Lack of evidence for Mycoplasma spp. in bulk tank milk of herds located in mid-western Colombia

J. Velasco–Bolaños, A. S. Jaramillo-Jaramillo, N. A. Villa–Arcila, S. Dufour, A. Ceballos-Márquez, S. Piepers
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Abstract

Mycoplasma spp. is reported as a highly contagious mastitis-causing bacteria in dairy cattle, without successful or low response to most common antibiotic treatments due to the lack of cell wall. In Colombia it has been reported in the Central Andean region during 2014. The aim was to estimate the prevalence of Mycoplasma spp. in bulk tank milk using microbiological and molecular diagnosis. A random longitudinal study en­rolling 220 commercial dairy farms located in four provinces of the mid-western region of Colombia from four pasteurizer companies was performed. Bulk tank milk samples were collected once monthly for three months period for determining somatic cell count (SCC) and microbiological and molecular diagnosis of Mycoplasma spp. cultures were done without pre-enrichment procedures directly in mycoplasma agar with cefoperazone to inhibit growth of opportunistic microorganisms, plates were incubated under 37° C and atmosphere of 10% CO2 and inspected during a 10d period. Molecular analysis was done by a multiplex PCR using specific primers targeting the 16S-23S rARN gene of Mycoplasma spp. and from non-pathogenic bacteria occasionally found in milk. LnSCC average of included dairy farms was 6.19 x103 cells/mL, Mycoplasma spp. was not isolated during microbiological cultures, and no DNA belonging to the species was detected by PCR in the 220 bulk tanks milk, with an estimated prevalence lower than 2.3%. This finding shows that there is not microbiological or molecular evidence that demonstrates the presence of the pathogen in the milk from the mid-western region of Colombia at herd level.  
在哥伦比亚中西部牛群的散装罐奶中缺乏支原体的证据
据报道,支原体是一种高度传染性乳腺炎引起的奶牛细菌,由于缺乏细胞壁,对大多数常见的抗生素治疗没有成功或反应低。据报道,哥伦比亚中部安第斯地区在2014年发生了这种情况。目的是通过微生物学和分子诊断来估计散装罐奶中支原体的流行率。对哥伦比亚中西部四个省的四家巴氏灭菌公司的220个商业奶牛场进行了随机纵向研究。每月采集一次散装罐奶样品,为期三个月,用于测定体细胞计数(SCC)以及支原体的微生物和分子诊断。培养物无需预富集程序,直接在支原体琼脂中用头孢哌酮进行,以抑制机会微生物的生长,将平板在37°C和10%CO2的气氛下孵育,并在10d期间进行检查。使用针对支原体属16S-23S rARN基因的特异性引物和偶尔在牛奶中发现的非致病菌,通过多重PCR进行分子分析。纳入奶牛场的LnSCC平均值为6.19 x103个细胞/mL,支原体属在微生物培养过程中未分离,在220个散装罐牛奶中未通过PCR检测到属于该物种的DNA,估计患病率低于2.3%。这一发现表明,没有微生物或分子证据表明哥伦比亚中西部地区的牛奶中存在群体水平的病原体。
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