酶标单抗阻断ELISA检测鸡白痢和鸡伤寒抗体

以辣根过氧化物酶标记的沙门氏菌O9单抗3-47-0与包被的鸡白痢沙门氏菌脂多糖抗原建立了一种抗体阻断EL ISA以检测鸡白痢和鸡伤寒沙门氏菌感染。该方法基于待检血清样品抑制酶标单抗与脂多糖的结合,以检测鸡白痢和鸡伤寒特异性抗体。对72份已知鸡白痢阳性血清(抑制率为(98.5±4.0)%)、54份已知鸡白痢阴性血清(抑制率为(15.3±8.0)%)、42份SPF鸡血清(抑制率为(0.8±7.2)%)检测的结果表明,该方法具有很强的区分能力。在人工感染试验中,从第2周开始,该方法能从全部鸡只中检测到特异性抗体,比全血玻板凝集试验(PAT)早1周时间。对344份种鸡血清的检测结果表明,单抗阻断EL ISA比PAT特异性好、敏感性高。另外,该方法也能检测出与鸡白痢和鸡伤寒沙门氏菌有共同抗原的沙门氏菌感染产生的抗体。     

Characteristics of systemic infection and host responses in chickens experimentally infected with Salmonella enterica serovar Gallinarum biovar Gallinarum

Salmonella enterica serovar Gallinarum biovar Gallinarum (S. Gallinarum) is a host-specific pathogen causing systemic infection in poultry, which leads to significant economic losses due to high mortality. However, little is known about the dynamic process of systemic infection and pathogenic characteristics of S. Gallinarum in chickens. In the present study, we developed an oral infection model that reproduces the pathology of S. Gallinarum and clarified the host immune response of the infected chickens. Chickens at 20 days of age orally inoculated at a dose of 10⁸ colony forming unit (CFU) showed typical clinical signs of fowl typhoid and died between 6 and 10 days post infection. The inoculated S. Gallinarum rapidly disseminated to multple organs and the bacterial counts increased in the liver and spleen at 3 days post infection. Pathological changes associated wirh inflammation in the liver and spleen became apparent at 4 days post infection, and increased expression of interferon (IFN)-γ and interleuikin (IL)-12 in the liver and spleen did not observed until 3 days post infection. These results indicate that S. Gallinarum rapidly spread to entire body through intestine, and the low-level of inflammatory responses in the liver during the early stage of infection may contribute to rapid, systemic dissemination of the bacteria. Our infection model and findings will contribute to the better understanding of the pathogenic mechanism of S. Gallinarum, and provide new insights into the prevention and control of fowl typhoid.     

部分省市鸡白痢和鸡伤寒的单抗阻断ELISA血清学调查

采用单抗阻断ELISA法对我国部分省市不同品种、不同类型的鸡群进行了鸡白痢、鸡伤寒的血清流行病学调查。结果，在2309份鸡血清样品中检出阳性359份，阳性率为15．5％，不同鸡场鸡群感染率悬殊较大，从0到54．5％不等。表明，我国鸡群中鸡白痢沙门菌、鸡伤寒沙门菌的流行呈现多样化。     

Salmonella Gallinarum field isolates and its relationship to vaccine strain SG9R

1. The aim of the present study was to determine if the 9R-strain of the Salmonella Gallinarum live vaccine was responsible for having fowl typhoid outbreaks in chicken flocks from both chicken and turkey breeders as well as to verify the antimicrobial resistance of the isolates from the outbreaks.

2. The triplex polymerase chain reaction, standard antimicrobial test, beta-lactamase genes identification and Ion Torrent PMG whole-genome sequence were used in the field isolates and in the vaccine strain of S. Gallinarum.

3. The 60 tested isolates were not from vaccine origin and manifested high resistance to drugs from macrolide and quinolone groups. Whole-genome sequencing (WGS) and single nucleotide polymorphism analysis on selected isolates for core genes from Salmonella enterica confirmed the wild origin of these isolates and showed two possible sources of S. Gallinarum in the studied outbreaks.

4. S. Gallinarum isolated from fowl typhoid outbreaks in the studied period were not caused by the use of the SG9R live vaccine. The source of strains sequenced was diverse.     

Detection and Molecular Characterization of Salmonella enterica Serovar Eppendorf Circulating in Chicken Farms in Tunisia

Salmonella enterica subsp. enterica serovar Eppendorf, with antigenic formula 1,4,12,[27]:d:1,5, is an infrequent serovar. However, 14% (20 of 142) of the isolates recovered during June–July 2012 in chicken farms in Tunisia belonged to S. Eppendorf. These isolates were analysed for resistance and virulence profiles. None of them were susceptible to all antimicrobials tested, while 70%, 60%, 50%, 50%, 20% and 5% were resistant to sulphonamides (sul1, sul2 and sul3), streptomycin (aadA1‐like), trimethoprim (dfrA1‐like), nalidixic acid (GyrA Asp₈₇→Asn and not identified), gentamicin (not identified) and ampicillin (bla_TEM‐1‐like). About 30% of the isolates showed decreased susceptibility to ciprofloxacin and carried the qnrB gene; 65% of the isolates were multidrug resistant and contained class 1 integrons with sul1 or sul3 in the 3′ conserved segment. The orgA, ssaQ, mgtC, siiD and sopB virulence genes located on SPI1 to SPI5 and the fimbrial bcfC gene were present in all isolates; the sopE1 and sodC1 carried by prophages were variably detected; however, the prophage gipA gene and the spvC gene of serovar‐specific virulence plasmids were absent. Altogether, ten resistance and three virulence profiles were identified. Typing of the isolates with XbaI‐ and BlnI‐PFGE supports a close relationship, although they appear to be evolving under selective pressure probably caused by antimicrobial use in chicken husbandry. As far as we know, this is the first study investigating the molecular bases of antimicrobial drug resistance, the virulence gene content and the PFGE profiles of S. Eppendorf. The epidemiological surveillance of this serovar would be necessary to evaluate its possible impact on human health, particularly in Tunisia and other African countries where it was already reported.     

鸡白痢、鸡伤寒沙门菌分子分型方法的验证

为验证鸡白痢、鸡伤寒沙门菌分子分型方法的准确性,本研究以3株鸡白痢、鸡伤寒沙门菌国际参考菌株和306株国内分离株为研究对象,采用5种已知的分子分型方法开展验证性实验。结果发现,基于特定基因可变区的2种分型方法具有良好的特异性,其检测结果与生化分型结果一致,而3种基于特定基因单核苷酸多态性的分型方法则出现假阳性或假阴性的结果。上述结果表明,鸡白痢、鸡伤寒沙门菌分子分型方法的准确性仍需进一步验证。     

鸡伤寒沙门菌的分离鉴定及药敏试验

为给鸡伤寒沙门菌病的防控提供理论依据与数据参考,采集疑似鸡伤寒病死鸡的肝、脾病料,进行细菌分离.根据细菌形态、培养特性、生化试验等鉴定出13株沙门菌.对分离到的菌株进行21种常用抗菌药物的敏感性试验,结果显示,分离株对阿米卡星、新霉素高度敏感,对卡那霉素、痢特灵中度敏感,而对其他药物则表现为耐药.将筛选出的阿米卡星对发...     

Comparative Efficacy of Various Toxoids Against Salmonellosis in Poultry

An attempt was made to develop a vaccine against salmonellosis in poultry by formalizing the Salmonella toxins (enterotoxin plus cytotoxins) that have been found to be the main virulent products of the organisms. Formalized (FT) and carbonated (CT) toxoids were prepared from partially purified toxins of Salmonella enterica subspecies enterica ser. Weltevreden (BM-1643) and S. enterica ser. Gallinarum (L-19/a). There was no mortality in birds vaccinated with formalized toxoid of serovar Weltevreden plus Freund's complete adjuvant (FCA) following homologous or heterologous (S. enterica ser. Gallinarum and S. enterica ser. Typhimurium) challenges. Protection ranged from 50% to 83.3% in the groups immunized with other preparations of S. enterica ser. Weltevreden, i.e. with FT without FCA or with CT with or without FCA. Formalized toxoid prepared from S. enterica ser. Gallinarum (FTSG) toxins given with FCA afforded 100% protection against homologous challenge, but not against heterologous serovars. In the control group, only 16.7% of the birds survived in a subgroup challenged with S. enterica ser. Gallinarum, and none withstood challenge with S. enterica ser. Weltevreden or S. enterica ser. Typhimurium. No untoward reactions were observed in any of the immunized groups. Thus, the vaccine was considered to be potent and safe.     

头孢噻呋对实验性感染鸡白痢沙门氏菌病的药效学研究

研究了头孢噻呋对实验性感染鸡白痢沙门氏菌的防治效果。结果显示，头孢噻呋体内疗效良好，治疗组与预防组的治愈率、有效率、相对增重率均不同程度高于感染对照组（P〈0.05或P〈0.01）；头孢噻呋的治疗效果高剂量组与中剂量组无显著差异（P〉0．05），高剂量组与低剂量组差异显著（P〈0．05）；头孢噻呋的中剂量组与头孢曲松、氨苄西林组的疗效相比均差异显著（P〈0．05或P〈0．01）。实验结果表明，头孢噻呋按0．1mg／只的剂量肌肉注射，每日一次，可对鸡白痢沙门氏菌病起到有效的防治效果。     

雏鸡白痢和大肠杆菌病的药物治疗试验

采用药敏试验测得的药物硫酸丁胺卡那霉素、氟苯尼考、硫酸安普霉素、盐酸环丙沙星、硫酸新霉素、盐酸多西环素对试验动物进行分组用药。试验鸡700只,每组100只。试验结果表明,应用药物治疗雏鸡白痢和大肠杆菌病的效果与药敏试验结果基本一致。     

Distribution of Salmonella Serovars and Antimicrobial Susceptibility from Poultry and Swine Farms in Central Vietnam

This study was conducted to estimate the prevalence of Salmonella spp. and their antimicrobial susceptibilities on poultry and swine farms, sampled in 2 regions in Central Vietnam. A total of 67 poultry farms and 46 swine farms were sampled in a period of 5 months (from September 2012 to January 2013). Salmonella spp. was prevalent in 46.3% and 71.7% of poultry and swine farms, respectively. Altogether, 99 non‐typhoidal Salmonella were isolated and the most common serovars were Salmonella Weltevreden (19%), followed by Salmonella Typhimurium (12%) and Salmonella 4,[5],12:i:‐ (11%). Overall, 71 of 99 (72%) Salmonella isolates were resistant to at least one of the 14 antimicrobial agents tested. Both in poultry and swine farms, high levels of resistance were observed for ampicillin, chloramphenicol, ciprofloxacin, sulphamethoxazole and tetracycline. The presence of Salmonella isolates from poultry and swine farms which were resistant to different classes of antimicrobials suggests that alternative control measures to antimicrobials should be implemented. Moreover, an effective policy should be promoted to encourage a prudent use of these agents in animal farming in Vietnam.     

制定养殖场免疫程序应考虑的主要因素

免疫接种是预防控制传染病的有效方法,免疫程序合理与否直接决定到免疫效果,因此养殖场要按照因地制宜的原则,根据本地的疫病种类、疫病流行情况、抗体水平、免疫途径、疫苗种类与性质、种畜禽来源、饲养管理水平等综合因素,科学制定适合本场的免疫程序。     

华东某省部分禽场重金属污染检测与分析

对华东某省部分家禽养殖场的水体、饲料及其粪便重金属含量进行了调查。共采集包括水样、饲料及粪样在内的49份样品,其中水样16份,饲料样24份,粪样9份。样品经微波消解处理,用原子吸收法测定了铜(Cu)、锌(Zn)、铅(Pb)、镉(Cd)、铬(Cr)含量,用电感耦合等离子质谱法测定砷(As)、汞(Hg)含量。结果表明:水样中各元素含量均符合地表水环境质量标准中三类水质要求;部分饲料样品中铬和砷含量超过饲料卫生标准要求,超标率分别为33.3%和8.3%,其中铬元素最高含量达622.5mg/kg,砷元素含量最高为5.8mg/kg;粪样中除铜元素超标外,其余元素均在标准范围内。研究提示少数家禽养殖场存在一定的重金属污染,应加强对其饲料中铜、砷、铬的含量控制。     

Molecular Detection of Avian Pathogens in Poultry Red Mite (Dermanyssus gallinae) Collected in Chicken Farms

Poultry red mite (PRM, Dermanyssus gallinae) is a blood-sucking ectoparasite as well as a possible vector of several avian pathogens. In this study, to define the role of PRM in the prevalence of avian infectious agents, we used polymerase chain reaction (PCR) to check for the presence of seven pathogens: Avipox virus (APV), Fowl Adenovirus (FAdV), Marek’s disease virus (MDV), Erysipelothrix rhusiopathiae (ER), Salmonella enterica (SE), Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG). A total of 159 PRM samples collected between 2004 and 2012 from 142 chicken farms in 38 prefectures in Japan were examined. APV DNA was detected in 22 samples (13.8%), 19 of which were wild-type APV. 16S ribosomal RNA (16S rRNA) of MS was detected in 15 samples (9.4%), and the mgc2 gene of MG was detected in 2 samples (1.3%). Eight of 15 MS 16S rRNA sequences differed from the vaccine sequence, indicating they were wild-type strains, while both of the MG mgc2 gene sequences detected were identical to the vaccine sequences. Of these avian pathogen-positive mite samples, three were positive for both wild-types of APV and MS. On the other hand, the DNAs of ER, SE, FAdV and MDV were not detected in any samples. These findings indicated that PRM can harbor the wild-type pathogens and might play a role as a vector in spreading these diseases in farms.     

广西玉林市规模化禽场禽流感病毒检测

为了解广西玉林市2020年规模禽场禽流感病毒感染状况,采用荧光RT-PCR方法,对广西玉林市7个县(市、区)42个规模化禽场采集的1260份禽喉/泄殖腔棉拭子样品进行了通用型禽流感病毒核酸检测(荧光PCR),并对检测为阳性的样本进行H5、H7亚型(双重荧光PCR)和H9亚型(荧光PCR)分型鉴定.结果显示:在42个规模...     

种畜禽场疫病防控与净化现状分析

伴随着我国畜牧业的快速发展,疫病防控难度日益增加,成为制约畜牧业健康发展的重要因素。改变传统疫病防控模式、实施疫病净化是综合提升养殖场生物安全水平和动物抗病能力的有效手段,是推动养殖业持续向好发展的重要基础。本文就种畜禽场疫病净化现状进行分析并提出建议。     

麻鸡白痢沙门氏菌的分离鉴定和中药对其耐药性的消除作用

通过菌落形态、革兰氏染色、生化试验、运动力检查、全血平板凝集试验以及动物致病性试验对死亡麻鸡组织中分离的细菌进行了鉴定,确定为鸡白痢沙门氏菌。通过药敏试验证明,麻鸡白痢沙门氏菌对阿莫西林、氨苄西林、卡那霉素、氟苯尼考、利福平高度敏感,对头孢噻肟、头孢拉啶、头孢曲松纳、庆大霉素、泰乐菌素、硫红霉素、粘杆菌素、氧氟沙星、诺氟沙星、环丙沙星以及恩诺沙星不敏感。麻鸡白痢沙门氏菌分别与双黄连和三黄片两种中药作用后,大多数抗菌药物的抑菌环都扩大0.6~6.3 mm,部分药物的敏感性发生了变化。     

Detection of Salmonella in Chicken Embryos in a Breeding Poultry Field of Chengdu

In a poultry farm of Chengdu city,Sichuan province,isolation,identification and drug sensitivity test of Salmonella were conducted from 156 dead embryos.In this experiment,we used Salmonella chromogenic medium,biochemical identification of enterobacteriaceae,trisaccharide iron experiment,Salmonella polyvalent serum and 16S rRNA PCR to identify suspected strains,PCR identification of isolated Salmonella were conducted with 6 kinds of virulence genes.The results concluded that Salmonella separation rate was 15.4%(24/156),including Salmonella typhi accounted for 58.3%(14/24); Detection rates of fimY,invA and mgtC virulence genes were all 100%;The isolated Salmonella were resistant to most clinical drugs to Salmonella.     

成都某种禽场鸡胚沙门氏菌的检测

对四川省成都市某种禽场的156个死胚进行沙门氏菌的分离鉴定及药物敏感性检测.本试验采用沙门氏菌显色培养基、肠杆菌科生化鉴定管、三糖铁试验、沙门氏菌多价血清和16S rRNA PCR鉴定5种方法对疑似菌株进行鉴定,并用6种毒力岛基因将分离的沙门氏菌进行PCR鉴定.结果显示,沙门氏菌的分离率为15.4%(24/156),其中伤寒沙门氏菌占58.3%(14/24);fimY、invA和mgtC毒力基因的检测率均为100%;本试验分离菌对大部分沙门氏菌临床药物表现出明显的耐药性.