Milk microbiome and bacterial load following dry cow therapy without antibiotics in dairy cows with healthy mammary gland.
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ABSTRACT:
SUBMITTER: Bonsaglia ECR
PROVIDER: S-EPMC5556035 | biostudies-other | 2017 Aug
REPOSITORIES: biostudies-other
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Project description:Mastitis is an inflammatory disease of the mammary gland, caused by the invasion of microorganism on this site, associated with an altered immune response. Recent studies in this field hypothesize that the origin of these pathogens can also be from the gastrointestinal tract, through the entero-mammary pathway in relation to an increase in gut permeability. In this study, we wanted to investigate if inflammatory status of the mammary gland is related to an alteration of gut permeability. The microbiome of feces, blood and milk of lactating cows, recruited on the basis of the total somatic cell count and of the percentage of polymorphonuclear neutrophils and lymphocytes, was studied. Cows were divided into healthy (G), at risk of mastitis (Y) and with mastitis (R) classifications. The bacterial DNA was extracted and the V3 and V4 regions of 16S rRNA sequenced. Moreover, the quantification of total bacteria was performed with quantitative real-time PCR. A non-parametric Kruskal-Wallis test was applied at the phylum, family and genera levels and beta biodiversity was evaluated with the unweighted UniFrac distance metric. Significant differences between groups were found for the microbial composition of feces (Clostridiaceae, Turicibacteriaceae for family level and Clostridium, Dorea, SMB53 and Turicibacter for genus level), blood (Tenericutes for phylum level and Mycoplasma for genus level) and milk (OD1 and Proteobacteria for phylum level, Enterobacteriaceae and Moraxallaceae for family level and Olsenella and Rhodococcus for genus level). The beta biodiversity of feces and blood did not change between groups. Significant differences (p < 0.05) were observed between the beta diversity in milk of G group and Y group and between Y group and R group. The number of taxa in common between feces, blood and milk were 8 at a phylum, 19 at a family and 15 at a genus level. From these results, the bacterial crossing from gut to milk in cows was not confirmed but remained hypothetical and deserves further investigation.
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Project description:Synergistic regulation among microRNAs (miRNAs) is important to understand the mechanisms underlying the complex molecular regulatory networks in goats. Goat milk fat synthesis is driven by a gene network that involves many biological processes in the mammary gland. These biological processes are affected by several miRNAs rather than a single miRNA. Therefore, identifying synergistic miRNAs is necessary to further understand the functions of miRNAs and the metabolism of goat milk fat synthesis. Using qRT-PCR, we assessed the expression of 11 miRNAs that have the potential to regulate milk fat synthesis in the goat mammary gland. Six of these miRNAs exhibited expression during the lactation cycle. Additionally, we also found that prolactin, the key hormone that regulates lactation, promotes the expression of four miRNAs (miR-23a, miR-27b, miR-103, and miR-200a). Further functional analysis showed that overexpression of all four miRNAs by using recombinant adenovirus in goat mammary gland epithelial cells can affect gene mRNA expression associated with milk fat synthesis. Specifically, elevated miR-200a expression suppressed the mRNA expression of genes involved in fat droplet formation. To analyze the synergistic regulation among these four miRNAs (miR-23a, miR-27b, miR-103, and miR-200a), we used the Pearson correlation coefficient to evaluate the correlation between their expression levels in 30 lactating goats. As a result, we found a strong correlation and mutual regulation between three miRNA pairs (miR-23a and miR-27b, miR-103 and miR-200a, miR-27b and miR-200a). This study provides the first experimental evidence that miRNA expression is synergistically regulated in the goat mammary gland and has identified the potential biological role of miRNAs in goat milk fat synthesis. The identification of synergistic miRNAs is a crucial step for further understanding the molecular network of milk fat synthesis at a system-wide level.
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