气肿疽梭菌延边样的分离鉴定

牛、羊等反刍动物的气肿疽病是由气肿疽梭菌感染引起的严重危害畜牧业发展的一种疾病,本研究从延边州延吉市采集疑似气肿疽梭菌感染的牛肝脏、脾脏、肌肉等组织进行气肿疽梭菌的分离鉴定。结果表明:经病原学检查、生化鉴定、动物试验及分子生物学鉴定,本试验成功对气肿疽梭菌进行了分离,该菌株细胞毒素、cct A基因与Gen Bank上已发表的气肿疽梭菌ATCC 10092菌株的同源性达到99.0%,此结果为该地区气肿疽病的免疫和微生态防治提供了重要的参考依据。     

气肿疽及防控措施

气肿疽又被称为黑腿病,为气肿疽梭菌引起的一种急性败血症的传染病,呈散发或地方性流行,气肿疽梭菌在深层肌肉产酸产气导致气肿坏疽为特征。主要通过春秋两季气肿疽疫苗注射来防控气肿疽传染病。2016年4月7日,宁洱县勐先镇黄寨村箐头组鲁明文、鲁升2户养殖户的黄牛发生疫病,在饲养的27头黄牛中发病7头,死亡5头。经普洱市动物卫生监督所、宁洱县动物疫病预防控制中心和勐先镇农业技术服务中心技术人员通过黄牛发病的流行病学调查、现场病理解剖、采样送实验室检测诊断为牛气肿疽病。及时采取牛气肿疽病的防控治疗措施,有效控制了疫情的流行蔓延,按期回访没有出现黄牛发病死亡情况,减少了养殖户的经济损失,巩固了本地山区养牛业的健康发展。     

牛气肿疽暴发流行的诊疗

气肿疽是气肿疽梭菌引起反刍动物的一种急性发热性传染病，主要侵害黄牛，呈散发或地方流行性，以败血症及深层肌肉发生气肿性坏疽为特征。1952年-1954年，湖南省部分地区曾发生本病，1955年起每年进行预防接种，至1974年疫情已被完全控制，2002年7月，湖南省湘南某村暴发本病，病死率达45．6％。     

山区散养放牧牛气肿疽病的诊断及防制措施

气肿疽病又称为黑腿病或鸣疽,是一种急性败血性传染病,主要由气肿疽梭菌引起的,自然条件下气肿疽主要侵害黄牛,也严重影响水牛、奶牛、牦牛、犏牛的生存环境和质量,再加上地理位置和条件的特殊性,牛气肿疽病发生率也不断增加,为山区散养牛养殖户带来危害和巨大经济损失。本文从山区散养放牧牛气肿疽病的病因、症状及诊断方法介绍入手,重点阐述切实可行的防制措施。     

气肿疽灭活苗的制造

气肿疽是气肿疽梭菌引起的牛的急性热性、败血性传染病。病的特征是在肌肉丰满部位发生气性炎性水肿，按压患部常有捻发音，病早期有跛行，后期皮肤发黑故又称“爬腿瘊”、“黑腿病”、“鸣疽”。本病分布于世界各地，我国很早就有发生，延边地区建国前时有发生，但从５０...     

牛气肿疽的诊断与防治

气肿疽又称黑腿病或呜病。主要是牛的一种气肿疽梭菌引起的急性、热性传染病,其特征为肌肉丰满部位发生炎性气性肿胀,并常有跛行。主要侵害1～3岁的黄牛,2008年6月,     

一起牛气肿疽病的防治

正牛气肿疽又称黑腿病,是由气肿疽梭菌引起牛发生的一种急性败血性传染病。其主要特征是在肌肉丰满处出现气性肿胀,触诊有捻发音。本文以澜沧县南岭乡下南现村下新寨组饲养的本地黄牛突然发病,迅速死亡,5 d内有6户农户饲养的6头黄牛发病死亡为例,通过流行病学调查和观察黄牛的发病情况、临床症状、尸体剖检,确诊为牛气肿疽传染病。1发病情况     

与气肿疽棱菌有关的牛心包炎和脑膜炎的病理学变化

两年多来,研究了29例牛的气肿疽梭菌感染。14例仅有肌炎病变,8例同时有肌炎和纤维素性心包炎,6例仅有纤维素性心包炎,余1例仅见化脓性脑膜炎病变。应用荧光抗体技术在所有病变中均鉴定出气肿疽梭菌。气肿疽梭菌(Colstridium Chauvoei)是世界上很多地区牛坏疽性肌炎(黑腿病)的一种已被_(aa)充认识的病原。肌肉病损已有详细描述,但有关气肿疽梭菌在其他组织引起病损的详细描述却很少,至今尚无死于气肿疽梭菌感染的动物脑和脊髓研究方面的报告。     

敦化地区黄牛气肿疽的防治与诊断

气肿疽是由气肿疽梭菌引起的主要感染牛的一种急性发热性传染病,不直接传播呈散发或地方性流行。主要特征为肌肉肿胀有捻发音,其发病率、死亡率都很高。2006年敦化市辖区内相邻4个乡镇相继发生牛的气肿疽病,现将诊断和治疗情况     

牛气肿疽的防治体会

<正>1病原体气肿疽梭菌为两端钝圆的粗大杆菌,长2~8μm,宽0.5~0.6μm。有周口鞭毛,能运动,无荚膜,在机体外均可形成芽孢,位于菌体中央或稍于一侧,在病料中一般为直形,单在或成对排列,能产生不耐热的外毒素,气肿疽梭菌的繁殖体对热和消毒药的抵抗力不大,52℃30min就能使其丧失传染能力,但气肿疽梭菌的繁殖体在病死动物的肌肉内形成芽孢的抵抗力较强,能     

Diagnostic Report of Clostridium chauvoei Infection of Cattle in Yanbian of Jinlin Province

The study was aimed to rapidly diagnose two ill and dead cattle in three bay town of Yanji city in Jilin province.We collected liver,spleen,heart,muscle and other tissues to diagnose Clostridium chauvoei infection by epidemiological investigation,clinical autopsy,microscopic obsevation,bacterial culture observation,biochemical experiment,molecular biology diagnosis and animal experiment.The muscles of the ill and dead cattle contacted with crepitus,and the section had a lot of blood and bubble flowing out,microscopic obsevation found bacillus with obtuse at both ends and spores,the bottom of the tube had loose and white precipitate in the broth,biochemical experiments showed that the isolate had acid-production and gas-production reactions which was unique to anaerobic bacteria causing Clostridium chauvoei infection,and 501 bp fragment was amplified by PCR.The results showed that two ill and dead cattle were casued by Clostridium chauvoei infection in Yanbian area,and proved the existence of Clostridium chauvoei.The test separated Clostridium chauvoei Yanbian strain for the frist time,which provided important reference basis for the prevention and treatment of Clostridium chauvoei infection,and laid the important foundation for the further study of drug and immune preventions of this disease.     

Isolation,Identification and Pathogenicity Research of a Strain of Clostridium perfringens from Goat

A pathogenic bacteria was isolated from a dead goat,and it was identified as goat-pathogenic type A Clostridium perfringens by differential culture,biochemical test,molecular biological technology and animal pathogenicity test.Analysis results of toxin genes showed that this bacteria contained both α and β2 toxin genes.Animal pathogenicity test result showed that this bacteria was highly pathogenic to mice,and the same bacteria was isolated from dead mice as which was isolated from the dead goat,so it was certain that type A Clostridium perfringens was the main pathogenic bacteria which caused the goat dead.The results would provide scientific basis for the treatment and prevention of the disease caused by Clostridium perfringens in this goat farm.     

一株山羊魏氏梭菌的分离鉴定及其致病性研究

本试验从一例病死山羊组织内分离到一株致病性细菌,通过鉴别培养、生化试验、分子生物学试验和动物致病性试验,证明该病原菌为羊致病性A型魏氏梭菌;毒素基因分析结果显示,该菌同时含有α和β2两种毒素基因;动物致病性试验结果表明,该菌对昆明小鼠具有较强的致病性,并从试验致死的昆明小鼠病料中分离到了与病死山羊病原相一致的细菌,从而确定A型魏氏梭菌为引起该山羊死亡的主要病原菌。本试验结果为该羊场魏氏梭菌病的治疗和预防提供了科学依据。     

奶牛产气荚膜梭菌的分离鉴定与药物敏感性分析

本研究旨在对从奶牛内脏、组织病料样品中分离得到产气荚膜梭菌并进行鉴定分型,为后续疫苗研究提供材料,并通过药敏试验筛选出较为敏感的药物以针对性的指导牧场对牛群进行治疗并提出有效的防治隔离建议。2019年山东21家牧场部分奶牛出现腹泻、便血及短期内死亡等现象,对部分死亡奶牛进行剖检,并送检64份内脏、组织病料样品进行CP培养初筛,对初筛阳性菌株用生化鉴定及PCR分型鉴定;以16S rDNA技术对CP菌株进行同源性分析;采用E-test法测定分离菌对7种抗菌药物（红霉素、左氟沙星、利奈唑胺、万古霉素、克林霉素、四环素、利福平）的最低抑菌浓度（micro inhibition concentration,MIC）,筛选出敏感药物。8份样品血琼脂培养基细菌培养出现灰白色菌落,梭菌显色培养基培养为橘红色梭菌典型特征菌落;生化鉴定结果符合产气荚膜梭菌特征;PCR分型结果显示5株为A型、2株C型和1株E型;16S rDNA分析得出8株分离菌与产气荚膜梭菌同源性最高可达100%;8株CP菌株对7种药物敏感,其中2株对克林霉素、利福平更为敏感,3株对红霉素及利奈唑胺敏感,1株对左氟沙星更敏感。对有病牛的2家牧场推荐使用林可酰胺类、利福霉素类及大环内酯类药物进行防治,对牧场防治效果及时追踪,对于没有治疗价值的牛立即扑杀,无害化处理,改善饲养条件,减小饲养密度。     

鸡源高产淀粉酶丁酸梭菌的分离鉴定

为了获得产淀粉酶的丁酸梭菌,本研究从鸡小肠表面黏液中进行丁酸梭菌的分离与筛选,首先对样品进行80℃水浴10 min除去非芽孢菌后,然后接种到TSN培养基进行菌株的分离与筛选。对分离到的菌株进行菌落形态、显微形态初步观察,然后对疑似丁酸梭菌的菌株进行产酸能力和淀粉酶酶活检测,最后对既产酸又高产淀粉酶的菌株进行生理生化鉴定和16S rDNA序列分析。结果表明分离到一株革兰氏阳性厌氧菌C.B1,菌体呈短杆状,能形成圆形或椭圆形芽孢,生理生化结果也符合丁酸梭菌的基本特征,16S rDNA序列长度为1450 bp,与丁酸梭菌的同源性高达99%以上,因此确定该分离菌株为丁酸梭菌,为进一步开发新的微生态制剂奠定了基础。     

腐败梭菌无毒重组α毒素的表达与免疫保护性分析

本研究旨在获得腐败梭菌无毒重组α毒素,并评价其免疫保护性。对已知的腐败梭菌α毒素编码基因进行优化设计和人工合成,获得了缺少第212-222位共11个氨基酸编码序列的基因片段（GCSA_Δ11）。将该基因克隆至原核表达载体pET-30a（+）中进行表达与纯化。利用Western blot方法检测获得的重组α毒素（rCSA_Δ11）与腐败梭菌天然毒素抗血清的反应性,并采用小鼠检测其毒力。随后,以rCSA_Δ11制备疫苗免疫家兔,按照《中华人民共和国兽药典》（2015年版）规定的方法检测家兔血清的中和抗体效价。结果表明,rCSA_Δ11可溶表达比例可达46%,且能与腐败梭菌天然毒素抗血清反应。进一步的试验结果显示,rCSA_Δ11丧失了对小鼠的毒力,0.1 mL的一免兔血清可中和8~12个小鼠最小致死量（MLD）的腐败梭菌天然毒素;二免兔血清可中和32~45个小鼠MLD的腐败梭菌天然毒素。1个家兔MLD的腐败梭菌天然毒素攻毒后,对照组家兔4/4死亡,免疫组家兔得到了100%（4/4）的保护。综上表明,无毒性的rCSA_Δ11保留了良好的免疫保护性,从而为腐败梭菌病基因工程亚单位疫苗的研制提供了重要的试验数据。     

Diagnosis and Treatment of Serotype A Pasteurella multocida Infection in a Dairy Farm

The assay was aimed to clarify the reasons why dairy cattle became ill with the symptoms including fever,breathing difficulty,salivation and subcutaneous emphysema at neck or chest in a dairy cow farm at Hubei province,even part of dairy cows were dead for exhaustion.The disease was diagnosed by laboratory methods,and treatment scheme was proposed.The samples from heart,liver,lung or trachea tissues were collected,then M.bovis and pathogenic bacteria were isolated,respectively.Biochemical characteristics,pathogenicity test and drug sensitivity analysis were performed on the isolated pathogenic bacteria.RNA was extracted from bovine serum,then bovine ephemeral fever virus was identified by RT-PCR.The results showed that nothing grew in PPLO medium,and the isolated bacteria could form indole,but did not ferment glycogen and inositol in the biochemical tests.The isolated pathogenic bacteria was conformed as serotype A P.multocida by PCR while bovine ephemeral fever virus of RT-PCR result was negative.The pathogen could kill mice and the pathogenic bacteria could be isolated from heart blood of the dead mice in pathogenicity test.The isolated bacteria was sensitive to cefoperazone or levofloxacin in drug sensitivity analysis.In conclusion,the disease was diagnosed as bovine serotype A P.multocida infection.Some treatments including choosing sensitive antibiotic drugs based on the drug sensitivity tests,holding sick cattle under quarantine treatment and isolating suspected cattle should be taken.Moreover,some complex measures such as keeping the air flowing,cleaning up in time,improving the management,and preventing stress from congestion,cold and long distance transport were very important to prevent and control serotype A P.multocida infection in dairy cows.     

某牛场奶牛A型多杀性巴氏杆菌的分离鉴定

湖北某奶牛场牛群爆发体温升高、呼吸困难、流涎及颈胸皮下气肿等症状的疾病,部分发病牛衰竭死亡,为确诊牛场牛群发病原因并提出防控方案,本试验采集死亡牛的心脏、肝脏、肺脏及气管组织进行病原菌的分离纯化及PCR鉴定,并对鉴定的病原菌进行生化特性鉴定、致病性试验和药物敏感性分析;同时提取患牛血清RNA,开展牛流行热病毒的RT-PCR鉴定。结果显示,病料在类胸膜肺炎固体培养基上不生长,生化特性鉴定显示能形成靛基质、不发酵肝糖和肌醇;牛流行热病毒RT-PCR扩增阴性;所分离的病原菌经16S rRNA及牛A型多杀性巴氏杆菌特异性引物PCR扩增阳性;致病性试验显示该病原可致死小鼠,且能从死亡小鼠体内分离到感染菌;药敏试验结果显示该病菌对头孢哌酮、左氧氟沙星敏感,其他20种临床常见药物表现耐药。综上所述,该牛场患病牛确诊为牛A型多杀性巴氏杆菌感染,建议根据药敏试验结果选择敏感药物,对患病牛进行隔离治疗,疑似患病牛隔离观察,同时加强通风,及时清理污物并消毒,改善饲养管理,避免拥挤、寒冷及长途运输等应激因素。     

甘肃、宁夏地区奶牛衣原体血清流行病学调查及风险因素分析

本研究旨在查明甘肃与宁夏地区奶牛衣原体的感染情况并分析影响其感染的风险因素。本试验采用间接血凝试验(IHA)方法检测了甘肃榆中(751份)、宁夏青铜峡(450份)和宁夏吴忠(456份)3个地方总计1 657份奶牛血清样品,并应用流行病学调查及统计学方法对影响奶牛衣原体感染的因素进行了分析。流行病学调查结果显示,奶牛衣原体抗体总阳性率为29.33%;应用logistic回归分析评估奶牛衣原体感染的风险因素,结果显示年龄和胎次不是显著风险因素(P>0.05),而地区因素是影响奶牛衣原体感染的风险因素(P<0.05)。奶牛衣原体抗体滴度最高达1:1 024。结果表明,甘肃和宁夏地区奶牛衣原体普遍流行。因此,应当提高对调查地区奶牛衣原体感染的重视,采取适当的综合控制方法和有效的管理措施以防控奶牛衣原体病,以保证奶牛养殖业的经济效益。     

Seroprevalence Investigation and Risk Factors Analysis of Chlamydia Infection in Dairy Cattle in Gansu and Ningxia Areas

The objective of the present investigation was to examine seroprevalence of Chlamydia in dairy cattle in Gansu and Ningxia areas, Northwest China, and to analyze the factors affecting Chlamydia infection in dairy cattle.Indirect hemagglutination assay (IHA) was used to detect antibodies against Chlamydia in 1 657 dairy cattle serum samples from Gansu and Ningxia areas.Epidemiological investigation and statistical methods were used to analyze the data.The results showed that the overall seroprevalence was 29.33%, logistic regression analysis was applied to evaluate the risk factors of Chlamydia infection in dairy cattle, and the results indicated that age and numbers of pregnancy of dairy cattle were not the significant risk factors (P>0.05), and were left out of the final model, however, region was considered as the main risk factor associated with Chlamydia infection (P<0.05).The highest titer was 1:1 024.In conclusion, the results of the present survey indicated the widespread of Chlamydia infection in Gansu and Ningxia areas.In order to ensure the economic benefit of dairy farming, we should pay more attention to Chlamydia infection in dairy cattle, and integrated control strategies and efficient management measures should be implemented to prevent and control Chlamydia infection in dairy cattle.