• Asian-Australasian Journal of Animal Sciences
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• v.19 no.8
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• pp.1120-1126
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• 2006

Dry matter (DM) degradation of leaves from Quercus cercis, Quercus libari, Quercus branti, and Quercus coccifera was determined using two different techniques: (i) in vitro gas production and (ii) the nylon bag degradability technique. In vitro gas production in the presence or absence of PEG and in situ DM disappearance were measured at 3, 6, 12, 24, 48, 72 and 96 h. In situ and in vitro DM degradation kinetics were described using the equation y = a+b ($1-e^{-ct}$). At all incubation times leaves from Quercus branti incubated with or without PEG gave significantly higher gas production than the other oak leaves except for 3 and 6 h incubation when leaves from Quercus branti without PEG supplementation only gave higher gas production than Quercus cercis and Quercus coccifera. At all incubation times except at 3, 6 and 12 h the DM disappearance from Quercus branti was significantly higher than the other species. Generally, PEG supplementation considerably increased the gas production at all incubation times and estimated parameters such as gas production rate ($c_{gas}$), gas production (ml) from the quickly soluble fraction ($a_{gas}$), gas production (b) from the insoluble fraction, potential gas production (a+b). However, all oak leaves did not give the same response to the PEG supplementation. Although the increase in gas production at 96 h incubation time was 8.9 ml for Quercus libari the increase was 5.5 ml for Quercus coccifera. It was concluded that except at early incubation times the relationships between the two methodologies seem to be sufficiently strong to predict degradability parameters from gas production parameters obtained in the presence or absence of PEG.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.7
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• pp.1049-1056
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• 2007

This study was conducted to compare the methane ($CH_4$) production estimated by in vivo (sulfur hexafluoride tracer technique ($SF_6$)) with that of two in vitro rumen simulation (RUSITEC) and gas production (IVGPT)) techniques. Four adult dry Holstein cows, aged $7.4{\pm}3.0$ years and weighing $697{\pm}70$ kg, were used for measuring methane production from five diets by the $SF_6$ technique. The experimental diets were alfalfa hay ($D_1$), corn silage + soybean meal (SBM) (910: 90, $D_2$), Italian rye grass hay +SBM (920: 80, $D_3$), rice straw +SBM (910: 90, $D_4$) and Sudan grass hay +SBM (920: 80, $D_5$). Each diet was individually fed to all 4 cows and 5 feeding studies of 17 d each were conducted to measure the methane production. In the RUSITEC, methane production was measured from triplicate vessels for each diet .In vitro gas production was measured for each of the diets in triplicate syringes. The gas produced after 24 and 48 h was recorded and gas samples were collected in vacuum vials and the methane production was calculated after correction for standard temperature and pressure (STP). Compared to the $SF_6$ technique, estimates of methane production using the RUSITEC were lower for all diets. Methane production estimated from 24 h in vitro gas production was higher (p<0.001) on $D_1$ as compared to that measured by $SF_6$, whereas on $D_2$ to $D_5$ it was lower. Compared to $SF_6$, methane production estimated from 48 h in vitro gas production was higher on all diets. However, methane estimated from the mean of the two measurement intervals (24+48 h/2) in IVGPT was very close to that of $SF_6$ (correlation 0.98), except on $D_1$. The results of our study confirmed that IVGPT is reflective of in vivo conditions, so that it could be used to generate a database on methane production potential of various ruminant diets and to examine strategies to modify methane emissions by ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.1
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• pp.51-61
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• 2016

In order to provide recommendations on the most useful forage species to smallholder farmers, eleven grass and eleven legume forages grown in Abomey-Calavi in Republic of Benin were investigated for nutritive value (i.e. chemical composition and energy content) and fermentation characteristics (i.e. gas and volatile fatty acid production, organic matter degradability). The in vitro gas production technique was used, incubating the forages for 120 h under anaerobic condition with buffalo rumen fluid. Compared to legume, tropical grass forages showed lower energy (8.07 vs 10.57 MJ/kg dry matter [DM]) and crude protein level (16.10% vs 19.91% DM) and higher cell wall content (neutral detergent fiber: 63.8% vs 40.45% DM), respectively. In grass forages, the chemical composition showed a quite high crude protein content; the in vitro degradability was slightly lower than the range of tropical pasture. The woody legumes were richer in protein and energy and lower in structural carbohydrates than herbaceous plants, however, their in vitro results are influenced by the presence of complex compounds (i.e. tannins). Significant correlations were found between chemical composition and in vitro fermentation characteristics. The in vitro gas production method appears to be a suitable technique for the evaluation of the nutritive value of forages in developing countries.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.1
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• pp.36-45
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• 2014

The objective of this study was to determine the effects of protein sources and roughage (R) to concentrate (C) ratio on in vitro fermentation parameters using a gas production technique. The experimental design was a $2{\times}5$ factorial arrangement in a completely randomized design (CRD). Factor A was 2 levels of protein sources yeast fermented cassava chip protein (YEFECAP) and soybean meal (SBM) and factor B was 5 levels of roughage to concentrate (R:C) ratio at 80:20, 60:40, 40:60, 20:80, and 0:100, respectively. Rice straw was used as a roughage source. It was found that gas production from the insoluble fraction (b) of YEFECAP supplemented group was significantly higher (p<0.05) than those in SBM supplemented group. Moreover, the intercept value (a), gas production from the insoluble fraction (b), gas production rate constants for the insoluble fraction (c), potential extent of gas production (a+b) and cumulative gas production at 96 h were influenced (p<0.01) by R:C ratio. In addition, protein source had no effect (p>0.05) on ether in vitro digestibility of dry matter (IVDMD) and organic (IVOMD) while R:C ratio affected the IVDMD and IVOMD (p<0.01). Moreover, YEFECAP supplanted group showed a significantly increased (p<0.05) total VFA and $C_3$ while $C_2$, $C_2:C_3$ and $CH_4$ production were decreased when compared with SBM supplemented group. In addition, a decreasing R:C ratio had a significant effect (p<0.05) on increasing total VFA, $C_3$ and $NH_3$-N, but decreasing the $C_2$, $C_2:C_3$ and CH4 production (p<0.01). Furthermore, total bacteria, Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminococcus albus populations in YEFECAP supplemented group were significantly higher (p<0.05) than those in the SBM supplemented group while fungal zoospores, methanogens and protozoal population remained unchanged (p>0.05) as compared between the two sources of protein. Moreover, fungal zoospores and total bacteria population were significantly increased (p<0.01) while, F. succinogenes, R. flavefaciens, R. albus, methanogens and protozoal population were decreased (p<0.01) with decreasing R:C ratio. In conclusion, YEFECAP has a potential for use as a protein source for improving rumen fermentation efficiency in ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.4
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• pp.526-532
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• 2006

The aim of this study was to evaluate the chemical composition, in vitro DM degradability, ME and OMD of alfalfa-maize silage mixtures in comparison to pure maize and alfalfa silages, and to test the existence of associative effects of ensiling alfalfa forage with whole-crop maize using the in vitro gas production technique. Ensiling alfalfa with whole-crop maize had a significant (p<0.001) effect on chemical composition, pH, in vitro DM degradability, OMD and estimated ME values of mixtures. DM content of the resultant silages significantly increased with increasing proportion of whole-crop maize in the mixtures, whereas the pH value, crude protein (CP), acid detergent fibre (ADF) and ash contents of mixtures decreased due to the dilution effect of whole-crop maize which was low in CP, ADF and ash. The pH values of all alfalfa-maize silage mixtures were at the desired level for quality silage. Gas production of alfalfa-maize silage mixtures at all incubation times except 12 h increased with increasing proportion of whole-crop maize. When alfalfa was mixed with whole-crop maize in the ratio 40:60, ME and OMD values were significantly (p<0.001) higher than other silages. Maximum gas production ($A_{gas}$) ranged from 65.7 to 78.1 with alfalfa silage showing the lowest maximum gas production. The results obtained in this study clearly showed that maximum gas production increased with increased percentage of whole-crop maize in the silage mixtures (r = 0.940, p<0.001). It was concluded that ensiling alfalfa with whole-crop maize improved the pH, OMD and ME values. However, trials with animals are required to see how these differences in silage mixtures affect animal performance.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.10
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• pp.1404-1410
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• 2012

In vitro experiments were conducted to evaluate the suitability of several mixtures of high tanniniferous non legumes with low tanniniferous legumes on in vitro gas production (IVGP), dry matter degradation, Ammonia-N, methane production and microbial population. Eight treatments were examined in a randomized complete block design using four non-legumes and two legumes (Carallia integerrima${\times}$Leucaena leucocephala (LL) (Trt 1), C. integerrima${\times}$Gliricidia sepium (GS) (Trt 2), Aporosa lindeliyana${\times}$LL (Trt 3), A. lindeliyana${\times}$GS (Trt 4), Ceiba perntandra${\times}$LL (Trt 5), C. perntandra${\times}$GS (Trt 6), Artocarpus heterophyllus${\times}$LL (Trt 7), A. heterophyllus${\times}$GS (Trt 8). The condensed tannin (CT) content of non legumes ranged from 6.2% (Carallia integerrima) to 4.9% (Ceiba perntandra) while the CT of legumes were 1.58% (Leucaena leucocephala) and 0.78% (Gliricidia sepium). Forage mixtures contained more than 14% of crude protein (CP) while the CT content ranged from 2.8% to 4.0% respectively. Differences (p<0.05) were observed in in vitro gas production (IGVP) within treatments over a 48 h period dominated by C. perntandra${\times}$G. sepium (Trt 6). The net gas production (p<0.05) was also high with Trt6 followed by A. heterophyllus${\times}$L. leucocephala (Trt 7) and A. heterophyllus${\times}$G. sepium (Trt 8). Highest (p>0.05) NH3-N (ml/200 mg DM) production was observed with the A. heterophyllus${\times}$G. sepium (Trt 8) mixture which may be attributed with it's highest CP content. The correlation between IVGP and CT was 0.675 while IVGP and CP was 0.610. In vitro dry matter degradation (IVDMD) was highest in Trt 8 as well. Methane production ranged from 2.57 to 4.79 (ml/200 mg DM) to be synonimous with IVGP. A higher bacteria population (p<0.05) was found in C. perntandra${\times}$G. sepium (Trt 6) followed by Artocarpus heterophyllus+G. sepium (Trt 8) and the same trend was observed with the protozoa population as well. The results show that supplementing high tannin non leguminous forages by incremental substitution of legume forage increased gas production parameters, NH3-N, IVDMD and microbial population in the fermentation liquid. Methane production was not significantly affected by the presence of CT or different levels of CP in forage mixtures. Among non legumes, Ceiba perntandra and Artocarpus heterophyllus performed better in mixture with L. leucocephala and G. sepium.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.2
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• pp.240-247
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• 2012

Dietary supplementation with conventional linted cottonseed hulls (LCSH) is a common practice in livestock production all over the world. However, supplementation with mechanically delinted cottonseed hulls (DCSH) and cottonseed linter residue (CLR) is uncommon. Cottonseed by-products, including LCSH, DCSH and CLR, were assessed by chemical analysis, an in situ nylon bag technique, an in vitro cumulative gas production technique and in vitro enzyme procedure. The crude protein (CP) content of CLR (302 g/kg dry matter (DM)) was approximately 3 times that of LCSH and 5 times that of DCSH. The crude fat content was approximately 3 times higher in CLR (269 g/kg DM) than in LCSH and 4 times higher than in DCSH. Neutral detergent fibre (311 g/kg DM) and acid detergent fibre (243 g/kg DM) contents of CLR were less than half those of DCSH or LCSH. Metabolisable energy, estimated by in vitro gas production and chemical analyses, ranked as follows: CLR (12.69 kJ/kg DM)>LCSH (7.32 kJ/kg DM)>DCSH (5.82 kJ/kg DM). The in situ degradation trial showed that the highest values of effective degradability of DM and CP were obtained for CLR (p<0.05). The in vitro disappearance of ruminal DM ranked as follows: CLR>LCSH>DCSH (p<0.05). The lowest digestibility was observed for DCSH with a two-step in vitro digestion procedure (p<0.05). The potential gas production in the batch cultures did not differ for any of the three cottonseed by-product feeds. The highest concentration of total volatile fatty acids was observed in CLR after a 72 h incubation (p<0.05). The molar portions of methane were similar between all three treatments, with an average gas production of 22% (molar). The CLR contained a higher level of CP than did LCSH and DCSH, and CLR fermentation produced more propionate. The DCSH and LCSH had more NDF and ADF, which fermented into greater amounts of acetate.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.6
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• pp.797-805
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• 2014

The effect of concentrate mixtures with crude protein (CP) levels 10%, 13%, 16%, and 19% and diets with roughage to concentrate ratios 80:20, 60:40, 40:60, and 20:80 (w/w) were determined on dry matter (DM) and organic matter (OM) digestibility, and fermentation metabolites using an in vitro fermentation technique. In vitro fermented attributes were measured after 4, 24, and 48 h of incubation respectively. The digestibility of DM and OM, and total volatile fatty acid (VFA) increased whereas pH decreased with the increased amount of concentrate in the diet (p<0.001), however CP levels of concentrate did not have any influence on these attributes. Gas production reduced with increased CP levels, while it increased with increasing concentrate levels. Ammonia nitrogen ($NH_3$-N) concentration and microbial CP production increased significantly (p<0.05) by increasing CP levels and with increasing concentrate levels in diet as well, however, no significant difference was found between 16% and 19% CP levels. Therefore, 16% CP in concentrate and increasing proportion of concentrate up to 80% in diet all had improved digestibility of DM and organic matter, and higher microbial protein production, with improved fermentation characteristics.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1278-1285
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• 2008

Measurement of gas produced during in vitro fermentation was used to investigate the fermentation characteristics and interactions of rice straw, wheat straw or maize stover mixed with alfalfa or clover at proportions of 100:0, 75:25, 50:50, 25:75 and 0:100, respectively. Cumulative gas production was recorded at 2, 4, 8, 12, 16, 24 and 48 h of incubation, and the Gompertz function was used to describe the kinetics of gas production. In vitro dry matter and organic matter disappearances (IVDMD and IVOMD) were determined after 48 h incubation. The rate of gas production of clover was higher (p<0.05) than that of rice straw, wheat straw, maize stover and alfalfa when straws and hays were incubated separately. Increasing the proportion of alfalfa in the straw-alfalfa mixtures increased (p<0.05) the rates, but not the maximum volume of gas production. However, both rate and the maximum volume of gas production were increased (p<0.01) as the proportions of clover increased in the straw-clover mixtures. The cumulative gas production at 48 h, IVDMD and IVOMD showed no consistent interaction effects between different mixtures of cereal straws and hays. The extent of interactive effects was affected by the types of cereal straw, legume hay and their proportions in the mixture. The appropriate combination for the mixture of rice straw or maize stover with leguminous hays was 75:25 and 25:75, respectively. The better combination occurred at a proportion of 50:50 for the mixture of wheat straw and alfalfa. We conclude that the suitable proportion of low-quality straw and high quality legume hay combination should be considered in the ration formulation system of ruminants according to the extent of positive interactive effects.