牛传染性胸膜肺炎抗体间接ELISA检测方法的建立

旨在建立一种检测牛传染性胸膜肺炎（CBPP）血清抗体的间接ELISA方法,用于该病的监测。利用生物信息学软件对CBPP的病原丝状支原体丝状亚种国内分离株Ben-1株的全基因组进行分析,选取脂蛋白rP0308作为包被抗原,通过一系列反应条件的优化,建立了基于rP0308蛋白的间接ELISA抗体检测方法,并对其性能进行评价。结果显示该方法的敏感性为92%,特异性为96%,与牛支原体、牛鼻支原体、无乳支原体、口蹄疫、牛病毒性腹泻、牛传染性鼻气管炎和牛结核分枝杆菌等阳性血清均不发生交叉反应,批内变异系数在2.41%~6.03%,批间变异系数在2.94%~6.59%,重复性良好。利用本方法和商品化的cELISA试剂盒分别对实验室保存的1 648份临床样品进行检测,该方法与商品化试剂盒的阴性符合率为89.1%,阳性符合率为79.2%,总符合率为88.7%。以上研究结果表明,本研究建立的间接ELISA方法具有良好的敏感性、特异性和重复性,具有良好的临床应用前景。     

猪传染性胸膜肺炎诊断方法研究进展

猪胸膜肺炎是国际上公认的危害现代养猪业的五大疾病之一。在我国,其血清阳性率呈上升趋势。研究快速、科学、准确的诊断方法对于本病的监测和控制具有很大的意义。本文从病原学、血清学和分子生物学等几个方面对该病的诊断研究进展作了综述。     

Contagious Bovine Pleuropneumonia in Tanzania: Current Status

CBPP reappeared in Arusha, Northern Tanzania in 1990, having been introduced from Kenya. The disease spread rapidly to Mara region through rustling of sick or infected animals. In November 1992, an unrelated outbreak occurred in Kagera, having spread from Southern Uganda. Up to the end of December 1994, the disease appeared to be confined to Kagera and Arusha. In January 1995, CBPP was observed in Morogoro region, south of the central railway line. Thereafter, the disease spread through western Tanzania. More recently, further disease has occurred in the Southern Highlands and Central regions. The contaminated area now stretches roughly between latitudes 1° and 9°S and longitudes 30° and 37°E, with a cattle population of about 10 million. The direct losses incurred as a result of animal mortality, and vaccination campaign and disease surveillance costs have been assessed at over US$11 million. Indirect losses resulting from chronic disease are much more difficult to assess but are believed to be even higher. Control of the disease has been through restricting animal movements and a mass vaccination campaign. Uncontrolled animal movement during transhumance, trade, cattle thefts and vaccination breakthroughs facilitated the spread of the disease.     

山羊传染性胸膜肺炎研究进展

山羊传染性胸膜肺炎是由山羊支原体山羊肺炎亚种引起的一种山羊常见的高度接触性呼吸道传染病,有传染快速,发病率、死亡率高的特点,给山羊养殖业带来巨大损失。本文从病原学、流行病学、临床症状以及诊断与防控方面对山羊传染性胸膜肺炎进行综述,帮助读者更加全面、深入的了解本病,以期为山羊传染性胸膜肺炎的诊断和防治提供新思路。     

Prevalence of contagious bovine pleuropneumonia (CBPP) in northern Nigeria

An abattoir study on the prevalence of contagious bovine pleuropneumonia (CBPP) in five cattle-producing states of Nigeria from 1988 to 1997 was carried out. A total of 1,936,015 slaughtered cattle was examined for characteristic CBPP lung lesions. The overall lesion-based prevalence of CBPP was 0.29% (95% CI 0.24, 0.35). The prevalence varied significantly (P<0.05) by state but not across the years. A total of 279 CBPP outbreaks occurred and overall vaccination coverage was only 9.7%, both varied over the years and across the states. The reasons for inadequate vaccination coverage for CBPP as well as the need for re-establishment of a national CBPP control programme are suggested.     

Danofloxacin (Advocin) reduces the spread of contagious bovine pleuropneumonia to healthy in-contact cattle

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides SC (MmmSC), is one of the most important diseases of cattle in Sub-Saharan Africa. The live T1/44 vaccine is normally used for its control but produces only transient protection and gives rise to adverse reactions. The present study evaluated the efficacy of danofloxacin (2.5% Advocintrade mark, Pfizer Ltd.) for the treatment of naturally infected cattle and in the prevention of CBPP transmission to in-contact cattle. Adult cattle, taken from a natural outbreak, were placed into two groups of 10 animals and kept on a research farm in paddocks 50m apart. One group was treated with 2.5mg/kg danofloxacin on days 0, 1 and 2; the other group were saline treated. On day 2, 10 CBPP-free, seronegative cattle were placed in contact with each of the two groups. All cattle were monitored for 3.5 months. No differences were seen in clinical improvement of the CBPP-affected cattle treated with danofloxacin compared with the untreated CBPP-affected cattle with approximately half of each group being withdrawn because of CBPP or showing CBPP lesions at post mortem examination. Clinical scores of the two groups were also similar. However cattle kept in contact with the danofloxacin-treated CBPP-affected animals showed significantly fewer lesions, less mortality and fewer animals were seropositive (P<0.02) and had reduced clinical scores (P<0.001) compared to cattle kept in contact with untreated CBPP-affected cattle. MmmSC was also isolated from fewer contact controls kept with the treated group. These findings could have important implications for the control of CBPP in Africa.     

由丝状霉形体山羊亚种引起的羊传染性胸膜肺炎诊断方法的研究进展

羊传染性胸膜肺炎对养羊业造成了一定的经济损失,笔者综述了由丝状霉形体山羊亚种引起的羊传染性胸膜肺麦的诊断方法,旨在为控制和预防该病奠定基础.     

猪传染性胸膜肺炎与猪肺疫二联亚单位疫苗的免疫效果评价

为了评价猪传染性胸膜肺炎与猪肺疫二联亚单位疫苗(由多杀性巴氏杆菌外膜蛋白H (OmpH)质粒导入胸膜肺炎放线杆菌(APP)菌影制备)的免疫效果,于某规模化猪场选取30头无上述病原菌和抗体的4周龄(w)仔猪,随机分为安全评估组和试验组进行免疫,试验组包括APP组、OmpH组、APP+OmpH联苗组,分别于4、6w肌肉注射...     

A competitive ELISA for the specific diagnosis of contagious bovine pleuropneumonia (CBPP)

A competitive ELISA, using a specific monoclonal antibody, was designed to detect antibodies to Mycoplasma mycoides subsp. mycoides SC, the agent of contagious bovine pleuropneumonia. One monoclonal antibody was found suitable for such a test, ‘117/5', it does not cross-react with any of the other mycoplasma species tested, furthermore, its binding is inhibited by positive sera. The cutoff, 50% of inhibition, was determined using a set of negative sera from CBPP-free areas. The sensitivity was controlled with sera from artificially infected animals as well as from sera from areas where CBPP is enzootic. In both cases, cELISA compared favorably with CFT. The precocity of detection was similar but cELISA detected more positives and the positive titers seemed to persist longer than in the case of CFT. Lysis of the antigen used to coat the ELISA plates reduced the variability of fixation and improved the repeatability of the test. A field evaluation is now in progress which will determine the true sensitivity and specificity of the test and also check if antibodies are detected after vaccination.     

Characterization of the in vitro core surface proteome of Mycoplasma mycoides subsp. mycoides, the causative agent of contagious bovine pleuropneumonia

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides (Mmm) is a severe cattle disease, present in many countries in sub-Saharan Africa. The development of improved diagnostic tests and vaccines for CBPP control remains a research priority. Polyacrylamide gel electrophoresis and mass spectrometry were used to characterize the Triton X-114 soluble proteome of nine Mmm strains isolated from Europe or Africa. Of a total of 250 proteins detected, 67 were present in all strains investigated. Of these, 44 were predicted to be lipoproteins or cytoplasmic membrane-associated proteins and are thus likely to be members of the core in vitro surface membrane-associated proteome of Mmm. Moreover, the presence of all identified proteins in other ruminant Mycoplasma pathogens were investigated. Two proteins of the core proteome were identified only in other cattle pathogens of the genus Mycoplasma pointing towards a role in host–pathogen interactions. The data generated will facilitate the identification and prioritization of candidate Mycoplasma antigens for improved control measures, as it is likely that surface-exposed membrane proteins will include those that are involved in host–pathogen interactions.     

丝状霉形体丝状亚种SC型脂蛋白LppQ N末端基因克隆与序列分析

脂蛋白LPPQ是丝状霉形体丝状亚种SC型(MmmSC)非洲株、欧洲株和疫苗株所特有的。LPPQ N末端域具有良好的免疫原性，在牛体内可诱导产生强大、特异、早期、持续的免疫反应。本研究根据已发表的LPPQ基因序列设计引物，用Pyrobest^TM高保真DNA聚合酶从MmmSC HVRI X株中扩增出了LPPQ N末端基因序列，并进行了克隆与序列测定。核苷酸序列比较结果显示，HVRI X株的LPPQ N末端基因序列与国外发表的序列同源性为99．7％，由其推导的氨基酸序列同源性为99，1％，为脂蛋白LPPQ N末端基因体外表达奠定了基础。     

丝状支原体丝状亚种SC型脂蛋白Q的N末端定点突变及表达载体的构建

根据已发表的丝状支原体丝状亚种SC型（MmmSC）LppQ基因序列设计引物，从MmmSC HVRIx株中扩增出了LppQN末端基因，并将其分别克隆到pUC18和pUC19上，利用一步重叠延伸PCR突变方法将其66位的TGA突变为TGG，经克隆与序列测定证实突变成功后，将已突变的LppQN末端基因插入原核表达载体pET32a的多克隆位点，成功地构建了LppQN末端基因原核表达载体pET32a-LppQ，为其下一步体外表达奠定了基础。     

Cellular and extracellular vesicular origins of miRNAs within the bovine ovarian follicle

The ovarian follicle components must provide an ideal environment to ensure the success of reproductive processes, and communication between follicular cells is crucial to support proper oocyte growth. Recently, it has been demonstrated that the presence of extracellular vesicles (EVs) carrying microRNAs (miRNAs) in follicular fluid represents an important autocrine and paracrine communication mechanism inside the ovarian follicle. In this study, we tested the hypothesis that the miRNA content of EVs isolated from ovarian follicular (granulosa and cumulus–oocyte complexes) cell‐conditioned culture media is dependent upon cell type. We initially screened bovine granulosa cells (GCs) and cumulus–oocyte complexes (COCs), as well as their derived EVs for 348 miRNAs using real‐time PCR, and detected 326 miRNAs in GCs and COCs cells and 62 miRNAs in GCs and COCs EVs. A bioinformatics analysis of the identified cell‐specific and differentially expressed miRNAs predicted that they likely modulate important cellular processes, including signalling pathways such as the PI3K‐Akt, MAPK and Wnt pathways. By investigating the origins of miRNAs within the follicular fluid, the results of this study provide novel insights into follicular miRNA content and intercellular communication that may be of invaluable use in the context of reproductive technologies, diagnostic of ovarian‐related diseases and/or the identification of biomarkers for oocyte and embryo quality.     

Animal Health and Disease Control in the Usangu Wetland of Southwestern Tanzania

The Usangu Wetland in the Southern Highlands of Tanzania has always been a major livestock production area. This paper describes the physical and social enviroment of these Plains before presenting a short history of the veterinary services in the area. The main part of the paper examines, through historical records and interviews with livestock owners and administrative officials, the history of the major diseases affecting livestock.