Oultram Joanne W H, Ganda Erika K, Boulding Sarah C, Bicalho Rodrigo C, Oikonomou Georgios
Department of Livestock Health and Welfare, Institute of Veterinary Science, University of Liverpool , Neston , UK.
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University , Ithaca, NY , USA.
Front Vet Sci. 2017 Mar 10;4:36. doi: 10.3389/fvets.2017.00036. eCollection 2017.
Mastitis is one of the most costly diseases affecting the dairy industry, and identification of the causative microorganism(s) is essential. Here, we report the use of next-generation sequencing of bacterial 16S rRNA genes for clinical mastitis diagnosis. We used 65 paired milk samples, collected from the mastitic and a contralateral healthy quarter of mastitic dairy cattle to evaluate the technique as a potential alternative to bacterial culture or targeted PCR. One large commercial dairy farm was used, with one trained veterinarian collecting the milk samples. The 16S rRNA genes were individually amplified and sequenced using the MiSeq platform. The MiSeq Reporter was used in order to analyze the obtained sequences. Cattle were categorized according to whether or not 1 of the 10 most abundant bacterial genera in the mastitic quarter exhibited an increase in relative abundance between the healthy and mastitic quarters equal to, or exceeding, twofold. We suggest that this increase in relative abundance is indicative of the genus being a causative mastitis pathogen. Well-known mastitis-causing pathogens such as and spp. were identified in most cattle. We were able to diagnose 53 out of the 65 studied cases and identify potential new mastitis pathogens such as and , which are difficult to identify by bacterial culture because of their fastidious nature.
乳腺炎是影响乳制品行业成本最高的疾病之一,确定致病微生物至关重要。在此,我们报告了利用细菌16S rRNA基因的新一代测序技术进行临床乳腺炎诊断。我们使用了65对牛奶样本,这些样本分别采自患乳腺炎奶牛的患侧乳腺和对侧健康乳腺,以评估该技术作为细菌培养或靶向PCR潜在替代方法的可行性。我们选取了一个大型商业奶牛场,由一名经过培训的兽医采集牛奶样本。利用MiSeq平台对16S rRNA基因进行单独扩增和测序。使用MiSeq Reporter分析所得序列。根据患侧乳腺中10个最丰富细菌属中的某一个在健康乳腺和患侧乳腺之间的相对丰度是否增加到两倍或超过两倍,对奶牛进行分类。我们认为这种相对丰度的增加表明该属是引起乳腺炎的病原体。在大多数奶牛中都鉴定出了诸如 和 等众所周知的引起乳腺炎的病原体。我们能够诊断出65例研究病例中的53例,并鉴定出潜在的新的乳腺炎病原体,如 和 ,由于它们苛求的特性,通过细菌培养很难鉴定。