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通用基因芯片对奶牛乳房炎主要致病菌的检测 总被引：1，自引：0，他引：1
应用生物信息学手段和查阅文献资料设计了金黄色葡萄球菌、大肠埃希菌、无乳链球菌、绿脓杆菌、停乳链球菌、乳房链球菌6种奶牛乳房炎主要致病菌的通用引物和金黄色葡萄球菌、大肠埃希菌、绿脓杆菌的寡核苷酸探针及无乳链球菌、停乳链球菌、乳房链球菌的特异引物,并用这3种特异引物扩增片段的纯化产物作为这3种链球菌的检测探针。在引物对样品中细菌的相应基因片段扩增的同时进行靶基因的生物素标记,扩增的产物与硝酸纤维素膜上的探针进行杂交,酶联、显色后根据芯片扫描仪的判读结果来确定奶牛乳房炎致病菌感染的种类。结果表明,建立的以16S rDNA为对象的基因芯片技术可以快速的检测出以上6种细菌,整个检测过程需要6h-7h,灵敏度高,特异性好,能快速的对奶牛乳房炎的主要致病菌做出诊断。 相似文献
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奶牛乳房炎一直是困扰养牛业的顽疾。诱发奶牛乳房炎的致病菌通常包括金黄色葡萄球菌、无乳链球菌等致病菌,停乳链球菌、乳房链球菌、大肠埃希菌等环境致病菌以及表皮葡萄球菌、腐生葡萄球菌、模仿葡萄球菌等机会性致病菌。噬菌体是一类细菌依赖性的病毒,可通过内溶素抑制肽聚糖的合成,或利用穿孔素-内溶素系统水解肽聚糖。在体外试验中,噬菌体及其内溶素可溶解、抑制细菌生物膜,裂解乳房炎致病菌;在体内试验中,可清除感染动物体内的乳房炎致病菌,对奶牛乳房炎具有治疗效果。因此,噬菌体及其内溶素在奶牛乳房炎治疗方面具有良好的应用前景。 相似文献
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为研究西藏拉萨市林周县牦牛乳理化指标和乳房炎主要病原菌,采集牦牛乳样品100份,开展乳理化指标和主要病原菌检测。采用CMT法筛选隐性乳房炎病例,针对筛选出隐性乳房炎阳性乳样20份进行病原菌分离鉴定,共分离出11种52株细菌,通过鉴定,其中包含4株凝固酶阴性葡萄球菌,10株金黄色葡萄球菌,2株停乳链球菌,2株乳房链球菌,1株粪链球菌,8株无乳链球菌,6株大肠埃希菌,4株真菌。乳房炎63.5%的病例是由链球菌、葡萄球菌和大肠埃希菌引起,结果证实,链球菌、葡萄球菌和大肠埃希菌是导致林周县牦牛乳房炎的主要病原菌。 相似文献
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为了解某奶牛场乳房炎主要致病菌和合理用药,该试验对乳房炎奶牛取样1 15份,开展了分离鉴定和药敏试验.结果表明,该场主要致病菌为金黄色葡萄球菌、无乳链球菌、停乳链球菌、大肠埃希菌、乳房链球菌和沙门菌;主要病原菌均对头孢喹诺和左氧氟沙星高度敏感. 相似文献
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《动物医学进展》2016,(12)
为了解吉林省各地区奶牛乳房炎发病情况,以便于科学合理防治该病,对长春市、吉林市、白城市以及延吉等多个大型奶牛场200份乳样进行了细菌分离与鉴定,并对其主要致病菌进行敏感药物筛选试验。结果显示,导致吉林省奶牛乳房炎发生的主要病原菌为大肠埃希菌、金黄色葡萄球菌、无乳链球菌以及停乳链球菌等,在检出的267株病原菌中,金黄色葡萄球菌98株,占36.70%;无乳链球菌65株,占24.34%;停乳链球菌57株,占21.35%;大肠埃希菌43株,占16.10%;其他菌4株,占1.5%。4种病原茵对丁胺卡那霉素、环丙沙星、恩诺沙星、头孢噻肟以及万古霉素均高度敏感,但对青霉素、链霉素、红霉素、庆大霉素等具有较强的耐药性,因此丁胺卡那霉素、环丙沙星、恩诺沙星、头孢噻肟以及万古霉素可以作为吉林省奶牛乳房炎治疗的首选药物。 相似文献
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"乳康2号"对奶牛细菌性乳房炎的疗效研究 总被引：2，自引：0，他引：2
用＂乳康2号＂治疗临床型乳房炎奶牛37头。结果表明,治愈28头,有效6头,无效3头,总有效率为91.89%,平均治疗时间4.7d。对采集的37头奶牛治疗前的42个乳区奶样,进行细菌学检查,有33个乳区检出与乳房炎有关的病原菌,占78.57%。检出病原菌5种,主要为无乳链球菌、停乳链球菌、金黄色葡萄球菌、乳房链球菌和大肠埃希菌。停药后采集33个乳区奶样,结果有14个乳区细菌转阴,细菌转阴率为42.42%。在转阴的病原菌中,大肠埃希菌转阴率最高（100%）,其次是混合感染（60.00%）和乳房链球菌（50.00%）。 相似文献
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2023年3月,青海省西宁市某牦牛养殖场部分牦牛出现乳房炎,为确定发病原因,从该场采集确诊乳房炎奶牛乳样15份,进行细菌分离、纯化致病菌及PCR鉴定,并采用纸片扩散法检测部分致病菌对抗菌药物的敏感程度。结果发现,从13份乳样中共分离获得23株细菌,分别为无乳链球菌(n=7)、金黄色葡萄球菌(n=8)、大肠埃希氏菌(n=4)、粪肠球菌(n=3)和蜡样芽胞杆菌(n=1)。对无乳链球菌、金黄色葡萄球菌和大肠埃希氏菌进行药敏试验,结果发现,3种病原菌对链霉素、环丙沙星、氧氟沙星、头孢噻呋和头孢拉定高度或中度敏感。研究结果提示,无乳链球菌、金黄色葡萄球菌和大肠埃希氏菌是该场牦牛乳房炎的主要致病菌,且头孢类抗生素和恩诺沙星等可作为首选药物用于该病的防治。 相似文献
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奶牛乳房炎主要致病菌16S rDNA基因芯片检测方法的建立 总被引：1，自引：0，他引：1
本研究建立了快速检测奶牛乳房炎主要致病茵的基因芯片方法,试验以奶牛乳房炎4种主要致病菌的16SrDNA基因作为基因芯片检测的靶片段,设计和筛选通用性引物和特异性探针,对通用引物进行荧光标记、PCR扩增、芯片杂交和信号扫描分析,根据杂交信号强度和聚类分析结果,判定奶牛乳房炎感染的致病菌种类。实验结果显示：以16SrDNA为靶基因检测奶牛乳房炎主要致病菌（无乳链球菌、肺炎克雷伯氏菌、奇异变形杆菌、金黄色葡萄球菌）的基因芯片方法,可以快速、特异、准确地对这4种试验菌株进行检测和鉴定。该检测方法的建立将为流行病学调查、食品卫生监督、乳房炎的防控等提供快速、有效的检测方法,具有良好的市场应用前景。 相似文献
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多重PCR快速检测奶牛乳房炎3种主要病原体 总被引：10，自引：0，他引：10
奶牛乳房炎是引起奶牛业经济损失的一种重要疫病,目前还没有快速、特异检测奶牛乳房炎主要致病原的方法。本试验根据金黄色葡萄球菌、无乳链球菌、大肠杆菌各自保守的16S或23S rRNA基因序列,合成了3对特异性引物,建立了三重PCR检测方法。特异性试验表明,该方法对所有参与测试的金黄色葡萄球菌、无乳链球菌和大肠杆菌都能扩增出各自的阳性条带,而对所有参与测试的对照菌株则不能扩增出任何条带。敏感性试验表明该方法能检测到4个菌的金黄色葡萄球菌、无乳链球菌和2个菌的大肠杆菌。对送检的乳房炎奶样36份直接进行PCR检测,金黄色葡萄球菌阳性7份,无乳链球菌阳性2份,大肠杆菌阳性6份。 相似文献
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Accurate identification of mastitis pathogens is often compromised when using conventional culture-based methods. Here, we report a novel, rapid assay tested for speciation of bacterial mastitis pathogens using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time PCR amplification of 16S rRNA gene fragment, spanning the variable region V5 and V6 was performed with a resulting amplicon of 290bp. First, a library was generated of melt curves of 9 common pathogens that are implicated in bovine mastitis. Six of the isolates, Escherichia coli, Streptococcus agalactiae, Klebsiella pneumoniae, Streptococcus uberis, Staphylococcus aureus and Mycoplasma bovis, were type strains while the other 3, Arcanobacterium pyogenes, Corynebacterium bovis and Streptococcus dysgalactiae, were bovine mastitis field isolates. Four of the type strains, E. coli, S. agalactiae, K. pneumoniae and S. aureus, were found to be of human origin, while the other 3 type strains were isolated from bovine infections. Secondly, the melt curves and corresponding amplicon sequences of A. pyogenes, E. coli, S. agalactiae, S. dysgalactiae, K. pneumoniae, S. uberis and S. aureus were compared with 10 bovine mastitis field isolates of each pathogen. Based on the distinct differences in melt curves and sequences between human and bovine isolates of E. coli and K. pneumoniae, it was deemed necessary to select a set of bovine strains for these pathogens to be used as reference strains in the HRMA. Next, the HRMA was validated by three interpreters analyzing the differential clustering pattern of melt curves of 60 bacterial cultures obtained from mastitis milk samples. The three test interpreters were blinded to the culture and sequencing results of the isolates. Overall accuracy of the validation assay was 95% as there was difficulty in identifying the streptococci due to heterogeneity observed in the PCR amplicons of S. uberis. The present study revealed that broad-range real-time PCR with HRMA can be used as a powerful, fast and low-cost tool for the differentiation of clinically important bacterial mastitis pathogens. 相似文献
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We previously showed that Staphylococcus aureus cells adhered mainly to an elongated cell type, present in cultures of bovine mammary gland cells. Moreover, we showed that this adhesion was mediated by binding to fibronectin. The same in vitro model was used here, to study adhesion of other important mastitis pathogens. Like the S. aureus strains, the Streptococcus dysgalactiae strains adhered mainly to elongated cells, which seemed to be mediated by fibronectin binding. In contrast, Streptococcus uberis strains adhered mainly to cubic cells. Since the cubic cells did not express fibronectin and S. uberis cells bound fibronectin less efficiently, the adhesion of S. uberis cells was independent of fibronectin binding. Streptococcus agalactiae strains adhered poorly to both cell types. The specificity and efficiency of adhesion of Escherichia coli strains was strongly strain dependent. None of the S. agalactiae and E. coli strains tested was able to bind fibronectin efficiently. The results suggest that the different mastitis pathogens have different target cell specificities and use different mechanisms to adhere to cells of the bovine mammary gland. 相似文献
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Antimicrobial resistance in streptococcal species isolated from bovine mammary glands 总被引：1，自引：0，他引：1
Streptococcal species isolated from dairy cows with clinical mastitis were obtained from mastitis research workers in Florida, Louisiana, New York, Vermont, Washington, and West Virginia. Seventy-one streptococcal isolates were tested, including 39 strains of Streptococcus agalactiae, 21 strains of S dysgalactiae, and 11 strains of S uberis. The minimal inhibitory concentration of erythromycin, lincomycin, oxytetracycline, penicillin, spectinomycin, streptomycin, and tetracycline was determined for each isolate. Differences were not detected among strains with respect to geographic origin. None of the strains was resistant to penicillin. Lincomycin was the next most effective antimicrobial, with only 2 resistant strains of each streptococcal species. There were no differences among the streptococcal species with respect to resistance to either penicillin or lincomycin. Streptococcus uberis was more likely to be resistant to erythromycin than were S agalactiae and S dysgalactiae (P less than 0.02). Streptococcus agalactiae and S uberis had similar distributions for resistance to oxytetracycline, tetracycline, spectinomycin, and streptomycin. Strains of S dysgalactiae were more likely to have intermediate resistance to oxytetracycline and streptomycin than were strains of S agalactiae and S uberis, which were highly resistant to oxytetracycline and streptomycin (P less than 0.001). Differences were not detected among the streptococcal species with respect to resistance to spectinomycin. Resistance to multiple antimicrobials was observed in all streptococcal species tested. Although S dysgalactiae appeared to have a greater percentage of strains (73%) that were resistant to multiple antimicrobials than did S agalactiae (31%) or S uberis (45%), differences were not statistically significant. 相似文献
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A total of 1140 clinical cases of mastitis, with at least one inflamed quarter, were reported on 125 farms with somatic cell counts in bulk milk less than 150,000/ml. The average annual incidence was 17.9 cases per 100 cows and ranged from none to 80 cases per 100 cows. The microorganisms most frequently isolated were Escherichia coli (16.2 per cent), coagulase negative staphylococci (13.0 per cent), Staphylococcus aureus (9.6 per cent) and Streptococcus uberis (8.0 per cent). Only two cases of Streptococcus agalactiae were found. As the incidence of clinical mastitis increased, the proportion of S aureus also increased, while the proportions of E coli, S uberis and Streptococcus dysgalactiae remained about the same. Most of the clinical cases of mastitis occurred in early lactation, in November, December and January. However, after correction for the number of calvings per month, the incidence of mastitis was highest in the early summer months. 相似文献
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奶牛乳腺炎病原菌的分离鉴定及耐药性分析 总被引：8，自引：1，他引：8
2 0 0 2～ 2 0 0 3年采集上海地区 32个规模化奶牛场 433份奶样 ,共分离 2 54株细菌。其中包括金黄色葡萄球菌 48/2 54 (1 8 9% )、表皮葡萄球菌 34/2 54 (1 3 4% )、大肠杆菌 2 8/2 54 (1 1 0 % )、肺炎 /溶血巴氏杆菌 1 1 /2 54 (4 3 % )、无乳链球菌 2 5/2 54 (9 8% )、停乳链球菌 1 6/2 54(6 3 % )、乳房链球菌 1 5/2 54 (5 9% )及酵母菌 8/2 54 (5 9% )。表明葡萄球菌性乳腺炎仍占主导地位。药敏试验结果表明 ,分离菌株对临床常用的多种抗菌药物都产生了耐药性 ,耐药菌株的出现和抗生素的残留给乳腺炎的防治及食品安全带来了隐患 相似文献
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采集具有临床型乳腺炎奶牛的奶样,经细菌的分离培养、纯培养和生化试验,以确定奶牛临床型乳腺炎的致病菌种类。结果发现患临床型乳腺炎奶牛的致病菌和占分离菌株的百分率为:腐生葡萄球菌19.7%、金黄色葡萄球菌16.5%、兽疫链球菌17.3%、停乳链球菌9.4%、乳房链球菌3.9%、无乳链球菌1.6%、产气肠杆菌17.3%、聚团肠杆菌4.7%、奇异变形杆菌3.0%、大肠杆菌3.0%、蜡样芽胞杆菌3.0%,样品中单纯感染与混合感染的百分率分别为66.0%和27.8%。 相似文献