Eradication of lice in cattle.

The purpose of this field study was to develop and evaluate eradication as a strategy to control lice in cattle. Thirty-three herds of cattle were selected and observed during a period of two and a half years. Before eradication, biting lice (Damalinia bovis) were present in 94% of the herds and 27% of the animals. Sucking lice (Linognathus vituli) were present in 42% of the herds and 5% of the animals. These levels were very similar to those reported from other countries in Northern Europe. The eradication strategy was successful in 28 of 33 herds, but lice were still present in 5 herds 3 to 6 months after treatment. Biting lice were present in all these 5 herds, sucking lice were present in 3 herds. During the next 12 months, nine of the 28 herds were reinfected with lice. Six herds were reinfected with just biting lice, 2 herds with just sucking lice and one herd was reinfected with both. There was no significant difference between the 2 louse species regarding the risk of unsuccessful eradication or reinfection. The only significant risk factor for reinfection was either purchase of livestock or use of common pasture, combined with failure in pre-treatment of newly introduced animals.     

水牛牛巴贝斯虫体外培养的研究：Ⅱ.冷冻血清的应用和筛选

应用冷冻血清对１株采自自然感染的水牛牛巴贝斯虫进行了长达７２ｄ的体外连续培养，共继代２０次，培养７２ｈ红细胞染虫率最高达１４．１％，平均为８％～１０％。培养２０ｄ和３０ｄ的牛巴贝斯虫经液氮保存复苏后，接种于去脾水牛犊均引发了严重的牛巴贝斯虫病，从而说明已建立了水牛牛巴贝斯虫的体外连续培养，且经培养后的牛巴贝斯虫致病力没有改变。本试验利用６头份的冷冻健康水牛血清同时进行培养，结果发现，并非所有的健康水牛血清均适合于体外培养牛巴贝斯虫。这一发现对建立水牛牛巴贝斯虫的体外培养和研究水牛牛巴贝斯虫病均具有重要意义     

The redox potential of growing cultures of Streptococcus bovis Orla-Jensen compared with other facultative anaerobes.

The changes of redox potential were measured in growing cultures of three strains of Streptococcus bovis, together with three strains of Staphylococcus aureus and one strain of each of Lactobacillus plantaram, Lactobacillus casei, and Eschericia coli. It was found that both S. aureus and E. coli could reduce the redox potential of the growth medium to very low values (between —400 mv and —600 mv), whereas the streptococci and lactobacilli were able to cause only slight or insignificant changes of the redox potential. Respirometric measurements confirmed that the capacity of oxygen consumption of S. bovis was very small compared to that of E. coli and S. aureus. On this basis the authors conclude that S. bovis in all probability is unable to contribute significantly to maintenance of the low redox potential of its natural habitat, the rumen. This function must be carried out by other bacteria, such as enterobacteria or staphylococci, which are capable of performing a true, aerobic respiration.     

应用基因分型技术研究牛结核分枝杆菌分子流行病学

牛分枝杆菌是引起牛结核病(bTB)的病原体,侵染宿主广泛,可感染多种家畜和野生动物,对人类和动物健康构成巨大危害。随着多年不懈的研究,有关分枝杆菌分子流行病学的知识积累不断增加。作者对近年来应用新的基因分型技术研究牛结核分枝杆菌分子流行病学方面的一些研究进展。这些技术包限制性片段长度多态性分析(RFLP)、PCR介导的间隔区寡核苷酸分析(spoligotyping)、数目可变串联重复位点(VNTR)等。主要用于对人、牛、家畜以及野生动物的结核病分子流行病学进行分析和监测。另外,还利用一系列敏感的基于PCR的技术从分枝杆菌杆菌复合体(NTM)中对菌型进行鉴别。对分枝杆菌感染分子流行病学的全面了解,有助于对本病扩散传播机制的深入研究。可为制定科学防治结核病的措施做出贡献。     

Expression,Purification and Activity Evaluation of Mb1230 Gene of Mycobacterium bovis

To explore the value of Mb1230 protein of Mycobacterium bovis in the diagnosis of bovine tuberculosis,we obtained the Mb1230 gene by PCR and constructed recombinant plasmid pET-22b-Mb1230.Recombinant Mb1230 protein was obtained by IPTG induction and purified by affinity chromatography.The activity of the recombinant protein was evaluated by TST test,IGRA test and indirect ELISA.The size of the recombinant protein matched with the theoretical value proved by SDS-PAGE;Western blotting result showed that the recombinant protein could react with mouse anti-His antibody,and had specific band;The results of TST test,IGRA test and indirect ELISA test also showed the recombinant protein had antigenic activity.The results indicated the recombinant protein Mb1230 had good B cell and T cell activity,so,it had the potential application in the diagnosis of bovine tuberculosis.     

Chronic tuberculosis caused by Mycobacterium bovis in a domestic donkey in Central Europe

Bovine tuberculosis is a contagious and zoonotic disease of animals and humans. In Europe, the number of reported cases of tuberculosis has decreased. Equidae are relatively rarely infected even in endemic areas. The presented report describes a case of chronic Mycobacterium bovis tuberculosis in a 30-year-old female donkey. The donkey initially presented with persistent lymphadenopathy; however, as the disease progressed, weight loss became apparent. To the authors' knowledge, this is the second confirmed case of tuberculosis in a donkey in Europe.     

28株牛源分枝杆菌的耐药基因突变分析

为明确分离自宁夏、甘肃和新疆地区的28株牛源分枝杆菌的耐药基因突变情况,通过采用16S rRNA、MPT64和多位点PCR方法进行基因分型鉴定,L-J比例法对其进行药敏试验,随后用DNA测序技术对10株耐药菌和4株敏感菌进行耐药基因检测及突变分析。分型鉴定结果表明,28株分离株均属于结核分枝杆菌复合群(MTBC),其中26株为牛分枝杆菌,2株为人型结核分枝杆菌。药敏试验结果表明,28株MTBC中18株对4种一线抗结核药物敏感,10株耐药。耐药菌株中,1株对利福平耐药;7株对异烟肼、利福平和链霉素三重耐药;2株对异烟肼、利福平、链霉素和乙胺丁醇四重耐药。耐药基因突变位点分析结果表明,rpoB 378位点的突变率为10.0%;katG 463位点的突变率为100.0%;rrs 311位点的突变率为11.1%;embB 378位点的突变率为100.0%;oxyR-ahpC和rpsL位点无突变。提示宁夏、甘肃和新疆地区奶牛场中MTBC主要以牛分枝杆菌为主,且存在耐多药情况,耐药突变位点以katG 463和embB 378为主。     

Improved detection of Mycobacterium tuberculosis and M. bovis in African wildlife samples using cationic peptide decontamination and mycobacterial culture supplementation

In South Africa, mycobacterial culture is regarded as the gold standard for the detection of Mycobacterium tuberculosis complex (MTBC) infection in wildlife even though it is regarded as “imperfect.” We compared a novel decontamination and mycobacterial culture technique (TiKa) to the conventional mycobacterium growth indicator tube (MGIT) system using known amounts of bacilli and clinical samples from MTBC-infected African buffaloes (Syncerus caffer), white rhinoceros (Ceratotherium simum), and African elephants (Loxodonta africana). Use of the TiKa-KiC decontamination agent on samples spiked with 10,000 to 10 colony forming units (cfu) of M. bovis (SB0121) and M. tuberculosis (H37Rv) had no effect on isolate recovery in culture. In contrast, decontamination with MGIT MycoPrep resulted in no growth of M. bovis samples at concentrations < 1,000 cfu and M. tuberculosis samples < 100 cfu. Subsequently, we used the TiKa system with stored clinical samples (various lymphatic tissues) collected from wildlife and paucibacillary bronchoalveolar lavage fluid, trunk washes, and endotracheal tube washes from 3 species with known MTBC infections. Overall, MTBC recovery by culture was improved significantly (p < 0.01) by using TiKa compared to conventional MGIT, with 54 of 57 positive specimens versus 25 of 57 positive specimens, respectively. The TiKa mycobacterial growth system appears to significantly enhance the recovery of MTBC members from tissue and paucibacillary respiratory samples collected from African buffaloes, African elephants, and white rhinoceros. Moreover, the TiKa system may improve success of MTBC culture from various sample types previously deemed unculturable from other species.     

Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

牛分枝杆菌抗原MPB70、MPB83和ESAT-6的融合表达及重组蛋白的初步应用

应用PCR方法从牛分枝杆菌Vallee株基因组中扩增获得mpb70、mpb83和esat-6三个目的基因片段。采用重叠延伸剪接技术(splicing by overlap extension,SOE)获得融合基因mpb70-mpb83后,将mpb70-mpb83和esat-6串连于同一表达载体pET32a( )中得到重组质粒pET70-83-E6。转化BL21(DE3)大肠杆菌感受态后,经IPTG诱导以可溶的形式表达融合蛋白。用Ni2 亲合层析的方法纯化该融合蛋白。Western blot分析显示:该融合蛋白能与抗牛分枝杆菌阳性血清发生特异性反应,而与牛副结核病阳性血清不反应。用该纯化蛋白初步建立了间接ELISA方法,并检测了117份临床血清样本(其中67份为PPD阳性牛血清),阳性率为39.32%(46/117)份,与PPD皮试诊断的符合率为82.05%(96/117)。