTS: Salmonella typhimurium was isolated from the mesenteric lymph nodes of 2 foals (2/102, 1.96% of the horses). Salmonella organisms were not isolated from the mesenteric lymph nodes of adult horses. CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of Salmonella infections in horses of our study (1.96%) suggests that the results of cross-sectional surveys, using bacteriologic culture to determine prevalence of Salmonella infection, should be interpreted with caution. Prevalence of Salmonella infections determined in a single facility may not reflect the prevalence of Salmonella-infected horses in the general population; furthermore, obtaining a Salmonella isolate from a horse does not establish that the horse is a chronic Salmonella carrier.  相似文献   

14.
1. When the contents of 4 or more naturally-contaminated intact eggs were combined, the isolation rate of Salmonella enteritidis was improved by extending incubation at 37 degrees C from 24 to 48 h before sub-culture. 2. The isolation rate of salmonellas from raw homogenised whole egg was significantly increased by the inclusion of Novobiocin and Cefsulodin in the primary culture media. 3. Rappaport Vassiliadis broth was found to be superior to Selenite as a selective enrichment medium.  相似文献   

15.
We established the PCR detection system specific to Salmonella species using Salmonella enterotoxin gene (stn). The detection limit was one bacterial cell per one gram of fecal and minced-meat samples using enrichment procedure by Tripticase soy broth or Salmonella enrichment broth, respectively. We concluded that this PCR system is useful for the practical application in the field of the public hygiene.  相似文献   

16.
Three different selective enrichment media, Rappaport-Vassiliadis broth (RV), selenite broth (SB) and Müller-Kauffmann tetrathionate broth (MKTB), in combination with plating on modified brilliant green agar (BGA), were compared for the isolation of Salmonella from samples of pig feces. These conventional methods were also compared with a new ELISA kit in conjunction with RV and SB enrichment. Of the conventional methods, enrichment in RV had a higher sensitivity and selectivity than SB and MKTB. Recovery of S. typhimurium from MKTB was significantly poorer than recovery of other serotypes. The combination of RV enrichment and ELISA was as good as the conventional method involving RV enrichment, with a similar high sensitivity and specificity.  相似文献   

17.
The aim of the study was to give an account of the epidemic of abortions in sheep caused by Salmonella enterica ssp. enterica serovar Abortusovis, which occurred in Dalmatia, south Croatia, in winter 2003-2004. Five sheep flocks with rate of abortion ranging from 22% to 38% during the last-third of gestation were examined. Salmonella Abortusovis was isolated from 13 vaginal smears and two fetuses. Direct inoculation was found to be superior to pre-enrichment and enrichment in selective broth for Salmonella Abortusovis isolation. The isolates were biochemically identified, and characterized by serotyping and polymerase chain reaction based on the amplification of the IS200 sequence specific for Salmonella Abortusovis. A fragment of 900 bp was detected in all Salmonella Abortusovis isolates. The sensitivity testing of the isolates, carried out by the disk diffusion method and the determination of the minimal inhibitory concentrations, resulted in a high sensitivity to almost all antimicrobials used. Only two isolates were moderately sensitive to oxytetracycline, whereas one isolate showed resistance to streptomycin. Campylobacter fetus ssp. fetus and Listeria monocytogenes were excluded as causative agents of abortion in sheep by culture testing, and brucellosis, leptospirosis, Q fever and chlamydiosis by serological testing.  相似文献   

18.
To attempt the rapid detection of Salmonella enterica, we have coupled a culture procedure with PCR amplification of the genus-specific invE/invA genes. The method was applied to different kinds of samples from the poultry industry and evaluated by using hydrolyzed feather meal, meat meal, litter and viscera, all experimentally inoculated with a known number of Salmonella followed by cultivation in selenite--cystine broth prior to the PCR reaction. The expected 457bp specific DNA fragment could be amplified from dilutions containing as few as 5.7CFU, indicating that the PCR technique can be successfully coupled with culture in an enrichment broth to distinguish Salmonella species from other enteric bacteria present in samples from the poultry industry. Tetrathionate broth proved to be a much better enrichment media compared to selenite-cystine when the presence of Salmonella was evaluated by PCR in 1-day-old chicks experimentally infected with known numbers of Salmonella. Samples included cecal tonsils and viscera, collected at 48h and 7 days postinfection. The PCR technique was more sensitive in detecting infected animals than the standard microbiological procedure, which detected only 47% of all PCR positive samples.  相似文献   

19.
A rapid and readily available DNA probe kit was developed for the detection of Salmonella spp. This kit utilized the colorimetric DNA/rRNA sandwich hybridization method in microtiter wells. Within 3 hr Salmonella spp. in selective enrichment broth cultures were detected by the DNA probe kit. The kit effectively identified all of 187 strains of Salmonella tested and yielded no false-positive reactions in the examination of 674 pure cultures of non-salmonellae. The DNA probe kit could detect 10(5) cfu/ml in pure culture. A total of 379 naturally contaminated samples (raw chicken meat, liquid egg, animal feeds, poultry feces and frozen foods) were tested, both by the standard culture method and the DNA probe kit. The 169 of these samples were culture positive and 210 were culture negative. The sensitivity of the DNA probe kit was 98.2% (166/169) and the specificity was 99.5% (209/210). These results show that the DNA probe kit is a useful tool to examine a large number of various samples for contamination by Salmonella spp. in food and livestock industry.  相似文献   

20.
The selective L-PALCAMY differential enrichment broth, the Listeria enrichment broth of the International Dairy Federation, Oxford Listeria selective agar, and PALCAM Listeria selective agar were comparatively examined in the cultural isolation of Listeria spp. from ten ruminant brains. The L-PALCAMY medium proved to be superior to the IDF broth in both selectivity and productivity for Listeria spp. in the brain samples, which were also contaminated with other bacteria. The Oxford and PALCAM agars corresponded in their productivity for Listeria spp. The latter, however, was more selective than the Oxford agar. Bacterial counts of up to 1.2 x 10(9) CFU/g of brain stem sample were made from Listeria monocytogenes (L.m.), and up to 6.2 x 10(4) CFU/g from Listeria innocua. A total of 164 brains from ruminants showing CNS disturbances and/or pathoanatomical CNS alterations were examined using L-PALCAMY medium, and Oxford and PALCAM agar. L.m. could be isolated from 29 of the brains, and Listeria innocua from five. Cultural isolation of both Listeria spp. occurred in one brain. Of 27 brains containing L.m., which were also examined using cold enrichment, L.m. was isolated in 59.3% of the cases with direct culture, in 81.5% of the cases using selective warm enrichment, and in 77.8% of the cases by means of selective cold enrichment. Five cases each were identified solely by cold or warm enrichment, respectively. In investigations of further 69 ruminant brains the number of brains shown to contain L.m. could be increased from seven to 13 by means of selective cold enrichment for three months.  相似文献   

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