乳房炎奶牛金黄色葡萄球菌毒素基因的检测及PFGE分型研究     

王新  韦艺媛  张静  周婷  梁珍娟  杨保伟  席美丽  夏效东  孟江洪  俞英 《畜牧兽医学报》2011,42(7)

作者针对临床及亚临床乳房炎奶牛乳汁中金黄色葡萄球菌分离株的毒素基因进行检测和脉冲场凝胶电泳(PFGE)基因分型,比较2种类型乳房炎金黄色葡萄球菌分离株的差异.无菌法采集奶样,采用国际标准方法从中分离金黄色葡萄球菌,用多重PCR方法扩增nuc基因和mecA基因以确证金黄色葡萄球菌(SA)和耐甲氧西林金黄色葡萄球菌(MRSA).进一步用PCR方法检测SA的各种毒素基因(SEs、ETs、TSST 1和PVL基因等).利用限制性内切酶Sma Ⅰ对SA基因组DNA进行酶切和PFGE分析,最后利用BioNumerics软件进行聚类分析.结果:19.3％(23/119)的临床乳房炎奶样和14.8％(26/176)的亚临床乳房炎奶样确定为金黄色葡萄球菌阳性样品,分别从中分离鉴定出43株和26株金黄色葡萄球菌,其中临床乳房炎分离株中有5株为mecA基因阳性.临床乳房炎奶牛奶样中检测到SA的SEA、SEB、SED、SEJ和PVL毒素基因,检出率分别为3.8％(1株)、11.5％(3株)、19.2％(5株)、7.7％(2株)和31.2％(10株);亚临床乳房炎奶牛乳样中仅检测到SA的SEA和PVL毒力基因,检出率分别为7.0％(3株)和84.1％(37株).表明临床与亚临床乳房炎奶牛乳汁中SA菌株携带的毒素基因不一样,SEs可能是临床乳房炎菌株的重要致病基因,PVL可能是亚临床乳房炎菌株的重要致病基因.69株SA使用Sma Ⅰ酶切分型后,可分为7个大簇、50个基因型,来源相同的SA分型后大部分位于同一簇内.临床乳房炎奶牛乳汁中检测到MRSA菌株,PVL基因在亚临床乳房炎中的检出率为临床乳房炎的2.7倍.PFGE方法能较好的区分临床乳房炎和亚临床乳房炎的SA分离菌株.  相似文献   

Hepatozoon spp Infections in the United States     

Kelly E. Allen  Eileen M. JohnsonSusan E. Little  DVM  PhD 《Veterinary Clinics of North America: Small Animal Practice》2011,41(6):1221-1238

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几株桑天牛成虫肠道优势细菌的分离与鉴定   总被引：2，自引：1，他引：1  

袁秀洁  唐秀光  李会平  黄大庄  王达 《蚕业科学》2011,37(2):181-186

为了研究桑天牛成虫肠道微生态环境,探索破坏桑天牛营养利用等功能的生物防治新途径,对健康桑天牛成虫肠道的优势细菌进行分离鉴定。选择平皿上出现3个以上的菌落作为肠道优势细菌进行分离纯化,共获得6个菌株。6个菌株均呈杆状,革兰染色和氧化酶反应均呈阴性,在25～35℃、pH 5.5～9.5的环境中生长良好,但各菌株的菌落形态及多种生化性状有差异。通过菌体形态观察、染色反应、生理生化特征分析及16S rDNA碱基序列测定与同源性分析,鉴定1、2、3号菌株分别为产酸克雷伯氏菌(Klebsiella oxytoca)、大肠埃希氏菌(Escherichia coli)、荧光假单胞菌(Pseudomonas fluorescens),4、5、6号菌株分别属于不动杆菌属(Acinetobacter)、沙雷氏菌属(Serratia)、肠杆菌属(Enterobacter)。  相似文献   

牛附红细胞体单管套式PCR检测方法的建立   总被引：1，自引：1，他引：0  

邢莹  郑秀红  贾立军  薛书江  张守发 《畜牧与兽医》2011,43(4)

为快速、准确地检测出牛附红细胞体,根据GenBank上发表的牛温氏附红细胞体(Mycoplasma wenyonii)l6S rRNA基因序列(登录号AF016546)设计合成内外2对引物,建立了牛附红细胞体单管套式PCR检测方法,并进行了特异性、敏感性及应用试验。结果:建立的牛附红细胞体单管套式PCR检测方法扩增的片段大小为361 bp,与GenBank中相应序列同源性为98%,该方法扩增不出牛瑟氏泰勒虫、新孢子虫、布氏杆菌及牛无乳链球菌等基因片段,检测出标准模板DNA的最小量为1.22 fgμ/L。通过对吉林省延边地区66份血液样本的临床检测显示,单管套式PCR检出率28.8%,高于常规PCR的21.2%和鲜血压片镜检的12.1%,具有准确、特异、敏感等优点,是检测牛附红细胞体的一种新型、可靠的诊断技术。  相似文献   

细菌16SrDNA基因芯片的构建及应用     

林居纯  曹三杰  蒋红霞  曾振灵 《中国兽医杂志》2011,47(4)

本试验为建立能检测兽医临床重要病原菌的基因芯片方法,采用通用引物扩增菌株16S rDNA V1-V3区,制备16SrDNA PCR产物基因芯片,对5种兽医临床微生物进行检测.结果显示,制备的基因芯片能特异性地检测金黄色葡萄球菌、链球菌和鸡毒支原体,以及这3种菌株混合样品,但对大肠杆菌及沙门菌检测结果不理想.基因芯片检测灵敏度为3μg/L.  相似文献   

除臭菌株对NH_3和H_2S释放及物质转化的影响     

刘春梅  徐凤花  曹艳花  齐婧媛  杜俊杰 《农业环境保护》2011,(3):585-590

将通过高温和氨水逐级驯化筛选的除臭菌株用于牛粪堆肥,研究其对NH3和H2S释放量及物质转化的影响。其中菌株B1与A1除臭效果最好,NH3总释放量较对照分别降低68.59%和61.00%,NH4＋-N仅增加4.47%和7.19%,NO3-N略有提高,而有机氮分别增加28.99%和27.42%,说明除臭菌株能将无机态氮转化为有机氮,使全氮含量提高19.81%和18.80%,有效地减少了氮素损失;H2S总释放量较对照分别降低89.69%和86.88%,硫酸盐增加40.77%和36.49%,说明菌株可有效促进H2S向硫酸盐转化,减少H2S挥发,使全硫含量提高29.05%和22.64%。相关分析显示NH3释放量与H2S释放量、pH呈极显著正相关,与全氮、有机氮和硫酸盐呈负相关;H2S释放量与pH呈显著正相关,而与全硫和硫酸盐呈负相关。添加与未添加除臭菌株的NH3和H2S释放量经方差分析差异达到显著水平,表明添加除臭菌株可有效地控制牛粪堆肥NH3和H2S释放,保留氮素和硫素养分。  相似文献   

铅锌矿区重金属污染对微生物数量及放线菌群落结构的影响     

李小林  颜森  张小平  韦成健 《农业环境保护》2011,(3):468-475

采集四川省汉源县富泉乡万顺铅锌矿区5个不同重金属浓度的土壤样品,进行了微生物数量及放线菌多样性的研究。经分离、纯化得到43株不同的放线菌,然后对其进行BOXAIR-PCR和16SrDNAPCR-RFLP分析。结果表明,铅锌矿区重金属复合污染对土壤微生物数量有较大的影响,随着铅锌矿区重金属污染程度的加剧,土壤微生物的总数下降。相关性分析表明,重金属含量与细菌数量呈极显著负相关（P〈0.01）,与放线菌数量、真菌数量呈显著负相关（P〈0.05）。供试菌株的16SrDNA用HaeⅢ、HinfⅠ和TaqⅠ酶切后具有32种遗传图谱类型。BOXAIR-PCR的聚类结果表明在86%的水平上,所有菌株分为10个遗传类型,结果基本与16SrDNAPCR-RFLP聚类差异不大。来源于高重金属的含量样品的菌株基本聚在一起,可能是重金属含量影响了放线菌的分布。同时,16SrDNA序列聚类分析结合系统发育树分析表明链霉菌属是汉源铅锌矿区主要的放线菌属并且具有遗传多样性。  相似文献   

The under-recognized dominance of Verrucomicrobia in soil bacterial communities     

Bergmann GT  Bates ST  Eilers KG  Lauber CL  Caporaso JG  Walters WA  Knight R  Fierer N 《Soil biology & biochemistry》2011,43(7):1450-1455

Verrucomicrobia are ubiquitous in soil, but members of this bacterial phylum are thought to be present at low frequency in soil, with few studies focusing specifically on verrucomicrobial abundance, diversity, and distribution. Here we used barcoded pyrosequencing to analyze verrucomicrobial communities in surface soils collected across a range of biomes in Antarctica, Europe, and the Americas (112 samples), as well as soils collected from pits dug in a montane coniferous forest (69 samples). Data collected from surface horizons indicate that Verrucomicrobia average 23% of bacterial sequences, making them far more abundant than had been estimated. We show that this underestimation is likely due to primer bias, as many of the commonly used PCR primers appear to exclude verrucomicrobial 16S rRNA genes during amplification. Verrucomicrobia were detected in 180 out of 181 soils examined, with members of the class Spartobacteria dominating verrucomicrobial communities in nearly all biomes and soil depths. The relative abundance of Verrucomicrobia was highest in grasslands and in subsurface soil horizons, where they were often the dominant bacterial phylum. Although their ecology remains poorly understood, Verrucomicrobia appear to be dominant in many soil bacterial communities across the globe, making additional research on their ecology clearly necessary.  相似文献   

Land-use history has a stronger impact on soil microbial community composition than aboveground vegetation and soil properties     

Kamlesh Jangid  Mark A. Williams  Thomas M. Schmidt  William B. Whitman 《Soil biology & biochemistry》2011,43(10):2184-2193

The response of soil microbial communities following changes in land-use is governed by multiple factors. The objectives of this study were to investigate (i) whether soil microbial communities track the changes in aboveground vegetation during succession; and (ii) whether microbial communities return to their native state over time. Two successional gradients with different vegetation were studied at the W. K. Kellogg Biological Station, Michigan. The first gradient comprised a conventionally tilled cropland (CT), mid-succession forest (SF) abandoned from cultivation prior to 1951, and native deciduous forest (DF). The second gradient comprised the CT cropland, early-succession grassland (ES) restored in 1989, and long-term mowed grassland (MG). With succession, the total microbial PLFAs and soil microbial biomass C consistently increased in both gradients. While bacterial rRNA gene diversity remained unchanged, the abundance and composition of many bacterial phyla changed significantly. Moreover, microbial communities in the relatively pristine DF and MG soils were very similar despite major differences in soil properties and vegetation. After >50 years of succession, and despite different vegetation, microbial communities in SF were more similar to those in mature DF than in CT. In contrast, even after 17 years of succession, microbial communities in ES were more similar to CT than endpoint MG despite very different vegetation between CT and ES. This result suggested a lasting impact of cultivation history on the soil microbial community. With conversion of deciduous to conifer forest (CF), there was a significant change in multiple soil properties that correlated with changes in microbial biomass, rRNA gene diversity and community composition. In conclusion, history of land-use was a stronger determinant of the composition of microbial communities than vegetation and soil properties. Further, microbial communities in disturbed soils apparently return to their native state with time.  相似文献   

基于6S模型的山区环境星影像大气校正研究     

韩杰  杨磊库  李家国  孟庆岩  李慧娜 《水土保持通报》2011,36(4)

针对山区地形复杂,高程变化较大的特点,利用6S辐射传输模型建立查找表,结合研究区DEM对影像进行了大气校正。试验结果表明,校正后的影像清晰度、对比度提高了,消除了大气对NDVI的负面影响。  相似文献