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1.
新疆地区规模化奶牛场牛支原体流行病学调查 总被引：2，自引：0，他引：2
为了调查新疆地区规模化奶牛场牛支原体的感染情况,采用牛支原体间接ELISA和特异性PCR检测牛血清中的牛支原体抗体及病料中牛支原体核酸,共检测9个地区15个规模化奶牛场437份血清,肺脏、关节液及鼻腔黏液44份。结果显示,血清抗体阳性率为76.43%（334/437）,其中脐带血抗体阳性率为40.00%（4/10）;病料阳性率为40.91%（18/44）。检测结果表明,新疆地区大部分奶牛场存在牛支原体感染,部分奶牛场发生牛支原体肺炎及关节炎病例,并存在垂直传播的风险。牛支原体感染可能成为危害新疆规模化奶牛场犊牛健康的主要疫病之一。 相似文献
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牛支原体疫苗的研究进展 总被引：2，自引：2，他引：0
牛支原体（Mycoplasma bovis,M.bovis）是一种造成全世界范围内育肥牛和奶牛多种疾病综合征的重要病原体。临床症状主要包括长期性肺炎及多发性关节炎（CPPS）、呼吸道疾病综合征（BRD）、乳腺炎及生殖器官疾病。牛支原体能感染多种组织和器官,也能从健康的牛体内分离,是威胁畜牧业生产的主要病原体。由于临床上抗生素治疗效果不佳,预防或控制牛支原体感染最好的选择是研发有效的商业可用的疫苗。牛支原体疫苗的研究已历经多年,虽然存在很多问题,但也取得了一定进展。文章对牛支原体弱毒疫苗、灭活疫苗及亚单位疫苗的研究进展进行了总结,并讨论了疫苗设计的优化方案,为合理设计和研发有效的牛支原体防控技术提供参考。 相似文献
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牛支原体（M. bovis）是牛的重要病原体,能够引起牛的乳腺炎、关节炎和肺炎等疾病。牛支原体已在全世界传播,对全球范围内的养牛业产生了重大不利影响。目前,由于牛支原体的耐药性有增强的趋势,同时缺乏有效的疫苗和治疗方法,这严重阻碍了牛支原体感染的控制。预防、控制甚至清除牛支原体最好的方法是使用有效的疫苗。虽然目前我国还没有获得新兽药证书的牛支原体疫苗,但相关的研究已取得了一定的进展。本文根据多年来牛支原体疫苗研究的相关资料,从牛支原体感染与免疫机制、灭活疫苗、弱毒活疫苗、基因工程疫苗四个方面进行综述,以期为今后牛支原体疫苗的研制提供思路及参考。 相似文献
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[目的]为了解凯里市A奶牛场牛呼吸道疫病的流行情况。[方法]本次采用酶联免疫吸附试验（ELISA）对142 份奶牛血清样品进行牛呼吸道合胞体病毒（BRSV）、牛病毒性腹泻（BVDV）、牛传染性鼻气管炎（IBRV）、牛支原体感染抗体检测。[结果]凯里市A奶牛场4 种呼吸道疾病病原普遍存在,BRSV阳性率为41.5%（59/142）;BVDV阳性率为40.1%（57/142）;IBRV阳性率为45.0%（64/142）;支原体阳性率为33.8%（48/142）。犊牛4 种呼吸疾病单一感染率为15.1%~45.5%;BVDV感染最严重,为45.5%;成年牛单一感染率为33.8%~45.1%。此外,奶牛场存在IBRV、BRSV、BVDV、支原体多种病原体混合感染,二重感染率为18.3％~30.2％;BVDV+IBRV感染率最高,为30.2%;三重感染以BVDV+支原体+IBRV感染率最高,为19.7%;四重感染率为9.9%。[结论]凯里市A奶牛场存在4 种疫病,有不同程度的病原体感染,应加强对上述病原体的流行控制,提高奶牛场管理水平,减少奶牛场疫病发生和经济损失。 相似文献
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《畜牧与兽医》2016,(6):20-24
为了筛选一种可靠的PCR方法作为实验室日常快速检测支原体的手段,利用实验室保存的17种55株支原体和13种16株细菌,对5种已发表的支原体属特异性PCR方法进行了评估和筛选。结果显示,方法 4(使用引物对RNA5和MGSO扩增1 021 bp的16S rRNA基因片段)具有良好的特异性,能够完全区分支原体与非支原体细菌,以牛支原体为扩增对象,最低能够检出10 pg牛支原体基因组DNA以及浓度为100 ccu/m L的牛支原体培养物。利用该PCR方法和支原体分离方法检测人工感染牛支原体样品和临床采集样品73份,二者符合率为93.15%。结果表明方法4可作为实验室支原体常规快速检测方法。 相似文献
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[目的]为了了解山东省滨州地区及周边规模化牛场支原体流行的基本情况,[方法]对该区域14个规模化奶牛场和肉牛场进行了血清采集,采用牛支原体IgG抗体ELISA检测方法进行牛支原体血清抗体检测,并使用SPSS 20进行卡方统计学检验。[结果]山东省规模化牛场的牛支原体IgG抗体总阳性率为77.86%,奶牛场的阳性率为74.29%,肉牛场的阳性率为81.43%,奶牛场牛支原体IgG阳性率与肉牛场的统计学差异不显著（P>0.05）;奶牛场不同场区,肉牛场不同场区之间,牛支原体的IgG阳性率差异显著（P<0.05）。[结论]山东省滨州地区及周边规模化牛场存在不同程度的牛支原体感染,牛支原体感染可能成为危害国内规模化奶牛场奶牛健康的主要疫病之一。 相似文献
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试验旨在研究牛支原体（Mycoplasma bovis,M.bovis）武威株二氢硫辛酰胺转乙酰酶（PDHc-E2）基因序列特征及其在牛支原体细胞中的位置。参照GenBank中牛支原体HB0801株pdhc基因（登录号：CP002058.1）设计引物,应用PCR扩增获得牛支原体武威株pdhc基因,在测序及序列分析的基础上,应用Overlap PCR完成点突变后将其克隆至pET-28a（+）中,构建原核表达载体pET-pdhc。pET-pdhc转化大肠杆菌Rosetta（DE3）感受态细胞后经IPTG诱导获得融合蛋白,将纯化蛋白免疫新西兰兔制备多抗血清,应用iELISA和Western blotting对牛支原体武威株PDHc-E2在细胞内的分布进行初步研究。结果显示,牛支原体武威株pdhc基因CDS全长735 bp,编码244个氨基酸,与国内牛支原体分离株HB0801、Hubei-1、CQ-W70、NM2012等基因序列完全一致,与国际标准株PG45同源性为99.2%,与无乳支原体（M.agalactiae）同源性为90.9%~91.2%,与加利福尼亚支原体（M.californicum）ST6株的同源性仅为78.4%,基因序列非常保守;通过Overlap PCR将该基因中4个编码色氨酸的TGA密码子突变为TGG,且完成点突变后的基因在大肠杆菌中成功表达,重组蛋白大小约为29 ku,主要以可溶性形式存在,iELISA结果显示,重组蛋白PDHc-E2具有较高的免疫原性,可刺激新西兰兔产生高水平的抗体,血清效价高达1：100 000;亚细胞定位结果表明,制备的多抗血清与重组蛋白PDHc-E2、牛支原体全菌蛋白、牛支原体膜蛋白、牛支原体胞浆蛋白均能发生特异性结合,说明该蛋白在牛支原体细胞膜和细胞质中均有分布,为膜相关蛋白,但在细胞质中的分布多于细胞膜。本研究结果为进一步研究牛支原体的生物学功能提供了理论依据。 相似文献
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试验旨在确定牛支原体P48基因的免疫原性，为进一步筛选牛支原体免疫保护性基因奠定基础。本研究以牛支原体新疆分离株为研究对象，运用Overlap PCR方法扩增得到点突变后的牛支原体新疆分离株P48基因，构建原核表达载体pET-32a （+）-P48，转化大肠杆菌BL21（DE3）感受态细胞，在诱导剂ITPG的诱导下获得重组蛋白P48，纯化后的重组P48蛋白免疫BALB/c小鼠制备多克隆抗体，运用Western blotting和ELISA方法验证其反应原性和免疫原性。结果表明，试验成功构建原核表达载体pET-32a （+）-P48，重组蛋白P48大小约为66 ku，纯化后的牛支原体P48重组蛋白免疫小鼠后可产生良好的免疫反应，血清抗体滴度达到较高水平（D450 nm值为1.126）。Western blotting结果显示，抗牛支原体P48重组蛋白的鼠血清与牛支原体P48重组蛋白及牛支原体全菌蛋白抗原均能产生明显的抗原抗体反应，表明P48重组蛋白具有良好的免疫原性与反应原性，可作为牛支原体新型疫苗的候选基因，且牛支原体新疆分离株P48基因与国内外5株牛支原体P48基因的同源性很高，亲缘关系较近。 相似文献
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Application of an indirect ELISA to milk samples to identify cows with Mycoplasma bovis mastitis 总被引：1，自引：0，他引：1
Byrne WJ Ball HJ Brice N McCormack R Baker SE Ayling RD Nicholas RA 《The Veterinary record》2000,146(13):368-369
An indirect ELISA was used to detect antibodies to Mycoplasma bovis in milk samples collected from a herd with M bovis mastitis. Antibodies were detected in samples from nine cows which had developed clinical M bovis mastitis. Milk from only three consistently antigen-negative cows tested positive for M bovis antibodies. These results indicate the potential value of the indirect ELISA for the detection of cows which have recently developed M bovis mastitis during the early stages of an outbreak. 相似文献
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Punyapornwithaya V Fox LK Hancock DD Gay JM Wenz JR Alldredge JR 《Preventive veterinary medicine》2011,98(1):74-78
The objective was to determine the incidence and transmission of mycoplasma mastitis in the hospital pen in a dairy herd of 650 lactating cows after a hospital pen was established following an outbreak of this disease. Mycoplasma mastitis status was monitored for 3 months through repeated collection of milk samples from cows with clinical mastitis (CM) and from bulk tank milk. During the outbreak 13 cows were diagnosed with Mycoplasma bovis CM, 1 cow with Mycoplasma sp. mastitis and 8 cows showed signs of arthritis, 3 of which were confirmed as having M. bovis arthritis. M. bovis isolates from cows with CM, arthritis and bulk tank milk had indistinguishable chromosomal digest pattern fingerprints. Incidence rates of M. bovis CM cases in the milking and hospital pens were 0.01 and 1.7 cases per 100 cow-days at risk. Approximately 70% of cows with M. bovis CM became infected within 12 days of entering the hospital pen. Transmission of M. bovis in the hospital pen occurred as 3 episodes. Each episode corresponded to the introduction of a cow with M. bovis CM from a milking pen. Evidence indicates that cows with M. bovis CM from milking pens were the source of transmission of the disease in the hospital pen and thus their presence in the hospital pen appeared to be a risk factor for transmission of M. bovis mastitis in this single case study herd. 相似文献
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Development of a real-time PCR for detection of Mycoplasma bovis in bovine milk and lung samples. 总被引：1，自引：0，他引：1
Hugh Y Cai Patricia Bell-Rogers Lois Parker John F Prescott 《Journal of veterinary diagnostic investigation》2005,17(6):537-545
A real-time polymerase chain reaction (PCR) assay using hybridization probes on a LightCycler platform was developed for detection of Mycoplasma bovis from individual bovine mastitis milk and pneumonic lung tissues. The detection limit was 550 colony forming units (cfu)/ml of milk and 650 cfu/25 mg of lung tissue. A panel of bovine Mycoplasma and of other bovine-origin bacteria were tested; only M. bovis strains were positive, with a melting peak of 66.6 degrees C. Mycoplasma agalactiae PG2 was also positive and could be distinguished because it had a melting peak of 63.1 degrees C. In validation testing of clinical samples, the relative sensitivity and specificity were 100% and 99.3% for individual milks and 96.6% and 100% for the lung tissue. Using M. bovis real-time PCR, the M. bovis culture-positive milk samples were estimated to contain between 5 x 10(4) and 7.7 x 10(8) cfu/ml and the M. bovis culture-positive lungs between 1 x 10(3) and 1 x 10(9) cfu/25 mg. Isolation, confirmed with the real-time PCR and colony fluorescent antibody test, showed that at the herd level, the proportion of samples positive for M. bovis isolation in mastitis milk samples submitted to the Mastitis Laboratory, Animal Health Laboratory, University of Guelph, Ontario, Canada, was 2.4% (5/201). We conclude that this probe-based real-time PCR assay is a sensitive, specific, and rapid method to identify M. bovis infection in bovine milk and pneumonic lungs. 相似文献
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Ghazaei C 《Journal of the South African Veterinary Association》2006,77(4):222-223
Mycoplasmas are an important and economically significant cause of mastitis in dairy cows in various parts of the world. The organisms are highly contagious, with the main reservoir of infection originating from cows with subclinical mastitis. In 1998 the 1st cases of bovine mastitis due to Mycoplasma bovis were diagnosed in Ardabil State, Iran. An investigation was carried out with the aim of establishing the extent of mycoplasma infections in dairy cows in Ardabil State. Milk samples obtained from 80 cows with clinical mastitis were cultured in the laboratory for the presence of mycoplasmas. Similarly, 48 bulk-tank milk samples were examined for the presence of mycoplasmas. A modified Hayflick broth was used to isolate the mycoplasmas and an immunoperoxidase test used for the species identification of the isolates. Mycoplasma bovis was isolated from 39 (48.75%) of the clinical mastitis samples and from 48 of the bulk-tank milk samples tested. This indicated that mycoplasma udder infections were more prevalent in dairy cows in Ardabil State than previously thought. 相似文献
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Passchyn P Piepers S De Meulemeester L Boyen F Haesebrouck F De Vliegher S 《Research in veterinary science》2012,92(2):219-220
Mycoplasma bovis (M. bovis) is a highly infectious pathogen of cattle causing pneumonia, polyarthritis, otitis, and less frequently, subcutaneous abscesses, abortions and meningitis. Ineffective drugs treatments, culling of infected cows and loss of milk production can lead to significant economic loss on dairy farms. The early detection of cows excreting M. bovis bacteria to prevent mastitis outbreaks is warranted. Reports suggest that the risk of M. bovis mastitis is higher in larger dairy herds. The objective of this study is to estimate the herd-level prevalence of M. bovis in Flanders, Belgium by culturing bulk tank milk samples taken from dairy farms. Three bulk tank milk samples per dairy herd were taken over four weeks, with collection intervals of two weeks. Culturing was done after pre-incubation using modified Hayflicks media to increase the chances of recovery of bacteria. For the identification of M. bovis, tDNA intergenic spacer PCR was used. In three herds (1.5%) of the 200 herds sampled, M. bovis was isolated from one of the three consecutive bulk tank milk samples. We conclude that in Flanders in 2009 at least 1.5% of the dairy herds had one or more cows excreting M. bovis in the milk. The frequent monitoring of bulk tank milk to detect the presence of M. bovis, especially in expanding herds on farms that often purchase replacement animals, should be encouraged in order to detect the presence of M. bovis and to monitor the success of control procedures following an outbreak of mycoplasmal mastitis in the herd. 相似文献
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Detecting Mycoplasma bovis in milk by enzyme-linked immunosorbent assay, using monoclonal antibodies 总被引：1，自引：0，他引：1
J T Boothby R Mueller D E Jasper C B Thomas 《American journal of veterinary research》1986,47(5):1082-1084
Enzyme-linked immunosorbent assay, using monoclonal antibodies, was used to detect Mycoplasma bovis in milk samples from a dairy experiencing an epizootic of mastitis. This method was specific (100%) for M bovis. Broth enrichment increased the sensitivity from 65% to 86%, compared with standard culture methods. 相似文献
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Mycoplasma species and related organisms isolated from ruminants in Britain between 1990 and 2000 总被引：1，自引：0，他引：1
Between 1990 and 2000, more than 1600 mycoplasmas and the related acholeplasmas were identified from ruminant animals by the Mycoplasma Group at the Veterinary Laboratories Agency--Weybridge. Mycoplasma bovis was the most commonly identified pathogen, mostly from pneumonic calves but occasionally from cattle with mastitis and arthritis. Mycoplasma canis was first isolated in Britain in 1995 from pneumonic calves and the number of isolates increased to 18 per cent of the total mycoplasmas isolated from cattle in 1999. The ELISA for antibodies to M. bovis detected 1971 positive samples (22 per cent) among 8959 serum samples, mainly from pneumonic cattle. Other mycoplasmas identified included Mycoplasma dispar from the lungs of cattle with respiratory disease, and Mycoplasma bovigenitalium from the reproductive tract of cows with vulvovaginitis and infertility. Mycoplasma bovirhinis and Acholeplasma species were found commonly but are thought to be more opportunistic than pathogenic. In sheep and goats, the majority of Mycoplasma species isolated were identified as Mycoplasma ovipneumoniae from pneumonic sheep, Mycoplasma conjunctivae from sheep with keratoconjunctivitis, and the ubiquitous Mycoplasma arginini. 相似文献
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Development of a novel biochip for rapid multiplex detection of seven mastitis-causing pathogens in bovine milk samples 总被引：2，自引：0，他引：2
Kuo-Hua Lee Jai-Wei Lee Shih-Wen Wang Lu-Yuan Liu Mei-Fen Lee Shih-Te Chuang Yih-Min Shy Chu-Li Chang Ming-Che Wu Chau-Hwa Chi 《Journal of veterinary diagnostic investigation》2008,20(4):463-471
To efficiently prevent and treat bovine mastitis and minimize its effect on the dairy industry, a sensitive, rapid, and specific test is required for identifying the mastitis-causing pathogens. In this study, a biochip capable of detecting 7 common species of mastitis-causing pathogens, including Corynebacterium bovis, Mycoplasma bovis, Staphylococcus aureus, and the Streptococcus spp. S. agalactiae, S. bovis, S. dysgalactiae, and S. uberis, within 6 hr was developed. The technique is based on DNA amplification of genes specific to the target pathogens and consists of 4 basic steps: DNA extraction of bacteria, polymerase chain reaction, DNA hybridization, and colorimetric reaction. To examine the accuracy and specificity of this biochip, a preliminary test with 82 random quarter milk samples were analyzed and compared with results from conventional microbiological methods conducted simultaneously. Results from all but 1 sample analyzed by the biochip were in agreement with those analyzed by bacteriology. The biochip could be a feasible tool for rapidly diagnosing mastitis-causing pathogens in milk and providing information for a more effective treatment to cure mastitis. 相似文献
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H Bocklisch S Kreusel A Bry? H Pfützner 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1991,38(5):385-390
Twelve freshly lactating ewes were experimentally infected with 2 Mycoplasma (M.) bovis strains via the teat canal in the left udder. The M. bovis infection produced a febrile clinical mastitis in all infected animals. M. bovis could be re-isolated regularly from the experimentally infected udder halves and the infection spread to the other halves. Some contact animals and 4 suckling lambs became naturally infected. Antibody titres were detected by means of the indirect hemagglutination test in blood sera 2 to 3 weeks post infectionem. The pathological lesions were similar to those of the M. bovis mastitis of cows. By the end of the trial the ewes had recovered from the clinical mastitis. 相似文献
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Isolation of Mycoplasma bovis from bovine clinical mastitis cases in Northern Greece 总被引：1，自引：0，他引：1
Filioussis G Christodoulopoulos G Thatcher A Petridou V Bourtzi-Chatzopoulou E 《Veterinary journal (London, England : 1997)》2007,173(1):215-218
Mycoplasma bovis was detected in 18/219 (8.2%) quarter milk samples collected from cases of bovine clinical mastitis in Northern Greece between November 1997 and March 1999. The cases occurred in 2/37 (5.4%) of the herds examined. The micro-organism was isolated from bulk milk samples (BTS) from the two positive herds but was not isolated from 111 composite milk samples collected from clinically healthy cows from all 37 herds. Isolates were identified as M. bovis by polymerase chain reaction (PCR) assay. Other micro-organisms were also isolated from the M. bovis positive samples. The M. bovis-positive cows had all been imported into Greece from other European countries. 相似文献