Review on microbial metabolomics of probiotics and pathogens:Methodologies and applications

In recent years,microbial metabolomics,a new field that has attracted wide attention,provides a map of metabolic pathways and clarifies the interaction mechanism between microorganisms and hosts.Many microorganisms are found in the human intestine,oral cavity,vagina,etc.Probiotics could maintain the good health of the host,while pathogens and an imbalance of bacterial flora lead to a series of diseases of the body and mind.Metabolomics is a science for qualitative and quantitative analysis of all metabolites in an organism or biological system,which could provide key information to understand the related metabolic pathways and associated changes.This approach analyzes the final products of cellular regulatory processes,the level of which can be regarded as the ultimate response of the biological system to genetic or environmental changes.Microbial metabolomics has been widely used in different research fields,such as microbial phenotypic classification,mutant screening,metabolic pathways,microbial metabolic engineering,fermentation engineering monitoring and optimization,microbial environmental pollution,and so on.However,there are only a few reviews on microbial metabolomics of probiotics and pathogens.This review summarizes the main methodologies,including sample preparation,identification of metabolites,data processing,and analysis.Recent applications in microbial metabolomics of probiotics and pathogens are also described.This paper first summarized the research progress and application of microbial metabolomics from two aspects:probiotics and pathogenic bacteria.Probiotics and pathogenic bacteria do not exist independently most of the time;hence,these were reviewed in the research field of coexistence of probiotics and pathogenic bacteria,which was subdivided into important microbial research fields closely related to human health,including the human gut,oral cavity,food,and nutrition-related microorganisms.Then,the main problems and trends associated with microbial metabolomics are discussed.

Multi‑omics reveals that the host‑microbiome metabolism crosstalk of differential rumen bacterial enterotypes can regulate the milk protein synthesis of dairy cows

Background Dairy cows’lactation performance is the outcome of the crosstalk between ruminal microbial metabo-lism and host metabolism.However,it is still unclear to what extent the rumen microbiome and its metabolites,as well as the host metabolism,contribute to regulating the milk protein yield(MPY).Methods The rumen fluid,serum and milk of 12 Holstein cows with the same diet(45%coarseness ratio),parity(2–3 fetuses)and lactation days(120–150 d)were used for the microbiome and metabolome analysis.Rumen metabolism(rumen metabolome)and host metabolism(blood and milk metabolome)were connected using a weighted gene co-expression network(WGCNA)and the structural equation model(SEM)analyses.Results Two different ruminal enterotypes,with abundant Prevotella and Ruminococcus,were identified as type1 and type2.Of these,a higher MPY was found in cows with ruminal type2.Interestingly,[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae(the differential bacteria)were the hub genera of the network.In addition,differential ruminal,serum and milk metabolome between enterotypes were identified,where the cows with type2 had higher L-tyrosine of rumen,ornithine and L-tryptophan of serum,and tetrahydroneopterin,palmitoyl-L-carnitine,S-lactoylglutathione of milk,which could provide more energy and substrate for MPY.Further,based on the identi-fied modules of ruminal microbiome,as well as ruminal serum and milk metabolome using WGCNA,the SEM analysis indicated that the key ruminal microbial module1,which contains the hub genera of the network([Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae)and high abundance of bacteria(Prevotella and Ruminococcus),could regulate the MPY by module7 of rumen,module2 of blood,and module7 of milk,which contained L-tyrosine and L-tryptophan.Therefore,in order to more clearly reveal the process of rumen bacterial regulation of MPY,we established the path of SEM based on the L-tyrosine,L-tryptophan and related components.The SEM based on the metabolites suggested that[Ruminococcus]gauvreauii group could inhibit the energy supply of serum tryptophan to MPY by milk S-lactoylglutathione,which could enhance pyruvate metabolism.Norank_f_Ruminococcaceae could increase the ruminal L-tyrosine,which could provide the substrate for MPY.Conclusion Our results indicated that the represented enterotype genera of Prevotella and Ruminococcus,and the hub genera of[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae could regulate milk protein synthesis by affecting the ruminal L-tyrosine and L-tryptophan.Moreover,the combined analysis of enterotype,WGCNA and SEM could be used to connect rumen microbial metabolism with host metabolism,which provides a fundamental understanding of the crosstalk between host and microorganisms in regulating the synthesis of milk composition.