辽西地区鸡大肠杆菌分离鉴定、药敏及IHA诊断液制备

本试验从辽西地区部分养鸡场采集具有典型临诊特征的鸡大肠杆菌病的病、死鸡病料,进行细菌分离培养和鉴定,分离到89株鸡大肠杆菌。通过血清型鉴定,鉴定出血清型的82株,血清型9种,分别为O₁、O₂、O₈、O₁₅、O₁₈、O₃₅、O₇₈、O₁₁₄、O₁₄₁,其中O₁₈、O₃₅型鸡大肠杆菌为辽西地区优势流行血清型。将分离纯化的O₃₅型鸡大肠杆菌超声裂解后,加入致敏双醛化处理过的红细胞,制备O₃₅型鸡大肠杆菌间接血凝诊断液,验证其特异性、敏感性和稳定性,完成诊断液的标准化。同时采用Kirby-Bauer纸片法,选择12种标准药敏片对此89株分离菌株进行药敏试验。结果表明,间接血凝诊断液特异性和敏感性好,质量稳定。而药敏试验结果显示,各地区均有耐药菌株的出现,仅对氟苯尼考、头孢噻呋、左旋氧氟沙星、盐酸沙拉沙星这4种药物敏感性较高。     

鹅大肠杆菌油佐剂灭活疫苗的研制

从湛江地区疑似大肠杆菌病的病死鹅组织中分离鉴定2株致病性大肠杆菌O₈₆K₆₁和O₄₄K₇₄,以此制备油佐剂灭活疫苗。用上述疫苗接种20日龄雏鹅,35日龄加强免疫一次,分别于27、35、42、50日龄检测免疫鹅血清抗体水平,并进行攻毒保护试验。结果表明,制备的大肠杆菌油佐剂灭活疫苗安全可靠,免疫后15、30 d后,免疫鹅血清抗体水平分别达到1∶32和1∶128;且免疫鹅能抵抗10¹⁰/mL的致病性大肠杆菌攻击,对照组鹅全部发病,有明显临床症状,从发病鹅组织中分离到致病性大肠杆菌。     

广西中小规模猪场大肠杆菌病流行情况调查

从广西23家中小规模猪场采集病料,分离、鉴定出致病性大肠杆菌56株,并对其进行血清型鉴定和耐药性检测,结果鉴定出43株致病性大肠杆菌,分属于16种血清型,其中以O₁₃₈、O₁₈、O₈₅、O₉和O₂₁为优势血清型,占定型菌株的69.76%;用20种常用抗菌药进行药物敏感性试验,发现高敏的药物有头孢拉啶、头孢唑啉、头孢曲松、头孢西丁、多黏菌素B、新霉素、壮观霉素和阿米卡星;而不敏感的药物有青霉素、阿莫西林、强力霉素、卡那霉素、万古霉素、林可霉素。说明这些猪场大肠杆菌的耐药性十分严重,应加强生物安全措施等进行综合防控。     

肉鹅大肠杆菌病的病原分离与鉴定

本试验旨在研究肉鹅大肠杆菌的生化特性、药敏情况、血清型和致病性。2010年12月江苏省某肉鹅场鹅群出现呼吸困难,死亡增多现象,对病死鹅剖检发现以气囊炎和肺部坏死为主,无菌采集病料分离到一株细菌,经分离培养、染色镜检、生化特性鉴定,确定为大肠杆菌,微量凝集试验表明其血清型为O₂。经过肺结节压片镜检,没有发现霉菌的菌丝;检测健康鹅和患病鹅各10只的鹅新城疫抗体,发现其抗体滴度比较高且整齐度较好,两者之间的抗体无明显差异。对分离菌株进行药敏试验,结果表明对氟苯尼考等7种药物敏感,对先锋Ⅴ等16种药物已产生了耐药性。将分离到的菌株接种21日龄雏鹅进行动物回归试验,引起80%死亡率,说明该分离菌株对雏鹅具有较强的致病性。     

一株兔溶血性大肠杆菌的分离鉴定

从发病家兔脏器中分离到一株细菌,进行了初步鉴定。结果表明,该菌为致病性大肠杆菌,在血平板上呈β溶血,对小鼠和家兔毒力强,能致其发病死亡,血清型为O₁₁₉,对多数抗生素具有耐药性,只对庆大霉素和丁胺卡那霉素敏感。     

盐城地区鹅大肠杆菌的致病性试验

从盐城地区临诊上有典型鹅大肠杆菌病变的病、死鹅和鹅炕坊的死胚蛋中分离培养出256株大肠杆菌。为了解这些鹅大肠杆菌的致病性情况,用不同地区、不同血清型的51株分离株做致病性试验。所测分离株对1日龄鹅均有致病性。3个优势血清型分离株中,对1日龄鹅高度致病者占77.8%,中度致病者占16.7%。低度致病者占5.5%。试验表明我们所获得的大肠杆菌分离株均为致病性。     

盐城地区鹅大肠菌的致病性试验

从盐城地区临诊上有典型鹅大肠杆菌病变的病、死鹅和鹅炕坊的死胚蛋中分离培养出256株大肠杆菌。为了解这些鹅大肠杆菌的致病性情况,用不同地区、不同血清型的51株分离株做致病性试验。所测分离株对1日龄鹅均有致病性。3个优势血清型分离株中,对1日龄鹅高度致病者占77.8%,中度致病者占16.7%。低度致病者占5.5%。试验表明我们所获得的大肠杆菌分离株均为致病性。     

大肠杆菌与肠道沙门菌混合感染朗德鹅的诊断分析

为确诊某朗德鹅养殖场大批雏鹅出现腹泻、急性死亡的病因,采集患病雏鹅心脏、肝脏、肾脏等病料接种胰蛋白胨大豆琼脂(TSA)、麦康凯和伊红美蓝等细菌培养基,进行细菌分离培养、革兰染色、16S rRNA测序和系统发育分析,并对分离菌进行小鼠致病性试验和抗菌药物药敏分析。结果显示:鉴定得到1株肠道沙门菌(Salmonella enterica)和2株大肠杆菌(Escherichia coli);3株分离菌对小鼠均具有较强的致病力;药物敏感性试验显示分离株对呋喃唑酮、复方新诺明、多黏菌素B敏感,对诺氟沙星、氨苄西林、环丙沙星等抗菌药物具有多重耐药性。诊断分析结果为朗德鹅养殖场雏鹅大肠杆菌病和沙门菌病诊断、治疗和预防提供了依据。     

盐城地区鹅源大肠杆菌的致病性试验

从盐城地区临诊上有典型鹅大肠杆菌病变的病、死鹅和鹅炕坊的死胚蛋中分离培养出256株大肠杆菌。为了从盐城地区临诊上有典型鹅大肠杆菌病变的病、死鹅和鹅炕坊的死胚蛋中分离培养出256株大肠杆菌。为了解这些鹅源大肠杆菌的致病性情况,用不同地区、不同血清型的51株分离株做致病性试验。所测分离株对1日龄鹅均有致病性。3个优势血清型分离株中,对1日龄鹅高度致病者占77.8%,中度致病者16.7%,低度致病者占5.5%。     

鹅源鸭疫里默氏杆菌的分离和鉴定

为研究鹅源鸭疫里默氏杆菌的生化特性、药敏情况、血清型和致病性,从扬州郊区发病鹅场分离到1株细菌,经分离培养、染色镜检、生化特性鉴定、玻片凝集试验,确定为Ⅱ型鸭疫里默氏杆菌。对病料进行病毒分离试验,经过血凝试验和琼脂扩散试验,没有发现鹅新城疫病毒、禽流感病毒和小鹅瘟病毒。动物致病性试验结果表明,鸭疫里默氏杆菌分离菌株可以通过静脉注射、皮下注射和滴鼻3种途径感染鹅,出现100%的死亡率,说明分离株对扬州鹅具有很强的致病性,同时提示在鹅的免疫计划中也应该充分考虑到鸭疫里默氏杆菌的致病和传播。     

鹅源致病性大肠杆菌O86K61 和O44K74亚型的分离与鉴定

从湛江养鹅场发生的疑似大肠杆菌病病死鹅无菌采集病料,分离鉴定出8株大肠杆菌,其中,O86K61 4株,O44K74 3株,1株未定型,上述菌株均不产生内毒素;药敏试验结果表明,大部分菌株对阿米卡星高度敏感,对恩诺沙星、先锋霉素、磺胺嘧啶、诺氟沙星、强力霉素呈交叉耐药;对万古霉素、新霉素、呋喃唑酮中度敏感;对阿莫西林、链霉素、卡那霉素、庆大霉素、四环素不敏感。上述结果提示,同一发病鹅场存在着多种致病性大肠杆菌的血清型,并存在多重耐药。     

内蒙古中东部地区不同来源大肠杆菌耐药性调查

本研究旨在调查内蒙古赤峰、通辽地区不同来源的大肠杆菌对抗生素类药物的耐药情况。采用K-B法分析来源于鹅、牛和猪等患病动物分离出的46株大肠杆菌对13种常用的抗生素类药物的耐药性;鹅源和猪源大肠杆菌对盐酸多西环素的耐药率最高,分别达95.2%和93.3%;牛源大肠杆菌对盐酸沙拉沙星、恩诺沙星和盐酸多西环素的耐药率最高,达80.0%。多重耐药性结果表明,患病动物的大肠杆菌以6耐（23.9%）为主;牛源大肠杆菌则以7耐（40%）最多。总体耐药谱型以恩诺沙星/氟苯尼考/盐酸多西环素/阿莫西林/硫酸新霉素为主（24.4%）。不同来源的大肠杆菌对抗生素类药物均产生了严重的耐药性,因此不但要求养殖人员学会合理使用抗生素类药物,同时研究开发逆转耐药性的新药物或对抗生素类药物有增敏效用的新药物十分必要。     

Investigation on Antibiotics Resistance of Escherichia coli Isolated fromDifferent Sources in the Middle East Region of Inner Mongolia

The study was aimed to research the antibiotics resistance of Escherichia coli (E.coli) isolated from different sources of Chifeng and Tongliao areas in Inner Mongolia.Using K-B method, we analysed the resistance situation of 46 strains E.coli isolated from goose, bovine and porcine to thirteen kinds of antibiotic drugs. The resistance rates of E.coli isolated from goose and porcine had the highest resistance to doxycycline hydrochloride drug, which were 95.2% and 93.3%,respectively. The resistant rate of E.coli isolated from bovine had the highest resistance to sarafloxacinhydrochloride,enrofloxacin and hydrochloric acid doxycycline, which was 80.0%. Multi-resistance results showed that the prevalence of E.coli in diseased animals with 6 resistance mainly(23.9%);Bovine source with 7 resistance (40%). The resistance type in general was enrofloxacin/florfenicol/doxycycline/amoxicillin/neomycin sulfate. E.coli from different sources of diseased animals to antibiotics had a serious resistance. So,it is not only necessary for farmers to learn to use antibiotics rationally, but also to develop new drugs that are resistant to drug resistance or new drug allergic to antibiotics.     

种鹅大肠杆菌分离鉴定及耐药性分析

为了解种鹅致病性大肠杆菌病的病原及其药物敏感性,2012年从广东地区患病种鹅器官和粪便中分离鉴定了62株大肠杆菌,采用琼脂稀释法测定了大肠杆菌对17种抗菌药物的敏感性。结果发现大肠杆菌耐药性严重,对大部分药物的耐药率超过60.0%,仅对头孢西丁、头孢他啶和黏菌素较敏感。联合药敏试验结果显示多西环素和氟苯尼考联用对耐药大肠杆菌产生协同抑菌作用,FIC指数为0.375～0.75。采用脉冲场凝胶电泳（PFGE）法对24株大肠杆菌进行分型,结果分为17种不同的PFGE型别。大肠杆菌系统进化分群结果显示43.5%属于系统进化组群的D组,其次是A组和B1组,分别为34.8%和21.7%,B2组未检测到。通过对种鹅大肠杆菌耐药性、PFGE分型及其系统发育群进行综合分析,为临床用药提供了参考依据。     

吉林白鹅大肠杆菌的分离鉴定及药敏试验

为了指导临床合理使用药物 ,有效防治鹅大肠杆菌病 ,采取病鹅的肝、心血作细菌的分离培养 ,经革兰氏染色、镜检和多种糖发酵试验以及VP、MR试验 ,鉴定该细菌为大肠杆菌 ;采用试管液体二倍稀释法进行药敏试验 ,测定了 10种药物对鹅大肠杆菌分离株的最小抑菌浓度 ,从中筛选出最有效的抗大肠杆菌药物。     

产蛋鹅大肠杆菌的分离鉴定和免疫效果观察

通过对某养鹅专业户病死产蛋鹅的病原分离,革兰氏染色镜检,生化试验,药敏实验。结果表明:分离菌为大肠杆菌,对头孢噻肟、阿米卡星、壮观霉素等高度敏感,对青霉素、四环素等产生耐药。分离菌制成蜂胶灭活苗免疫该专业户产蛋鹅后,发病率和死亡率明显下降,具有较好的免疫效果。     

Serum resistance and virulence of Escherichia coli isolated from turkeys

Twenty-five strains of Escherichia coli isolated from turkeys were characterized for their serum resistance and virulence. An in vitro bactericidal assay was used to determine the serum resistance of E coli. Virulence was determined by survival time after IV inoculation of each strain into 3-week-old turkeys. Serum-resistant E coli strains were generally found to be virulent for turkeys, whereas serum-sensitive E coli strains were avirulent. Of the 25 strains, 18 strains were placed in the 2 categories of serum-resistant/virulent and serum-sensitive/avirulent. Five strains were serum-resistant and avirulent, and 2 strains were serum-sensitive and virulent. Serum resistance appears to be an important determinant of virulence for E coli in turkeys; however, the requirement for other virulence factors, in addition to serum resistance, was suggested by the finding that 5 serum-resistant strains were avirulent in turkeys.     

Adhesive properties and iron uptake ability in Escherichia coli lethal and nonlethal for chicks

Escherichia coli that are virulent for poultry usually result in a respiratory disease, which is frequently followed by a general infection. Adhesiveness of E. coli to epithelial cells and iron-uptake ability of E. coli could be involved in different steps of the disease. These properties were studied in 59 E. coli strains originating from poultry, with reference to lethality for day-old chicks. Adhesive properties were found in 64% of the lethal strains and in only 23% of the nonlethal strains. The ability to grow in limited iron conditions was strongly correlated with lethality. Fifty-two percent of the lethal E. coli strains, but none of the nonlethal strains, possessed both adhesive and iron-uptake abilities. It is suggested that these two properties play a role in the virulence of E. coli for poultry.     

Occurrence and characterization of verocytotoxigenic Escherichia coli (VTEC) strains from dairy farms in Trinidad

A cross-sectional study was conducted to determine the prevalence and characteristics of verocytotoxigenic Escherichia coli (VTEC) on 25 dairy farms each located in Waller field and Carlsen field farming areas in Trinidad. On each selected farm, faecal samples were collected from milking cows, calves and humans; rectal swabs were obtained from pet farm dogs; bulk milk was sampled as well as effluent from the milking parlour. Escherichia coli was isolated from all sources on selective media using standard methods. Isolates of E. coli were subjected to slide agglutination test using E. coli O157 antiserum, vero cell cytotoxicity assay to detect verocytotoxin (VT) and heat labile toxin (LT) production, the polymerase chain reaction (PCR) to detect VT genes, and the dry spot test to screen for E. coli O157 and non-O157 strains. In addition, faecal samples from animal and human sources were tested for VT genes using PCR. Of a total of 933 E. coli isolates tested by the slide test, eight (0.9%) were positive for the O157 strain. The vero cell cytotoxicity assay detected VT-producing strains of E. coli in 16.6%, 14.6%, 3.2% and 7.1% of isolates from cows, calves, farm dogs and humans respectively (P < 0.05; chi(2)). For LT production, the highest frequency was detected amongst isolates of E. coli from calves (10.8%) and the lowest (0.0%) amongst isolates from humans and bulk milk (P < 0.05; chi(2)). Of the 61 VT-producing isolates by vero cell cytotoxicity assay tested by PCR, the VT, LT and eae genes were detected in 62.3%, 4.9% and 1.6% respectively (P < 0.05; chi(2)). Amongst the 45 E. coli isolates that were VT positive (vero cell) or VT-gene positive by PCR, 2.2%, 2.2%, 4.4% and 6.7% belonged to non-O157 strains O91, O111, O103 and O157, respectively, as determined by the Dry spot test. Detection of VTEC strains in milk and dairy animals poses a health risk to consumers of milk originating from these farms. In addition, the demonstration of VTEC strains in humans, VT gene in faecal samples and E. coli isolates as well as non-O157 VTEC strains of E. coli are being documented for the first time in the country.     

60株大肠杆菌的分离与致病性鉴定

大肠杆菌(Escherichia coli,E.coli)是鸡肠道常在菌,部分菌株具有致病性。鸡大肠杆菌病就是由大肠杆菌中的某些致病性菌株引起的鸡感染性疾病。致病性大肠杆菌均携带毒力岛基因ChuA。为鉴定分离自临床病例的鸡大肠杆菌的致病性,本试验采用PCR法检测ChuA基因进行分子生物学鉴定,60株大肠杆菌中有31株属于致病性大肠杆菌,总阳性率为51.67%。