Effect of rice bran extract on in vitro rumen fermentation and methane production.

This study was done to investigate which components of rice bran (RB) are involved in the inhibition of methanogenesis by fractionating the rice bran and adding it to a rumen in vitro culture system. The RB extract obtained using ethanol and water was screened in an in vitro fermentation system. The experimental treatment conditions were as follows: a control group containing a substrate without supplements; substrates with 0.06 g of RB; 0.6 mL of ethanol; 0.6 mL of distilled water (DW); 0.6 mL of ethanol-soluble fraction (ESF); 0.06 g of ethanol-insoluble rice bran (EIRB); 0.6 mL of water-soluble fraction (WSF); and 0.06 g of water-insoluble rice bran (WIRB). Based on the result of the analysis, the addition of ESF significantly decreased CH4 and CH4 /g dry matter digested, methanogen population (p < 0.05), while gas and dry matter digestibility (DMD) were comparable with the control group. Total short-chain fatty acid (SCFA), and proportion of propionate were reduced, and the proportion of butyrate was increased by the addition of ethanol and ESF (p < 0.05). This result suggests that the supplementation of 10% ESF can substantially reduce methane production in vitro without a negative effect on substrate digestibility.

Nutrient digestibility, fecal output of fractionated proteins, and ruminal fermentation parameters of goats fed a diet supplemented with spent green tea and black tea leaf silage.

Spent green and black tea leaf silage (GTS and BTS, respectively) was offered as a protein supplement to goats to examine in vivo digestibility, nitrogen balance, urinary excretion of purine derivatives, and ruminal fermentation. Four castrated goats were fed a basal diet supplemented with alfalfa hay cube (AHC), GTS, or BTS in a 4 × 4 Latin square design. Digestibilities of various nutrients except for nitrogen (N) fraction were unaffected by the type of supplement. Digestibility of acid detergent insoluble N (ADIN) in BTS treatment was a negative value and significantly lower than those in other treatments. Urinary N output and retained N were not significantly affected by the diets. The fecal output of neutral detergent insoluble nitrogen (NDIN) and ADIN in the BTS treatment was significantly higher than those in other treatments. Urinary excretion of purine derivatives was not affected by the treatments. Ruminal NH3 -N concentration in AHC and GTS treatments were not significantly different, but that in the BTS treatment was significantly lower than others. These results indicated that GTS is substitutable for AHC as a protein supplement, whereas BTS was able to bind proteins tightly in the digestive tract, which lowered ruminal N degradability and increase fecal N output.

Effects of the supplementation of a calcium soap containing medium-chain fatty acids on the fecal microbiota of pigs, lactating cows, and calves.

Medium-chain fatty acids (MCFAs) have antialgal, antibacterial, antifungal, antiprotozoan, and antiviral activities. However, antibacterial activities of MCFAs in the hindgut of pigs and cattle are still unknown. We report the effects of the supplementation of MCFAs on fecal bacteria of pigs, lactating cows, and Japanese Black calves. Lactobacillus spp., Bifidobacterium spp., eaeA(+) Escherichia coli, Salmonella spp., Campylobacter jejuni, and Clostridium perfringens in the feces of animals were quantified by real-time PCR assay. There was no significant increase or decrease in Lactobacillus spp. and Bifidobacterium spp. in the three animals. In the pig feces, eaeA(+) E. coli was reduced to less than a third in the treatment group (P < 0.01). C. jejuni in the pig feces was also significantly less in the treatment group compared with the control (P < 0.01). In the lactating cow, eaeA(+) E. coli was reduced to one fifth of that in the control (P < 0.01). Salmonella spp. was halved in calf feces (P < 0.01). Thus, a reduction in Gram-negative pathogenic bacteria was observed. In conclusion, supplementation of a MCFA calcium soap in the diet would be beneficial to growing pigs, lactating cow, and calves by reducing pathogenic bacteria.

Combination Effects of Plant Extracts Rich in Tannins and Saponins as Feed Additives for Mitigating in Vitro Ruminal Methane and Ammonia Formation.

The objective of this experiment was to test the effects of combining plant extracts rich in tannins and saponins at varying proportions on in vitro ruminal methane and ammonia formation. Tannins were extracted from Swietenia mahogani leaves and saponins from Sapindus rarak fruits with various solvents. The extracts obtained with the most efficient solvents (tannins: 75% water and 25% methanol; saponins: pure methanol) were then used in vitro. The treatments consisted of two substrate types (high-forage (HF) or high-concentrate (HC) diets) and five extract combinations (tannins: saponins, 1:0, 3:1, 1:1, 1:3, and 0:1) added at 2 mg/mL in incubation liquid. In vitro incubation was performed in four runs, with each treatment being represented with two replicates per run. The addition of plant extracts rich in tannins and saponins, either individually or in combination, decreased the methane proportion of total gas in both the HF (p < 0.05) and HC (p < 0.05) diets. The effects of the plant extracts rich in tannins and saponins were generally additive in mitigating methane emissions. Favorable associative effects between the extracts were observed in the ammonia concentration, both in the HF (p < 0.001) and HC (p < 0.01) diets and in the methane proportion of total gas, with a 1:3 mixture of tannins and saponins added to the HC diet (p < 0.05).

Yokukansan-induced reduction of ethanol preference is associated with antagonism of 5-HT3 receptors.

BACKGROUND: We recently reported that alkaloids in Uncaria hook (a constituent of yokukansan) contribute to antagonism of 5-HT3 receptors. Many studies have reported that 5-HT3 receptor antagonists reduce alcohol preference. However, the effect of yokukansan on alcohol preference is not clear. PURPOSE: This study was performed to investigate the direct effect of yokukansan on alcohol preference and the effect of 5-HT3 receptors on the preference. STUDY DESIGN: We examined ethanol preference effected by yokukansan. Next, we analyzed the contribution of 5-HT3 receptors to the effect of yokukansan. METHODS: Ethanol preference was measured using the two-bottle preference test in mice fed with or without yokukansan diet. Next, the contribution of 5-HT3 receptors to ethanol preference was investigated using 5-HT3 receptor-deficient mice. RESULTS: Reduction of ethanol preference by yokukansan was not observed using 5-HT3 receptor deficient mice. CONCLUSION: Yokukansan contributes to reduced ethanol preference and antagonism of 5-HT3 receptors is associated with the effect.

Nutritive evaluation of spent green and black tea leaf silages by in vitro gas production characteristics, ruminal degradability and post-ruminal digestibility assessed with inhibitory activity of their tannins.

Spent tea leaf contains high levels of crude protein, suggesting that it may be used as an alternative source for ruminant feeding. We assessed the nutritive characteristics of spent green tea leaf silage (GTS) and black tea leaf silages (BTS) in comparison with soybean meal (SBM) and alfalfa hay cube (AHC) using in vitro assay. The effects of tannin on the nutritive characteristics were also evaluated by adding polyethylene glycol (PEG) as a tannin-binding agent. The amount of gas production was greater for SBM, followed by AHC, GTS, and BTS. A significant improvement in gas production upon addition of PEG was observed only for BTS. Ruminal protein degradability and post-ruminal digestibility was higher for SBM, followed by AHC, GTS, and BTS. The presence of PEG significantly increased ruminal degradability and post-ruminal protein digestibility for GTS and BTS, but not for AHC. The increment of protein digestibility by PEG was much greater for BTS than for GTS, indicating that GTS tannins suppress protein digestibility slightly, whereas BTS tannins do so strongly. According to these results, GTS but not BTS has a potential as an alternative to AHC as a ruminant feedstuff.

Yokukansan contains compounds that antagonize the 5-HT3 receptor.

BACKGROUND: We recently focused on alkaloids in Uncaria hook (a constituent of the Kampo medicine, yokukansan) and identified the pharmacological action of geissoschizine methyl ether on several G protein-coupled receptors. However, the functions of other identified alkaloids in Uncaria hook, including hirsutine, hirsuteine, rhynchophylline, isorhynchophylline, corynoxeine, isocorynoxeine, are not clear. PURPOSE: To evaluate the effect of seven alkaloids in Uncaria hook (hirsutine, hirsuteine, rhynchophylline, isorhynchophylline, corynoxeine, isocorynoxeine and geissoschizine methyl ether) on the hydroxytryptamine type-3 (5-HT3) receptor ion channel. STUDY DESIGN: We examined the effect of these alkaloids on the current of human 5-HT3 receptors expressed in Xenopus laevis oocytes. METHODS: The human 5-HT3A subunit alone for the 5-HT3A receptor, or 5-HT3A and 5-HT3B subunits for the 5-HT3AB receptor, were expressed in Xenopus laevis oocytes. The 5-HT current was measured with or without alkaloid application using a two-electrode voltage clamp. RESULTS: Each alkaloid, except for geissoschizine methyl ether, weakly inhibited the 5-HT-mediated 5-HT3A and/or 5-HT3AB receptor current, but co-application of these seven alkaloids inhibited the current strongly. CONCLUSION: Each alkaloid contributes to antagonism of the 5-HT3 receptor.

Identification, expression analysis, and functional characterization of peptide YY in chickens (Gallus gallus domesticus).

Peptide YY (PYY) functions as a postprandial satiety signal in mammals. However, the genomic information and physiological roles of chicken PYY have not yet been clarified, although PYY peptide was isolated from chicken intestines in 1992. In this study, we identified a full-length complementary DNA (cDNA) sequence encoding the chicken PYY precursor. The deduced amino acid sequence of chicken PYY was completely consistent with the previously identified peptide sequence. PYY mRNA was abundantly expressed in the small intestine compared with the large intestine. PYY mRNA levels in the jejunum were significantly higher during ad libitum feeding compared with fasting, suggesting that intestinal PYY expression is altered in response to nutritional status in chicks. Intravenous administration of PYY significantly suppressed food intake in chicks. Furthermore, neuropeptide Y receptor Y2, a possible target of PYY, was expressed in various brain regions including the appetite-regulating centers in chicks. This is the first evidence that the intestinal hormone PYY may function as an anorexigenic hormone in chicks.

Membrane-Permeable Calpain Inhibitors Promote Rat Oral Mucosal Epithelial Cell Proliferation by Inhibiting IL-1α Signaling.

To standardise regenerative medicine using cultured cells, the use of serum-free, chemically defined media will be necessary. We have reported that IL-1α inhibits the growth of epithelial cells in culture and that recombinant IL-1 receptor antagonist (IL-1RA) significantly promotes epithelial cell growth in no feeder layer condition. In this study, we examined inhibitors of calpain, a cysteine proteinase that plays crucial roles in various cellular functions, including IL-1α maturation and secretion. The culturing of epithelial cells in serum-free media supplemented with a membrane-permeable calpain inhibitor significantly promoted growth while suppressing IL-1α maturation and secretion. By contrast, non-membrane-permeable calpain inhibitor treatment did not have these effects. Interestingly, immunoblotting analysis revealed that immature, untruncated, IL-1α expression was also downregulated by cell-permeable calpain inhibitor treatment, and the difference in IL-1α gene expression increased from day 2 to day 6. Although IL-1RA has been reported to promote epithelial cell growth, we detected no synergistic promotion of epithelial cell growth using a calpain inhibitor and IL-1RA. These findings indicate that calpain inhibitors promote epithelial cell proliferation by inhibiting IL-1α maturation at an early phase of epithelial cell culture and by suppressing the positive feedback-mediated amplification of IL-1α signalling.

Effect of monensin withdrawal on rumen fermentation, methanogenesis and microbial populations in cattle.

This study was designed to obtain information on the residual influence of dietary monensin on ruminant fermentation, methanogenesis and bacterial population. Three ruminally cannulated crossbreed heifers (14 months old, 363 ± 11 kg) were fed Italian ryegrass straw and concentrate supplemented with monensin for 21 days before sampling. Rumen fluid samples were collected for analysis of short chain fatty acid (SCFA) profiles, monensin concentration, methanogens and rumen bacterial density. Post-feeding rumen fluid was also collected to determine in vitro gas production. Monensin was eliminated from the rumen fluid within 3 days. The composition of SCFA varied after elimination of monensin, while total production of SCFA was 1.78 times higher than on the first day. Methane production increased 7 days after monensin administration ceased, whereas hydrogen production decreased. The methanogens and rumen bacterial copy numbers were unaffected by the withdrawal of monensin.

Downregulation of Pgc-1α expression by tea leaves and their by-products.

Previous studies indicate that muscle Pgc-1α expression governs the proportion of muscle fibre types. As a first step in using diet to manipulate the proportion of muscle fibre types by using Pgc-1α expression, the present study investigates the modulation of Pgc-1α expression by feedstuffs. A luciferase-based Pgc-1α reporter construct (Pgc-1α(-2553)-luc) that contains the mouse Pgc-1α promoter (-2553 to +78 bp) was prepared. A screen of ethanol extracts from 33 feedstuffs indicated that oolong tea and roasted green tea extracts decreased Pgc-1α(-2553)-luc expression in C2C12 myoblasts. The transcriptional repression of Pgc-1α by tea leaf extracts was reproduced in hepatic HepG2 cells. We further examined the effects of the alcohol extracts of tea waste and its silage on Pgc-1α transcription; the tea waste silage extract inhibited Pgc-1α transcription. Treatment with the extracts of raw tea leaves, tea waste and tea waste silage effectively decreased Pgc-1α mRNA levels during myogenesis of myosatellite cells. The present results suggest that tea leaves and their by-products could be used to modulate proportions of muscle fibre types.

Xanthine oxidase inhibitory activity and hypouricemic effect of aspalathin from unfermented rooibos.

Rooibos is rich in flavonoids such as aspalathin, which is a unique C-glycosyl dihydrochalcone, that is used as a traditional herbal tea. This study was designed to evaluate the in vitro xanthine oxidase (XOD) inhibitory activity of the aspalathin-rich fraction (ARF) and purified aspalathin from rooibos. The hypouricemic effects of the ARF and aspalathin on hyperuricemic mice were also assessed. The ARF was prepared from aqueous extract of unfermented rooibos leaves and stems, and it was collected by column chromatography; the aspalathin content in this fraction was 21.4%. The ARF and aspalathin inhibited XOD in a dose-dependent manner. The concentrations of the ARF and aspalathin required to inhibit XOD at 50% (IC50 ) were 20.4 µg/mL (4.4 µg/mL aspalathin equivalents) and 4.5 µg/mL, respectively. Lineweaver-Burk plot analysis indicated that aspalathin was a competitive inhibitor of XOD, and the inhibition constant (Ki) was 3.1 µM. In hyperuricemic mice induced by inosine-5'-monophosphate, treatment with the ARF and aspalathin significantly suppressed the increased plasma uric acid level in a dose-dependent manner. The suppressed plasma uric acid level in mice could be attributed to the XOD inhibitory activity of the ARF and aspalathin. Further study is required to determine the effect of aspalathin or its metabolites on XOD activity in vivo.