• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1649-1657
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• 2006

In order to find an alternative source of inoculum to caecal content for studying the fermentation activity of rabbit hindgut, caecal content and faeces of 25 hybrid Hyla rabbits were used as inocula for an in vitro gas production trial. About 1 g of three substrates (dehydrated alfalfa meal, dehydrated beet pulp, barley) was weighed, in quadruplicate per inoculum, in 120 ml bottles; 75 ml of anaerobic medium and 4 ml of reducing solution were added and bottles were placed at $39^{\circ}C$. Caecal content and faeces were diluted respectively 1:2 (CI) and 1:8 (FI) with anaerobic medium and were introduced in the respective bottles (10 ml). Gas production was recorded 20 times at 2-24 h intervals throughout fermentation (96 h). The fermentation characteristics (i.e. degraded organic matter, OMd; potential gas production, A; fermentation rate, Rmax; time at which it is reached, Tmax; pH, volatile fatty acid, VFA) were studied by inoculum and feedstuffs. The feedstuffs, according to their chemical composition, showed very different fermentation characteristics. In particular, OMd, A and Rmax allowed feedstuff classification as follows: barley>beet pulp>alfalfa. The inocula differ (p<0.05) in Tmax, were higher for CI (15.53 vs. 11.96 h) and in VFA production. In particular, CI produced higher levels of acetate (38.9 vs. 33.4 mM/g OM incubated, p<0.01) and isobutyrate (0.72 vs. 0.42, p<0.01) but less propionate (7.1 vs. 10.3, p<0.01) and butyrate (11.3 vs. 14.0, p<0.01). However, the trend of gas production, similar for the inocula according to the fermented substrate, and the good regression equation to estimate some caecal fermentation parameters from faeces suggest that, after standardisation, the faeces could be used as an alternative inoculum for gas tests in rabbit.

• Journal of Animal Science and Technology
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• v.62 no.1
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• pp.52-57
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• 2020

In rumen in vitro experiments, although nitrogen gas (N₂) flushing has been widely used, its effects on rumen fermentation characteristics are not clearly determined. The present study is the first to evaluate the effects of N₂ flushing on rumen fermentation characteristics in in vitro batch culture system by comparing with new applicable non-metabolizable gas: argon (Ar). The rumen fluid was taken from two Korean native heifers followed by incubation for 3, 9, 12, and 24 h with N₂ or Ar flushing. As a result, in all incubation time, N₂ flushing resulted in higher total gas production than Ar flushing (p < 0.01). Additionally, in N₂ flushing group, ammonia nitrogen was increased (p < 0.01). However, volatile fatty acids profiles and pH were not affected by the flushing gases (p > 0.05). In conclusion, the present study demonstrated that N₂ flushing can influence the rumen nitrogen metabolism via increased ammonia nitrogen concentration and Ar flushing can be used as a new alternative flushing gas.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.75-82
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• 2008

In vivo and in vitro digestibility of 6 diets with a forage to concentrate ratio (F/C) ranging from 100 to 50:50 (diet 1: all hay, diet 2: 90:10, diet 3: 80:20, diet 4: 70:30, diet 5: 60:40, diet 6: 50:50) were investigated using 6 buffaloes in a $6{\times}6$ Latin square design. For the in vivo trial, the individual faeces of buffaloes were collected 3 times per day for 7 days. Individual pooled faeces and samples of each diet were analysed for chemical composition and insoluble acid ash (AIA) contents in order to estimate the coefficient of apparent digestibility (ADC). On the last day of the in vivo trial a sample of faeces was collected from each animal and used as inoculum for the in vitro test, using the gas production technique (IVGPT). The in vivo organic matter digestibility (ADC) rose as the percentage of concentrate increased up to the 70:30 (F/C) diet (67.01, 73.03, 78.06 and 79.05, respectively for diets 1, 2, 3 and 4); the other two diets (60:40 and 50:50 F/C) unexpectedly did not follow this trend (75.11 and 79.06, respectively for diet 5 and 6). However, these data agree with the results of the in vitro trial. The ADC was positively correlated with the dOM (p<0.001), but not with the gas production at different times; cumulative gas production recorded at the end of incubation (OMCV) showed an irregular trend and was not closely correlated to degraded OM. Estimation of in vivo digestibility from in vitro fermentation data was acceptable, despite leaving room for improvement.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.7
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• pp.937-945
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• 2014

Green and black tea by-products, obtained from ready-made tea industry, were ensiled at $10^{\circ}C$, $20^{\circ}C$, and $30^{\circ}C$. Green tea by-product silage (GTS) and black tea by-product silage (BTS) were opened at 5, 10, 45 days after ensiling. Fermentation characteristics and nutrient composition, including tannins, were monitored and the silages on day 45 were subjected to in vitro ruminal fermentation to assess anti-nutritive effects of tannins using polyethylene glycol (PEG) as a tannin-binding agent. Results showed that the GTS and BTS silages were stable and fermented slightly when ensiled at $10^{\circ}C$. The GTS stored at $20^{\circ}C$ and $30^{\circ}C$ showed rapid pH decline and high acetic acid concentration. The BTS was fermented gradually with moderate change of pH and acid concentration. Acetic acid was the main acid product of fermentation in both GTS and BTS. The contents of total extractable phenolics and total extractable tannins in both silages were unaffected by storage temperatures, but condensed tannins in GTS were less when stored at high temperature. The GTS showed no PEG response on in vitro gas production, and revealed only a small increase by PEG on $NH_3$-N concentration. Storage temperature of GTS did not affect the extent of PEG response to both gas production and $NH_3$-N concentration. On the other hand, addition of PEG on BTS markedly increased both the gas production and $NH_3$-N concentration at any ensiled temperature. It can be concluded that tannins in both GTS and BTS suppressed rumen fermentation, and tannins in GTS did more weakly than that in BTS. Ensiling temperature for both tea by-products did not affect the tannin's activity in the rumen.

• Animal Bioscience
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• v.34 no.4
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• pp.584-593
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• 2021

Objective: The objective of this study was to compare fenugreek (FG) with alfalfa (Alf) in ruminal fermentation and methane (CH4) production in vitro. Methods: Whole-plant FG harvested at 11- and 15-wk and Alf harvested at early and mid-bloom maturities, alone or as 50:50 mixture of FG and Alf at the respective maturity, were assessed in a series of 48-h in vitro batch culture incubations. Total fermentation gas and methane gas production, dry matter (DM) disappearance, volatile fatty acids, microbial protein and 16S RNA gene copy numbers of total bacteria and methanogens were determined. Results: Compared to early bloom Alf, FG harvested at 11-wk exhibited higher (p<0.05) in vitro DM and neutral detergent fibre disappearance, but this difference was not observed between the mid-bloom Alf and 15-wk FG. Regardless plant maturity, in vitro ruminal fermentation of FG produced less (p<0.001) CH4 either on DM incubated or on DM disappeared basis than that of Alf during 48-h incubation. In vitro ruminal fermentation of FG yielded similar amount of total volatile fatty acids with higher (p<0.05) propionate percentage as compared to fermentation of Alf irrespective of plant maturity. Microbial protein synthesis was greater (p<0.001) with 11-wk FG than early bloom Alf as substrate and 16S RNA gene copies of total bacteria was higher (p<0.01) with 15-wk FG than mid-bloom Alf as substrate. Compared to mid-bloom Alf, 15-wk FG had lower (p<0.05 to 0.001) amount of 16S RNA methanogen gene copies in the whole culture during 48-h incubation. Conclusion: In comparison to Alf, FG emerges as a high quality forage that can not only improve rumen fermentation in vitro, but can also remarkably mitigate CH4 emissions likely due to being rich in saponins.

• Journal of Animal Environmental Science
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• v.17 no.3
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• pp.171-180
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• 2011

The researchers have tried to reduce ruminal methane gas ($CH_4$) and to convert it into beneficial nutrient for several decades. This study was conducted to screen the methane-reducing vegetables among lettuce, hot pepper, spring onion, onion, turmeric, sesame leaf, garlic, radish sprout, leek and ginger nutritiously on the in vitro ruminal fermentation. The heat-treated vegetables at the 10% of substrate (timothy) were used to reduce methane production on the in vitro anaerobic experiment of 0, 6, 12, 24 and 48 h incubation time. Total gas production, pH, ammonia, $H_2$, $CO_2$, $CH_4$, and volatile fatty acid (VFA) were measured as indicators of in vitro fermentation product containing methane gas. All treatments except garlic showed a tendency to increase in total gas production. The result of ammonia showed that garlic and hot pepper affected rumen bacteria concerned protein metabolism and that lettuce and spring onion increased ammonia production. Garlic decreased $CH_4$ production in inverse proportion to $H_2$. Lettuce, spring onion, onion, garlic, radish sprout, leek and ginger increased propionate of VFA. Garlic balanced the ruminal fermentation in the pH, $H_2$, $CH_4$, acetate and propionate. This results showed that methane production at in vitro study was inhibited by heat-treated garlic supplementation. In conclusion, this study suggests that ruminal fermentation covering methane production might be controled by proper vegetables.

• Journal of Animal Science and Technology
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• v.60 no.11
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• pp.26.1-26.9
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• 2018

Background: This experiment aimed at assessing polyphenol-rich plant biomass to use in complete feed making for the feeding of ruminants. Methods: An in vitro ruminal evaluation of complete blocks (CFB) with (Acacia nilotica, Ziziphus nummularia leaves) and without (Vigna sinensis hay) polyphenol rich plant leaves was conducted by applying Menke's in vitro gas production (IVGP) technique. A total of six substrates, viz. three forages and three CFBs were subjected to in vitro ruminal fermentation in glass syringes to assess gas and methane production, substrate degradability, and rumen fermentation metabolites. Results: Total polyphenol content (g/Kg) was 163 in A. nilotica compared to 52.5 in Z. nummularia with a contrasting difference in tannin fractions, higher hydrolysable tannins (HT) in the former (140.1 vs 2.8) and higher condensed (CT) tannins in the later (28.3 vs 7.9). The potential gas production was lower with a higher lag phase (L) in CT containing Z. nummularia and the component feed block. A. nilotica alone and as a constituent of CFB produced higher total gas but with lower methane while the partitioning factor (PF) was higher in Z. nummularia and its CFB. Substrate digestibility (both DM and OM) was lower (P < 0.001) in Z. nummularia compared to other forages and CFBs. The fermentation metabolites showed a different pattern for forages and their CFBs. The forages showed higher TCA precipitable N and lower acetate: propionate ratio in Z. nummularia while the related trend was found in CFB with V. sinensis. Total volatile fatty acid concentration was higher (P < 0.001) in A. nilotica leaves than V. sinensis hay and Z. nummularia leaves. It has implication on widening the forage resources and providing opportunity to use forage biomass rich in polyphenolic constituents in judicious proportion for reducing methane and enhancing green livestock production. Conclusion: Above all, higher substrate degradability, propionate production, lower methanogenesis in CFB with A. nilotica leaves may be considered useful. Nevertheless, CFB with Z. nummularia also proved its usefulness with higher TCA precipitable N and PF. It has implication on widening the forage resources and providing opportunity to use polyphenol-rich forage biomass for reducing methane and enhancing green livestock production.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.529-535
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• 2007

The nutritive values of leaves of Citrus grandis, Citrus aurantium, Citrus oranges, Citrus limon, and Citrus deliciosa were evaluated by chemical composition and in vitro gas production techniques. There were significant (p<0.001) differences among citrus species in terms of chemical composition. Crude protein (CP) contents ranged from 123.0 to 148.3 g/kg DM. Neutral detergent fibre (NDF) and acid detergent fibre (ADF) contents were varied with species in the range 219.4-355.4 and 215.0-278.8 g/kg DM respectively. Condensed tannin (CT) contents were ranged from 5.9 to 10.2 g/kg DM. The PEG addition significantly (p<0.001) increased the gas production and some estimated parameters of citrus tree leaves. However, species showed variable responses to polyethylene glycol (PEG) treatment. There were also significant (p<0.001) differences among species in terms of gas production and estimated parameters. The OMD and ME contents of citrus leaves without PEG supplementation were ranged from 66.5 to 73.3% and 9.8 to 10.9 MJ/kg DM respectively. The improvement in gas production, organic matter digestibility (OMD) and metabolizable energy (ME) with PEG emphasized the negative effect of tannins on digestibility. The increase (%) in the estimated OMD and ME contents ranged from 5.5 to 9.8% and 5.7 to 10.2% respectively. All citrus tree leaves studied in this experiment have potential nutritive values indicated by high crude protein content, OMD, ME and low fiber values.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.1
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• pp.73-79
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• 2016

The objective was to evaluate the effect of fodder tree species (FTS) with condensed tannin contents: Cordia elaeagnoides, Platymiscium lasiocarpum, Vitex mollis, and Haematoxylon brasiletto, on in vitro methane ($CH_4$) production at 24 h post incubation. The analysis was performed using the in vitro gas production technique, with three levels of inclusion/species: 600, 800, and 1,000 mg and with 4 replicates/species/level of inclusion. The substrate was incubated at $39^{\circ}C$, and the gas and $CH_4$ production were recorded at 4, 8, 12, and 24 h post incubation. The data collected was analyzed through Pearson correlation, polinomial regression and fixed effects models. There were negative correlations between FTS-total gas volume (r = -0.40; p<0.001); FTS-volume of $CH_4$ produced (r = -0.40; p<0.001) and between the inclusion level-volume of $CH_4$ produced (r = -0.20; p<0.001). As well as a positive correlation between hours post incubation-total gas volume (r = 0.42; p<0.001) and between hours post incubation-volume of $CH_4$ produced (r = 0.48; p<0.001). The FTS: C. elaeagnoides, V. mollis, and H. brasiletto have potential, in the three inclusion levels analyzed, to reduce $CH_4$ emission on in vitro trials (>32.7%), taking into account the total $CH_4$ production at 24 h of the forage used as reference (Avena sativa). It's suggested that C. elaeagnoides-according to its crude protein, neutral detergent fiber, and condensed tannins content- is the best alternative within the FTS analyzed, for feeding ruminants and for the control of $CH_4$ emissions during the dry season.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.4
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• pp.533-540
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• 2006

In this study, the possibility of green tea waste (GTW) as a new ingredient of byproducts-mixed silage was investigated. Characteristics of GTW were low in dry matter (DM) content (20%), and high in crude protein (30 to 36%) and tannins (8.5%). The GTW was added to mixed silages composed of tofu cake, rice straw and rice bran that are locally available in Japan. In experiment 1, the effect of GTW addition to silage made from various patterns of byproducts mixture based on tofu cake was studied. In experiment 2, the effect of GTW addition and storage temperature on fermentation characteristics, nutrient contents and in vitro ruminal gas production of byproducts-mixed silages were examined. In experiment 1, GTW addition on tofu cake accelerated acetic, propionic and butyric acid accumulation in the silage. When rice straw was mixed with tofu cake, DM content was increased from 47 to 56%, lactic acid was the main acid and the pH was decreased below 4.2. In this case, GTW addition to those mixtures did not affect acid concentrations of the silage. In experiment 2, GTW addition to the byproducts mixture increased lactic acid concentration, decreased the pH and DM loss of the silages. In GTW treatments, tannin concentration was lower in the silage stored at $30^{\circ}C$ than $15^{\circ}C$. Addition of GTW into the silage also increased in vitro ruminal gas production. It was concluded that addition of GTW into byproducts-mixed silage enhanced lactic acid fermentation when there were insufficient materials for lactic acid production. Utilization of GTW as an ingredient in mixed silages would be effective in enhancing fermentation characteristics, lowering tannin content and in vitro ruminal gas production.