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This study was conducted to determine the response of the reproductive hormones and the mTOR/AKT pathway in the ovaries of post-partum dairy cows with the dietary rumen-protected glucose (RPG) supplementation. Twelve Holstein cows were randomly assigned to two groups (n = 6 / group): control group (CT) and RPG group. Cows were fed with the diets from 1 week prepartum to 2 weeks postpartum, and euthanized on d 22 postpartum. Blood samples were collected from cows in each group on d 1, 7 and 14 after calving. The plasma hormones were determined using a bovine specific enzyme linked immimosorbent assay method. The gene expressions of the reproductive hormone receptors and mTOR/AKT pathway in the ovarian tissues were detected using real-time polymerase chain reaction, Western blot, and immunohistochemistry, respectively. The results showed that the RPG addition significantly increased the plasma LH, E2, and P4 concentrations on d14 after calving. The mRNA and protein expressions of the estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), 17β-hydroxysteroid dehydrogenase (17β-HSD), FSHR, LHR and cytochrome p450 (CYP17A1) were upregulated, while the mRNA and protein expression of steroidogenic acute regulatory protein (StAR) was downregulated in cows of the RPG group compared with the CT group. Immunohistochemical analysis identified higher expressions of the FSHR and LHR in the ovaries of RPG-fed cows versus CT cows. Furthermore, the ratios of the protein expressions of p-AKT to total AKT and p-mTOR to total mTOR ratio were significantly increased in the ovaries of RPG-fed cows compared with the CT group cows. The current results indicated that dietary RPG supplementation regulated the secretion of gonadal hormones, and stimulated the expressions of hormone receptors and the mTOR/AKT pathway in the ovaries of early postpartum dairy cows, and might beneficial for ovarian activity recovery in the post-calving dairy cows.

Ramie (Boehmeria nivea) is noted for the production of a large biomass that has a high protein content and is rich in antioxidants. It may thus serve as a high-quality forage material to replace alfalfa and improve the meat quality of farmed animals. In this study, we evaluated the carcass characteristics and meat quality of goats when 0, 35%, 75%, and 100% of dietary alfalfa was replaced with ramie. Crude protein content (linear, P

Tea tree oil (TTO) is a plant-derived additive with anti-inflammatory, bactericidal, and growth-promoting properties. However, little is known about the effects of TTO on intestinal amino acid transport and immune function in goats. Twenty-four Ganxi goats (initial body weight of 13.5 ± 0.70 kg) were randomly allotted two treatments and fed either control (CON) or CON+TTO (0.2 ml/kg) diet. The addition of TTO to the diet significantly decreased (

Maintaining muscle quality throughout life is crucial to human health and well-being. Muscle is the most extensive form of protein storage in the human body; skeletal muscle mass is determined by the balance between muscle protein synthesis (MPS) and muscle protein breakdown (MPB). MPB provides amino acids needed by various organs; however, excessive MPB, especially with aging, may cause loss of muscle mass and a decline in motor function, even threatening life. The turnover of muscle protein is vital to the health of humans. Thus, although the study of MPS and MPB has theoretical and practical significance, the network that controls MPS is very complicated and we cannot discuss both MPS and MPB in a single review. Therefore, the aim of this review is to discuss the regulation of MPS, especially by amino acids. Amino acids regulate protein synthesis in cell and animal models, but compelling evidence for amino acids promoting protein synthesis in human muscles is ambiguous. Studies on the stimulation of human MPS by branched-chain amino acids (BCAAs) have been inconsistent. Amino acids other than BCAAs such as threonine and tryptophan may also have MPS-stimulating effects, and alternatives to BCAAs, such as β-hydroxy-β-methyl butyrate and branched-chain keto acids are also worthy of further investigation. Amino acids coordinate protein synthesis and degradation through the mechanistic target of rapamycin complex 1 (mTORC1); however, the amino acid-mTORC1-protein synthesis pathway is complex, and new insights into amino acid control continue to emerge. Understanding how amino acids control MPS is of forward-looking significance for treating muscle mass loss during human aging.

Alfalfa silage is one of the main roughages in the production of dairy cow, which can provide nutrition with high quality to improve milk quality and production. Sucrose additions have been widely used to improve the silage quality. In this study, the effects of sucrose on the fermentation quality and bacterial communities of alfalfa silage were investigated here using 0, 0.5, and 1% sucrose ensiling treatments for 15, 30, and 60 days. The ensiling time significantly decreased the crude fiber content and increased the ammonia nitrogen, acetic acid content, and the relative abundance of Enterococcus in the silages. The 1% sucrose-treated silage at 60 days had the lowest neutral detergent fiber acid, acid detergent fiber, and crude fiber content and the highest relative feed value. Moreover, sucrose-treated silage contained less acetic acid, propionic acid, and butyric acid, and had a lower pH than the controls for each duration. Enterobacteriaceae, Klebsiella, and Enterococcus were the dominant genera in all groups, and the relative abundance of Enterococcus and Lactobacillus was higher in the 1% sucrose-treated group than in the control. These results suggested that sucrose supplementation could improve alfalfa silage quality and increase its beneficial bacterial content.

The study investigated the effect of alfalfa hay substituted with ramie silage on the expression of apoptotic genes in the gastrointestinal tract of goats. Thirty-two goats were randomly allocated into four groups, in which the alfalfa was substituted with ramie at 0%, 35%, 75%, and 100% levels, respectively. In the rumen, the mRNA expression of Bax was significantly up-regulated (

Mulberry and pigeonpea trees are two widely distributed trees in southwest regions of China, and grow well in summer and winter season respectively. The present study is to investigate the nutritive value of mulberry and pigeonpea leaves by using in vitro ruminal fermentation. Pigeonpea leaves were harvested in February, October, November and December, while mulberry leaves were harvested in May, July, September, October. Comparisons of chemical composition, kinetics of in vitro gas production, methane production, dry matter degradation and VFA production of pigeonpea and mulberry leaves were performed by using in vitro ruminal batch culture system. Pigeonpea and mulberry leaves had crude protein varying from 186 to 208 and 209 to 269 g/kg dry matter, respectively, and neutral detergent fiber ranging from 395 to 456 and 252 to 336 g/kg dry matter, respectively. In vitro fermentation indicated that pigeonpea leaves had lower in vitro dry matter degradation and theoretical maximum of gas production, but greater initial fractional rate of degradation and acetate to propionate ratio in comparison with mulberry leaves. Meanwhile, harvest time influenced the acetate to propionate ratio of both leaves. In conclusion, pigeonpea and mulberry leaves have protein content larger than 180 g/kg dry matter, and can be alternative protein resource for ruminants. In addition, harvest time significantly affect the fermentation pathway of these two leaves.

Supplementation of rumen-protected glucose (RPG) may be a useful dietary strategy to improve energy balance in transition dairy cows. The present study was performed to investigate whether supplementation with RPG could improve milk production and reduce inflammation or infection during the transition period in cows. Twelve Holstein cows (age, 4–5 yrs; bodyweight, 515 ± 42 kg; milk yield, 16.1 ± 3.7 kg/d)in late gestation were allotted to one of 2 treatments. The treatments were a basal diet plus 200 g/d of RPG (450 g/kg of glucose, 450 g/kg of coating fat and 100 g/kg of water) or a basal diet plus 90 g/d of coating fat (CON). Treatments were applied from 1 wk before to 2 wk after calving. Animal performance data were recorded, and the overall tract apparent digestibility of prepartum or postpartum cows was determined. Blood was sampled on d −4, 0, 7, and 14 to examine the plasma biochemical parameters and proinflammatory cytokines. The RPG supplementation increased the milk yield, but did not change the dry matter intake. Compared with cows in the CON group, the plasma total protein concentration was lower in the RPG group. Cows in the RPG group had greater plasma albumin concentrations but lower globulin and interleukin-8 concentrations during the transition period. The RPG supplementation tended to increase the plasma alanine aminotransferase concentration but did not change the plasma aspartate aminotransferase concentration. Dietary RPG supplementation increased the non-esterified fatty acid concentration and tended to decrease the plasma glucose concentration. In conclusion, these results indicate that supplementation with 200 g RPG per day improves milk production and reduces inflammatory markers but might have compromised the liver function of the transition cows in this study.

Zinc supplementation in the diet of goats affects pancreas development in offspring. However, the impact of maternal inorganic and organic zinc supplementation in offspring is poorly defined. In this study, 14 late-pregnant goats were assigned at random to the zinc sulfate group (ZnSO

This study investigated changes in oxidative stress and the relevant mechanisms in the liver of goats fed a high-concentrate (HC) diet for 5 weeks. Twelve goats were randomly assigned to a low-concentrate (concentrate-to-forage = 55:45, LC, n = 6) or HC diet (concentrate-to-forage = 90:10, n = 6), with dry matter as the base. Enzyme activity assays, real-time polymerase chain reaction and western blotting were used to evaluate antioxidant parameters and gene expression in the liver. Superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), total nitric oxide synthase activity and total antioxidant capacity (T-AOC) in the liver declined (p < .01) in HC-fed goats compared to those in LC-fed goats. The mRNA levels of GPX1, CAT and SOD1 were down-regulated (3.69, 47.37 and 27.61%, respectively) in HC-fed goats compared to those in LC-fed goats. Furthermore, glutathione S-transferase M1 (GSTM1) was upregulated (466.35%, p < .01) in the liver of HC-fed goats. The mRNA and protein levels of the nuclear factor E2-related factor 2 (NRF2) and total glucocorticoid receptor (GR) declined (p < .05) in HC-fed goats (by 28.57, 33.1, 30.85 and 34%, respectively). However, the nuclear translocation of GR increased (p < .05; by 44.75%) in HC-fed goats. Negative correlations were detected for hepatic nuclear GR protein expression with hepatic CAT activity and GPx activity. In conclusion, feeding an HC diet to goats for 5 weeks suppressed NRF2-dependent antioxidant responses and enhanced GR nuclear translocation in the liver.HighlightsFeeding goats high-concentrate diets suppresses NRF2-dependent antioxidant responses in liver, downregulating NRF2 and altering SOD1, GPx and CAT activity.This suppression of NRF2-dependent antioxidant responses is associated with GR activation in the liver of goats. Feeding goats high-concentrate diets suppresses NRF2-dependent antioxidant responses in liver, downregulating NRF2 and altering SOD1, GPx and CAT activity. This suppression of NRF2-dependent antioxidant responses is associated with GR activation in the liver of goats.

To evaluate the inhibitory effect of chlorogenic acid on the forming of type 2 diabetes mellitus (T2DM), using Sprague Dawley (SD) rats, a recognized T2DM model induced by high-fat high-sucrose diet (HFSD) and streptozotocin (STZ). Thirty female SD rats were assigned equally to three groups randomly: normal control with standard commercial (NC), chlorogenic acid treatment with HFSD and chlorogenic acid (90mg/kg, CA), and diabetes model with HFSD (DM). Upon treatment with chlorogenic acid, suppression of the onset of diabetes, reduced serum glucose and insulin concentrations, improved glucose tolerance and increased body weight and visceral fat weight were observed. Serum triglyceride, total cholesterol, low density lipoprotein levels, and kidney and pancreas morphology were significantly ameliorated. Chlorogenic acid also inhibited the mRNA levels of hepatic G-6-Pase and up-regulated the mRNA levels of skeletal muscle GLUT4. Our results indicated that before the onset of diabetes, chlorogenic acid had an inhibitory effect against the forming of T2DM induced by HFSD and STZ through regulating the glucose and lipid metabolism.

The objective of this study was to investigate the variations in nutrient digestibility, rumen fermentation, serum biochemical parameters, and performance resulting from an increase in the proportion of ramie (Boehmeria nivea) substitution for alfalfa in the diets of goats. In a completely randomized block design, thirty-two Liuyang black goats were divided into two blocks according to body weight (BW) (light, BW: 16.7 ± 1.72 kg; and heavy, BW: 26.6 ± 2.95 kg). Then, the sixteen goats in each block were further randomly divided into four subgroups that randomly accepted four treatments, i.e., alfalfa was replaced by ramie at 0%, 35%, 70% and 100%. The results showed that the apparent digestibility of gross energy (GE) (linear, p 0.05) by the inclusion of ramie. The concentration of ammonia nitrogen (NH3-N) in the rumen fluid decreased (linear, p

Simple Summary A rapid recovery of the uterus is important in order to shorten the interval between pregnancies. An insufficient availability of glucose in early lactation can delay the uterine recovery. However, little information is available on dietary rumen-protected glucose addition on the uterine involution and insulin-like growth factor system in the endometrium of dairy cows during the early post-natal period. In this study, dairy cows were fed with 0 or 200 g rumen-protected glucose twice every day from −7 ± 2 to 14 d during the post-natal period. The results indicated that dietary rumen-protected glucose addition promoted the proliferation of endometrial cells in postpartum dairy cows by stimulating the insulin-like growth factor system and rapamycin complex 1 pathway, and might be beneficial for uterine recovery. Abstract This study aimed to elucidate the effects of a dietary rumen-protected glucose (RPG) addition on uterine involution through the analysis of an insulin-like growth factor (IGF) system and associated pathways in the post-natal endometrium. Twelve Holstein cows were assigned equally to two groups: a control group (CT) and an RPG group (200 g of RPG per cow per day). The plasma content of insulin-like growth factor 1 (IGF1) was determined by using the ELISA method. Expressions of IGF members, the matrix metalloproteinase, protein kinase B (AKT)/mechanistic target of rapamycin complex1 (mTOR) signaling pathway, and cell proliferation factors (proliferating cell nuclear antigen (PCNA) and Ki67) were detected using real-time polymerase chain reaction, Western blot, immunohistochemistry, and immunofluorescence, respectively. The results showed that the positive cells of PCNA and Ki67 were increased in the endometrium of RPG versus CT cows. The RPG addition significantly increased the plasma IGF1 level 14 d after delivery. The mRNA expressions of the IGF family members (IGF1, IGF2, type 1 IGF receptor (IGF1R) and IGF-binding proteins (IGFBP1, IGFBP2, IGFBP4 and IGFBP5)) were upregulated, and mRNA expressions of matrix metalloproteinase MMP3 and MMP9 were downregulated in cows from the RPG group compared with the CT group. Meanwhile, the protein expressions of IGF1, IGF2, IGF1R, IGFBP1 and IGFBP4 were upregulated in cows from the RPG group compared with the CT group. Immunohistochemical analysis identified a positive response for IGF1R and IGF2R in the endometrium of RPG versus CT cows. Furthermore, the RPG supplementation increased the protein expressions of phosphorylated (p)-AKT to total AKT and p-mTOR to total mTOR ratio in the endometrium. The current results indicated that the RPG supplementation promoted the proliferation of endometrial cells by stimulating the IGFs and mTOR/AKT pathway in the early post-natal endometrium of dairy cows.

Sufficient amino acid is necessary to the growth of animals. The experiment investigated the effects of dietary unprotected methionine (Met) and lysine (Lys) sources on nutrient digestion, nitrogen utilization and mRNA expression of related genes in growing goats fed a maize stover-based diet. Twenty Liuyang black weather goats were used and assigned randomly to 4 dietary treatments: (1) the control (basal) diet (NS:), (2) control diet with a supplementation of unprotected DL-Met (SM), (3) control diet with a supplementation of unprotected L-Lys-HCI (SL), (4) control diet with a supplementation of unprotected DL-Met and L-Lys-HCI (SML). The experiment showed that there were no differences of DMI (dry matter intake), fecal output, apparent digestibility, energy balance and N balance (P > 0.05). Adding Lys decreased serum threonine, isoleucine, phenylalanine, total amino acids (P

The 4 experimental diets, all containing rapeseed oil, consisted of one control and 3 other diets supplemented with 3 different levels of commercial oenological extracts tannins (i.e. 3, 6 and 9 g/kg dry matter (DM), respectively). The effect of the added tannins on the fatty acid profiles of ruminal fluid was tested in vitro, and 20 multiparous Liuyang black nannies [average body weight (BW): 26.3 ± 0.5 kg] at week 6 of lactation were used in an in vivo experiment. For the in vitro experiment, the inclusion of dietary tannins did not ( P > 0.05) affect fatty acid contents in the culture fluid in vitro, except the percentage of total conjugated linoleic acid (CLA) tended to increase (quadratically, P = 0.075) with increasing tannins level. Increasing incubation time decreased the percentage of total cis C17:1 ( P = 0.001), cis -11 C18:1 ( P cis -12 C18:1 ( P trans -11 C18:1 ( P = 0.016), total cis C18:1 ( P P cis -9, cis -12 C18:2 n-6 linoleic acid (LA) ( P P UFA) ( P = 0.001). In the in vivo experiment, concentration of C16:1 in goat milk linearly increased ( P = 0.032) and that of cis -11 C18:1 tended (quadratic effect, P = 0.072) to increase with increasing levels of tannins. Tannin inclusion did not affect ( P > 0.05) the content of protein, lactose, ash, solid not fat and total solids in the goat milk, but higher levels of tannins reduced milk fat content ( P = 0.005), compared with the control diet. Compared with the control, tannins inclusion decreased concentrations of cis -11 C20:1 ( P = 0.022) and total n-9 monounsaturated fatty acids (MUFA) ( P = 0.017) in milk at the 3 g/kg DM inclusion rate, decreased C11:0 ( P = 0.012) and C13:0 ( P = 0.029) at 6 g/kg DM, but increased cis -9 C18:1 ( P = 0.043) and C22:6 n-3 docosahexenoic acid (DHA) ( P = 0.013) at the 9 g/kg DM inclusion rate. Concentrations of cis -14 C18:1, cis -9, cis -12 C18:2 n-6 LA, total cis MUFA, PUFA, n-3 and n-6 PUFA in milk linearly increased ( P

Four levels of Momordica charantia polysaccharide (MCP) supplements (0, 0.1, 0.3, 0.6 mg/ml) were designed to investigate the effects of MCP on ruminal fermentation and cellulolytic bacteria in vitro. The pH, ammonia-N (NH3-N) and volatile fatty acids (VFA) were measured at 6, 24, 48 h, whilst the cellulolytic bacteria population was determined at 6 and 24 h. The 0.6 mg/ml MCP inclusion decreased the theoretical maximum of gas production and the half-life. The NH3-N concentration was decreased by MCP at all doses at 24 h. The MCP inclusion increased the concentration of total VFA at 24 and 48 h and the acetate to propionate ratio, the molar proportion of isovalerate at 6 h, while decreased that of isobutyrate at 24 h and that of isovalerate, valerate at 24 and 48 h, respectively. The relative abundances of Ruminococcus albus and Ruminococcus flavefaciens were decreased at 6 h, while that of Butyrivibrio fibrisolvens was increased at all times of incubation and that of Fibrobacter succinogenes reached the greatest value at 0.6 mg/ml MCP supplementation at 24 h. This study demonstrated that the MCP had the ability to enhance the total VFA production, modulate the rumen fermentation pathway and influence the number of cellulolytic bacteria population.

Background In developing countries, maternal undernutrition is the major intrauterine environmental factor contributing to fetal development and adverse pregnancy outcomes. Maternal nutrition restriction (MNR) in gestation has proven to impact overall growth, bone development, and proliferation and metabolism of mesenchymal stem cells in offspring. However, the efficient method for elucidation of fetal bone development performance through maternal bone metabolic biochemical markers remains elusive. Methods We adapted goats to elucidate fetal bone development state with maternal serum bone metabolic proteins under malnutrition conditions in mid- and late-gestation stages. We used the experimental data to create 72 datasets by mixing different input features such as one-hot encoding of experimental conditions, metabolic original data, experimental-centered features and experimental condition probabilities. Seven Machine Learning methods have been used to predict six fetal bone parameters (weight, length, and diameter of femur/humerus). Results The results indicated that MNR influences fetal bone development (femur and humerus) and fetal bone metabolic protein levels (C-terminal telopeptides of collagen I, CTx, in middle-gestation and N-terminal telopeptides of collagen I, NTx, in late-gestation), and maternal bone metabolites (low bone alkaline phosphatase, BALP, in middle-gestation and high BALP in late-gestation). The results show the importance of experimental conditions (ECs) encoding by mixing the information with the serum metabolic data. The best classification models obtained for femur weight (Fw) and length (FI), and humerus weight (Hw) are Support Vector Machines classifiers with the leave-one-out cross-validation accuracy of 1. The rest of the accuracies are 0.98, 0.946 and 0.696 for the diameter of femur (Fd), diameter and length of humerus (Hd, Hl), respectively. With the feature importance analysis, the moving averages mixed ECs are generally more important for the majority of the models. The moving average of parathyroid hormone (PTH) within nutritional conditions (MA-PTH-experim) is important for Fd, Hd and Hl prediction models but its removal for enhancing the Fw, Fl and Hw model performance. Further, using one feature models, it is possible to obtain even more accurate models compared with the feature importance analysis models. In conclusion, the machine learning is an efficient method to confirm the important role of PTH and BALP mixed with nutritional conditions for fetal bone growth performance of goats. All the Python scripts including results and comments are available into an open repository at https://gitlab.com/muntisa/goat-bones-machine-learning.

Changes in the environment such as high-altitude hypoxia (HAH) high-altitude hypoxia can lead to adaptive changes in the blood system of mammals. However, there is limited information about the adaptation of Holstein dairy cows introduced to high-altitude areas. This study used 12 multiparous Holstein dairy cows (600 &plusmn, 55 kg, average three years old) exposed to HAH conditions in Nyingchi of Tibet (altitude 3000 m) and HAH-free conditions in Shenyang (altitude 50 m). The miRNA microarray analysis and iTRAQ proteomics approach (accepted as more suitable for accurate and comprehensive prediction of miRNA targets) were applied to explore the differences in the plasma proteomic and miRNA profiles in Holstein dairy cows. A total of 70 differential miRNAs (54 up-regulated, Fold change (FC) FC &gt, 2, and 16 down-regulated, FC &lt, 0.5) and 226 differential proteins (132 up-regulated, FC &gt, 1.2, and 94 down-regulated, FC &lt, 0.8) were found in the HAH-stressed group compared with the HAH-free group. Integrative analysis of proteomic and miRNA profiles demonstrated the biological processes associated with differential proteins were the immune response, complement activation, protein activation, and lipid transport. The integrative analysis of canonical pathways were most prominently associated with the APR signaling (z = 1.604), and LXR/RXR activation (z = 0.365), and FXR/RXR activation (z = 0.446) pathways. The current results indicated that Holstein dairy cows exposed to HAH could adapt to high-altitude hypoxia by up-regulating the APR, activating the LXR/RXR and FXE/RXR pathways.

This study investigated the in vitro fermentation of three cereal straws (corn stover, wheat straw, and rice straw) with culture media containing two nonionic surfactants (alkyl polyglycoside and Tween 80) and variable surface tension (42, 48, and 54 mN/m) in a 2 × 3 factorial arrangement. The in vitro dry matter degradation of rice straw and corn straw linearly (P

Quorum sensing (QS) is a type of microbe-microbe communication system that is widespread among the microbial world, particularly among microorganisms that are symbiotic with plants and animals. Thereby, the cell-cell signalling is likely to occur in an anaerobic rumen environment, which is a complex microbial ecosystem. In this study, using six ruminally fistulated Liuyang black goats as experimental animals, we aimed to detect the activity of quorum sensing autoinducers (AI) both in vivo and in vitro and to clone the luxS gene that encoded autoinducer-2 (AI-2) synthase of microbial samples that were collected from the rumen of goats. Neutral detergent fiber (NDF) and soluble starch were the two types of substrates that were used for in vitro fermentation. The fermented fluid samples were collected at 0, 2, 4, 6, 8, 12, 24, 36, and 48 h of incubation. The acyl-homoserine lactones (AHLs) activity was determined using gas chromatography-mass spectrometer (GC-MS) analysis. However, none of the rumen fluid extracts that were collected from the goat rumen showed the same or similar fragmentation pattern to AHLs standards. Meanwhile, the AI-2 activity, assayed using a Vibrio harveyi BB170 bioassay, was negative in all samples that were collected from the goat rumen and from in vitro fermentation fluids. Our results indicated that the activities of AHLs and AI-2 were not detected in the ruminal contents from six goats and in ruminal fluids obtained from in vitro fermentation at different sampling time-points. However, the homologues of luxS in Prevotella ruminicola were cloned from in vivo and in vitro ruminal fluids. We concluded that AHLs and AI-2 could not be detected in in vivo and in vitro ruminal fluids of goats using the current detection techniques under current dietary conditions. However, the microbes that inhabited the goat rumen had the potential ability to secrete AI-2 signaling molecules and to communicate with each other via AI-2-mediated QS because of the presence of luxS .

The objective of this study was to determine whether 13 C labeling occurs in goats fed diets with different natural levels of 13 C enrichment and, if so, to use 13 C as a label to determine true amino acid (AA) digestibility. Thirty-six Xiangdong black goats with initial body weights of 7.5 ± 0.5 kg were selected as experimental animals and fed dietary treatments consisting of wheat, barley, and soybeans (C3 diet) or corn, sorghum grain, and sugar cane (C4 diet). The 13 C abundance of the amino acid fraction (AAF) of the C3 and C4 diets had average delta 13 C values of −28.57‰ and −12.93‰, respectively. Three goats/treatment were slaughtered on days 1, 28, 56 and 84 during the labeling phase, and the AAFs of organs were analyzed for 13 C abundance. 13 C in the blood AAF increased (−21.70‰, −22.65‰, −23.59‰, and −24.43‰, respectively) with increasing feeding durations in goats fed the C3 diet and decreased (−21.13‰, −16.01‰, −13.78‰, and −12.03‰, respectively) in goats fed the C4 diet. Longissimus dorsi, liver and wool AAFs showed similar trends to those observed in the blood. Each tissue showed a significant dietary treatment effect ( P P 13 C abundance. 13 C abundance of the blood AAF was used as an index of endogenous protein labeling. Apparent and true protein digestibilities in the forestomach, whole intestine and whole digestive tract were not different ( P > 0.05) between the diet treatment groups. The levels of true protein digestibility in the forestomach, whole intestine and whole digestive tract in goats fed the C4 diet tended to be higher than for those in goats fed the C3 diet. On average, the true digestibilities of protein, essential AAs and nonessential AAs were 5.30%, 4.28% 3.17% (forestomach), 5.20%, 5.39% 3.75% (whole intestine) and 3.01%, 4.97%, 4.17% (whole digestive tract) higher, respectively, than the apparent digestibilities of these components in goats fed the C3 diet. In summary, goat tissue protein was labeled with 13 C using different diets, and the results show that 13 C can be used to determine true AA digestibility.

This study was conducted to investigate the effects of Momordica charantia saponin (MCS) on ruminal fermentation of maize stover and abundance of selected microbial populations in vitro. Five levels of MCS supplements (0, 0.01, 0.06, 0.30, 0.60 mg/mL) were tested. The pH, NH3-N, and volatile fatty acid were measured at 6, 24, 48 h of in vitro mixed incubation fluids, whilst the selected microbial populations were determined at 6 and 24 h. The high dose of MCS increased the initial fractional rate of degradation at t-value = 0 (FRD0) and the fractional rate of gas production (k), but decreased the theoretical maximum of gas production (V F) and the half-life (t0.5) compared with the control. The NH3-N concentration reached the lowest concentration with 0.01 mg MCS/mL at 6 h. The MSC inclusion increased (p

Gas produced was measured based on headspace pressures in both manual and automated in vitro incubation systems, which were always vented at pre-set fixed times ( FT ) and at a pre-set threshold of fixed pressure ( FP ) respectively. This study was to investigate these two venting procedures (VP) on kinetics of total gas, methane ( CH 4 ) and hydrogen gas ( gH 2 ), substrate degradation and volatile fatty acid ( VFA ) production. Corn grain, rice straw, rice grain and tall fescue were selected for in vitro ruminal batch incubation. Headspace pressure of FT procedure was manually released at incubation times being 2, 4, 8, 12, 20, 28, 36, 48 and 72 h in manual incubation system, while headspace pressure of FP procedure was automatically released at threshold pressure being 9 kPa/bottle in automated incubation system. The FT procedure had particular high venting headspace pressure (> 40 kPa) at some incubation times for corn grain and rice grain. The FT procedure had a lower gas volume ( P P P =0.01) substrate degradation and increased partial dissolution of CO 2 under higher venting headspace pressure. The FP procedure had higher ( P =0.016) fractional rate of CH 4 production than FT procedure. The FT procedure had lower final asymptotic gH 2 volume ( P =0.01) and fractional rate of gH 2 formation ( P =0.002), and higher propionate production ( P =0.05), in comparison with FP procedure. The higher propionate production under FT procedure indicated that more H 2 was entering to VFA production, leading to less gH 2 released into the headspace of bottle. In summary, the rate of gas, CH 4 and gH 2 generation could be different between VP and VT procedure, and higher venting headspace pressure caused by FT procedure altered kinetics of gas, CH 4 and gH 2 volume, substrate degradation and VFA production.

Protein nutrition is particularly important for the self-renewal processes of gastrointestinal epithelial cells. The self-renewal of cells is inseparable from the interaction between apoptosis and autophagy. However, there are few reports on the relationship between different nitrogen sources and apoptosis/autophagy. In this study, the relative protein expression of Bcl-2-associated X protein(Bax), caspase-3, and p62 was significantly higher (p &lt, 0.05), while that of Bcl-xl, Bcl-2, Beclin1, and Microtuble-associated protein light chain 3 (LC3-II) was significantly lower (p &lt, 0.05), in the NH4Cl group in comparison with the NH4Cl + 4-phenylbutyric acid (4PBA) group. In addition, the relative protein expression of Bax and caspase-3 was significantly higher (p &lt, 0.05), while that of Bcl-2 and Bcl-xl was decreased significantly (p &lt, 0.05), in the NH4Cl + 3-Methyladenine (3-MA) group and the methionine (Met) + 3-MA group in comparison with the NH4Cl group. Furthermore, the relative protein expression of Beclin1 and LC3B-II was significantly lower (p &lt, 0.05), while that of p62 was significantly higher (p &lt, 0.05), in the NH4Cl + Z-VAD-FMK group and the Met + Z-VAD-FMK group in comparison with the NH4Cl group. In conclusion, our results suggested that endoplasmic reticulum (ER) stress played a critical role in the crosstalk between apoptosis and autophagy induced by NH4Cl and Met. Autophagy had a more obvious ameliorative effect on ruminal epithelial cell apoptosis after treatment with nonprotein nitrogen than after treatment with protein nitrogen. These findings may reveal the molecular mechanism of apoptosis and autophagy induced by nonprotein nitrogen and protein nitrogen.

Sufficient amino acid (AA) transport is essential to ensure the normal physiological function and growth of growing animals. The processes of AA sensing and transport in humans and murine animals, but rarely in goats, have been arousing great interest recently. This study was conducted to investigate the messenger RNA expression patterns of lysophosphatidic acid receptor 5 (LPAR5), guanine nucleotide-binding protein α-transducing 3 (GNAT3) and important partial AA transporters in digestive tracts, metabolic organs and muscles of growing goats. The results showed that these genes were widely expressed in goats, and had different expression patterns. LPAR5, GNAT3, solute carrier (SLC38A2), SLC7A7, SLC7A1 and SLC3A1 were rarely expressed in the rumen, but were highly expressed in the abomasum and intestine which are the main sites of AA absorption. GNAT3, SLC38A1, SLC38A2, SLC6A19, SLC7A7 and SLC7A1 showed comparatively high expression in the pancreas and the vital digestive glands, and the relatively high expression of these nine genes were noted in the tibialis posterior, the active muscle in energy metabolism. The correlation analysis showed that there were certain positive correlation among most genes. The current results indicate that the AA sensing and transport occur extensively in the abomasum and small intestine, metabolic organs and muscle tissues of ruminants, and that related genes have tissue specificity.

This study was performed in a 2 × 4 factorial arrangement to explore and compare the effects of inclusion of two live Bacillus additives (B. licheniformis and B. subtilis) at four doses (0, 0.25 × 10(7), 0.50 × 10(7) and 0.75 × 10(7) colony-forming units (cfu)) on in vitro gas production kinetics, fiber degradation, methane production and ruminal fermentation characteristics of maize stover and rice straw by mixed rumen microorganisms in dairy cows. The pH, concentrations of ammonia nitrogen (NH3-N) and isovalerate were increased (P

This study was conducted to evaluate the effects of the dietary ratio of ruminal degraded protein (RDP) to ruminal undegraded protein (RUP) and the dry matter intake (DMI) on the intestinal flows of endogenous nitrogen (N) and amino acids (AA) in goats. The experiment was designed as a 4×4 Latin square using four ruminally, duodenally and ileally cannulated goats. The treatments were arranged in a 2×2 factorial design; two ratios of RDP to RUP (65:35 and 45:55, RDP1 and RDP2, respectively) and two levels at 95% and 75% of voluntary feed intake (DMI1 and DMI2, respectively) were fed to the goats. There were no significant differences in the N intake, duodenal flow of total N, undegraded feed N, microbial N, endogenous N or ileal flow of endogenous N, but the duodenal and ileal flow of endogenous N numerically decreased by approximately 22% and 9%, respectively, when the feed intake changed from DMI1 (0.63 kg/d) to DMI2 (0.50 kg/d). The dietary ratio of RDP to RUP had significant effects (p0.05) on the ileal flows of endogenous leucine, phenylalanine and cysteine. The present results implied that the duodenal flows of endogenous N and AA decreased when the dietary RDP to RUP ratio and DMI decreased, and the flow of endogenous AA at the ileum also decreased when the DMI decreased but increased with decreasing RDP to RUP ratios.

Information on the effects of different yeast species on ruminal fermentation is limited. This experiment was conducted in a 3×4 factorial arrangement to explore and compare the effects of addition of three different live yeast species (Candida utilis 1314, Saccharomyces cerevisiae 1355, and Candida tropicalis 1254) at four doses (0, 0.25×10(7), 0.50×10(7), and 0.75×10(7) colony-forming unit [cfu]) on in vitro gas production kinetics, fiber degradation, methane production and ruminal fermentation characteristics of maize stover, and rice straw by mixed rumen microorganisms in dairy cows. The maximum gas production (Vf), dry matter disappearance (IVDMD), neutral detergent fiber disappearance (IVNDFD), and methane production in C. utilis group were less (p

The objective of this study was to evaluate the effects of ammonia and amino acids (AA) supplementation on activities of ruminal enzymes involved in ammonia assimilation. In addition, the temporal changes of ruminal bacterial populations and enzyme activities during in vitro incubation were investigated. Rumen fluid from four ruminally fistulated goats was used in a 3×3 factorial arrangement of treatments with ammonia equivalent to 1, 4, and 15 mM ammonium chloride (NH4Cl), and with an added AA mixture (containing 992 g casein acid hydrolysate plus 1.4 g l -cysteine plus 8.68 g l -tryptophan) at 0, 1, and 15.5 g/L in the in vitro culture solution. Both ammonia and AA supplementation increased (P

An in vitro batch culture study was conducted to investigate (1) 13 C-labeled methane (CH 4 ) production efficiency (PE) which was produced from the reduction of 13 C-labeled sodium bicarbonate (NaH 13 CO 3 ) in media; and (2) the effects of NaHCO 3 and hydrogen (H 2 ) additions on ruminal fermentation, CH 4 production and methanogenic populations in a 4 × 3 factorial design with lucerne hays as substrates. Factors were NaH 13 CO 3 concentration in media (0, 11.9, 23.8 or 47.6 μmol/ml) and H 2 volume in headspace (0, 36 or 60 ml). The 13 CH 4 PE produced from NaH 13 CO 3 was linearly increased ( P 2 addition, but quadratically decreased ( P 13 CO 3 addition. Moreover, addition of H 2 quadratically increased ( P 4 production, apparent dry matter degradability (ADMD), total volatile fatty acids (VFA), molar proportion of acetate and populations of Methanosarcina barkeri ( M. barkeri ), Methanosphaera stadtmanae ( M. stadtmanae ), Methanobrevibacter ruminantium M1 ( M. ruminantium ) and Methanobrevibacter smithii (M. smithii ). Quadratic effects ( P 4 production, populations of M. barkeri , M. stadtmanae , M. ruminantium and M. smithii in response to NaHCO 3 addition. Additionally, the addition of NaHCO 3 linearly increased ( P M. barkeri was increased ( P = 0.023) by incubation time with the highest value being at 24 h, but no differences were observed for the M. stadtamanae , M. ruminantium and M. smithii gene copy numbers among the incubation time points. These results demonstrate that both CO 2 and H 2 concentrations affect methanogenesis in the rumen. Furthermore, the present study suggests that headspace hydrogen and media bicarbonate play roles in regulating feed fermentation, CH 4 production and methanogenic populations in vitro .

SUMMARYBatch cultures of mixed rumen micro-organisms were used to evaluate varying enzyme products with high xylanase activity (EPX), four of which were recombinant single xylanase activity developmental enzyme products (EPX1–EPX4, products of xylanase genes derived fromTrichoderma harzianum, Trichoderma reesei, OrpinomycesandAspergillus oryzae, respectively), for their potential to improvein vitroruminal fermentation of three forages [maize (Zea mays) stover (MS), rice (Oryza sativa) straw (RS) and Guimu No. 1 grass (Pennisetum americanum×Pennisetum purpureum, GM)]. The enzyme product EPX5, derived fromTrichoderma longibrachiatum, was used as a positive control that could improvein vitrofermentation of forages. Enzymes were supplied at dose rates of 0 (control), 20 (low), 50 (medium) and 80 (high) enzymic units of xylanase/g of dry matter (DM). There were no interactions between EPX and dose for the fermentation characteristics evaluated. Increasing EPX dose linearly increased gas production (GP) kinetic characters [i.e. asymptotic GP (VF), half time when GP is half of the theoretical maximum GP (t0·5), and initial fractional rate of degradation (FRD0)] and methane (CH4) production from RS and GM at 24 h, and increased degradability of DM at 24 h for MS and RS. A linear increase in degradability of neutral detergent fibre (NDF) of the three forages at 24 h was observed with increasing dose of EPX, but at 48 h only NDF degradability of RS was increased. There were differences in the effects of EPX on degradability of DM and NDF from RS at 24 h, with EPX4 having the highest and EPX1 having the lowest. In addition, increasing EPX dose linearly increased acetate proportion at 24 h and total volatile fatty acids (TVFA) at 48 h in MS. Increasing EPX dose linearly increased TVFA at 24 h, and ammonia-nitrogen (NH3-N) concentration at 48 h in RS. For GM, linear or quadratic effects of dose on acetate and butyrate concentration were observed at 24 and 48 h. The present study indicates that applying EPX to low-quality forages has the potential to improve rumen degradability and utilization. Furthermore, EPX from different sources differed in their effects when applied at the same dose rate, with the responses being forage-specific. For RS, the EPX derived fromA. oryzaeshowed the greatest positive effects on forage degradation; whereas for MS and GM, the source of micro-organism where EPX gene was derived did not affect the degradation, with little difference among the EPX evaluated.

BACKGROUND Post-ruminal digestion of fiber has received much less attention than its ruminal digestion. Using in vitro incubation techniques, the present study explored whether variations in fiber digestion occurred in different segments of the post-ruminal tract and whether fiber structure could influence its digestibility. A split plot design was conducted with gut segments (jejunum, ileum, cecum and colon) as main plot and substrates (neutral detergent fiber (NDF) and cellulose (CEL)) as subplot. RESULTS With the same substrate, the final asymptotic gas volume (VF), gas production at ti (), digestibility, microbial crude protein (MCP), total bacteria number (TBN), total short-chain fatty acids (TSCFA) and xylanase in incocula from the cecum and colon exceeded (P < 0.01) those in incocula from the jejunum and ileum, while the NH3-N in the former was less (P < 0.01). For the same gut segment, the digestion of CEL was superior to NDF, as reflected in greater VF, , maximum rate of gas production, digestibility, enzyme activities and SCFA but lower pH and NH3-N. CONCLUSION The current results imply that the intestinal contents from the cecum and colon have greater potential to digest fiber than those from the jejunum and ileum, and CEL is more easily digested in the post-ruminal tract than NDF. © 2013 Society of Chemical Industry

A series of batch cultures were conducted to investigate the effects of oleic acid (OA) on in vitro ruminal dry matter degradability (IVDMD), gas production, methane (CH4) and hydrogen (H2) production, and proportion of fatty acids. Rumen fluid was collected from fistulated goats, diluted with incubation buffer, and then incubated with 500 mg Leymus chinensis meal supplemented with different amounts of OA (0, 20, 40, and 60 mg for the CON, OA20, OA40 and OA60 groups, respectively). Incubation was carried out anaerobically at 39°C for 48 h, and the samples were taken at 12, 24 and 48 h and subjected to laboratory analysis. Supplementation of OA decreased IVDMD, the cumulative gas production, theoretical maximum of gas production and CH4 production, but increased H2 production. However, no effect was observed on any parameters of rumen fermentation (pH, ammonia, production of acetate, propionate and butyrate and total volatile fatty acid production). The concentrations of some beneficial fatty acids, such as cis monounsaturated fatty acids and conjugated linoleic acid (CLA) were higher (P < 0.05) from OA groups than those from the control group at 12 h incubation. In summary, these results suggest that the OA supplementation in diet can reduce methane production and increase the amount of some beneficial fatty acids in vitro.