PCR技术检测饲料中沙门氏菌的应用研究

用聚合酶链式反应穴PCR雪技术检测饲料中沙门氏菌,对已知被沙门氏菌污染的动物饲料的培养物均能检出阳性,说明PCR方法对检测饲料中沙门氏菌具有特异性高,且灵敏、快速等特点,适用于快速、准确地检测饲料中沙门氏菌的需要。     

北京郊区散养鸡及鸡蛋沙门氏菌带菌情况调查

目的：和方法采用菌落PCR法和细菌培养法调查北京郊区三个散养鸡场的鸡蛋和鸡粪样沙门氏菌的带菌情况,并对两种方法的检测结果进行比较。结果 PCR检测结果表明北京郊区散养鸡粪便沙门氏菌平均阳性率90.87%,蛋内沙门氏菌平均阳性率40%;细菌培养结果表明散养鸡鸡粪平均沙门氏菌阳性率82.3%,蛋沙门氏菌平均阳性率33.3%。结论与传统细菌培养方法相比,菌落PCR法在数值上可提高散养鸡鸡粪和鸡蛋沙门氏菌检出率。     

鸡肠炎沙门氏菌外膜蛋白F的原核表达及免疫原性分析

根据GenBank中收录的鸡肠炎沙门氏菌OmpF基因序列设计1对引物,以鸡肠炎沙门氏菌辽宁分离株基因组为模板,扩增鸡肠炎沙门氏菌OmpF基因,用IEDB Analysis Resource在线预测分析,该基因可能存在15个线性B细胞抗原表位;同时将该基因克隆至原核表达载体PGEX-4T-1中,构建重组表达质粒PGEX-4T-1-OmpF,转入大肠杆菌BL21中,获得OmpF重组蛋白。Western-blot检测结果表明,该重组蛋白的分子质量约为66 kD(含标签蛋白),能被鸡沙门氏菌阳性血清识别,初步证实该蛋白具有免疫原性,为鸡肠炎沙门氏菌亚单位疫苗的进一步研究和开发奠定了理论基础。     

A retrospective study on salmonella infection in Danish broiler flocks

A retrospective longitudinal study was conducted to identify risk factors associated with Salmonella enterica infection in Danish broiler production. The study was based on information in the antemortem database (AM database) where data were available for all broiler flocks slaughtered over the 2-year period from 1992 to 1993 in Denmark. The AM database contains information collected by the ante-mortem veterinarians, from the slaughterhouses, and from the salmonella examinations carried out at the National Veterinary Laboratory. The epidemiological unit was the individual broiler flock. The salmonella status of the flock was determined by examining the caecal tonsils from 16 3-week-old chickens from each flock. This procedure would detect a salmonella-infected flock, with a probability above 95%, if the prevalence is above 20%. Furthermore, the structure and quality of the collected data have been evaluated.

Fourteen variables were selected for analysis by multivariable logistic regression. An increased risk of salmonella infection in the broiler flocks was associated with the biggest hatcheries and feedmill, with an increasing number of houses on the farm, if the preceding flock was infected, and if the flock was reared in the autumn. Additionally, the main variables of the model were analysed by including a random effect at the house level. This resulted only in minor changes of the parameter estimates.     

进口动物性饲料中沙门氏菌的分离鉴定

通过常规分离培养鉴定技术，对上海口岸2001年1－6月份进口的动物性饲料498份进行了沙门氏菌的分离鉴定。结果共分离到沙门氏菌23株，分离率为4．62％（23／498）。其中，鱼粉164份，阳性6份，阳性率3．66％（6／164）；肉骨粉86份，阳性12份，阳性率为13．95％（12／86）；明虾壳27份，阳性5份，阳性率18．52％（5／27）；乳清粉和饲料添加剂类221份，阳性0份，阳性率0％。最终对21株沙门氏菌进行定型。     

Epidemiological Cut-off Values (ECOFFs) of Salmonella from Chicken Intestinal Tract for Three Animal-specific Antibiotics

In order to establish epidemiological cut-off values (ECOFFs) of Salmonella for three animal-specific antibiotics (florfenicol,apramycin and danofloxacin) in Guangdong province.A total of 166 strains of Salmonella isolates from chicken intestinal tract from veterinary clinics and farms in Guangdong province in 2017 were recruited in this study,the minimum inhibitory concentrations (MIC) of florfenicol,apramycin and danofloxacin were determined in triplicate for each bacterial strain using the agar dilution method on Mueller-Hinton agar plates according to the CLSI reference method (CLSI-M100-S26).ECOFFs were calculated for the MIC data sets by application of nonlinear regression analysis,NRI(normalized resistance interpretation) and ECOFFinder software.MIC distribution of florfenicol,apramycin and danofloxacin against Salmonella from 2 to >512,4 to >512 and 0.015 to 64 μg/mL,respectively.MIC₅₀ and MIC₉₀ for apramycin,florfenicol and danofloxacin were 256 and >512,16 and >512,0.5 and 16 μg/mL,respectively.MIC distribution of apramycin and florfenicol presented obvious unimodal shape,while that of danofloxacin was a discontinuous multi-peak.The ECOFFs of florfenicol,apramycin and danofloxacin were recommended to be 16,16 and 0.125 μg/mL,respectively.In present study,the ECOFFs of Salmonella from chicken intestinal tract for apramycin,florfenicol and danofloxacin were determined,which provided scientific basis for resistance surveillance.     

Cytokine response of porcine cell lines to Salmonella enterica serovar typhimurium and its hilA and ssrA mutants

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a facultative intracellular bacterium which can infect and colonize pigs. After contact with enterocytes and macrophages, S. Typhimurium induces production of cytokines thus triggering the innate immune response. In this study we evaluated the cytokine response of two porcine cell lines, IPI-2I and 3D4/31, of epithelial or macrophage origins, respectively, to the wild-type S. Typhimurium and its hilA and ssrA mutants. We observed that the 3D4/31 cell line essentially did not respond to S. Typhimurium infection when a medium with foetal calf serum was used. However when the 3D4 cell line was incubated overnight in the presence of porcine serum, it efficiently responded to the wild-type strain and the ssrA mutant but not to the noninvasive hilA mutant as measured by mRNA quantification of TNF-alpha, IL-8 and GM-CSF by the real-time RT-PCR. In IPI-2I, all the cytokines were also induced by the wild-type S. Typhimurium and the ssrA mutant although the induction of TNF-alpha was lower than that induced by the wild-type strain. The hilA mutant was unable to induce any of the cytokines tested. The ssrA mutant can therefore be considered as more suitable for further vaccine development as the stimulation of innate immune response is important for animal protection against a challenge with virulent strains.     

禽源沙门菌对四环素的耐药性及耐药基因调查分析

为了更好地了解禽源沙门菌对四环素的耐药性及耐药基因分布,从不同来源的家禽样品中分离沙门菌,调查其对四环素的耐药性以及耐药菌株中8种四环素耐药基因[tet(A)、tet(B)、tet(C)、tet(W)、tet(M)、tet(D)、tet(K)和tet(L)]的携带情况.结果表明,18.8%的沙门菌分离株对四环素耐药,健康成鸡分离株对四环素的耐药性明显高于雏鸡、死胚或病禽分离株;四环素耐药株中tet(A)、tet(B)和tet(M)基因的携带比例分别为73.1%、11.5%和3.8%,说明沙门菌对四环素的耐药机制以tet(A)和tet(B)基因介导的主动外排为主.本研究首次在对四环素耐药的沙门菌中检测到tet(M)基因,说明tet(M)基因在沙门菌对四环素的耐药性方面也具有潜在的作用.     

表达猪链球菌溶血素基因的减毒沙门氏菌的构建及鉴定

将猪链球菌溶血素（suilysin，SLY）基因克隆入原核表达栽体pBV220，将重组质粒再导入减毒鼠伤寒沙门氏菌SV4089株，经PCR和酶切鉴定，构建成携带猪链球菌溶血素基因的重组减毒鼠伤寒沙门氏菌。结果表明：该减毒株具有相对安全性；用酶切和PCR鉴定法证实在无抗生素存在的条件下携带重组质粒的减毒株比较稳定；SDS-PAGE显示SLY能在宿主菌中进行表达。该结果为进一步研究制备猪链球菌口服活疫苗奠定了基础。     

雏鸡沙门氏菌病的病原分离与鉴定

邯郸地区某县养殖场于2008年2月引进一批海兰白雏鸡共6500只,4日龄时部分雏鸡开始发病,6日龄时开始死亡,7d内共死亡1248只,死亡率达19.2%。剖检不同日龄死雏21只,活病雏13只,死雏呈现严重败血症病变,活雏心肌、肝、肺、肌胃等脏器出现黄白色或大小不等的灰白色坏死灶。取其卵黄、肝、脾、心血等病料进行病原分离培养,从获得的153个菌落中挑选可疑菌落27个,经细菌学鉴定,确认17株为鸡白痢沙门氏菌,抗原式为1,9,12∶-∶-,10株为德尔卑沙门氏菌(S.derby),抗原式为1,4,12∶f,g∶-。