Jinhe Kang

In vitro evaluation on neutral detergent fiber and cellulose digestion by post-ruminal microorganisms in goats.

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 (V(F)), gas production at t(i) (V(t(i)), 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 V(F), V(t(i)), 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.

Protein or energy restriction during late gestation alters fetal growth and visceral organ mass: an evidence of intrauterine programming in goats.

The objective of this study was to examine the effects of maternal protein or energy restriction during late gestation on postnatal fetal growth and visceral organ mass of goats. Eighty pregnant goats with similar age (2.0 ± 0.3 yr) and body weight (BW, 20.0 ± 1.0 kg before pregnancy) were assigned to 3 dietary treatments during late gestation: control (CON), 40% protein restricted (PR) and 40% energy restricted (ER) diets until parturition, after which offspring received normal diets for nutritional recovery. Kids were killed and visceral tissues were harvested at birth and week 6. Maternal protein or energy restriction decreased (P < 0.05) birth weight, and the weights of thymus, heart, abomasums, small intestine. The length of fetus from PR and ER were all decreased (P < 0.05) compared with that from control. When expressed relative to BW, thymus and small intestine for PR and ER still remained less (P < 0.05) than that for control. After 6 weeks of nutritional recovery, there was no difference (P = 0.91) in BW among groups; the kids from nutritional restriction groups showed a greater (P < 0.05) growth rate compared with kids from CON. Moreover, liver (only in ER, P < 0.10) and kidney (only in ER, P < 0.05) were proportionally increased to BW at week 6. The results indicate that maternal protein or energy restriction programs the fetal growth in goats, particularly the proportional responses of fetal organs relative to BW, including thymus, small intestine, kidney and liver.

Effects of maternal protein or energy restriction during late gestation on antioxidant status of plasma and immune tissues in postnatal goats

Maternal malnutrition can have temporary or long-lasting effects on development and physiological function of offspring. Our objective was to investigate whether maternal protein or energy restriction in late gestation affects the antioxidant status of plasma, immune organs (thymus and spleen), and natural barrier organs (jejunum) in neonatal goats and whether the effects could be reversed after nutritional recovery. Forty-five pregnant goats (Liuyang Blacks) of similar age (2.0 ± 0.3 yr) and BW (22.2 ± 1.5 kg at d 90 of gestation) were assigned to 3 dietary treatments during late gestation: control (ME = 9.34 MJ/kg and CP = 12.5%, DM basis), 40% protein restricted (PR), and 40% energy restricted (ER) until parturition, after which offspring received the normal diet for nutritional recovery. Plasma and tissues of kids were sampled to determine antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), and catalase (CAT)] and gene expression of antioxidant enzymes (Cu/Zn-SOD [SOD1], CAT, and peroxiredoxin 2 [PRDX2]). Maternal protein or energy restriction decreased (P < 0.05) SOD activities in plasma, liver, thymus, and spleen and SOD1 expression in thymus, and maternal energy restriction also decreased (P < 0.05) plasma GSH-Px activity and expressions of SOD1 and CAT in liver at birth. After nutritional recovery of 6 wk, SOD activities in thymus (both in PR and ER) and spleen (only in PR) were greater (P < 0.05), but CAT activity of thymus (both in PR and ER) and CAT expression (only in ER) were less (P < 0.01) than those in control. After nutritional recovery of 22 wk, SOD1 and PRDX2 expression in thymus (both in PR and ER) and SOD1 expression in liver (only in ER) were greater (P < 0.05) whereas CAT expression in thymus (both in PR and ER) was less (P < 0.001) than in control. The current results indicate that maternal protein or energy restriction can decrease the antioxidant capacity of the neonatal kids and result in an imbalance of SOD and hydrogen peroxide-inactivating systems in thymus, even after 6 or 22 wk of nutritional recovery.

Effects of maternal protein or energy restriction during late gestation on immune status and responses to lipopolysaccharide challenge in postnatal young goats

Knowledge of maternal malnutrition of ruminants and effects on development of the immune system of their offspring is lacking. A study was conducted to investigate the effects of maternal protein or energy restriction during late gestation on immune status of their offspring at different ages. Sixty-three pregnant goats (local breed, Liuyang black goat, 22.2 ± 1.5 kg at d 90 of gestation) were fed control (CON, ME = 9.34 MJ/kg and CP = 12.5%, DM basis), 40% protein restricted (PR), or 40% energy restricted (ER) diets from d 91 of gestation to parturition, after which all animals received an adequate diet for nutritional recovery. Plasma concentrations of complement components (C3, C4), C-reactive protein (CRP) and immunoglobulins (IgG and IgM), jejunum cytokines (IL-2, IL-6, and IL-10) expression levels and morphology in the offspring were measured. Additionally, plasma concentration of complement and IL-6, and cytokines expression levels in gastrointestinal tract obtained at 6 wk from young goats were assessed under saline or lipopolysaccharide (LPS) challenging conditions. Maternal PR or ER decreased (P < 0.05) plasma C3, C4, IgG, and IgM concentrations, and IL-2 and IL-6 mRNA expression in the jejunum from neonatal kids, but did not alter (P > 0.05) plasma CRP concentration. The IL-10 mRNA expression of jejunum from PR kids was also less (P < 0.01) than that from CON kids. Moreover, jejunum villous height (P < 0.10 in PR, P < 0.05 in ER) and crypt depth (P < 0.05 both in PR and ER) were reduced in neonatal kids from malnourished mothers. At 6 wk of age, there were no differences (P > 0.05) in any plasma or tissue immune parameters among the 3 treatments. However, when given a LPS challenge, ER and PR kids had greater (P = 0.02) IL-6 concentration compared with CON kids. Our results suggest that both PR and ER during late gestation induced short-term as well as long-lasting alterations on immune responses in their offspring, which may make the animals more susceptible to a bacterial pathogen challenge. The present findings expand the existing knowledge in immunological mechanisms responsible for the development of disease in later life.

Proteomic Analysis of Isolated Plasma Membrane Fractions from the Mammary Gland in Lactating Cows

The mammary gland of dairy cows is a formidable lipid-synthesizing machine for lactation. This unique function depends on the activities of plasma membrane (PM) proteins in mammary cells. Little information is known about the expression profiles of PM proteins and their functions during the lactating process. This study investigated the proteome map of PM fractions of mammary gland in lactating cows using 1D-Gel-LC-MS/MS and identified 872 nonredundant proteins with 141 unknown proteins, wherein 215 were PM-associated proteins. Most of the PM-associated proteins were binding, transport, and catalytic proteins such as annexin proteins, heat shock proteins, integrins, RAS oncogene family members, and S100 calcium binding proteins. The PM-associated pathways such as caveolae-mediated endocytosis, leukocyte extravasation, aldosterone signaling in epithelial cells, and remodeling of epithelial adherens junctions were also significantly over-represented. Proteomic analysis revealed the characteristics and predicted functions of PM proteins isolated from the lactating bovine mammary gland. These results further provide experimental evidence for the presence of many proteins predicted in the annotated bovine genome. The data generated here also provide a reference for the PM-related functional research in the mammary gland of lactating cows.

Effects of Momordica charantia polysaccharide on in vitro ruminal fermentation and cellulolytic bacteria

Abstract 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.

Effects of Momordica charantia Saponins on In vitro Ruminal Fermentation and Microbial Population

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 (VF) 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<0.001) the molar proportion of butyrate, isovalerate at 24 h and 48 h, and the molar proportion of acetate at 24 h, but then decreased (p<0.05) them at 48 h. The molar proportion of valerate was increased (p<0.05) at 24 h. The acetate to propionate ratio (A/P; linear, p<0.01) was increased at 24 h, but reached the least value at the level of 0.30 mg/mL MCS. The MCS inclusion decreased (p<0.05) the molar proportion of propionate at 24 h and then increased it at 48 h. The concentration of total volatile fatty acid was decreased (p<0.001) at 24 h, but reached the greatest concentration at the level of 0.01 mg/mL and the least concentration at the level of 0.60 mg/mL. The relative abundance of Ruminococcus albus was increased at 6 h and 24 h, and the relative abundance of Fibrobacter succinogenes was the lowest (p<0.05) at 0.60 mg/mL at 6 h and 24 h. The relative abundance of Butyrivibrio fibrisolvens and fungus reached the greatest value (p<0.05) at low doses of MCS inclusion and the least value (p<0.05) at 0.60 mg/mL at 24 h. The present results demonstrates that a high level of MCS quickly inhibits in vitro fermentation of maize stover, while MCS at low doses has the ability to modulate the ruminal fermentation pattern by regulating the number of functional rumen microbes including cellulolytic bacteria and fungi populations, and may have potential as a feed additive applied in the diets of ruminants.

Comparison of two live Bacillus species as feed additives for improving in vitro fermentation of cereal straws.

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 < 0.05), while the methane (CH4) production, ratio of acetate to propionate, and total volatile fatty acids (TVFA) concentration were decreased (P < 0.05) by the supplementation of B. licheniformis compared with that of B. subtilis. Adding B. licheniformis and B. subtilis raised (P < 0.05) or numerically raised the maximum gas production, while decreasing (P < 0.05) or numerically lowering pH and concentrations of most volatile fatty acids. The addition of B. licheniformis increased (P < 0.05) the NH3-N concentration but reduced CH4 production and ratio of acetate to propionate (P < 0.05), while the NH3 -N concentration was decreased (P < 0.05), and the CH4 production and ratio of acetate to propionate were increased by that of B. subtilis compared to the control. Results obtained in this research suggest that B. licheniformis would be preferred as a live Bacillus additive in comparison with B. subtilis, and its optimal dose should be 0.25 × 10(7) cfu/500 mg substrates.

Age and feeding system (supplemental feeding versus grazing) modulates colonic bacterial succession and host mucosal immune maturation in goats

The gut microbiome plays important roles in the regulation of gastrointestinal tract functional development and host mucosal immune maturation. This study was conducted to test the hypothesis that age and feeding system (supplemental feeding [Sup] vs. grazing [G]) could alter colonic bacterial diversity and host mucosal immune maturation. Thirty Liuyang black goat kids ( = 4) were slaughtered on d 0, d 7 (nonrumination), d 28, d 42 (transition), and d 70 (rumination). The colonic microbiota was profiled by Miseq sequencing of the 16S rRNA gene. Host colonic mucosal immune maturation was examined using mRNA level expression of Toll-like receptors (TLR), proinflammatory cytokines, and the Toll-IL-1R (TIR) domain-containing adaptor. A correlation analysis was conducted to elucidate the relationship between bacterial diversity and fermentation parameters and host immune maturation variables. The results showed that α diversity indexes ( < 0.05), abundances of genera ( = 0.003) and ( = 0.024), ( = 0.004), and ( = 0.046) mRNA expressions were lower for Sup than for G, whereas the abundance of genera and ( < 0.05) was greater for Sup than for G. Regardless of the feeding system, bacterial 16S rRNA gene copy number and α diversity indexes increased ( < 0.05), whereas Proteobacteria abundance decreased linearly from d 0 to 70 after birth ( = 0.026). At the genus level, dominated the first week and declined sharply afterward, whereas abundance was greatest on d 7. abundance decreased linearly ( = 0.021), whereas abundances of , , , , and increased with age ( < 0.05). These findings coincided with increased , , and myeloid differentiation factor 88 () mRNA expressions with age ( < 0.05). Finally, correlation analysis revealed that different genera participated in different roles in fermentation capacity and host mucosal immune maturation. Collectively, colonic bacterial diversity and host mucosal immune maturation are age related, and concentrate supplement could alter bacterial diversity and alleviate overinflammation responses.

Effects of dietary cellulase and xylanase addition on digestion, rumen fermentation and methane emission in growing goats.

The objective of this study was to evaluate the effectiveness of supplementation of cellulase and xylanase to diets of growing goats to improve nutrient digestibility, utilisation of energy and mitigation of enteric methane emissions. The experiment was conducted in a 5 × 5 Latin square design using five goats with permanent rumen fistulae and five treatments consisted of two levels of cellulase crossed over with two levels of xylanase plus unsupplemented Control. The cellulase (243 U/g) derived from Neocallimastix patriciarum was added at 0.8 and 1.6 g/kg dry matter intake (DMI) and the xylanase (31,457 U/ml) derived from Aspergillus oryzae was fed at 1.4 and 2.2 ml/kg DMI. There were no differences in apparent digestibility of organic matter, neutral detergent fibre, acid detergent fibre and rumen fermentation parameters (i.e. ammonia-nitrogen [N], volatile fatty acids) among all treatments. Dietary cellulase and xylanase addition did not influence energy and N utilisation. But compared to xylanase addition at the higher dose, at the low xylanase dose the retained N, the availability of retained N and digested N were increased (p < 0.01). Moreover, enzyme addition did not affect the enteric methane emission and community diversity of ruminal methanogens. The present results indicated that previous in vitro findings were not confirmed in ruminant trials.