猪大肠杆菌融合菌株R2灭活苗的研制及免疫试验

为了研究猪大肠杆菌融合菌株R2蜂胶灭活苗的免疫效果,试验以猪大肠杆菌原生质体融合菌株R2为菌种制备蜂胶灭活苗,采用肌肉注射方式免疫SPF小白鼠,以间接ELISA方法检测血清中IgG抗体水平,同时用2种不同血清型的猪大肠杆菌亲本菌株与免疫血清做玻片凝集试验,并对小白鼠进行攻毒保护试验。结果表明:该疫苗免疫性能较好,在免疫小白鼠后的第28,35天左右机体内的抗体水平达到最高值,融合菌株R2蜂胶灭活苗可对2种不同血清型的猪大肠杆菌亲本菌株的攻毒试验产生保护,而且安全性能良好,是一种效果较好的菌苗。说明融合菌株R2蜂胶灭活苗作为一种新型疫苗防控猪大肠杆菌病将成为一种可能。     

大肠杆菌菌苗的研制

对苗用菌株的灭活时间和菌苗免疫条件进行筛选的条件下,用甘肃省大肠杆菌的代表血清型和筛选的大肠杆菌原生质体融合菌株研制大肠杆菌菌苗,并进行菌苗安全性试验。正交试验试验结果表明:菌苗的最佳组合是抗原含量是4亿/mL和佐剂含量是10 mg/mL。苗用菌株的灭活试验结果表明:鸡苗的灭活条件是1%甲醛作用96 h;猪苗的灭活条件是2%甲醛作用48 h。     

致病性大肠杆菌苗用菌株的原生质体融合的研究

对筛选的5株大肠杆菌(E2:O2、E6:O78、E7:O8、E9:O9、E22:O138)进行药敏试验以及致病性大肠杆菌原生质的制备、融合和筛选试验。结果表明:①5株大肠杆菌对多种抗菌素存在抗药性,并且这种抗药性能进行培育,同时,在原生质的融合过程中能递进;②大肠杆菌原生质体选择的最佳组合是:溶菌酶2.0 g/L,处理时间是20 m in;③大肠杆菌的融合率在10-8左右,大肠杆菌之间的融合率为10-6左右;共选出3株融合菌株为E2-E6、E6-E7和E2-E22,各融合株的生化特征介于起源菌株的生化特征之间,有2株融合株的抗原特征含原菌株各自的抗原成分。     

大肠杆菌菌苗的免疫研究

关于大肠杆菌菌苗的免疫研究,在国外已进行了很多工作,我国也有这方面的报导。但对新生幼驹大肠杆菌病的免疫研究,目前国内报导的还不多,我们在选定致病性大肠杆菌菌株的基础上,研制成大肠杆菌菌苗,对马进行了免疫试验研究工作,现将结果报告如下。实验材料 1.菌株来源:系由长春和牡丹江地区七个单位内死亡的幼驹、     

禽巴氏杆菌、大肠杆菌、沙门氏菌三联油乳灭活苗的研制及试用

以多杀性巴氏杆菌 (C-481 )、致病性大肠杆菌 O85、O119、O18、鸡白痢沙门氏菌等菌株制备的禽巴氏杆菌 大肠杆菌 鸡白痢沙门氏菌的油乳与中药灭活菌苗 ,经实验室免疫试验和中间试验证明 ,该菌苗在免疫后第 1 4天即可产生坚强的免疫力。经攻毒试验证明 ,该菌苗的保护率为 93 % ,免疫期为 9个月 ,免疫效果普遍反应良好。该菌苗在 4℃可保存 1 8个月以上 ,室温阴暗处可保存 1 0个月以上。临床证明该菌苗能有效地控制禽巴氏杆菌病、大肠杆菌病与鸡白痢在鸡群中的流行 ,可进行推广应用。     

兔巴氏杆菌-大肠杆菌油乳与中药灭活苗的研制

以多杀性巴氏杆菌(C-481)、致病性大肠杆菌O85、O119、O18等菌株制备的兔巴氏杆菌-大肠杆菌油乳与中药灭活菌苗，经实验室免疫试验和中间试验证明，该菌苗在免疫后第14天即可产生坚强的免疫力；经攻毒试验证明，该菌苗的保护率为93％，免疫期为9个月，免疫效果良好。该菌苗在4℃以下可保存18个月以上，室温阴暗处可保存10个月以上。临床使用证明，该菌苗能有效的控制巴氏杆菌病与大肠杆菌病在兔群中的流行，可在生产中推广应用。     

兔巴氏杆菌大肠杆菌油乳与中药灭活苗的研制及试用

以多杀性巴氏杆菌(C—481)、致病性大肠杆菌O8s、O119、O18等菌株制备的兔巴氏杆菌 大肠杆菌油乳与中药灭活菌苗。经实验室免疫试验和中间试验证明。该菌苗在免疫后第14天即可产生坚强的免疫力。经攻毒试验证明，该菌苗的保护率为93％。免疫期为9个月，免疫效果普通反应良好。该菌苗在4℃可保存18个月以上。室温阴暗处可保存10个月以上。临床证明该菌苗能有效地控制巴氏杆菌病与大肠杆菌病在兔群中的流行，可推广应用。     

鸡大肠杆菌蜂胶灭活菌苗的制作方法及注意事项

选本地分离的禽致病性大肠菌菌株若干株加标准菌株,用LB培养基培养细菌。甲醛灭活后,制成鸡大肠杆菌蜂胶灭活菌苗。经试验,该蜂胶灭活菌苗安全且免疫效力好。     

大肠杆菌多价灭活苗免疫雏鸡抗体消长规律

随着集约化养鸡业的不断发展,鸡大肠杆菌病已成为影响养鸡业经济效益的主要疫病之一.生产中抗生素的广泛应用,使得许多大肠杆菌成为耐药菌株,使药物防治效果越来越差,再者大剂量使用抗生素造成的动物性产品药物残留问题也引起了人们的广泛关注.尽管致病性大肠杆菌的血清型繁多,但对于特定地区感染鸡群的主要菌株的血清型一般不会很多,这就为采用主要血清型致病性菌株研究制成菌苗进行免疫预防提供了可行的办法.本试验的目的是应用3种不同佐剂的大肠杆菌多价灭活疫苗免疫雏鸡后,在不同时间测定鸡体内抗体的变化规律,并在一定的时间利用与菌苗同血清型的致病性菌株进行攻毒试验,以确定不同疫苗的免疫保护期、抗体变化规律和免疫保护效果,为免疫预防该病提供理论依据.     

鸭疫里默杆菌与大肠杆菌融合株的构建

对鸭疫里默杆菌（YL1）和大肠杆菌（ZYD2）原生质体的制备、再生、融合条件进行优化,成功获得了64株具有双亲菌株耐药性,能够同时凝集兔抗ZYD2血清和兔抗YL1血清,且能够稳定遗传的融合菌株;通过培养特性、菌落特征、融合菌株形态染色特性、生化试验等鉴定,筛选出4株典型融合菌株DL4、DL5、DL8、DL29。攻毒试验表明,ZYD2的半数致死量为1.5×107,YL1的半数致死量为5.0×107,4株典型融合菌株的半数致死量分别为2.8×108、1.7×108、1.0×109、4.4×109,融合菌株毒性比亲本菌株毒性减弱。免疫保护试验和免疫鸭抗体水平测定结果证实,融合菌株具有良好免疫原性。     

Preparation and testing of polyvalent EDTA-Na-extract vaccine from Escherichia coli strains pathogenic to Swine]

Monovalent vaccines were prepared of E. coli strains with pathogenicity to swine by means of a technique described elsewhere. A polyvalent vaccine was obtained by mixing the monovalent vaccines. This polyvalent vaccine was tested by criteria usually applied to vaccine of E. coli strains with pathogenicity to man, and it exhibited the same quality characteristics.     

肉鸡大肠杆菌多价灭活蜂胶苗的研制

从分离鉴定的49株大肠杆菌中筛选出5株致病力强、免疫原性好、具有优势血清型的大肠杆菌作为制苗菌株，经培养灭活后，以蜂胶为佐剂，研制出的肉鸡大肠杆菌多价灭活蜂胶疫苗，使用安全，免疫效果好。用该苗免疫后150d的保护率为100％，无不良反应。该苗4C下保存1年不失效。田间试验结果表明，该苗性能好，免疫后对肉仔鸡、肉种鸡的生产性能无不良影响，鸡群大肠杆菌病的死亡率明显下降。     

犊牛腹泻大肠杆菌疫苗的研制与应用

根据南京地区犊牛大肠杆菌腹泻流行病学调查结果,用分离鉴定的犊牛腹泻菌株O78、O26、O15大肠杆菌作菌种,制备油乳灭活疫苗,接种初生犊牛预防犊牛大肠杆菌腹泻。结果表明制备的大肠杆菌油乳灭活疫苗接种初生犊牛能产生良好的免疫保护。     

产蛋鸡卵黄性腹膜炎病原分离鉴定及防治试验

从25个鸡场疑似卵黄性腹膜炎的病料中分离到了18株大肠杆菌和2株沙门氏菌，经大肠杆菌O血清型鉴定，用优势血清型菌株分别接种改良的血清-葡萄糖-肉汤培养基所研制的多价油乳剂灭活苗，在开产前接种鸡群预防产蛋鸡卵黄性腹膜炎，预防效果良好，而采取多价油乳剂灭活苗预防接种配合活菌制剂益生素的使用，预防效果更佳。使用优势血清型菌株筛选的敏感药物硫酸丁胺卡那霉素、强力霉素同时配合清瘟败毒散和VC对卵黄性腹膜炎鸡群进行治疗，治疗效果良好。     

鹅大肠杆菌的分离鉴定及药敏试验

为了对鹅大肠杆菌的临床治疗提供指导并为制备大肠杆菌自家灭活疫苗奠定基础,采集疑似病例的病料,用麦康凯培养基分离细菌,对分离的细菌进行形态学鉴定、生化鉴定、血清型鉴定、致病性试验及药敏试验。结果显示,所分离到的菌落呈粉红色、光滑湿润、边缘整齐;革兰氏染色为阴性小杆菌,两端钝圆,多单个存在;共鉴定出15株大肠杆菌,其中4株O₃₅型,3株O₇₈型,3株O₂₆型,2株O₇型,1株O₁₀₉型,1株O₁₄₁型,1株O₂型;动物试验表明所鉴定的菌株对小鼠和雏鹅均有较高的致死率;体外抑菌试验结果发现强力霉素、阿莫西林对该场的鹅致病性大肠杆菌具有较强的抑制作用。本研究为有效防制鹅大肠杆菌病提供了理论依据,对于控制规模化鹅场大肠杆菌等细菌性疾病具有重要的应用价值。     

Differential effects of clathrin and actin inhibitors on internalization of Escherichia coli and Salmonella choleraesuis in porcine jejunal Peyer's patches

Peyer's patches constitute both an inductive immune site and an enteropathogen invasion route. Peyer's patch mucosae from porcine jejunum were mounted in Ussing chambers, and either Salmonella choleraesuis vaccine strain SC-54 or non-pathogenic rodent and porcine Escherichia coli strains contacted the Peyer's patch mucosa for 90 min. Internalized bacteria were quantified by a gentamicin resistance assay. Monodansylcadaverine (300 microM, luminal addition), an inhibitor of clathrin-mediated endocytosis, significantly inhibited internalization of both E. coli strains relative to tissues untreated with the inhibitor; internalization of SC-54 was unaffected. The actin-disrupting agent cytochalasin D (10 microM, luminal addition), inhibited internalization of pig-adapted E. coli but not that of rodent-adapted E. coli or SC-54. Internalization of SC-54 and non-pathogenic E. coli in Peyer's patches appears to occur through different cellular routes.     

Expression of Escherichia coli antigens in Salmonella typhimurium as a vaccine to prevent airsacculitis in chickens

Avian pathogenic Escherichia coli strains are associated with a variety of extraintestinal poultry diseases, including airsacculitis, colisepticemia, and cellulitis. A number of E. coli serotypes are associated with these diseases, although the most prevalent serotype is O78. Fimbrial proteins expressed by these strains appear to be important virulence factors, including type 1 fimbriae, P fimbriae, and curli. We have been working to develop an effective vaccine to protect chickens against these diseases. We have previously shown that an attenuated Salmonella typhimurium strain expressing O78 lipopolysaccharide provides protection against challenge with an O78 avian pathogenic E. coli strain. In this work, we have constructed an attenuated S. typhimurium that expresses both the O78 lipopolysaccharide and E. coli-derived type 1 fimbriae. In these studies, chickens were vaccinated at day of hatch and again at 2 wk of age. Birds were challenged at 4 wk of age. We found that the vaccine candidate provided significant protection against airsacculitis as compared to untreated controls or birds vaccinated with an attenuated S. typhimurium that did not express any E. coli antigens. In a separate experiment, challenged vaccinates showed significant weight gain compared to challenged nonvaccinates. We were not able to demonstrate protection against E. coli O1 or O2 serotype challenge, nor against challenge with wild-type S. typhimurium.     

豫北地区肉鸡大肠杆菌分离鉴定及其蜂胶灭活苗的研究

从豫北地区部分肉鸡场送检的病料中分离出了24株大肠杆菌，确定2l株菌分属8个血清型，其中02，074，01，078为主要血清型，分别占分离株的28．57％，23．8l％，19．04％，9．52％。药敏试验结果表明，所分离的24株大肠杆菌对丁胺卡那霉素、头孢曲松、头孢噻呋、氟苯尼考最敏感，可作为防治肉鸡大肠杆菌病的首选药物。从中筛选出3株致病力强、免疫原性好、具有优势血清型的大肠杆菌和标准菌株作为制苗菌株。以蜂胶为佐剂，研制出的肉鸡大肠杆菌多价灭活蜂胶疫苗，使用安全，免疫效果好。用该苗免疫后150d的保护率为100％，无不良反应。该苗4℃下保存1年不失效。田间试验结果表明，该苗性能好。免疫后对肉仔鸡、肉种鸡的生产性能无不良影响，鸡群大肠杆菌病的死亡率明显下降。     

Antibacterial activity of antisera against homologous and heterologous Escherichia coli of porcine origin.

Fourteen enteropathogenic and five nonenterotoxigenic Escherichia coli strains isolated from pigs were used for producing antisera in rabbits and pigs. These antisera were used in an vitro test system for antibacterial activity against homologous and heterologous porcine E. coli strains. Antibacterial titres were determined against the homologous strains and the percent reduction in CFU/ml caused by a 1/200 dilution of the sera against heterologous strains was determined. The results indicated that following immunization the antibacterial activity of serum against homologous and heterologous strains was significantly increased. This activity did not appear to be influenced by O and K antigen relationships among the organisms or by enterotoxigenicity of the vaccine strains. When antiserum produced against a combination of three enteropathogenic E. coli was tested against 20 strains a wider spectrum of heterologous antibacterial activity was obtained than with antiserum produced against any individual strain. The results indicate the existence in E. coli strains of porcine origin of common antigenic determinants not related to the serological formula and that a selected combination of strains can be expected to induce antibacterial acitivity against a wide variety of serological types of porcine enteropathogenic E. coli.     

Course of infection and immune responses in the respiratory tract of IBV infected broilers after superinfection with E. coli

Colibacillosis results from infection with avian pathogenic Escherichia coli bacteria. Healthy broilers are resistant to inhaled E. coli, but previous infection with vaccine or virulent strains of Infectious Bronchitis Virus (IBV) predisposes birds for severe colibacillosis. The aim of this study was to investigate how IBV affects the course of events upon infection with E. coli. Broilers were inoculated with IBV H120 vaccine virus or virulent M41 and challenged 5 days later with E. coli 506. A PBS and E. coli group without previous virus inoculation were included. Sections of trachea, lung and airsacs were stained for CD4, CD8, gammadelta-TCR, alphabeta1-TCR, and for macrophages (KUL-01) and both pathogens. Changes in the mucociliary barrier of trachea, lung and airsacs did not predispose for bacterial superinfection. The disease in the lungs of the E. coli group and both IBV/E. coli groups was similar. Lesions in the airsacs were more pronounced and of longer duration in the IBV/E. coli groups. The immunocytological changes differed substantially between the E. coli group and both IBV/E. coli groups. In trachea, lungs and airsacs the CD4+ and CD8+ populations were significantly larger than in the E. coli and PBS groups. In the lungs and the airsacs the macrophages were more numerous in the IBV/E. coli and the E. coli groups than in the PBS group. The presence of high numbers of T cells and macrophages in IBV infected birds most likely induced an altered immune response, which is responsible for the enhanced clinical signs of colibacillosis.