Abstract

Bamboo leaf extracts, with high content of flavonoids and diverse biological activities, are used in animal husbandry. Increasing evidence has suggested an association between the bovine physiology and the udder microbiome, yet whether the microbiota and the metabolites of milk affect the mammary gland health or the milk quality remains unknown. In this study, we provide a potential mechanism for the effects of bamboo leaf extracts on milk microbiota and metabolites of dairy cows. Twelve multiparous lactating Chinese Holstein dairy cows were randomly separated into two groups: basal diet as the control group (CON, n = 6) and a diet supplemented with 30 g/d bamboo leaf extract per head as antioxidants of bamboo leaf (AOB) group (AOB, n = 6) for 7 weeks (2-week adaptation, 5-week treatment). Milk samples were collected at the end of the trial (week 7) for microbiome and associated metabolic analysis by 16S ribosomal RNA (rRNA) gene sequencing and liquid chromatography-mass spectrometry (LC-MS). The results showed that the milk protein was increased (p < 0.0001) and somatic cell count (SCC) showed a tendency to decrease (p = 0.09) with AOB supplementation. The relative abundance of Firmicutes was significantly decreased (p = 0.04) while a higher relative abundance of Probacteria (p = 0.01) was seen in the group receiving AOB compared to the CON group. The AOB group had a significantly lower relative abundance of Corynebacterium_1 (p = 0.01), Aerococcus (p = 0.01), and Staphylococcus (p = 0.02). There were 64 different types of metabolites significantly upregulated, namely, glycerophospholipids and fatty acyls, and 15 significantly downregulated metabolites, such as moracetin, sphinganine, and lactulose in the AOB group. Metabolic pathway analysis of the different metabolites revealed that the sphingolipid signaling pathway was significantly enriched, together with glycerophospholipid metabolism, sphingolipid metabolism, and necroptosis in response to AOB supplementation. Several typical metabolites were highly correlated with specific ruminal bacteria, demonstrating a functional correlation between the milk microbiome and the associated metabolites. These insights into the complex mechanism and corresponding biological responses highlight the potential function of AOB, warranting further investigation into the regulatory role of specific pathways in the metabolism.

Highlights

  • Flavonoids and their analogs have been investigated as feed additives for improving milk production of dairy cows and to increase their growth and development [1, 2], because of their antibacterial and antioxidant properties [3], which have attracted wide attention

  • Gentisic acid was negatively correlated with Staphylococcus and Aerococcus but positively correlated with Massilia. These results indicated that in response to dietary antioxidants of bamboo leaf (AOB) supplementation, milk microbiota was associated with significant changes in metabolite levels, especially those related to the sphingolipid metabolism

  • We showed that pregnanediol was significantly increased by 7.17-fold in the AOB group, which supports the idea that AOB can stimulate milk production, and account for the negative correlation between pregnanediol and Staphylococcus

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Summary

Introduction

Flavonoids and their analogs have been investigated as feed additives for improving milk production of dairy cows and to increase their growth and development [1, 2], because of their antibacterial and antioxidant properties [3], which have attracted wide attention. Bamboo leaf extracts are rich in flavonoids, with a large group of naturally occurring polyphenolic compounds existing either as free aglycones or glycosidic conjugates, based on the saturation level consisting of an A and C ring or a B-ring substitution pattern [1]. These characteristic structures caused flavonoids to be targeted as key compounds with great potential in animal husbandry. More research needs to be done to understand the mechanism of the effects of bamboo leaf extract on the milk microbial composition and the metabolite profiles

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