奶牛源坏死梭杆菌四川株分离鉴定及生物学特性分析

从四川规模化奶牛场患腐蹄病的奶牛蹄部分离坏死梭杆菌,采用常规实验室检测方法对其进行分离鉴定,并进行坏死梭杆菌分离条件、保存条件、实验动物致死和感染试验以及药敏试验。结果:不同的分离方法导致坏死梭杆菌的分离率不同,坏死梭杆菌在不同的保存条件下存活时间不同;攻毒后死亡小鼠注射部位形成脓肿;绵羊蹄部感染坏死梭杆菌后蹄部灌脓,形成脓肿;药敏试验表明坏死梭杆菌对林可霉素、洁霉素、强力霉素、阿莫西林、氨苄青霉素、头孢唑啉高度敏感,对头孢拉定、青霉素中度敏感,对卡那霉素、氧氟沙星、四环素、链霉素耐药。     

奶牛腐蹄病的防治措施

正奶牛腐蹄病是一种兽医临床的常见病,主要是因为奶牛蹄部感染坏死厌氧丝状杆菌而持续炎症。一般在多雨时节发生,这种腐蹄病虽然有非常低的死亡率,但由于生病奶牛的蹄部已经变形、持续跛行等造成运动困难,对机体的使用年限产生的严重影响,而且生产性能降低,严重影响了养殖农户的收入。1原因解析(1)造成奶牛腐蹄病的病原菌主要由于坏死梭杆菌、节瘤拟杆菌,还有一些粪弯杆菌、脆弱类杆菌、酵母菌、梭菌、产     

源于奶牛蹄部感染的牛源污蝇解壳杆菌的分离鉴定与药敏试验

为确定导致江苏某奶牛场蹄部感染的病原,采集发病奶牛蹄部感染病灶的病料进行细菌分离培养,使用MAL-DI Biotyper微生物快速鉴定系统进行细菌鉴定,并对分离菌株进行药物敏感性试验.结果显示,病灶中的分离株为污蝇解壳杆菌(Wohlfahrtiimonas chitiniclastica),血平板上呈β型溶血.药敏试验...     

PCR法诊断奶牛腐蹄病坏死梭杆菌的研究与应用

试验根据坏死梭杆菌白细胞毒素基因序列的特异性,设计1对引物FLP并建立了PCR检测方法.试验结果表明,引物FLP具有相对较高的特异性和敏感性,其敏感性达到了48.5pg.对现地采集奶牛蹄部病料进行PCR检测,发现引物FLP具有很好的特异性,能够准确诊断出坏死梭杆菌的存在.     

牛腐蹄病的防治

<正>腐蹄病为牛群常见病,典型症状为跛行、喜卧,检查蹄部可见变形、趾间皮肤发红、红肿,蹄冠呈红紫色,湿热肿痛。随着病情加剧,蹄部组织脓肿,出现炎性化脓区。此病是非致死病,但是,严重影响养殖效益,造成一定的经济损失。1病因分析1.1病原体感染病原体主要包括结节状类杆菌、产黑色素类杆菌、脆弱类杆菌、坏死杆菌,此外,螺旋体、粪弯杆菌、梭杆菌、球菌、酵母菌及其他一些条件致病菌。坏死梭杆菌是从患蹄中最常分离到的     

奶牛腐蹄病的治疗

正奶牛腐蹄病是常发病之一,其发病率较高,有报道显示其发病率约为20%,有的牛场其发病率可能更高。该病又称为趾间腐烂、蹄间蜂窝织炎,是由坏死梭杆菌或其他致病菌混合感染受损蹄部,引起动物趾间皮肤及深层软组织坏死的一种高度接触性传染病,以奶牛蹄部受损组织的化脓坏死性分解、腐败恶臭和角质受到破坏为主的外科疾病特征,是引起奶牛跛行的主要原因之一。一般后蹄发生比例高于前蹄,而且天     

奶牛腐蹄病坏死杆菌分离鉴定及白细胞毒素基因lktA1序列分析

为分离纯化奶牛腐蹄病坏死杆菌,分析其与其他菌株的亲缘关系,本研究利用坏死杆菌白细胞毒素特异性引物,对奶牛腐蹄病病牛蹄部拭子样品进行了PCR检测,利用厌氧培养基对PCR检测阳性样品进行了坏死杆菌的分离培养,以分离的坏死杆菌基因组DNA为模板,对白细胞毒素基因进行了克隆和序列分析。结果显示,9份奶牛腐蹄病病牛蹄部拭子样品PCR检测结果均为阳性,对其中一份样品中的坏死杆菌进行分离培养,获得了纯培养物,命名为bFR13-1。坏死杆菌bFR13-1菌株白细胞毒素基因测序结果显示,与GenBank已发表的H05、A25和B35菌株的白细胞毒素基因在核苷酸水平的同源性分别为98.40%、98.35%和90.79%,推导氨基酸的同源性分别为97.7%、97.6%和89.0%。进化树分析结果显示,坏死杆菌bFR13-1菌株白细胞毒素与H05菌株的同源性最高,bFR13-1菌株与H05菌株和A25菌株呈较近亲缘关系。结果表明,不同坏死杆菌分离株的白细胞毒素呈现一定的变异性,这种变化是否与坏死杆菌致病性相关,值得深入研究。     

梅花鹿致病性源坏死梭杆菌的分离与鉴定

为确定湖北省恩施自治州某鹿场致患病鹿蹄腐烂的病原菌,本研究于患病梅花鹿的前蹄病健结合处采集病料并对其进行细菌分离培养,最终分离到一株厌氧菌。对该细菌进行了形态观察、生化试验、16S rRNA基因序列测定、细菌生化鉴定仪鉴定、药物敏感性试验以及小鼠致病力试验等。结果显示该细菌在显微镜下呈革兰氏阴性、丝状、长短不一的杆菌。16S rRNA基因序列测定结果显示该分离菌株与多株坏死梭杆菌参考株的同源性均为99%,将其命名为HNFnf;经Vitek细菌鉴定仪鉴定该细菌为坏死梭杆菌;药敏试验结果显示,该菌对青霉素、头孢类等中度敏感,对阿米卡星、链霉素、庆大霉素耐药。本研究为临床治疗由坏死梭杆菌引起的反刍动物"腐蹄病"与"肝脓肿"提供参考依据。     

羊腐蹄病的防治

羊腐蹄病是由坏死梭形杆菌和结瘤偶蹄杆菌感染而引起的以局部组织发炎、坏死为特征的一类传染病，因常侵犯蹄部，故称腐蹄病。     

牛坏死杆菌病的防治

随着舍饲奶牛业的快速发展，牛坏死杆菌病的发病率电有逐年上升的趋势，给奶牛养殖带来了不小的经济损失，该病是由坏死梭杆菌引起的一种慢性传染病，以蹄部、皮肤、消化道黏膜坏死为特征，一般多见于5∽10月份。     

Development of enzyme-linked immunosorbent assays for the detection of Fusobacterium necrophorum antibody in animal sera

Enzyme-linked immunosorbent assays (ELISA) for the detection of Fusobacterium necrophorum antibody in the sera of rabbits, cattle, and sheep were developed, using a ribosome-rich extract (RRE) from F necrophorum as the antigen. Test character, including optimal antigen dilution and substrate incubation periods, was established, using rabbit, bovine, and ovine antisera produced against RRE from isolates of F necrophorum. Rabbit antisera produced against 7 other species of bacteria were used to test the specificity of the F necrophorum RRE antigen. Cross-reactivity was not detected. Sera from 50 feedlot cattle were examined with the bovine ELISA. Of the 50 samples, 43 (88%) were positive for F necrophorum antibody. The ELISA developed in this study were sensitive and specific and appear to be readily adaptable to serologic investigations of F necrophorum.     

Laminitis and interdigital dermatitis and heel horn erosion. A European perspective

Laminitis is one of the most important claw disorders in dairy herds. Nutrition, calving, burdening of the lateral claw of the rear feet, and hereditary susceptibility are all contributing factors. Interdigital dermatitis in cattle may be a result of infection by Bacteroides nodosus and Fusobacterium necrophorum. If this infection becomes chronic, heel horn erosion is its consequence.     

Fusobacterium equinum possesses a leukotoxin gene and exhibits leukotoxin activity

Fusobacterium equinum, a gram negative, rod-shaped and an obligate anaerobic bacterium is a newly described species. The organism is associated with necrotic infections of the respiratory tract in horses that include necrotizing pneumonia, pleuritis and paraoral infections. The species is closely related to F. necrophorum that causes liver abscesses in cattle and sheep, calf-diphtheria in cattle, and foot-rot in sheep and cattle. Leukotoxin, an exotoxin, is an important virulence factor in bovine strains of F. necrophorum. Our objective was to examine strains (n=10) of F. equinum for leukotoxin (lktA) gene and its toxic effects on equine leukocytes. Southern hybridization and partial DNA sequencing revealed that all the 10 strains had the lktA gene with greater similarities to F. necrophorum subsp. necrophorum. The secreted leukotoxin was detected in the culture supernatant and its biological activity was determined by viability assays with equine polymorphonuclear cells (PMNs) using flow cytometry. While culture supernatants of four strains (E1, E7, E9, and E10) were highly toxic to equine PMNs; strain E5 was moderately toxic and the remaining strains (E2, E3, E4, E6, and E8) were only mildly toxic. Our data indicated that F. equinum isolates had lktA gene and its product was toxic to equine leukocytes. Therefore, leukotoxin may be an important virulence factor in F. equinum infections.     

The role of Fusobacterium necrophorum and Bacteroides melaninogenicus in the aetiology of interdigital necrobacillosis in cattle

When cultures of known pathogenic strains of Fusobacterium necrophorum, isolated either from cattle or sheep were injected through the interdigital skin of cattle typical lesions of interdigital necrobacillosis were produced. The inclusion of Bacteroides melaninogenicus in the inoculum did not appear to contribute to the development of lesions.     

Isolation,Identification and Sequence Analysis of Leukotoxin Gene lktA1 of Fusobacterium necrophorum of Cow Footrot

For separation and purification of Fusobacterium necrophorum of cow footrot, and analysis of genetic relationship with other strains, the hoof ministry swab samples were detected by PCR based on specific primers of leukotoxin gene, and genomic DNA were isolated from PCR positive samples of Fusobacterium necrophorum culturing in anaerobic medium.The genes of leukotoxin were cloned and sequenced.The results showed that nine of hoof ministry swab samples were all PCR positive samples, and we obtained Fusobacterium necrophorum pure culture from one of the samples which named bFR13-1.The gene sequencing results indicated that the homologies of leukotoxin gene nucleotide sequence of bFR13-1 strain compared with H05, A25 and B35 strains from GenBank were 98.40%, 98.35% and 90.79%, respectively, and the homologies of deduced amino acid sequence were 97.7%, 97.6% and 89.0%, respectively.Phylogenetic tree analysis results showed that leukotoxin gene of Fusobacterium necrophorum bFR13-1 and H05 had high homology and bFR13-1, H05 and A25 showed a close genetic relationship.The result indicated that leukotoxin showed variability between different Fusobacterium necrophorum isolated strains, and it was worth to study whether this change and pathogenicity of Fusobacterium necrophorum were related.     

Efficacy of vaccination against Fusobacterium necrophorum infection for control of liver abscesses and footrot in feedlot cattle in western Canada

A randomized and blinded field trial was carried out to evaluate the efficacy of a Fusobacterium necrophorum bacterin for control of liver abscesses and footrot under commercial feedlot conditions in western Canada. Half of the vaccinated and half of the unvaccinated control animals had ad libitum access to a forage-based (ALF) growing diet. The other half of each group was limit-fed a grain-based (LFG) growing diet. The overall prevalence of A and A+ liver abscesses in this trial was 16.7%. A strong association was found between diet group and presence of A or A+ liver abscessation at slaughter. Diet group modified the effect of vaccination on the prevalence of liver abscesses at slaughter, and on the incidence of footrot during the feeding period. The odds that a vaccinated animal in the ALF group would have an A or A+ liver abscess at slaughter were less than 1/3 the odds that an unvaccinated animal in the same diet group would have an A or A+ liver abscess at slaughter (OR = 0.27, [95% CI: 0.07 to 1.02], P = 0.05). The overall incidence of footrot in this trial was 6.5%. The odds that a vaccinated animal in the ALF group would be treated for footrot were less than 1/5 the odds that an unvaccinated animal in the same group would be treated for foot-rot (OR = 0.18, [95% CI: 0.04 to 0.82], P = 0.03). Within the LFG group there were no differences between vaccinated and unvaccinated animals in the odds of an animal being treated for footrot, or in the odds of having an A or A+ liver abscess score at slaughter. This trial suggests that vaccination against F. necrophorum infection may have applications to decrease the prevalence of severe liver abscesses at slaughter and decrease footrot treatments in certain diet situations.     

Failure to induce in rabbits effective immunity to a mixed infection of Fusobacterium necrophorum and Corynebacterium pyogenes with a combined bacterin.

Failure to induce in rabbits effective immunity to a mixed infection of Fusobacterium necrophorum and Corynebacterium pyogenes with a combined bacterin. Onderstepoort Journal of Veterinary Research, 44 (4), 253--2;6 (1977). Rabbits were immunized with alum-precipitated, oil adjuvant and an untreated bacterin composed of F. necrophorum and C. pyogenes. Immunized rabbits were challenged intradermally with a mixture of F. necrophorum and C. pyrogenes. Immunized rabbits were challenged intradermally with a mixture of F. necorphorum and C. pyrogenes. Initially a low level of initial transient resistance could be demonstrated but a solid immunity could not be established.     

A microaerophilic coccus in pyogenic infections of ruminants

Pyogenic infections of cattle, sheep and goats were examined for the presence of a Gram positive bacterium that has been designated "microaerophilic coccus" by other workers. The bacterium was found to be involved in a range of disease processes, including foot and soft tissue abscesses, mastitis, pericarditis and pyometra in cattle, joint and foot abscesses in sheep and foot abscesses in goats. The characteristic feature of the bacterium was its satellitic growth around colonies of other organisms. The microaerophilic coccus was usually part of a mixed flora, which included Corynebacterium pyogenes, Fusobacterium necrophorum, Peptostreptococcus indolicus and Bacteroides sp.     

Immunohistochemical and ultrastructural identification of Fusobacterium necrophorum subsp. necrophorum in bovine fatal necrotizing glossitis

A 37-day-old male Japanese black calf showing marked salivation and leucocytosis died and was examined the tissues histologically. Histological lesions were characterized by severe focal necrotic glossitis on the ventral side of the root of the tongue. Immunohistochemically, Fusobacterium necrophorum subsp. necrophorum antigen was detected in the necrotic tissues and its distribution corresponded to that of the gram-negative, nonsporeforming, long filamentous organisms. Ultrastructural similarities between the organism and F. necrophorum subsp. necrophorum, but not subsp. funduliforme were observed. These findings clearly demonstrated that the fatal necrotic glossitis was caused by F. necrophorum subsp. necrophorum. This is the first report of bovine fatal necrotizing glossitis with leucocytosis caused by F. necrophorum subsp. necrophorum infection, and this organism may be an important fatal pathogen in calves with glossal lesions.