致犊牛腹泻产肠毒素大肠杆菌及其检测

大肠杆菌是动物肠道中的正常菌群之一,可以分为致病性和非致病性两大类。在通常情况下,大肠杆菌并不引起疾病,因此人们最初认为大肠杆菌是动物肠道中的正常菌群。直到1885年Escherich首次从婴儿腹泻物中分离出大肠杆菌后,才开始认识到其致病性。致泻性大肠杆菌是引起人畜腹泻的致病性大肠杆菌的通称,主要包括肠致病性大肠杆菌(EPEC),肠出血性大肠杆菌(EH EC),肠侵袭性大肠杆菌(EIEG)和肠产毒性大肠杆菌(ETEC)。而肠毒素型大肠杆菌(En-terotoxigenic E.coli,ETEC)是引起婴幼儿、哺乳动物腹泻的重要病原菌是目前研究的热点之一。ET…     

四种致泻性大肠杆菌多重PCR检测方法的建立及在大熊猫上的应用

致泻性大肠杆菌可引起大熊猫血水样便、休克，严重者可危及生命。为准确鉴定4种致泻性大肠杆菌病原，本研究通过对引物浓度、退火温度、反应条件等进行优化，建立了能同时检测EPEC和EHEC的多重PCR方法以及ETEC和EAEC的多重PCR方法。结果得出：两种多重PCR方法特异性强，仅对特有的毒力基因进行扩增，对其他10种大熊猫源病原菌的扩增结果均为阴性；敏感性较高，细菌基因组DNA检测中EHEC和EPEC的检测下限为2.71×10⁴拷贝/μL,EAEC的检测下限为3.23×10⁴拷贝/μL,ETEC的检测下限为3.23×10³拷贝/μL；菌液检测中EPEC和EHEC的检测下限为1.85×10⁴CFU/mL,ETEC的检测下限为2.9×10³CFU/mL,EAEC的检测下限为2.62×10⁴CFU/mL；不同时间段重复8次，均扩增出对应的目的条带，证明重复性好。利用多重PCR和单一PCR方法分别检测110份样品，结果得出esc V基因阳性率为1.82%(2/110)...     

内蒙古羊屠宰场致泻性大肠杆菌和沙门氏菌污染状况分析

为了解内蒙古地区羊屠宰场各屠宰环节致泻性大肠杆菌和沙门氏菌的携带情况,评估是否存在污染风险,选取不同规模屠宰场,用棉拭子法分别采集不同屠宰环节样品349份,36 h内进行试验;应用PCR方法分离鉴定病原菌,对鉴定为致泻性大肠杆菌的分离株进行16S r DNA测序并构建系统发育树。结果显示:从刚宰杀的羊胴体表面和环境中共分离到致泻性大肠杆菌12株,其中11株携带毒力基因elt,1株携带毒力基因eae和elt;大中型屠宰场(1.21%)的致泻性大肠杆菌阳性率低于小型屠宰场(5.43%),不同规模屠宰场的预冷间均未检出致泻性大肠杆菌和沙门氏菌;从屠宰环境中分离出1株携带inv A基因的沙门氏菌,阳性率为0.29%;经高压冲洗和排酸预冷等屠宰工艺后,在羊胴体表面均未发现致泻性大肠杆菌和沙门氏菌。     

仔猪腹泻致病性大肠杆菌分型鉴定及耐药性分析

为了解贵州省规模化养猪场腹泻仔猪致病性大肠杆菌流行情况及耐药性变化,本研究运用凝集试验、PCR和药敏纸片琼脂扩散法等方法对分离的78株致病性大肠杆菌进行血清型、毒力基因及耐药性分析。结果显示,78株致病性大肠杆菌以O138、O87血清型为主,占定型菌株的60.8%;其中62株致病性大肠杆菌检出毒力基因,检出率为79.5%,可分为8种毒力基因类型,分属肠致病性大肠杆菌(EPEC)、肠产毒性大肠杆菌(ETEC)和肠聚集性大肠杆菌(EAEC),毒力基因eaeA、elt和escV检出率较高,分别为38.5%、28.2%和21.8%;分离到的致病性大肠杆菌对β-内酰胺类药物高度耐药,均为多重耐药株,耐药种类可达8种以上。结果表明,当前贵州省规模化养猪场腹泻仔猪致病性大肠杆菌的毒力基因检出率较高且基因型复杂,耐药性严重。本试验结果可为规模化养猪场防控仔猪腹泻提供基础资料及理论依据。     

大肠杆菌耐热性肠毒素的特性及检测

大肠杆菌为肠道中最常见的一类细菌。依据它的致病性大致可分为:肠致病性大肠杆菌(Enter-opathogenic E.coli,EPEC),侵袭性大肠杆菌(Enteroinvasive E.eoli,EiEC)、产毒素性大肠杆菌(Enterotoxigenic E.coli,ETEC)及大肠埃希氏菌(E.coli)。其中ETEC可产生不耐热性肠毒     

多重PCR方法鉴定5类牛源致泻性大肠埃希菌

为建立鉴定5类牛源致泻性大肠埃希菌(DEC)多重PCR检测方法,对陕西关中奶牛场犊牛粪便中分离的大肠埃希菌进行检测。以EAEC(astA、aggR、pic)、EHEC(stx1、stx2)、EPEC(escV、bfpB)、EIEC(invE)和ETEC(lt、stp、sth)5类DEC中11种毒力基因为目标,uidA基因为阳性对照,建立两体系(体系Ⅰ和Ⅱ)多重PCR检测方法,通过改变引物浓度、退火温度等检测条件进行优化,并对其特异性和灵敏度进行检测。结果显示,体系Ⅰ检测EAEC、EHEC,体系Ⅱ检测EPEC、EIEC、ETEC,除astA引物浓度为0.5μmol/L,其余均为0.3～0.4μmol/L;退火温度以59℃最佳;ETEC的检测下限最低(1.09×10³ CFU/mL),其余依次为EHEC(1.27×10³ CFU/mL)、EIEC(2.43×10³ CFU/mL)、EPEC(5.71×10³ CFU/mL)、EAEC(7.98×10³ CFU/mL);经验证该方法只...     

仔猪腹泻致病性大肠杆菌分型鉴定及耐药性分析

为了解贵州省规模化养猪场腹泻仔猪致病性大肠杆菌流行情况及耐药性变化,本研究运用凝集试验、PCR和药敏纸片琼脂扩散法等方法对分离的78株致病性大肠杆菌进行血清型、毒力基因及耐药性分析。结果显示,78株致病性大肠杆菌以O138、O87血清型为主,占定型菌株的60.8%;其中62株致病性大肠杆菌检出毒力基因,检出率为79.5%,可分为8种毒力基因类型,分属肠致病性大肠杆菌（EPEC）、肠产毒性大肠杆菌（ETEC）和肠聚集性大肠杆菌（EAEC）,毒力基因eaeA、elt和escV检出率较高,分别为38.5%、28.2%和21.8%;分离到的致病性大肠杆菌对β-内酰胺类药物高度耐药,均为多重耐药株,耐药种类可达8种以上。结果表明,当前贵州省规模化养猪场腹泻仔猪致病性大肠杆菌的毒力基因检出率较高且基因型复杂,耐药性严重。本试验结果可为规模化养猪场防控仔猪腹泻提供基础资料及理论依据。     

3种致泻性大肠埃希氏菌多重PCR检测方法的研究

根据毒力因子、致病机理及遗传特性，国际上将致泻性大肠埃希氏菌（E．coli）分为肠致病性E．coli（EPEC）、肠出血性E．coli（EHEC）、肠侵袭性E．coli（EIEC）、肠产毒性E．coli（ETEC）、肠集聚性E．coli（EAggEC）。其中ETEC菌株通过LT和ST致病基因的作用引起腹泻。EPEC的致病性与LEE毒力岛上的eaeA毒力基因有关。EIEC的侵袭性是因为在侵袭性质粒上含有ipaH毒素基因。     

单克隆抗体在大肠杆菌诊断中的应用

大肠杆菌的致病性可分为产肠毒素性大肠杆菌(ETEC),肠道致病性大肠杆菌(EPEC),肠道侵袭性大肠杆菌(EIEC)和肠道出血性大肠杆菌四大类。其中ETEC是引起幼畜腹泻的主要病原体,它的致病性取决于ETEC在宿主小肠上皮细胞上定居的能力和产生肠毒素的能力,两者缺一不可。ETEC定居于小肠上皮细胞能力是因为具有定居因子或称为粘着素,现已发现的动物源性粘着素主要为K_(99),K_(88),F_(41),987P。另外,ETEC还能     

哈尔滨市部分鲜奶中致泻性大肠埃希氏菌的分离与鉴定

为了解哈尔滨市鲜奶中致泻性大肠埃希氏菌(E.coli)的病原学特征,分别在10家奶站和牛场采集鲜奶共550份,通过菌落形态、生化特性、血清型及动物实验等一系列的鉴定,确认为致泻性大肠埃希氏菌的为5株.其中3株为肠致病性E.coli(EPEC),2株为肠侵袭性E.coli(EIEC).     

Heartworm in dogs in Canada in 1984

In late December 1984, 1853 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1984 and 35% of them responded. Veterinarians reported that 97,794 dogs were blood-tested to check for microfilariae and 1417 dogs (1.45% of those tested) were found with heartworm. Another 34 dogs were amicrofilaremic, but were diagnosed as having heartworm disease, to give the total number diagnosed in 1984 as 1451 (1.48%). Heartworm was reported from all provinces except Prince Edward Island and Newfoundland but most (1310) of the cases were in Ontario. In Quebec, 126 cases were reported mostly from west of Montreal.

Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 27% of the cases were observed with clinical signs of heartworm disease and 72% had a history of not having left Canada. Southwestern Ontario continued to be the primary focus of the infection.

     

Heartworm in dogs in Canada in 1983

In late December 1983, 2 800 veterinarians across Canada were sent a questionnaire in order to assess the status of heartworm disease in Canada in 1983 and 26% of them responded. Veterinarians reported that 59 504 dogs were blood-tested to check for microfilariae and 771 dogs (1.30% of those tested) were found with Dirofilaria immitis. Heartworm disease was diagnosed in all provinces except New Brunswick and Newfoundland but most (733) of the cases were in Ontario.

Heartworm disease was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 31% of the cases were observed with clinical signs of heartworm disease and 64% had a history of not having left Canada. Southwestern Ontario continues to be the focus of the infection and most of the dogs there had not left the province previously.

     

Heartworm in dogs in Canada in 1991

In late November 1991, 1883 clinics in Canada were sent a questionnaire to assess the status of Dirofilaria immitis in dogs in 1991 and there was a 60.0% response. There were 344,031 dogs tested for heart-worm (HW), 627 were found infected and the prevalence of HW infection was 0.18%. There were 417 dogs with HW in Ontario, 116 in Manitoba, 38 in Quebec, 53 in British Columbia, three in Alberta, and one in Nova Scotia. In British Columbia, all of the infected dogs but one were from the Okanagan valley which, as from 1991, is a new focus of infection in Canada. Most dogs with HW had not been on preventive medication in 1990, and the prevalence among dogs tested and unprotected was 0.59%. That prevalence was considerably higher in endemic areas. Companion dogs, over three years of age and maintained primarily outdoors in rural areas, were most frequently infected. One cat was diagnosed with D. immitis and 33 dogs had Dipetalonema reconditium.     

Heartworm in dogs in Canada in 1985

In late December 1985, 1485 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1985 and 44% of them responded. Veterinarians reported that 137,300 dogs were blood-tested to check for microfilariae and 1210 dogs were found with heartworm. Another 36 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number diagnosed in 1985 as 1247 (0.91%).

Heartworm was reported from all provinces except Prince Edward Island, Newfoundland and Saskatchewan but most (1126) of the cases were in Ontario. Southwestern Ontario continued to be the primary focus of the infection in Canada. From Quebec, 91 cases were reported mostly from and around Montreal. From Manitoba, 19 cases were reported from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 28% of the cases were observed with clinical signs of heartworm disease and 78% had a history of not having left Canada.

     

Heartworm in dogs in Canada in 1988

In late November 1988, 1581 small and mixed animal clinics and institutional veterinarians across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1988, and 46% of them responded. Veterinarians reported that 181,577 dogs were blood-tested for heartworm disease and 367 dogs were found with D. immitis microfilariae. Another 60 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1988 as 441 (0.24%).     

Heartworm in dogs in Canada in 1989

In late November 1989, 1732 clinics and institutional veterinarians were sent a questionnaire to assess the status of Dirofilaria immitis, and 51.7% responded. Of 247,716 dogs tested, 394 had D. immitis microfilariae and 51 were amicrofilaremic for a total of 445 cases and heartworm prevalence of 0.17%. Most (408) of these dogs had no preventive medication and the prevalence among dogs tested and unprotected was 1.01%. That prevalence was considerably higher in endemic areas. Thirty-seven dogs with heartworm had preventive medication. Heartworm was most frequent in companion dogs over three years of age maintained outdoors in rural areas. About 75% of the cases had never left Canada, 26% had clinical signs and 125 were not treated.

Heartworm was reported from British Columbia, Manitoba, Ontario, Quebec, Nova Scotia and Newfoundland, but 383 cases were in Ontario. South-western Ontario was the primary focus of infection. There were 33 cases in Quebec and 24 in Manitoba, mainly found in and around Metropolitan Montreal and Winnipeg respectively.

     

Heartworm in dogs in Canada in 1987

In late November 1987, 1246 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1987, and 50% of them responded. Veterinarians reported that 165,428 dogs were blood tested for heartworm disease and 511 dogs were found with D. immitis microfilariae. Another 78 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1987 as 589 (0.35%).     

Heartworm in dogs in Canada in 1986

In late December 1986, 1224 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1986; 46% of them responded. Veterinarians reported that 150,989 dogs were blood-tested for microfilariae and 869 dogs were found with heartworm. Another 65 dogs were amicrofilaremic but diagnosed with heartworm disease and one was found with heartworm at necropsy to give the total number diagnosed in 1986 as 935 (0.62%).

Heartworm was reported from Manitoba, New Brunswick, Ontario and Quebec, but most (810) of the cases were from Ontario. South-western Ontario continued to be the primary focus of the infection in Canada. There were 103 cases reported from Quebec, mostly from and around Montreal, and 21 cases from Manitoba, from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 33% of the cases were observed with clinical signs of heartworm disease and 81% had a history of not having left Canada.