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。     

鸡大肠杆菌多价蜂胶灭活苗的免疫效果试验

选择鸡大肠杆菌现地菌株制成蜂胶灭活苗，用该苗免疫肉仔鸡２９组共３１５００只，结果表明，该苗安全，免疫后无不良反应，免疫后１４天，３０天，４０天监测，抗体凝集价达１：３２并逐渐上升到１：２５６。其中有２８组共３０５００只鸡未发生大肠杆菌病；只有１组（１０００只鸡）有４％发生了大肠菌病，且由于漏免所致，采取２～３个免疫量紧急接种，３天后鸡群停止死亡，免疫的鸡生产性能不受影响，而且增重快，生长发育良好，     

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。     

辽宁省部分地区禽大肠杆菌血清型分布及禽大肠杆菌病防治措施初探

2001年7月至9月辽宁省动物防疫站从全省9个市病死禽中分离28株大肠杆菌，定型27株。其中10株O78型，占定型菌株的37％；4株O109型占定型菌株的14．8％：O45、O33和O4、18、142型各2株，分别占定型菌株的7．4％；O157、O1、O91、O32、P80、O26型各1株分别占定型菌株的3．7％。由此可见O78型为绝对优势菌株。辽宁省动物防疫站与山东宾州华宏生物制品有限责任公司合作，用辽宁省分离的大肠杆菌制成了大肠杆菌蜂胶灭活苗及大肠杆菌与新城疫二联蜂胶灭活苗。自2002年2月该疫苗应用于辽宁省禽大肠杆菌病预防，在肉仔鸡、商品蛋鸡、肉种鸡、蛋种鸡等各种禽类中应用，均收到理想效果。     

鸡大肠杆菌多价蜂胶灭活苗的研制

从分离鉴定的48株致病性大肠杆菌中,筛选出4株流行广泛、致病力强、抗原性好,且为不同血清型的大肠杆菌作为制苗菌株;以蜂胶这一新型佐剂作为免疫佐剂,研制出的鸡大肠杆菌多价蜂胶灭活菌苗,使用安全,免疫效果好,近期保护率为100％,免疫期为6个月,疫苗制作简单,用苗后鸡群无不良反应。田间应用后鸡群发病率、死亡率下降,增重快,产蛋率增加,饲料利用率提高。     

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。     

奶牛隐性乳房炎大肠杆菌蜂胶灭活苗的制备及其免疫效果研究

为了制备奶牛隐性乳房炎大肠杆菌蜂胶灭活疫苗,试验将菌液用甲醛灭活,以蜂胶为佐剂,灭活菌液与蜂胶佐剂按1∶1等体积混匀制成蜂胶佐剂灭活苗,并对灭活苗的物理性质、安全性及其免疫效果进行检测。结果表明:通过建立动物模型,小鼠接种0.25 mL灭活苗后没有出现不良反应,灭活苗具有较好的安全性且免疫效果好;通过攻毒试验,给小鼠注射奶牛隐性乳房炎大肠杆菌的生理盐水重悬液0.25 mL,灭活苗的保护率达87.5%。说明制备的大肠杆菌蜂胶灭活疫苗免疫效果良好。     

兔大肠杆菌病二价蜂胶灭活疫苗的研究

从分离鉴定的15株致病性大肠杆菌中，筛选出2株流行广泛，致病力强，免疫原性好，具有血清型（0103、085）代表性的大肠杆菌作为制苗菌株，以蜂胶为佐剂，研制出的兔大肠杆菌病二价蜂胶灭活疫苗，使用安全，免疫效果好，保护率达87％以下。该苗在4℃保存其为8个月以上。在黑龙江、辽宁两省免疫2万只家兔，1～2年的观察表明，用苗后兔群未出现不良反应，且用苗后仔兔成活率由50％左右提高到85％以上，疫苗使用不影响敏殖母兔的妊娠过程。     

不同剂型禽大肠杆菌疫苗免疫保护试验初探

测定了禽大肠杆菌Ｏ７８～１６６株全菌灭活Ａｌ（ＯＨ）３佐剂苗、超声波裂解Ａｌ（ＯＨ）３佐剂苗和全菌灭活油佐剂苗免疫对人工感染同一苗株的保护力。结果表明：全菌灭活Ａｌ（ＯＨ）３佐剂苗保护力最好，全菌灭活油佐剂苗免疫后的间接血凝抗体滴度最高。     

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。     

陕西省鸡胚源性大肠杆菌的分离鉴定

从陕西关中六个种鸡场的１９８只死胚及出壳弱雏中分离出９８株大肠杆菌,平均分离率为４９．５％。通过小鸡致病力试验,筛选出６３株具有致病力的菌株,占分离菌的６４．３％。生化鉴定表明,这些致病菌均符合大肠埃希氏菌的生化特性。血清型鉴定可确定６种,其中以Ｏ１、Ｏ２、Ｏ７８为主,分别占鉴定菌的４４．４％（２８／６３）、３３．３％（２１／６３）和９．５％（６／６３）,另三种为Ｏ２２、Ｏ７５、Ｏ８９共占８．０％（５／６３）。有３株未定型,占４．８％（３／６３）。用１６种抗生素进行药敏试验,对先锋霉素Ｖ和恩诺沙星高度敏感,对羧苄青霉素、氨苄青霉素、氯霉素、卡那霉素、磺胺嘧啶、新霉素、链霉素中度敏感。     

Protection of chickens against a lethal challenge of Escherichia coli by a vaccine containing CpG oligodeoxynucleotides as an adjuvant

Synthetic oligodeoxynucleotides (ODN) containing cytosine-phosphodiester-guanine (CpG) motifs (CpG-ODN) have been shown to be effective immunoprotective agents and vaccine adjuvants in a variety of bacterial, viral, and protozoan diseases in different animal species. The objective of this study was to compare the immune response of chickens to a killed Escherichia coli vaccine combined with oil in water emulsion or with CpG-ODN. Birds were vaccinated with killed E. coli antigens with either 10 or 50 microg of CpG-ODN on days 10 and 20 of age. At day 30, a virulent isolate of homologous E. coli was applied on a scratch site on the caudal abdominal region. Birds were examined for 10 days post-E. coli challenge, and pathologic and bacteriologic assessments were conducted on all birds that were either found dead or euthanized. The E. coli vaccine group that received no CpG-ODN had a survival rate of 65%. In contrast, groups that received the vaccine with CpG-ODN adjuvant had significantly higher survival rate of 92% (P < 0.01) with isolation of low numbers of E. coli from internal organs. Total IgG against E. coli antigens was highest in groups that received CpG-ODN as an adjuvant. Birds that received vaccine containing CpG-ODN had minimal inflammatory reaction without tissue necrosis at the injection site. Severe tissue necrosis was present in birds that received vaccine containing oil in water emulsion adjuvant. This study demonstrated that CpG-ODN is an effective vaccine adjuvant in chickens and results in minimal tissue destruction. This study is the first study in which CpG-ODN has been demonstrated to produce an adaptive immune response, at a significant level, against an extracellular bacterial infection in chickens.     

几种病毒与禽病原性大肠杆菌的人工联合感染

以2种剂量的低致病性禽流感病毒(lowly pathogenic avian influenza virus,LPAIV)，传染性支气管炎病毒（infectious bronchitis virus,IBV)疫苗株H120和H52，新城疫病毒（Newcastle disese viurs,NDV)Lasota株分别于气管内注射10日龄易感鸡，2d后，气管注射禽病原性大肠杆菌O37株（O78），连续观察5d，结果，除LPAIV单独感染组有6．25％的死亡率外，其余各病毒单独接种组均健活；大肠杆菌O37株单独接种组的死亡率为62．50％，较高剂量的LPAIV，IBV H120和H52，NDV Lasota株与大肠杆菌O37株有效强的协同致病作用，死亡率分别达到81.25%,100.00%,93.75%和87.50%，而较低剂量的上述病毒则无明显的协同作用，IBV，NDV疫苗株与大肠杆菌联合接种组的多数死亡鸡病程推迟。     

原场鸡大肠埃希氏菌多价灭活苗的制备及应用效果观察

用从豫北地区一家大型个体养鸡场分离到的致鸡大肠埃希氏菌病8个血清型菌株O1、O2、O78、O36、O11、O88和O50以及未知菌株的培养物,经0.4%的福尔马林灭活48h,制成原场大肠埃希氏菌多价灭活苗,含菌量为150亿/ml。分别免疫批10日龄鸡只,剂量0.2ml/只,使鸡只获得对原场大肠埃希氏菌株比较高的免疫力。该灭活苗的安全性和免疫原性良好,对同型菌株攻毒后对鸡只的保护率达90%以上,对鸡大肠埃希氏菌病有较强的预防作用。     

氟苯尼考长效制剂对鸡大肠杆菌病的疗效试验

用试管二倍稀释法测定了氟苯尼考与丁胺卡那霉素对大肠杆菌质控菌株ATCC25922,菌株K88、HD1047和临床分离的36株大肠杆菌的体外抗菌活性,进行了鸡大肠杆菌病的临床疗效试验.结果质控菌株和K88、HD1047株对氟苯尼考均敏感,36株临床分离菌株对氟苯尼考的敏感率为96.2%.在临床疗效试验中,氟苯尼考长效口服液和注射剂组的死亡率低于感染对照组和丁胺卡那霉素对照组(P《0.01),治愈率及有效率高于感染对照组和丁胺卡那霉素对照组(P《0.01),表明氟苯尼考长效制剂对鸡大肠杆菌病的治疗有很好的疗效并有较好的应用前景.     

Clinical encephalitis following avian encephalomyelitis vaccination in broiler breeder pullets

Twelve-to-fourteen-week-old broiler breeder pullets and cockerels from two different companies were found to have encephalitis 2 weeks after vaccination with avian encephalomyelitis (AE) vaccine. Histologic examination and virus isolation indicated that affected chickens had AE. All chickens had been vaccinated with either one of two serials of AE vaccine approximately 2 weeks before the onset of clinical signs.     

Immunogenicity of an Escherichia coli multivalent pilus vaccine in chickens

Immunogenicity of an oil-emulsified Escherichia coli multivalent pilus vaccine was evaluated in 4-week-old chickens. The vaccine contained 180 micrograms of pilus protein from each of serotypes O1 and O78 and 170 micrograms of pilus protein from serotype O2. Chickens were vaccinated twice subcutaneously at 4 and 6 weeks old and challenged via the posterior thoracic air sac with E. coli serotype O1, O2, or O78 2 weeks after the last vaccination. Unvaccinated challenged chickens suffered 8% to 26% mortality; no vaccinated chickens died. Vaccinated chickens had very mild gross lesions in the air sacs, livers, and pericardial sacs and eliminated E. coli more efficiently than the unvaccinated challenged chickens. The results showed that a multivalent pilus vaccine protects chickens against active respiratory infection.     

陕西某种鸡场大肠埃希菌血清型调查与灭活苗研制

为控制大肠埃希菌病的危害,对从陕西某种鸡场分离鉴定的20株鸡大肠埃希菌进行了血清型调查,结果O2型5株、O6型3株、O78型4株,未定型8株,表明鸡场的大肠埃希菌有了新的流行菌株O6出现。用定型的分离菌株研制的自家灭活菌苗免疫接种雏鸡,7 d后可检测到抗体,14 d产生免疫保护力。29日龄二免,90 d后抗体水平仍在5log2以上。119日龄三免,免疫后5个月抗体水平仍维持较高,但6个月后,抗体水平已在5 log2以下,失去保护力。通过3年的应用,该场种鸡大肠埃希菌病的平均死亡率由6.7%下降至1.6%,雏鸡死亡率由4.2%下降至1.8%,差异极显著(P<0.01)。     

Separation and Identification of Multi-drug Resistant and High Pathogenic Escherichia coli Causing Piglets Watery Diarrhea

Piglets watery diarrhea with high pathogenity and high mortality rate happened in many farms in Changchun area, the diarrhea piglets repeatedly tried a variety of medications,but no effect came out. From the onset of piglets intestinal wall lining,we had separated and identified some drugresistant and highly pathogenic E.coli strains.After the biochemical test,and cloning and sequencing of 16S rRNA of the strains,we compared them with the sequences in GenBank,the isolated strains had 99% homology with the 16S rRNA of E.coli sequence. After animal testing, we determined that this kind of E.coli could cause piglets watery diarrhea. Drug sensitivity test results showed that drugs commonly used in E.coli disease as treatments such as gentamicin,ofloxacin and ciprofloxacin were invalid.