Changes in the Microbiome in Yak Mastitis: Insights Based on Full-Length 16S rRNA Sequencing

Lihong Zhang, Hongcai Ma, Wenqiang Tang, Jiangyong Zeng, Md. F. Kulyar, Junjie Hu
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Abstract

Mastitis is an inflammation of the mammary gland that can be caused by various factors, including biological, chemical, mechanical, or physical. Microbiological culture, DNA techniques, and high-throughput next-generation sequencing have been used to identify mastitis-causing pathogens in various animal species. However, little is known about microbiota and microbiome changes linked to yak milk mastitis. This study aimed to characterize the milk microbiota of healthy and mastitis-infected yaks using full-length 16S rRNA sequencing. The results showed that the bacterial microbiota comprises 7 phyla, 9 classes, 20 orders, 39 families, 59 genera, and 72 species. Proteobacteria and Firmicutes were the predominant microbial communities, with lower abundances of Bacteroidota, Actinobacteriota, Acidobacteriota, and other minor groupings also observed. Proteobacteria dominated the clinical and subclinical mastitis groups (95.36% and 89.32%, respectively), in contrast to the healthy group (60.17%). Conversely, Firmicutes were more common in the healthy group (39.7%) than in the subclinical and clinical mastitis groups (10.49% and 2.92%, respectively). The predominant organisms found in the healthy group were Leuconostoc mesenteroides, Lactococcus piscium, Carnobacterium maltaromaticum, and Lactococcus raffinolactis. Low abundances of Staphylococcus aureus species were found in both subclinical and clinical mastitis groups, with Moraxella osloensis and Psychrobacter cibarius dominating the subclinical mastitis group and Pseudomonas fluorescens dominating the clinical mastitis group. An alpha diversity study revealed that the healthy group had a higher microbial diversity than the clinical and subclinical mastitis groups. According to beta-diversity analysis, the principal coordinate analysis identified that mastitis-infected samples significantly differed from healthy ones. The milk microbiota of healthy yaks is more varied, and specific prominent taxa within various groups can act as marker microorganisms for mastitis risk. The genera Leuconostoc and Lactococcus are promising candidates for creating probiotics.
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牦牛乳腺炎微生物组的变化:基于全长 16S rRNA 测序的见解
乳腺炎是一种乳腺炎症,可由生物、化学、机械或物理等多种因素引起。微生物培养、DNA 技术和高通量新一代测序技术已被用于鉴定各种动物物种中导致乳腺炎的病原体。然而,人们对与牦牛乳腺炎有关的微生物群和微生物组变化知之甚少。本研究旨在利用全长 16S rRNA 测序技术描述健康牦牛和乳腺炎感染牦牛乳汁微生物群的特征。结果显示,细菌微生物群包括 7 个门、9 个类、20 个目、39 个科、59 个属和 72 个种。蛋白质细菌和固缩菌是主要的微生物群落,类杆菌属、放线菌属、酸性杆菌属和其他小群的数量较少。蛋白菌在临床和亚临床乳腺炎组中占主导地位(分别为 95.36% 和 89.32%),而健康组则为 60.17%。相反,健康组(39.7%)中的真菌比亚临床和临床乳腺炎组(分别为 10.49% 和 2.92%)更常见。在健康组中发现的主要微生物是中肠亮球菌(Leuconostoc mesenteroides)、半球乳球菌(Lactococcus piscium)、麦芽肉杆菌(Carnobacterium maltaromaticum)和乳酸乳球菌(Lactococcus raffinolactis)。亚临床和临床乳腺炎组中的金黄色葡萄球菌含量都很低,亚临床乳腺炎组中主要是奥斯陆莫拉菌和西巴氏精神杆菌,临床乳腺炎组中主要是荧光假单胞菌。阿尔法多样性研究显示,健康组的微生物多样性高于临床和亚临床乳腺炎组。根据贝塔多样性分析,主坐标分析确定乳腺炎感染样本与健康样本存在显著差异。健康牦牛的牛奶微生物群更加多样,不同群中的特定突出类群可作为乳腺炎风险的标记微生物。Leuconostoc 属和 Lactococcus 属很有希望成为益生菌。
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