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Prevotella vs. Ruminococcus enterotypes drive divergent milk fat synthesis in dairy cows through rumen microbiota-host metabolic crosstalk

ABSTRACT:

Background: High-yield dairy cows are usually accompanied with lower milk fat (MF) rate, with huge variations in MF contents among individuals. The potential mechanisms of key microorganisms, their genes and metabolites participate in MF synthesis are still not well understood. By selecting 12 high-yielding (HIGH) and 12 low-yielding (LOW) cows from a large cohort of dairy cows, multi-omics technologies were applied to identify the hallmark microorganisms, metabolites, and metabolic pathways involved in MF synthesis.

Results: The LOW and HIGH cows under the same feeding and management conditions exhibited considerable phenotypic, microbial, functional, and metabolomic differences. The LOW and HIGH cows underwent acetic and propionic ruminal fermentation, respectively, with greater molar proportion of acetate and butyrate in the LOW cows, providing greater precursors for short-chain fatty acids (FAs) synthesis in mammary gland. The HIGH cows had greater molar proportion of propionate in rumen and contents of long-chain FAs and odd-chain FAs (OCFAs) in milk. Metagenomic analysis revealed that the HIGH and LOW cows possessed Prevotella- and Ruminococcus-driven enterotypes, respectively, associated with significantly lower abundances of Methanobacteriota and Methanobrevibacter_A in HIGH than that of the LOW cows. Moreover, 13 carbohydrate and 3 lipid metabolism pathways were greatly enhanced in HIGH cows, accompanied with greater abundances of CAZymes encoding genes and critical enzymes involved in fiber degradation and propionate synthetization. Metabolomic analysis revealed that the free FAs, glycerophospholipids and oxidized lipids in ruminal fluid, and bile acids, glycerides, and glycerophospholipids in serum were greater in HIGH cows; however, the contents of carnitines, being ubiquitously detected in ruminal fluid, serum, and milk, were consistently greater in LOW cows, suggesting the capability of being used as biomarkers for predicting the lactation potentiality.

Conclusions: The yield and MF composition between HIGH and LOW cows are microbiome-dependent, specifically greatly depend on the Prevotella- and Ruminococcus-driven enterotypes, respectively. The former enterotype is associated with higher milk efficiency and greater milk contents of OCFAs, which is more environmentally friend and healthier to human; whereas the later enterotype is characterized with greater methanogens and carnitines, which indicates a greater dietary energy loss and a poorer energy status of dairy cows.

INSTRUMENT(S): Liquid Chromatography MS - positive - HILIC, Liquid Chromatography MS - negative - HILIC

PROVIDER: MTBLS13211 | MetaboLights | 2025-10-28

REPOSITORIES: MetaboLights

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			Action	DRS
	a_MTBLS13211_LC-MS_negative_hilic_metabolite_profiling.txt	Txt
	a_MTBLS13211_LC-MS_positive_hilic_metabolite_profiling.txt	Txt
	i_Investigation.txt	Txt
	m_MTBLS13211_LC-MS_negative_hilic_metabolite_profiling_v2_maf.tsv	Tabular
	m_MTBLS13211_LC-MS_positive_hilic_metabolite_profiling_v2_maf.tsv	Tabular

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Project description:Bovine mastitis, the infection of the mammary gland which leads to great health and economic challenges for dairy farmers is accompanied by dramatic changes in the milk proteome. In this study of naturally occurring mastitis not only have the changes in the milk proteome been quantified in subclinical and clinical mastitis but simultaneous changes in the serum proteome have also been characterised and quantified. Milk and serum samples from healthy dairy cows (n=12) were compared to those of cows with subclinical (n=10) and clinical mastitis (n=112) using TMT label-based proteomic approach. The study included the milk and serum samples taken from thirty-two dairy cows ( kept on private farms located in Croatia. All cows were checked by physical examination. Somatic cells count (SCC) and mastitis test in milk samples were performed. According to the results, cows were assigned into three groups: Group I (control, n=10) consisted of healthy cows with SCC below 400,000 cells/ml on the monthly check-up and a negative mastitis test and without any clinical sign of mastitis. Group II (subclinical mastitis, n=12) comprised cows without clinical signs of mastitis but with SCC above 400,000 cells/ml on the monthly basis and a positive mastitis test at the time of sampling. Group III (clinical mastitis, n=10) consisted of cows with clinical signs of mastitis which include changes in milk appearance (flakes and clots in milk), different stages of udder inflammation (hyperemia, edema, pain, udder enlargement and elevated udder temperature) and disturbance of general health (depression, relaxed cold ears, dehydration, elevated body temperature, increased heart and respiratory rate, decreased ruminal contraction and decreased appetite). Blood samples were taken from v. coccygea and centrifuged at 3000 g for 15 min after clotting for two hours at room temperature. Serum samples were stored at -80°C until analysis. Milk samples were taken aseptically before the morning milking. First few streams were discarded. Teat ends were disinfected with cotton swabs soaked with 70% ethanol. Samples were taken into sterile tubes and transported to laboratory on ice within a few hours.

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Prevotella vs. Ruminococcus enterotypes drive divergent milk fat synthesis in dairy cows through rumen microbiota-host metabolic crosstalk

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