Effect of dietary peNDF levels on digestibility and rumen fermentation, and microbial community in growing goats.

Physically effective neutral detergent fiber (peNDF) is a concept that accounts for the particle length of NDF in diets, sustaining the normal chewing behavior and rumen fermentation of ruminants. Specifically, peNDF>1.18 is the commonest one that is calculated from NDF and the percentage of feed dry matter left on the 1.18, 8.00, and 19.00 mm sieves. This study aimed to investigate the effects of different levels of peNDF>1.18 on the rumen microbiome and its correlation with nutrient digestibility and rumen fermentation in goats. A total of 30 Lezhi black goats were randomized and blocked to five dietary treatments (n = 6). All the diets were identical in composition but varied in hay lengths, leading to the different peNDF>1.18 content of the diets: 32.97, 29.93, 28.14, 26.48, and 24.75%. The results revealed that the nutrient digestibility increased when dietary peNDF>1.18 levels decreased from 32.97% to 28.14%, with the highest digestibility at 28.14% peNDF>1.18 treatment, after which nutrient digestibility decreased with the decreasing of dietary peNDF levels. Ruminal NH3-N concentrations in the 29.93% and 28.14% groups were higher than that in the 24.75% group (p < 0.05). Ruminal microbial protein concentration was the highest in the 32.97% group (p < 0.05). Daily CH4 production in the 32.97% and 24.75% peNDF>1.18 treatments was lower than that in the 26.48% group (p < 0.05) and no differences were observed among other groups. The relative abundance of rumen fungi at the phylum and genus levels and archaea at the species were affected by dietary peNDF>1.18 content. In conclusion, decreasing dietary peNDF>1.18 levels within a certain range can improve nutrient digestibility and change the rumen microbial community structure of goats. Dietary peNDF>1.18 level should be 28.14% (roughage length around 1 cm) among the five levels for 4 months Lezhi black goats with the purpose of optimal nutrient digestibility.

Integration of transcriptomic and metabolomic analysis of the mechanism of dietary N-carbamoylglutamate in promoting follicle development in yaks.

Yak is the main livestock in the highlands of China. The low reproductive rate of yaks is a serious constraint on their production and utility. N-carbamylglutamate (NCG) can increase arginine synthesis in mammals and has been shown to improve reproductive performance. Twelve multiparous and simutaneous anoestrous female yaks were randomly divided into two groups, one of which was fed the basal diet (Control, n = 6), and the other was fed the basal diet supplemented with NCG at 6 g/day/yak (NCG, n = 6). All yaks were slaughtered on the 32nd day (the time predicted for the selection of the last wave of dominant follicles), and their ovarian tissues were collected and follicles were classified. NCG supplementation increased the number of large ovarian follicles (diameter > 10 mm), as well as caused significant changes in the transcriptional and metabolic levels in yak ovaries which due to the differential expression of 889 genes and 94 metabolites. Integrated analysis of the transcriptomics and metabolomics data revealed that the differentially expressed genes and differential metabolites were primarily involved in the process of energy metabolism, amino acid metabolic pathways, carbohydrate metabolic pathways, and lipid metabolic pathways. The highlighted changes were associated with amino acid synthesis and metabolism, ovarian steroid hormone synthesis, the pentose phosphate pathway, and the tricarboxylic acid cycle, suggesting that NCG supplementation may promote estrogen synthesis and help regulate follicular development by altering the pathways associated with glucose catabolism. The results present important clues for understanding the mechanisms by which NCG supplementation promotes follicular development in yaks. The findings of this study provide a basis for the development and application of NCG in optimizing animal reproduction, including yak reproductive performance, which may help optimize livestock management and uplift the pastoral economy.

Rumen Fermentation and Microbiome Responses to Enzymatic Hydrolysate of Cottonseed Protein Supplementation in Continuous In Vitro Culture.

This study aimed to evaluate the effect of enzymatic hydrolysate of cottonseed protein (ECP) on the kinetic of gas production, rumen fermentation characteristics, and microbial diversity in continuous in vitro culture with a single factorial design of supplementation with various concentrations of ECP or yeast culture. Treatments were control (without supplementation, CON), supplementation with 10 g/kg Diamond-V XP yeast culture of substrate (XP), and supplementation with 6, 12 and 18 g/kg ECP of substrate (ECP1, ECP2, ECP3), each incubated with 30 mL of buffered incubation fluids and 200 mg of fermentation substrate in graduated glass syringes fitted with plungers for 48 h. Compared with the CON treatment, supplementation of XP yeast culture increased the cumulative gas production at 12 and 24 h, the concentration of ammonia nitrogen (NH3-N) concentration at 24 and 36 h, the concentration of microbial protein (MCP) concentration at 24 and 48 h, the molar butyrate proportion at 12, 24, and 48 h, the molar valerate proportion at 48 h, and the ratio of non-glucogenic to glucogenic acids (p < 0.05). Compared with the CON treatment, the concentration of MCP and the molar propionate proportion at 12 h were higher in the ECP1 treatment (p < 0.05); the cumulative gas production at 2, 4, and 12 h, the concentration of NH3-N at 36 h and the molar valerate proportion at 48 h were higher in the ECP2 treatment (p < 0.05); the cumulative gas production at 2, 12, and 48 h, the concentration of NH3-N at 12 and 36 h, the concentration of MCP at 12, 36, and 48 h, the molar butyrate proportion at 12 and 48 h, and the molar valerate proportion at 48 h were higher in the ECP3 treatment (p < 0.05). Compared with the CON treatment, supplementation with XP yeast culture significantly altered the relative abundance of the phyla Firmicutes, Kiritimatiellaeota, and Proteobacteria, while supplementation with ECP had minimal effect on bacterial diversity. The prediction of bacterial functions showed that the main gene functions of rumen bacteria are associated with carbohydrate metabolism, amino acid metabolism, and membrane transport. The findings of this study suggest that ECP can be used as a superior feed ingredient for ruminants, the suitable level of ECP was 18 g/kg in vitro experiment.

Changes in Rumen Bacterial Community Induced by the Dietary Physically Effective Neutral Detergent Fiber Levels in Goat Diets.

Physically effective neutral detergent fiber (peNDF) is a concept that accounts for the particle length of NDF in a feed, sustaining the normal chewing behavior and rumen fermentation of ruminants. This study aimed to elucidate the effects of dietary peNDF on growth performance and bacterial communities in the rumen of goats through a high-throughput sequencing technique. A total of 30 male Lezhi black goats were randomly assigned to five groups, corresponding to five diets with identical compositions and nutrient levels but with varying forage lengths (the peNDF1.18 contents of the diets were 33.0, 29.9, 28.1, 26.5, and 24.8%, respectively). The whole trial lasted for 44 days. As results show, feed intake and average daily gain were highest when peNDF1.18 content was 26.5%, in which the papilla length of the dorsal sac in rumen was the highest. Chao1 and ACE indexes were similar among the treatments, while Shannon and Simpson indexes of the peNDF1.18 = 28.1% group were the highest (p < 0.05). As the level of dietary peNDF1.18 decreased, the dominant phylum transitioned from Bacteroidetes to Firmicutes. The top three dominant genera of rumen bacteria were Prevotella 1, Ruminococcaceae NK4A214 group, and Christensenellaceae R-7 group. They all showed a quadratic correlation with dietary peNDF1.18 level (p < 0.05). The relative abundance of Ruminococcaceae UCG-011 was positively correlated, while that of Prevotella 1 was negatively correlated, with amino acid metabolism and energy metabolism (p < 0.01). In conclusion, dietary peNDF level influenced goat growth performance, rumen development, and rumen bacterial community structures, and a peNDF1.18 level between 26.5 and 28.1% was considered optimal for goat diet.

Effect of hyperthermia on cell viability, amino acid transfer, and milk protein synthesis in bovine mammary epithelial cells.

The reduction of milk yield caused by heat stress in summer is the main condition restricting the economic benefits of dairy farms. To examine the impact of hyperthermia on bovine mammary epithelial (MAC-T) cells, we incubated the MAC-T cells at thermal-neutral (37°C, CON group) and hyperthermic (42°C, HS group) temperatures for 6 h. Subsequently, the cell viability and apoptotic rate of MAC-T cells, apoptosis-related genes expression, casein and amino acid transporter genes, and the expression of the apoptosis-related proteins were examined. Compared with the CON group, hyperthermia significantly decreased the cell viability (p < 0.05) and elevated the apoptotic rate (p < 0.05) of MAC-T cells. Moreover, the expression of heat shock protein (HSP)70, HSP90B1, Bcl-2-associated X protein (BAX), Caspase-9, and Caspase-3 genes was upregulated (p < 0.05). The expression of HSP70 and BAX (pro-apoptotic) proteins was upregulated (p < 0.05) while that of B-cell lymphoma (BCL)2 (antiapoptotic) protein was downregulated (p < 0.05) by hyperthermia. Decreased mRNA expression of mechanistic target of rapamycin (mTOR) signaling pathway-related genes, amino acid transporter genes (SLC7A5, SLC38A3, SLC38A2, and SLC38A9), and casein genes (CSNS1, CSN2, and CSN3) was found in the heat stress (HS) group (p < 0.05) in contrast with the CON group. These findings illustrated that hyperthermia promoted cell apoptosis and reduced the transport of amino acids into cells, which inhibited the milk proteins synthesis in MAC-T cells.

Enhanced supply of methionine regulates protein synthesis in bovine mammary epithelial cells under hyperthermia condition.

Recent evidence has shown that methionine (Met) supplementation can improve milk protein synthesis under hyperthermia (which reduces milk production). To explore the mechanism by which milk protein synthesis is affected by Met supplementation under hyperthermia, mammary alveolar (MAC-T) cells were incubated at a hyperthermic temperature of 42°C for 6 h in media with different concentrations of Met. While the control group (CON) contained a normal amino acid concentration profile (60 µg/mL of Met), the three treatment groups were supplemented with Met at concentrations of 10 µg/mL (MET70, 70 µg/mL of Met), 20 µg/mL (MET80, 80 µg/mL of Met), and 30 µg/mL (MET90,90 µg/mL of Met). Our results show that additional Met supplementation increases the mRNA and protein levels of BCL2 (B-cell lymphoma-2, an anti-apoptosis agent), and decreases the mRNA and protein levels of BAX (Bcl-2-associated X protein, a pro-apoptosis agent), especially at an additional supplementary concentration of 20 µg/mL (group Met80). Supplementation with higher concentrations of Met decreased the mRNA levels of Caspase-3 and Caspase-9, and increased protein levels of heat shock protein (HSP70). The total protein levels of the mechanistic target of rapamycin (mTOR) and the mTOR signalling pathway-related proteins, AKT, ribosomal protein S6 kinase B1 (RPS6KB1), and ribosomal protein S6 (RPS6), increased with increasing Met supplementation, and peaked at 80 µg/mL Met (group Met80). In addition, we also found that additional Met supplementation upregulated the gene expression of αS1-casein (CSN1S1), ß-casein (CSN2), and the amino acid transporter genes SLC38A2, SLC38A3 which are known to be mTOR targets. Additional Met supplementation, however, had no effect on the gene expression of κ-casein (CSN3) and solute carrier family 34 member 2 (SLC34A2). Our results suggest that additional Met supplementation with 20 µg/mL may promote the synthesis of milk proteins in bovine mammary epithelial cells under hyperthermia by inhibiting apoptosis, activating the AKT-mTOR-RPS6KB1 signalling pathway, and regulating the entry of amino acids into these cells.

Hepatic Injury Induced by Dietary Energy Level via Lipid Accumulation and Changed Metabolites in Growing Semi-Fine Wool Sheep.

Liver injury threatens the overall health of an organism, as it is the core organ of the animal body. Liver metabolism is affected by numerous factors, with dietary energy level being a crucial one. Therefore, the present study aimed to evaluate hepatic injury and to describe its metabolic mechanism in ruminants fed diets with different dietary energy levels. A total of 25 Yunnan semi-fine wool sheep were fed diets with five dietary metabolic energy levels and were randomly assigned to five groups as follows: low energy (LE), medium-low energy (MLE), medium energy (ME), medium-high energy (MHE), and high energy (HE). The results revealed that the average optical density (AOD) of lipid droplets in the LE, MLE, and HE groups was higher than that in the ME and MHE groups. The enzyme activity of alanine aminotransferase (ALT) was the lowest in the ME group. An increase in dietary energy level promoted the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity and altered the malondialdehyde (MDA) and protein carbonyl (PCO) concentration quadratically. In addition, both high and low dietary energy levels upregulated the mRNA abundance of proinflammatory cytokine interleukin (IL)-1ß, nuclear factor-kappa B (NF-κB), and tumor necrosis factor (TNF)-α. Metabonomic analysis revealed that 142, 77, 65, and 108 differential metabolites were detected in the LE, MLE, MHE, and HE groups, compared with ME group respectively. These metabolites were involved in various biochemical pathways, such as glycolipid, bile acid, and lipid metabolism. In conclusion, both high and low dietary energy levels caused hepatic injury. Section staining and metabonomic results revealed that hepatic injury might be caused by altered metabolism and lipid accumulation induced by lipid mobilization.

Effects of Premating Calcium and Phosphorus Supplementation on Reproduction Efficiency of Grazing Yak Heifers.

Reproductive efficiency is the main factor limiting yak production on the Tibet Plateau. The purpose of this study was to investigate the effect of supplementation with calcium chloride (CaCl) and monocalcium phosphate (MCP) for 30 days before breeding on body weight (BW) change, serum bone metabolism biomarkers, conception rate, and calving rate of grazing yaks. Ninety 3 year old yak heifers (153.05 ± 6.56 kg BW) were assigned to three treatments (n = 30 per treatment): grazing without supplementation (CONT), grazing plus calcium chloride supplementation (CaCl), and grazing plus monocalcium phosphate supplementation (MCP). Compared with the CONT group, supplementation with CaCl increased the serum concentrations of osteocalcin and decreased the alkaline phosphatase (ALP) levels (p < 0.05); supplementation with MCP increased the average daily gain (ADG), serum concentrations of phosphorus (P) and osteocalcin, conception rate, and calving rate (p < 0.05), whereas it decreased the serum concentrations of hydroxyproline, ALP, and calcitonin (p < 0.05). Both CaCl and MCP supplementation had no effect on serum calcium (Ca) concentration. The ADG, conception rate, and calving rate were higher in the MCP group than in the CaCl group (p < 0.05), while the serum concentrations of hydroxyproline and calcitonin were lower (p < 0.05). It could be concluded that premating supplementation with MCP increased the body weight gain and subsequent conception and calving rate of grazing yaks. Supplementation with MCP had a positive effect on body condition and bone metabolism, thus providing a better estrous condition for grazing yak heifers, which could contribute to enhancing reproduction efficiency.