• 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.25 no.3
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• pp.325-334
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• 2012

The objective of this study was to determine the roughage to concentrate (R:C) ratio with rain tree pod meal (RPM) supplementation on in vitro fermentation using gas production technique. The experiment design was a 6${\times}$4 factorial arrangement in a CRD. Factor A was 6 levels of R:C ratio (100:0, 80:20, 60:40, 40:60, 20:80 and 0:100) and factor B was 4 levels of RPM (0, 4, 8 and 12 mg). It was found that gas kinetic, extent rate (c) was linearly increased (p<0.01) with an increasing level of concentrate while cumulative gas production (96 h) was higher in R:C of 40:60. In addition, interaction of R:C ratio and RPM level affected $NH_3-N$ and IVDMD and were highest in R:C of 0:100 with 0, 4 mg of RPM and 40:60 with 8 mg of RPM, respectively. Moreover, interaction of R:C ratio and RPM level significantly increased total volatile fatty acids and propionate concentration whereas lower acetate, acetate to propionate ratios and $CH_4$ production in R:C of 20:80 with 8 mg of RPM. Moreover, the two factors, R:C ratio and RPM level influenced the protozoal population and the percentage of methanogens in the total bacteria population. In addition, the use of real-time PCR found that a high level of concentrate in the diet remarkably decreased three cellulolytic bacteria numbers (F. succinogenes, R. flavefaciens and R. albus). Based on this study, it is suggested that the ratio of R:C at 40:60 and RPM level at 12 mg could improve ruminal fluid fermentation in terms of reducing fermentation losses, thus improving VFA profiles and ruminal ecology.

• 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.26 no.6
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• pp.856-863
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• 2013

An in vitro gas production technique was used in this study to elucidate the effect of two strains of active live yeast on methane ($CH_4$) production in the large intestinal content of pigs to provide an insight to whether active live yeast could suppress $CH_4$ production in the hindgut of pigs. Treatments used in this study include blank (no substrate and no live yeast cells), control (no live yeast cells) and yeast (YST) supplementation groups (supplemented with live yeast cells, YST1 or YST2). The yeast cultures contained $1.8{\times}10^{10}$ cells per g, which were added at the rates of 0.2 mg and 0.4 mg per ml of the fermented inoculum. Large intestinal contents were collected from 2 Duroc${\times}$Landrace${\times}$Yorkshire pigs, mixed with a phosphate buffer (1:2), and incubated anaerobically at $39^{\circ}C$ for 24 h using 500 mg substrate (dry matter (DM) basis). Total gas and $CH_4$ production decreased (p<0.05) with supplementation of yeast. The methane production reduction potential (MRP) was calculated by assuming net methane concentration for the control as 100%. The MRP of yeast 2 was more than 25%. Compared with the control group, in vitro DM digestibility (IVDMD) and total volatile fatty acids (VFA) concentration increased (p<0.05) in 0.4 mg/ml YST1 and 0.2 mg/ml YST2 supplementation groups. Proportion of propionate, butyrate and valerate increased (p<0.05), but that of acetate decreased (p<0.05), which led to a decreased (p<0.05) acetate: propionate (A: P) ratio in the both YST2 treatments and the 0.4 mg/ml YST 1 supplementation groups. Hydrogen recovery decreased (p<0.05) with yeast supplementation. Quantity of methanogenic archaea per milliliter of inoculum decreased (p<0.05) with yeast supplementation after 24 h of incubation. Our results suggest that live yeast cells suppressed in vitro $CH_4$ production when inoculated into the large intestinal contents of pigs and shifted the fermentation pattern to favor propionate production together with an increased population of acetogenic bacteria, both of which serve as a competitive pathway for the available H2 resulting in the reduction of methanogenic archaea.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.3
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• pp.356-364
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• 2009

The current ruminant feeding models require parameterization of the digestion kinetics of carbohydrate fractions in feed ingredients to estimate the supply of nutrients from a ration. Using an automated gas production technique, statistically welldefined digestion rate of carbohydrate, including soluble carbohydrate, can be estimated in a relatively easy way. In this study, the gas production during in vitro fermentation was measured and recorded by an automated gas production system to investigate degradation kinetics of carbohydrate fractions of a wide range of ruminant feeds: corn silage, rice straw, corn, soybean hull, soybean meal, and cell mass from lysine production (CMLP). The gas production from un-fractionated, ethanol insoluble residue and neutral detergent insoluble residue of the feed samples were obtained. The gas profiles of carbohydrate fractions on the basis of the carbohydrate scheme of the Cornell Net Carbohydrate and Protein System (A, B1, B2, B3 and C) were generated using a subtraction approach. After the gas profiles were plotted with time, a curve was fitted with a single-pool exponential equation with a discrete lag to obtain kinetic parameters that can be used as inputs for modern nutritional models. The fractional degradation rate constants (Kd) of corn silage were 11.6, 25.7, 14.8 and 0.8%/h for un-fractioned, A, B1 and B2 fractions, respectively. The values were statistically well estimated, assessed by high t-value (>12.9). The Kd of carbohydrate fractions in rice straw were 4.8, 21.1, 5.7 and 0.5%/h for un-fractioned, A, B1 and B2 fractions, respectively. Although the Kd of B2 fraction was poorly defined with a t-value of 4.4, the Kd of the other fractions showed tvalues higher than 21.9. The un-fractioned corn showed the highest Kd (18.2%/h) among the feeds tested, and the Kd of A plus B1 fraction was 18.7%/h. Soybean hull had a Kd of 6.0, 29.0, 3.8 and 13.8%/h for un-fractioned, A, B1 and B2, respectively. The large Kd of fraction B2 indicated that NDF in soybean hull was easily degradable. The t-values were higher than 20 except for the B1 fraction (5.7). The estimated Kd of soybean meal was 9.6, 24.3, 5.0%/h for un-fractioned, A and B1 fractions, respectively. A small amount of gas (5.6 ml at 48 ho of incubation) was produced from fermentation of CMLP which contained little carbohydrate. In summary, the automated gas production system was satisfactory for the estimation of well defined (t-value >12) kinetic parameters and Kd of soluble carbohydrate fractions of various feedstuffs that supply mainly carbohydrate. The subtraction approach, however, should be applied with caution for some concentrates, especially those which contain a high level of crude protein since nitrogen-containing compounds can interfere with gas production.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.12
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• pp.1708-1716
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• 2013

The objective of the trial was to study the relationship between the methane ($CH_4$) production and the Cornell Net Carbohydrate and Protein System (CNCPS) carbohydrate fractions of feeds for cattle and the suitability of CNCPS carbohydrate fractions as the dietary variables in modeling the $CH_4$ production in rumen fermentation. Forty-five rations for cattle with the concentrate/roughage ratios of 10:90, 20:80, 30:70, 40:60, and 50:50 were formulated as feed samples. The Menke and Steingass's gas test was used for the measurement of $CH_4$ production. The feed samples were incubated for 48 h and the $CH_4$ production was analyzed using gas chromatography. Statistical analysis indicated that the $CH_4$ production (mL) was closely correlated with the CNCPS carbohydrate fractions (g), i.e. CA (sugars); $CB_1$ (starch and pectin); $CB_2$ (available cell wall) in a multiple linear pattern: $CH_4=(89.16{\pm}14.93)$ $CA+(124.10{\pm}13.90)$ $CB_1+(30.58{\pm}11.72)$ $CB_2+(3.28{\pm}7.19)$, $R^2=0.81$, p<0.0001, n = 45. Validation of the model using 10 rations indicated that the $CH_4$ production of the rations for cattle could accurately be predicted based on the CNCPS carbohydrate fractions. The trial indicated that the CNCPS carbohydrate fractions CA, $CB_1$ and $CB_2$ were suitable dietary variables for predicting the $CH_4$ production in rumen fermentation in vitro.

• 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.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.

• Animal Bioscience
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• v.35 no.9
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• pp.1367-1378
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• 2022

Objective: The present study was conducted to examine the gas production, fermentation characteristics, nutrient degradation, and methanogenic community composition of a rumen fluid culture with Broussonetia papyrifera (B. papyrifera) subjected to ensiling or steam explosion (SE) pretreatment. Methods: Fresh B. papyrifera was collected and pretreated by ensiling or SE, which was then fermented with ruminal fluids as ensiled B. papyrifera group, steam-exploded B. papyrifera group, and untreated B. papyrifera group. The gas and methane production, fermentation characteristics, nutrient degradation, and methanogenic community were determined during the fermentation. Results: Cumulative methane production was significantly improved with SE pretreatment compared with ensiled or untreated biomass accompanied with more volatile fatty acids production. After 72 h incubation, SE and ensiling pretreatments decreased the acid detergent fiber contents by 39.4% and 22.9%, and neutral detergent fiber contents by 10.6% and 47.2%, respectively. Changes of methanogenic diversity and abundance of methanogenic archaea corresponded to the variations in fermentation pattern and methane production. Conclusion: Compared with ensiling pretreatment, SE can be a promising technique for the efficient utilization of B. papyrifera, which would contribute to sustainable livestock production systems.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.10
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• pp.1424-1436
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• 2013

The objective of this study was to investigate the effect of banana flower powder (BAFLOP) supplementation on gas production kinetics and rumen fermentation efficiency in in vitro incubation with different ratios of roughage to concentrate in swamp buffalo and cattle rumen fluid. Two male, rumen fistulated dairy steers and swamp buffaloes were used as rumen fluid donors. The treatments were arranged according to a $2{\times}2{\times}3$ factorial arrangement in a Completely randomized design by using two ratios of roughage to concentrate (R:C; 75:25 and 25:75) and 3 levels of BAFLOP supplementation (0, 2 and 4% of dietary substrate) into two different kinds of rumen fluid (beef cattle and swamp buffalo). Under this investigation, the results revealed that the rumen ecology was affected by R:C ratio. The pH declined as a result of using high concentrate ratio; however, supplementation of BAFLOP could buffer the pH which led to an improvement of ruminal efficiency. BAFLOP supplementation affected acetic acid (C2) when the proportion of concentrate was increased. However, there were no effect on total volatile fatty acid (TVFA) and butyric acid (C4) by BAFLOP supplementation. The microbial community was affected by BAFLOP supplementation, especially the bacterial population. As revealed by real-time PCR, the populations of F. succinogenes and R. albus were reduced by the high concentrate treatments while that of R. flavafaciens were increased. The populations of three dominant cellulolytic bacteria were enhanced by BAFLOP supplementation, especially on high concentrate diet. BAFLOP supplementation did not influence the ammonia nitrogen ($NH_3$-N) concentration, while R:C did. In addition, the in vitro digestibility was improved by either R:C or BAFLOP supplementation. The BAFLOP supplementation showed an effect on gas production kinetics, except for the gas production rate constant for the insoluble fraction (c), while treatments with high concentrate ratio resulted in the highest values. In addition, BAFLOP tended to increase gas production. Based on this study, it could be concluded that R:C had an effect on rumen ecology both in buffalo and cattle rumen fluid and hence, BAFLOP could be used as a rumen buffering agent for enhancing rumen ecology fed on high concentrate diet. It is recommended that level of BAFLOP supplementation should be at 2 to 4% of total dry matter of substrate. However, in vivo trials should be subsequently conducted to investigate the effect of BAFLOP in high concentrate diets on rumen ecology as well as ruminant production.