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1.
本研究采用Nested-PCR方法,对于临床采集的感染空肠弯曲菌的病鸡组织样品进行了检测,研究结果表明,采用Nested-PCR方法检测空肠弯曲菌具有结果清晰,操作简便,快速等特点.研究还发现,空肠弯曲菌对鸡的致病性可能增强,应引起重视.  相似文献   

2.
部分省市鸡群空肠弯曲菌和结肠弯曲菌的流行状况分析   总被引:2,自引:0,他引:2  
本研究采用多重PCR方法对我国部分省市不同品种、不同类型的鸡群进行空肠弯曲菌和结肠弯曲菌流行病学调查。结果发现,在3132份鸡肛门棉拭样品中,检测出583份空肠弯曲菌阳性样品,平均阳性率18.61%;48份结肠弯曲菌阳性,平均阳性率1.53%。鸡饲养环境样品217份,6份空肠弯曲菌阳性,阳性率2.76%。不同鸡群感染率悬殊较大,空肠弯曲菌阳性率从0%-73.3%、结肠弯曲菌阳性率从0%-7.4%。30个鸡群中空肠弯曲菌阳性率为86.67%,结肠弯曲菌阳性率43.33%。本调查结果表明我国鸡群中空肠弯曲菌、结肠弯曲菌的流行呈现多样化。  相似文献   

3.
以空肠弯曲杆菌保守基因mapA和hipO基因为靶基因,建立空肠弯曲菌的双重PCR方法,该方法只对空肠弯曲菌有特异性,对大肠埃希菌、金黄色葡萄球菌等则不能扩增出特异性片段。利用该方法对从市场、养殖鸡和屠宰场的禽源空肠弯曲菌进行了分离和鉴定,平均分离率为35.8%,PCR鉴定结果与生化鉴定结果相一致,说明建立的双重PCR鉴定方法具有灵敏、特异的特点,可节省传统生化鉴定所耗费的时间,为空肠弯曲菌的检测提供新方法。  相似文献   

4.
本研究根据空肠弯曲菌及结肠弯曲菌的16s rRNA基因相同保守序列来设计引物,建立了一种PCR检测方法,该方法具有较强的特异性及灵敏性,其对空肠弯曲菌及结肠弯曲菌的最低检测限度为5 CFU/mL,它无需增菌培养过程可以直接对样品进行检验,在对鸡肉样品空肠弯曲菌及结肠弯曲菌的检测中取得了较好的效果.  相似文献   

5.
为了研究更快、更加准确分离空肠弯曲杆菌的方法,试验同时采用常规生化法对由12份猪盲肠内容物中分离的可疑菌进行检测,得到了6个阳性样品,经特异性PCR检测,2个样品可定性为空肠弯曲杆菌,检测时间为7 d;而对所得菌株直接进行PCR检测,检测结果相同,检测时间为2 d。结果表明:PCR可以快速、特异性地检测空肠弯曲杆菌,与常规生化检测法相比是一种更有效的方法。  相似文献   

6.
本研究应用多重PCR反应(mPCR)结合变性高效液相色谱(DHPLC)技术建立食品中空肠弯曲菌和结肠弯曲菌的快速检测方法.以编码嗜热弯曲菌属的16S rRNA基因、编码空肠弯曲菌的gyrA基因和编码结肠弯曲菌的cdt真基因为靶基因,选择3对引物,建立并优化了鉴别空肠弯曲菌和结肠弯曲菌的mPCR体系,扩增产物分别为287 bp、159 bp和173 bp.采用22株细菌验证了该mPCR具有特异性.mPCR检测的灵敏度在DNA水平上达到空肠弯曲菌10pg/μL、结肠弯曲菌1 pg/μL;在人工模拟污染样品起始污染浓度为1.5个/mL时,42℃微需氧条件下培养24 h即可被检出.在随机采集的172份冷冻鸡肉类样品中,检出了18份样品为空肠弯曲菌阳性,7份样品为结肠弯曲菌阳性.本研究建立的mPCR-DHPLC方法可特异、灵敏地实现对空肠弯曲菌和结肠弯曲菌的快速检测.  相似文献   

7.
了解库尔勒市牛源空肠弯曲菌流行及耐药性状况,为空肠弯曲菌的防控与临床用药提供理论基础。采用特异性选择培养基培养并结合PCR法对新疆库尔勒市及其周边牛养殖场采集的787份肛拭子样品进行空肠弯曲菌的分离鉴定,并采用纸片扩散法对牛源空肠弯曲菌分离株进行10类23种抗菌药物的耐药性检测。结果共分离到空肠弯曲菌38株,总分离率为4.83%(38/787); 38株空肠弯曲菌对克林霉素、四环素、氨苄西林、林可霉素、阿莫西林、头孢噻肟、头孢曲松表现高度耐药,耐药率均达80%以上;对美罗培南的敏感性最高,耐药率为0。研究表明,库尔勒市存在牛源空肠弯曲菌的流行隐患,空肠弯曲菌分离株对不同药物的耐药性存在差异,且耐药谱具有多样性,多重耐药情况普遍,应引起各部门的高度重视。  相似文献   

8.
科技动态     
正基因突变导致禽源弯曲菌对大环内酯类药物耐药为研究上海地区禽源弯曲菌对大环内酯类药物耐药状况及其耐药机制,中国农业科学院上海兽医研究所的研究人员采用微量琼脂稀释法测定了209株弯曲菌(空肠弯曲菌84株,结肠弯曲菌125株)对大环内酯类药物(红霉素和阿奇霉素)的最小抑菌浓度,采用错配PCR方法检测大环内酯类耐药弯曲菌的23S rRNA基因突变情况。药物  相似文献   

9.
对江苏省盐城国家级珍禽自然保护区丹顶鹤中空肠弯曲菌带菌情况进行监测,共采集泄殖腔标本80份,采用国标法,并用API Campy System进行了生化与分类鉴定,结合弯曲菌多重PCR检测方法,共检出弯曲菌17株,阳性率为21.67%,其中空肠弯曲菌10株。结果表明,丹顶鹤弯曲菌的感染与年龄有关,未成年丹顶鹤弯曲菌阳性率极显著地高于成年丹顶鹤;同时对部分菌株用琼脂扩散法进行药敏试验。  相似文献   

10.
以方阵滴定法确定了空肠弯曲菌抗血清、酶结合物的最佳工作浓度和孵育时间,从而立了快速检测空肠弯曲菌的间接ELISA。该试验可检出含8~10个空肠弯曲菌经24h培养的标本(含30万个菌/ml的肉汤培养物);不与沙门氏菌等15种对照菌及其混合液发生交叉反应。以鸡源和猪源空肠弯曲菌抗血清分别作为ELISA的第一抗体,对144份鸡泄殖腔粪便标本和116份猪直肠粪便标本的选择性增菌肉汤培养物进行了检测。其阳性率分别为83.3%和79.3%,可于28h内报告结果;常规法对上述标本的阳性检出率分别为85.4%和81%,但程序繁琐,需5~7d方能获得最终检查结果。两种方法对鸡、猪粪便标本检测结果的阳性符合率,分别为97.5%与97.9%。  相似文献   

11.
Consumption of poultry meat is associated with human Campylobacter and Salmonella infections. One way to control the presence of these bacteria in broiler flocks is to make chickens less susceptible for colonisation. Acidification of feed may be a tool to reduce the Campylobacter and Salmonella carriage in broiler chickens. In the present experiments an acidified feed with high levels of organic acid, 5.7% lactic acid and 0.7% acetic acid, was applied. In an in vitro experiment the reduction or growth of Campylobacter and Salmonella was measured after addition of 10(7)cfu of these bacteria into a conventional broiler feed, acidified feed and fermented feed, whereas the numbers of Salmonella increased in non-acidified feed. The number of Campylobacter decreased 2-3 (10)log cfu. In the acidified and fermented feed a complete reduction of Campylobacter was observed within 20 min, and a total Salmonella reduction started after 1h, and was complete after 2h. Subsequently, an in vivo experiment with 100 individually housed broiler chickens showed that chickens fed acidified feed were less susceptible to an infection with Campylobacter than were chickens fed conventional feed. The size of reduction was however limited. The susceptibility for Salmonella colonisation was not affected by acidified feed. It is concluded that the role for acidified feed in the control of Campylobacter and Salmonella is limited.  相似文献   

12.
Salmonella and Campylobacter are undesirable pathogens on poultry. Therefore the effect of fermented feed on the colonization in the gastro-intestinal tract of the chicken, the introduction of both bacteria in a chicken flocks, and the transmission between chickens was studied. Broilers that were fed with fermented feed were significantly less susceptible for Salmonella and Campylobacter than chickens on a standard chicken feed. The spread of Salmonella between broiler chickens was reduced. However, the results also showed, like for other known control measures, that this feed can not absolutely guarantee the absence of Salmonella and Campylobacter. Therefore fermented feed must be seen as one of the hurdles in a so called multiple hurdle strategy. The combination of different hurdles should prevent the introduction and transmission. The effect of fermented feed on Campylobacter and Salmonella is partially caused by the presence of high concentrations of organic acids. In chickens fed with liquid feed the acidic barrier in the first part of the GI-tract was clearly improved. Besides organic acids there are other changes in the GI-tract. Changes in colonization levels of indicator organisms, changes in levels of organic acids and an increased pH in ileum and ceacum. These changes indicate a stabilised GI-flora in fermented feed fed poultry. The research confirmed that by changes in the composition of the feed (carbohydrates, acids, or micro-organisms) the GI-health can be promoted and therewith can contribute to the control of food pathogens in farmed animals.  相似文献   

13.
An enzyme-linked immunosorbent assay (ELISA) was developed in a homologous system with bacterial ultrasonic-treated proteins as the antigen and antisera from chickens infected orally and subcutaneously with the strain Campylobacter jejuni serovar 6 (CJ 6). The cut-off level was determined using antisera from non-infected specific-pathogen-free chickens up to the age of 10 weeks. The suitability of the ELISA system was verified using antisera taken from chickens orally infected at the age of 4 weeks with CJ 1, 6, 28 or 36 or with Campylobacter coli serovar 28 (CC 28). The development of antibodies was monitored up to 6 weeks post-infection (p.i.). Sera from chickens infected with CJ 1, 6, 36 or CC 28 contained specific antibodies to Campylobacter, whereas in those infected with CJ 28 no specific antibodies were found. Distinct cross-reactions were observed between CJ 6, 28 and CC 28 antigens and their antisera 6 weeks p.i., while poor cross-reactions were found with antisera to CJ 1 and 28. Antibodies to strains of all heterologous serovars were successfully detected with an antigen pool comprised of CJ 1, 6 and 36 antigens. In 11 out of the 12 field sera obtained from 5- and 9-week-old broiler chickens suffering from campylobacteriosis, high specific antibody titres to Campylobacter jejuni were found.  相似文献   

14.
Thermophilic Campylobacter species, including Campylobacter jejuni and Campylobacter coli, are responsible for foodborne campylobacteriosis in humans and are increasingly resistant to fluoroquinolone (FQ) antimicrobials. The therapeutic use of FQ antimicrobial agents in food animal production, particularly in poultry, has become a concern for public health, because the practice may promote the emergence of FQ-resistant Campylobacter that can be transmitted to humans through the food chain. Recent studies have indicated that Campylobacter displays a hypermutable phenotype in response to in vivo treatment with FQ antimicrobials, resulting in the rapid emergence of resistant mutants. Distinct from other Gram-negative bacteria, the acquisition of FQ resistance in Campylobacter does not require stepwise accumulation of gyrA mutations and overexpression of efflux pumps, and is mainly mediated by single-step point mutations in gyrA in the presence of a constitutively expressed multidrug efflux pump, CmeABC. The simplicity of the resistance mechanisms may facilitate the rapid adaptation of Campylobacter to FQ treatment. The FQ-resistant Campylobacter mutants derived from chickens do not show a fitness cost in vivo and are ecologically competitive in the colonization of chickens even in the absence of antimicrobial selection pressure. These findings suggest that FQ-resistant Campylobacter may continue to persist regardless of antimicrobial usage, and highlight the need for extra effort to prevent the occurrence and spread of FQ-resistant Campylobacter in animal reservoirs.  相似文献   

15.
Vibrionic hepatitis is a disease of poultry which is characterised by the presence of focal lesions in the liver, usually 1-2mm in size and greyish-white in colour. The cause of the disease remains unclear, as do the reasons for its recent re-emergence. We examined the livers of commercial broiler chickens taken during processing and found Campylobacter spp. in both normal livers and those displaying signs indicative of focal hepatitis. Livers with signs of hepatitis had significantly more Campylobacter spp. present than those without and other bacterial genera were infrequently present. We were unable to replicate the disease in a healthy host following experimental infection with a Campylobacter jejuni strain isolated from a liver showing signs of focal hepatitis. However, a significant T cell response to C. jejuni was seen in the liver of Campylobacter infected birds. We conclude that the presence of Campylobacter spp. in the liver alone is not sufficient to cause vibrionic hepatitis, but that a predisposing factor, possibly within the host is required. We also provide evidence that chickens mount an adaptive T cell response to systemic C. jejuni.  相似文献   

16.
1. The frequency of thermophilic Campylobacter spp. on broiler carcases was determined during processing in a Southern Brazil slaughterhouse. Samples were collected after defeathering, evisceration, water chilling and freezing. In addition, samples were obtained from the water of the chiller tank and from the surface of equipment in direct contact with the chicken. 2. Samples (335) were analysed and 71.3% were positive for Campylobacter. The frequency of Campylobacter spp. on carcases rinsed in BPW and skin samples from carcases was 49 of 72 (68.0%) after defeathering, 50 of 72 (69.4%) after evisceration, 61 of 72 (84.7%) after chilling, and 46 of 72 (63.9%) after freezing. Campylobacter was positive for 21 of 23 (91.3%) samples in the chilling water and for 12 of 24 (50.0%) samples on the table surface. 3. The frequency of qualitative analysis for Campylobacter spp. was reduced in frozen chickens, but not during the slaughtering process. The use of drinking water alone as a decontaminant to reduce the incidence of Campylobacter spp. during slaughter is therefore not sufficient. Furthermore, to ensure food safety, chickens must be cooked properly before consuming.  相似文献   

17.
The main source for Campylobacter spp. transmission from the environment to broiler chickens is still unclear. One implicated reservoir for the organism has been untreated broiler drinking water. This study was conducted with broilers first using experimental conditions (isolation units) and second under commercial conditions. We compared the rate of intestinal colonization in chickens provided 2 to 5 parts per million (ppm) chlorinated drinking water in relation to the frequency of colonization in chickens given unsupplemented drinking water. No significant difference (P > 0.05) was detected in isolation frequency or level of Campylobacter spp. colonization in birds provided chlorinated drinking water and control birds provided water without supplemental chlorine. In the isolation unit experiments, 86.3% (69/80) of the control and 85.0% (68/80) of the treated birds were colonized at levels corresponding to an average of 10(5.2) and 10(5.1) log colony-forming units (cfu) Campylobacter spp./g of cecal contents, respectively. Additionally, two sets of paired 20,000 bird broiler houses, with and without chlorination (2-5 ppm chlorine), were monitored in a commercial field trial. Effectiveness of chlorination was judged by prevalence of Campylobacter spp. in fecal droppings (960 samples) taken from the flocks in treated and control houses. Birds from the control houses were 35.5% (175/493) Campylobacter spp. positive, while 45.8% (214/467) of the samples from the houses having chlorinated drinking water yielded the organism. Chlorination of flock drinking water at the levels tested in this study was not effective in decreasing colonization by Campylobacter spp. under commercial production practices presently used in the United States.  相似文献   

18.
Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in humans in the United States. Infectious bursal disease virus (IBDV) causes an immunosuppressive disease in young chickens. To analyze a possible role of IBDV-induced immunosuppression in colonization and shedding of C. jejuni, two experiments were conducted. In both experiments, group 1 consisted of noninoculated control chickens, groups 2 and 3 were inoculated with varying doses of C. jejuni, and groups 4 and 5 were inoculated initially with IBDV followed by doses of C. jejuni similar to groups 2 and 3. Campylobacter jejuni was recovered from the cloaca and cecum, but not the small intestines, from all chickens in groups 2 and 3. In groups 4 and 5, C. jejuni was recovered from the small intestines, cecum, and cloaca from all chickens. The amount (colony-forming units/sample) of C. jejuni recovered from chickens in groups 4 and 5 was significantly greater (P < 0.05) than the amount recovered from chickens in groups 2 and 3; and C. jejuni was also present sooner in these groups than in groups 2 and 3. Bursa samples from chickens in groups 4 and 5 were significantly smaller (P < 0.05) than in the other groups. Additionally, real-time polymerase chain reaction results for IBDV were positive in groups 4 and 5 and negative in all other groups. This study indicated that IBDV infection exacerbated colonization and shedding of C. jejuni, presumably through the immune suppression this virus causes in chickens. It highlights the need for further investigation into the role of immunosuppression in preharvest control strategies for food-borne disease-causing agents.  相似文献   

19.
Our objectives were to identify risk factors for contamination of French broiler flocks by Campylobacter. We used 75 broiler farms in western France. A questionnaire was administered to the farmers and samples of fresh droppings were taken to assess the Campylobacter status of the broiler flocks. 42.7% of the flocks were positive for Campylobacter spp. The risk of contamination of the broiler flocks by Campylobacter was increased in summer/autumn, in houses with static air distribution, when two or more people took care of the flock, in poultry farms with three or more houses and when the drinking water for the chickens was acidified. The presence of litter-beetles in the change room also increased the risk of contamination. The administration of an antibiotic treatment following a disease decreased the risk of a flock being contaminated by Campylobacter.  相似文献   

20.
An enzyme-linked immunosorbent assay (ELISA) was developed in a homologous system with bacterial ultrasonic-treated proteins as the antigen and antisera from chickens infected orally and subcutaneously with the strain Campylobacter jejuni serovar 6 (CJ 6). The cut-off level was determined using antisera from non-infected specific-pathogen-free chickens up to the age of 10 weeks. The suitability of the ELISA system was verified using antisera taken from chickens orally infected at the age of 4 weeks with CJ 1, 6, 28 or 36 or with Campylobacter coli serovar 28 (CC 28). The development of antibodies was monitored up to 6 weeks post-infection (p.i.). Sera from chickens infected with CJ 1, 6, 36 or CC 28 contained specific antibodies to Campylobacter, whereas in those infected with CJ 28 no specific antibodies were found. Distinct cross-reactions were observed between CJ 6, 28 and CC 28 antigens and their antisera 6 weeks p.i., while poor cross-reactions were found with antisera to CJ 1 and 28. Antibodies to strains of all heterologous serovars were successfully detected with an antigen pool comprised of CJ 1, 6 and 36 antigens. In 11 out of the 12 field sera obtained from 5- and 9-week-old broiler chickens suffering from campylobacteriosis, high specific antibody titres to Campylobacter jejuni were found.  相似文献   

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