共查询到19条相似文献，搜索用时 218 毫秒
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《中国兽医学报》2014,(8):1261-1266
利用THB(Todd-Hewitt Broth)固体培养基和色素试验培养基选择培养无乳链球菌,参照GenBank中无乳链球菌参考菌株(Accession:AF015927.1、JQ289582.1)16SrRNA和种属特异性基因cfb(CAMP因子)序列设计引物,对奶样中分离的12株疑似无乳链球菌进行鉴定。结果显示,经PCR扩增后,被检测的12株细菌均可扩增出预期大小的16S rRNA基因序列,而12株中有8株可以扩增出预期大小的cfb基因序列,条带单一,特异性好。序列BLAST显示,12株菌的16S rRNA基因序列与NCBI上报道的无乳链球菌相应序列高度同源(>99.0%),各分离菌株间的16S rRNA基因序列也高度同源(99.0%～100.0%);cfb基因序列与NCBI上已报道的无乳链球菌相应序列具有高度同源性(>99.0%),各菌株间cfb基因序列也高度同源(100.0%)。经选择培养与PCR鉴定结果可以确定12株疑似菌株中有8株为无乳链球菌。 相似文献
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《中国预防兽医学报》2016,(6)
为分离及种属鉴定引起牛乳腺炎相关的链球菌和肠球菌,本研究于兰州及周边地区采集疑似奶牛乳腺炎乳样382份,通过THB(Todd-Hewitt Broth)固体选择培养基初步分离到67株疑似链球菌或疑似肠球菌。参照已发表文献合成链球菌属16S r RNA和16S~23S r RNA间隔区基因引物序列,扩增分离菌株16S~23S r RNA间隔区序列,产物分别利用AluⅠ和RsaⅠ单酶切消化,并以参考菌株的16S~23S r RNA酶切图谱为参考,对分离株进行限制性片段多态性(RFLP)分类分析;再选取各RFLP类群的任一菌株,扩增其16S r RNA基因并测序,并经NCBI核酸数据库进行比对,结果显示67株疑似链球菌中有53株为粪肠球菌(79.1%)、3株为屎肠球菌(4.5%)、3株为肠道肠球菌(4.5%)、8株为无乳链球菌(11.9%)。结果表明肠球菌属细菌(粪肠球菌、肠道肠球菌、屎肠球菌)与兰州市及周边地区奶牛乳腺炎的发病紧密相关,肠球菌属细菌与链球菌属细菌16S r RNA基因同源性很高,但可以通过分子生物学方法准确区分。 相似文献
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青海地区奶牛无乳链球菌的分离与鉴定 总被引：1，自引：0，他引：1
无乳链球菌是导致奶牛乳房炎的常见病原菌之一,为了掌握青海地区无乳链球菌的流行情况及常用药物的敏感性,从而为本地区更好的预防与治疗奶牛乳房炎提供保障。本试验在青海地区16个奶牛场共采集258份乳房炎乳样,经革兰氏染色与绵羊鲜血琼脂培养基培养纯化,获得疑似链球菌137株;随后经玻片法血浆凝固酶试验、CAMP试验、生化试验与动物致病性试验对无乳链球菌进行鉴定,结果显示,137株链球菌中含有无乳链球菌42株,分离率为30.7%;经K-B纸片扩散法对无乳链球菌耐药性检测结果显示,该地区无乳链球菌主要对头孢类高度敏感,对氧氟沙星、磺胺类、氨苄青霉素与阿米卡星中度敏感。 相似文献
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对从黑龙江省部分奶牛场患乳房炎奶牛乳汁中分离的10株疑似链球菌进行了培养特性、菌落形态、染色特性、生化试验等研究，确定5株为无乳链球菌，3株为停乳链球菌，2株为乳房链球菌。溶血试验结果表明，10株链球菌中有8株出现溶血现象，其中无乳链球菌有3株呈β溶血，2株呈α溶血；停乳链球菌均呈α溶血；乳房链球菌为y溶血。动物致病性试验结果表明，停乳链球菌BHI培养液腹腔接种小白鼠0.6ml,18-24后致死率达100%;接种无乳链球菌的小白鼠均有不同程度的发病,但未见死亡；乳房链球菌对小白鼠无致病性.药敏试验结果表明、链球菌对临床上常用的青、链霉素及磺胺类药物均已产生不同程度的耐药。而红霉素、喹诺酮类药物则表现出较好的抑菌效果。 相似文献
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本试验利用Blue Light Broth "Nissui"和MRS培养基对门源县饲喂羊的青贮饲料中的菌种进行了分离分纯和16S rRNA序列通用引物PCR扩增和测序鉴定,分离分纯得到12株菌,其中8株为地衣芽孢杆菌,1株为单纯芽孢杆菌,2株为干酪乳杆菌,1株为副乳杆菌。 相似文献
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《中国奶牛》2015,(6)
通过对甘肃省部分地区奶牛场的323份乳腺炎乳样进行无乳链球菌的分离鉴定及耐药性分析,旨在为临床合理用药提供科学依据。采用6%~8%绵羊鲜血琼脂平板培养基、选择性培养基(B族链球菌显色培养基)、普通营养琼脂培养基,结合革兰染色、镜检,对可疑菌落分离纯化后进行生化试验,确定分离出无乳链球菌;选用临床常用的10种抗生素对分离株进行药敏试验。结果表明,55株无乳链球菌分离株对青霉素、头孢曲松钠、四环素表现出一定程度耐药,耐药率分别为63.64%,36.36%,34.55%,而对左氧氟沙星、庆大霉素、环丙沙星、甲砜霉素高度敏感,敏感率分别为100%、92.73%、85.45%、69.09%。 相似文献
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以取自四川省不同地区的牦牛粪便、肠道内空物为材料，用MRS琼脂双层培养基进行厌氧培养，分离到50株乳酸菌，经生化鉴定为嗜热链球菌（2株）、乳酸乳球菌（1株）、保加利亚乳杆菌（5株）、嗜粪乳杆菌（10株）、嗜酸乳杆菌（8株）、乳酸乳杆菌（9株）、肠乳杆菌（10株）、弯曲乳杆菌（5株）。采用乳酸菌16 SrDNA通用引物，对分离的8种菌的16S rDNA-段可变区序列进行扩增，均得到大小约470bp的产物；扩增产物经纯化、测序后与GenBank中标准菌株的核甘酸序列比较，同源性均大于97．5％，同源性分析与生化试验的结果是一致的。证实，牦牛肠道和粪便的乳酸菌较为丰富，且乳杆菌的数量较多，这可能与牦牛复杂的生长环境有关。 相似文献
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湖北地区奶牛乳房炎病原菌的分离鉴定与耐药性分析 总被引：15，自引：1，他引：15
本研究旨在分离鉴定湖北地区致奶牛乳房炎的病原菌，鉴定细菌的耐药特征．为奶牛乳房炎防治提供理论依据。从湖北两个主要奶牛小区的9个不同规模的奶牛场采集牛奶样本200份，经检测．其中3份为临床乳房炎样本，49份为隐性乳房炎。两地区隐性乳房炎的检出率分别为36．4％（20／55）与20％（29／145）。两地区乳房炎牛奶的细菌种类分布也各不同。其中武汉东西湖地区停乳链球菌的分离率最高，占33.3％（8／24）．而宜昌夷陵区以金黄色葡萄球菌分离率最高，占29％（9／31），其次为无乳链球菌，占25．8％（8／31）。从乳房炎牛奶中共分离到7种细菌共55株，其中金黄色葡萄球菌11株，表皮葡萄球菌10株，无乳链球菌10株，停乳链球菌14株，乳房链球菌2株，大肠埃希氏菌4株，克雷伯氏菌4株。分别对所有分离株进行12种药物的敏感性试验，结果表明所有菌株对先锋V、氧氟沙星均高度敏感．对临床常用的青霉素、链霉素具有较高的耐药性。 相似文献
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Bovine mastitis caused by Mycoplasma agalactiae subsp. bovis was first diagnosed in 16 of 55 cows in an Ontario herd in Feburary 1972. A total of 182 of 598 (30.4%) cows from 33 of 64 (51.5%) farms in widely separated areas of the province were culturally positive. Herd incidence varied from 15 to 40% with one closed herd having an incidence of 61%. Four herds were investigated culturally and serologically by the growth inhibition test for 15 months. In the acute phase the organism was present in the milk in extremely high numbers and could still be isolated from a few cows after eight to 12 months. The sera from 89.5% of the animals with clinical mycoplasma mastitis produced a zone of surface "film" and/or colony inhibition and some cows remained positive for six to 12 months. The disease was experimentally reproduced with a pure culture of the organism isolated from the milk of a cow from one of the herds. 相似文献
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牛源性无乳链球菌血清型分布及抗生素耐药性研究 总被引：2，自引：1，他引：1
本研究旨在查明牛源性无乳链球菌血清型分布及对常见抗生素的耐药情况,指导临床合理用药。对从中国部分地区奶牛场采集的临床型乳房炎病牛乳中分离鉴定出78株无乳链球菌地方菌株,制备沉淀反应抗原及6株标准血清型无乳链球菌单因子血清抗体,采用环状沉淀试验,对78株无乳链球菌地方菌株进行了血清学分型鉴定;同时采用K-B纸片法测定了这些菌株对抗生素的耐药情况。结果表明,引起奶牛乳房炎的无乳链球菌血清型主要为X型(60.26%),其次为Ⅲ型(10.26%)、R型(7.69%)、Ⅱ型(7.69%)和Ⅰb型(5.13%),Ⅰa型尚未发现。无乳链球菌对目前临床上使用的大部分抗生素,如头孢唑啉、头孢噻肟、丁胺卡那霉素、卡那霉素、庆大霉素、四环素、强力霉素、氟苯尼考、多黏菌素B、环丙沙星、氟哌酸和头孢他啶/棒酸均较敏感;但对氨苄青霉素、链霉素、恩诺沙星、阿莫西林/棒酸和复方新诺明,有一定的耐药性,其耐药率达50%~100%。本研究对进一步研制有效的药物及疫苗,指导临床合理用药具有重要的意义。 相似文献
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为了掌握辽宁地区奶牛乳房炎主要细菌性病原种类及致奶牛乳房炎大肠杆菌药物敏感性特征,本研究选取辽宁地区某大型奶牛场75头临床表现为乳房炎的奶样进行细菌培养与分离,通过生化方法对分离到的细菌进行鉴定,并对分离到的奶牛源大肠杆菌进行体外药物敏感性试验研究。结果发现,该场奶牛乳房炎细菌性病原主要为大肠杆菌、葡萄球菌和链球菌,其检出率分别为58.7%、64.0%和54.7%,存在二重感染和三重感染等混合感染的情况。大肠杆菌药物敏感性试验结果表明,该场分离株对磺胺类药物（耐药率>85%）及氯霉素类药物（耐药率>30%）耐药性较高,对氨苄西林（9.5%）、环丙沙星（9.5%）、头孢噻呋（7.1%）和氧氟沙星（4.8%）比较敏感。上述研究结果为该地区奶牛乳房炎的防制提供可靠的理论依据。 相似文献
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通过LMT法、乳汁pH检验法和体细胞直接计数等方法相结合,对冀东地区3个大型奶牛场、8个奶牛养殖户选取的1 021头奶牛进行隐性乳房炎流行病学调查与病菌分离鉴定。结果表明,隐性乳房炎发病率为60.63%(619/1021),乳房炎阳性乳样品中细菌分离率达88.21%(546/619)。从543头隐性乳房炎患牛的阳性乳区乳样中分得细菌共4类14种菌82株,其中葡萄球菌36株,占43.90%;链球菌33株,占40.24%;肠杆菌类8株,占9.76%;棒状杆菌5株,占6.10%。冀东地区奶牛隐性乳房炎主要病原菌为金黄色葡萄球菌和无乳链球菌。 相似文献
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YAO Wei YU Xue-wu CAO Dong ZHAO Xiao-tong GU Gui-bo CHEN Yao ZHANG Lei LIU Yao-chuan FENG Jun-ke 《中国畜牧兽医》2016,43(11):3080-3084
In order to understand the main bacterial pathogen species causing dairy cow mastitis in Liaoning, as well as the characteristics of drug sensitivity of the pathogenic E.coli,the milk samples from 75 dairy cows with clinical manifestations for mastitis in certain large-scale dairy farm in Liaoning were collected.The bacteria in milk were cultured and isolated with biochemical methods and in vitro drug sensitivity tests were processed with the isolated E.coli strains.The results showed that the main bacterial pathogen for dairy cow mastitis were E.coli（separation rate 58.7%),S.aureus（64.0%）and S.agalactiae（54.7%),and multiple infection including double and triple infection were identified.The drug sensitivity tests on the isolated E.coli indicated that the E.coli isolates were highly resistant to sulfonamides（resistance rate>85%）and chloramphenicol（resistance rate>30%),and they were relatively low resistant to ampicillin（9.5%),ciprofloxacin（9.5%),ceftiofur（7.1%）and ofloxacin（4.8%).The results was able to provide reliable theoretical basis for prevention and control of dairy cow mastitis in Liaoning area. 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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K H Hoblet G D Schnitkey D Arbaugh J S Hogan K L Smith P S Schoenberger D A Todhunter W D Hueston D E Pritchard G L Bowman 《Journal of the American Veterinary Medical Association》1991,199(2):190-196
Nine dairy herds (mean size, 149 cows) with bulk-tank milk somatic cell counts of less than 300,000 cells/ml and greater than 80% of cows with Dairy Herd Improvement Association linear somatic cell counts less than or equal to 4 were selected for study. Each herd was monitored for 12 consecutive months. Duplicate quarter-milk specimens were collected from each cow for bacteriologic culturing at beginning of lactation, cessation of lactation, and at the time of each clinical episode of mastitis. Streptococcus agalactiae was never isolated and Staphylococcus aureus was isolated from less than 1% of all quarters. There were 554 episodes of clinical mastitis. During the year of study, the incidence rate of clinical mastitis varied from 15.6 to 63.7% of cows among the 9 herds. Mean costs per cow per year in herd for mastitis prevention were: $10 for paper towels, $3 for nonlactating cow treatment, and $10 for teat disinfectants. Mean cost associated with clinical mastitis was $107/episode. Approximately 84% ($90) of the costs attributed to a clinical episode were associated with decreased milk production and nonsalable milk. Costs of medication and professional veterinary fees per clinical episode varied significantly among the 9 herds. Three of the herds did not have a veterinarian treat a clinical episode of mastitis during the year of study even though 2 of these herds had the first and third highest incidence rates of clinical mastitis. When calculated on a per cow in herd basis, mean costs of $40/cow/year were attributed to clinical mastitis.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献