Objective: Microbial exposure is critical to neonatal and infant development, growth and immunity. However, whether a microbiome is present in the fetal gut prior to birth remains debated. In this study, lambs delivered by aseptic hysterectomy at full term were used as an animal model to investigate the presence of a microbiome in the prenatal gut using a multiomics approach. Design: Lambs were euthanised immediately after aseptic caesarean section and their cecal content and umbilical cord blood samples were aseptically acquired. Cecal content samples were assessed using metagenomic and metatranscriptomic sequencing to characterise any existing microbiome. Both sample types were analysed using metabolomics in order to detect microbial metabolites. Results: We detected a low-diversity and low-biomass microbiome in the prenatal fetal gut, which was mainly composed of bacteria belonging to the phyla Proteobacteria, Actinobacteria and Firmicutes. Escherichia coli was the most abundant species in the prenatal fetal gut. We also detected multiple microbial metabolites including short chain fatty acids, deoxynojirimycin, mitomycin and tobramycin, further indicating the presence of metabolically active microbiota. Additionally, bacteriophage phiX174 and Orf virus, as well as antibiotic resistance genes, were detected in the fetal gut, suggesting that bacteriophage, viruses and bacteria carrying antibiotic resistance genes can be transmitted from the mother to the fetus during the gestation period. Conclusions: This study provides strong evidence that the prenatal gut harbours a microbiome and that microbial colonisation of the fetal gut commences in utero.

Background: Due to the high prevalence and complex etiology, bovine mastitis (BM) is one of the most important diseases to compromise dairy cow health and milk quality. The shift in milk compositions has been widely investigated during mastitis, but recent studies suggested that gastrointestinal microorganism also has a crucial effect on the inflammation of other peripheral tissues and organs, including the mammary gland. However, research focused on the variation of rumen inner-environment during mastitis is still limited. Therefore, the ruminal microbial profiles, metabolites, and milk compositions in cows with different udder health conditions were compared in the present study. Furthermore, the correlations between udder health status and ruminal conditions were investigated. Based on the somatic cell counts (SCC), California mastitis test (CMT) parameters and clinical symptoms of mastitis, 60 lactating Holstein dairy cows with similar body conditions (excepted for the udder health condition) were randomly divided into 3 groups (n = 20 per group) including the healthy (H) group, the subclinical mastitis (SM) group and the clinical mastitis (CM) group. Lactation performance and rumen fermentation parameters were recorded. And rumen microbiota and metabolites were also analyzed via 16S rRNA amplicon sequencing and untargeted metabolomics, respectively. Results: As the degree of mastitis increased, rumen lactic acid (LA) (P < 0.01), acetate, propionate, butyrate, valerate (P < 0.001), and total volatile fatty acids (TVFAs) (P < 0.01) concentrations were significantly decreased. In the rumen of CM cows, the significantly increased bacteria related to intestinal and oral inflammation, such as Lachnospiraceae (FDR-adjusted P = 0.039), Moraxella (FDR-adjusted P = 0.011) and Neisseriaceae (FDR-adjusted P = 0.036), etc., were accompanied by a significant increase in 12-oxo-20-dihydroxy-leukotriene B4 (FDR-adjusted P = 5.97 × 10^− 9) and 10beta-hydroxy-6beta-isobutyrylfuranoeremophilane (FDR-adjusted P = 3.88 × 10^− 10). Meanwhile, in the rumen of SM cows, the Ruminiclostridium_9 (FDR-adjusted P = 0.042) and Enterorhabdus (FDR-adjusted P = 0.043) were increased along with increasing methenamine (FDR-adjusted P = 6.95 × 10^− 6), 5-hydroxymethyl-2-furancarboxaldehyde (5-HMF) (FDR-adjusted P = 2.02 × 10^− 6) and 6-methoxymellein (FDR-adjusted P = 2.57 × 10^− 5). The short-chain fatty acids (SCFAs)-producing bacteria and probiotics in rumen, including Prevoterotoella_1 (FDR-adjusted P = 0.045) and Bifidobacterium (FDR-adjusted P = 0.035), etc., were significantly reduced, with decreasing 2-phenylbutyric acid (2-PBA) (FDR-adjusted P = 4.37 × 10^− 6). Conclusion: The results indicated that there was a significant shift in the ruminal microflora and metabolites associated with inflammation and immune responses during CM. Moreover, in the rumen of cows affected by SM, the relative abundance of several opportunistic pathogens and the level of metabolites which could produce antibacterial compounds or had a competitive inhibitory effect were all increased.

Summary: Early gut microbial colonization is important for postnatal metabolic and immune development. However, little is known about the effects of different feeding modes (suckling versus bottle‐feeding) or microbial sources on this process in farm animals. We found that suckled and bottle‐fed newborn lambs had their own distinct gut microbiota. Results from 16S rRNA gene sequencing and qPCR showed that, compared with suckling, bottle feeding significantly increased the abundances of Escherichia/Shigella , Butyricicoccus , and Clostridium XlVa, while significantly decreased the abundance of Clostridium XI. The higher levels of Escherichia/Shigella in bottle‐fed lambs suggest that artificial feeding may increase the number of potential pathogens and delay the establishment of the anaerobic environment and anaerobic microbes. Feeding modes also affected the direct transmission of bacteria from the mother and the environment to newborns. The SourceTracker analysis estimated that the early gut microbes of suckled lambs were mainly derived from the mother's teats (43%) and ambient air (28%); whereas those of bottle‐fed lambs were dominated by bacteria from the mother's vagina (46%), ambient air (31%), and the sheep pen floor (12%). These findings advance our understanding of gut microbiota in early life and may help design techniques to improve gut microbiota and health.

With the improvement in the intense genetic selection of dairy cows, advanced management strategies, and improved feed quality and disease control, milk production level has been greatly improved. However, the negative energy balance (NEB) is increasingly serious at the postpartum stage because the intake of nutrients cannot meet the demand of quickly improved milk production. The NEB leads to a large amount of body fat mobilization and consequently the elevated production of ketones, which causes metabolic diseases such as ketosis and fatty liver. The high milk production of dairy cows in early lactation aggravates NEB. The metabolic diseases lead to metabolic disorders, a decrease in reproductive performance, and lactation performance decline, seriously affecting the health and production of cows. Propylene glycol (PG) can alleviate NEB through gluconeogenesis and inhibit the synthesis of ketone bodies. In addition, PG improves milk yield, reproduction, and immune performance by improving plasma glucose and liver function in ketosis cows, and reduces milk fat percentage. However, a large dose of PG (above 500 g/d) has toxic and side effects in cows. The feeding method used was an oral drench. The combination of PG with some other additives can improve the effects in preventing ketosis. Overall, the present review summarizes the recent research progress in the impacts of NEB in dairy cows and the properties of PG in alleviating NEB and reducing the risk of ketosis.

Calcium propionate is a safe and reliable food and feed additive. It can be metabolized and absorbed by humans and animals as a precursor for glucose synthesis. In addition, calcium propionate provides essential calcium to mammals. In the perinatal period of dairy cows, many cows cannot adjust to the tremendous metabolic, endocrine, and physiological changes, resulting in ketosis and fatty liver due to a negative energy balance (NEB) or milk fever induced by hypocalcemia. On hot weather days, cow feed (TMR or silage) is susceptible to mildew, which produces mycotoxins. These two issues are closely related to dairy health and performance. Perinatal period metabolic disease significantly reduces cow production and increases the elimination rate because it causes major glucose and calcium deficiencies. Feeding a diet contaminated with mycotoxin leads to rumen metabolic disorders, a reduced reproductive rate (increased abortion rate), an increased number of milk somatic cells, and decreased milk production, as well as an increased occurrence of mastitis and hoof disease. Propionic acid is the primary gluconeogenic precursor in dairy cows and one of the safest mold inhibitors. Therefore, calcium propionate, which can be hydrolyzed into propionic acid and Ca²⁺ in the rumen, may be a good feed additive for alleviating NEB and milk fever in the perinatal period of dairy cows. It can also be used to inhibit TMR or silage deterioration in hot weather and regulate rumen development in calves. This paper reviews the application of calcium propionate in dairy cows.

Calcium propionate is one kind of good source for preventing and treating hypocalcemia and ketosis for dairy cows in early lactation. However, little is known about the effects of different feeding levels of calcium propionate on the ruminal bacterial community of early lactation dairy cows. This study aimed to explore the effects of different calcium propionate feeding levels on the ruminal fermentation and bacterial community composition of early lactation dairy cows. Twenty-four multiparous cows were randomly allocated into control (CON), low calcium propionate (LCaP), medium calcium propionate (MCaP), and high calcium propionate (HCaP) groups with six cows per group after calving. The CON group cows were fed the normal total mixed ration (TMR), and the cows of the LCaP, MCaP, and HCaP groups were fed TMR supplemented with 200, 350, and 500 g/day calcium propionate for 35 days after calving, respectively. The rumen fermentation parameters were measured every week, and the ruminal bacterial community composition of the last week was analyzed by 16S rRNA gene sequencing. Under the same diet, the rumen pH showed no difference among the four groups, but the content of microbial crude protein (MCP) and ammonia nitrogen quadratically decreased and linearly increased with calcium propionate supplementation, respectively. The feeding of calcium propionate linearly increased the concentrations of total volatile fatty acid (VFA), acetate, propionate, butyrate, iso-valerate, and valerate in the rumen. In all the treatment groups, Bacteroidetes, Firmicutes, and Proteobacteria were the dominant phyla, and Prevotella_1 and Succiniclasticum were the dominant genera in the rumen. Compared with the CON group, the addition of calcium propionate to the early lactation dairy cows quadratically improved the alpha diversity index of Chao1 estimator and observed species, but had little effect on the relative abundance of the major bacterial at phyla and genera level. These results suggested different levels of calcium propionate supplementation improved the rumen fermentation and the ruminal bacterial diversity but had little impact on the major ruminal bacterial community composition of dairy cows in early lactation.