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DNA微阵列分析和实时PCR作为研究幼年奶牛呼吸道支原体菌群的有用工具。

DNA microarray assay and real-time PCR as useful tools for studying the respiratory tract Mycoplasma populations in young dairy calves.

作者信息

Bottinelli Marco, Passamonti Fabrizio, Rampacci Elisa, Stefanetti Valentina, Pochiero Luca, Coletti Mauro, Rueca Fabrizio, Hyatt Doreene R, Schnee Christiane

机构信息

Veterinary Medicine Department, Università degli Studi di Perugia, Perugia, Italy.

Private Practitioner, Fiumicino (Rome), Italy.

出版信息

J Med Microbiol. 2017 Sep;66(9):1342-1349. doi: 10.1099/jmm.0.000571. Epub 2017 Sep 4.

Abstract

PURPOSE

With more than 120 species, the genus Mycoplasma is one of the largest taxa in the class Mollicutes, a group of micro-organisms that are characterized by apparent simplicity and to which important animal pathogens belong. Mycoplasmabovis is the most frequently identified pathogenic Mycoplasma in cattle; however, the prevalence of other Mycoplasma species living in calves' airways is poorly understood. The aim of this work was to characterize the respiratory tract mycoplasma populations in calves on one of the largest dairy farms in Italy using a real-time PCR assay and a DNA microarray assay.

METHODOLOGY

A total of 49 nasal swabs and 49 trans-tracheal aspirations from non-vaccinated veal calves were analysed. Genomic DNA was extracted from the samples and then tested using a real-time PCR targeting the oppD gene of M. bovis and a DNA microarray that was able to identify more than 70 Mycoplasma species.

RESULTS

Forty-two out of 49 calves tested positive for Mycoplasma spp. (85.7 %). None of the samples tested positive for M. bovis. A majority (73.5 %) of the 98 samples tested positive for M. dispar, while 8 samples tested positive for M. bovirhinis (8.2 %).

CONCLUSION

Our results expand our knowledge regarding the diversity of Mycoplasma populations in the respiratory airways of very young veal calves and add data regarding M. bovis prevalence in the Italian cattle population. However, the importance of these species in the respiratory diseases of calves still remains to be determined.

摘要

目的

支原体属有120多个物种,是柔膜菌纲中最大的分类群之一,柔膜菌纲是一类以明显简单为特征且包含重要动物病原体的微生物。牛支原体是牛中最常鉴定出的致病性支原体;然而,对生活在犊牛气道中的其他支原体物种的流行情况了解甚少。本研究的目的是使用实时PCR检测和DNA微阵列检测来鉴定意大利最大的奶牛场之一的犊牛呼吸道支原体种群特征。

方法

共分析了49头未接种疫苗的犊牛的鼻拭子和49份经气管抽吸物。从样本中提取基因组DNA,然后使用针对牛支原体oppD基因的实时PCR和能够鉴定70多种支原体物种的DNA微阵列进行检测。

结果

49头犊牛中有42头支原体检测呈阳性(85.7%)。没有样本检测出牛支原体呈阳性。98个样本中的大多数(73.5%)检测出副猪支原体呈阳性,而8个样本检测出牛鼻支原体呈阳性(8.2%)。

结论

我们的结果扩展了我们对极幼小犊牛呼吸道支原体种群多样性的认识,并增加了意大利牛群中牛支原体流行情况的数据。然而,这些物种在犊牛呼吸道疾病中的重要性仍有待确定。

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