• Asian-Australasian Journal of Animal Sciences
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• v.31 no.12
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• pp.1890-1896
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• 2018

Objective: As the climate changes, it influences ruminant's feed intake, nutrient digestibility, rumen methane production and emission. This experiment aimed to evaluate the effect of feeding Sweet grass (Pennisetum purpureum cv. Mahasarakham; SG) as a new source of good quality forage to improve feed utilization efficiency and to mitigate rumen methane production and emission. Methods: Four, growing crossbred of Holstein Friesian heifers, 14 months old, were arranged in a $4{\times}4$ Latin square design to receive four dietary treatments. Treatment 1 (T1) was rice straw (RS) fed on ad libitum with 1.0% body weight (BW) of concentrate (C) supplementation (RS/1.0C). Treatment 2 (T2) and treatment 3 (T3) were SG, fed on ad libitum with 1.0% and 0.5% BW of concentrate supplementation, respectively (SG/1.0C and SG/0.5C, respectively). Treatment 4 (T4) was total Sweet grass fed on ad libitum basis with non-concentrate supplementation (TSG). Results: The results revealed that roughage and total feed intake were increased with SG when compared to RS (p<0.01) while TSG was like RS/1.0C treatment. Digestibility of nutrients, nutrients intake, total volatile fatty acids (VFAs), rumen microorganisms were the highest and CH4 was the lowest in the heifers that received SG/1.0C (p<0.01). Total dry matter (DM) feed intake, digestibility and intake of nutrients, total VFAs, $NH_3-N$, bacterial and fungal population of animals receiving SG/0.5C were higher than those fed on RS/1.0C. Reducing of concentrate supplementation with SG as a roughage source increased $NH_3-N$, acetic acid, and fungal populations, but it decreased propionic acid and protozoal populations (p<0.05). However, ruminal pH and blood urea nitrogen were not affected by the dietary treatments (p>0.05). Conclusion: As the results, SG could be a good forage to improve rumen fermentation, decrease methane production and reduced the level of concentrate supplementation for growing ruminants in the tropics especially under global climate change.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.6
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• pp.818-823
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• 2008

The interaction of fibre degrading microbes and methanogens was studied using two forages, lucerne (Medicago sativa) hay and maize (Zea mays) hay, as substrate and 2-bromoethanesulphonic acid (BES) as an additive in an in vitro gas production test. Gas and methane production (ml/g dry matter) were significantly higher (p<0.05) on lucerne as compared to maize hay. Inclusion of BES in the incubation medium significantly suppressed methane emission irrespective of substrate. The population density of total bacteria, fungi, Ruminococcus flavefaciens and Fibrobacter succinogenes was higher, whereas that of methanogens was lower with maize hay as compared to lucerne as substrate. BES suppressed methanogen population by 7 fold on lucerene and by 8.5 fold on maize at 24 h incubation as estimated by real time-PCR. This suppression was accompanied by almost complete (>98% of control) inhibition of methanogenesis. The proportion of acetate decreased, whereas that of propionate increased significantly by inclusion of BES, resulting in narrowing of acetate to propionate ratio. In vitro true digestibility (IVTD) of lucerne was significantly higher as compared to maize but BES inclusion did not affect IVTD.

• Journal of Life Science
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• v.22 no.10
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• pp.1324-1329
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• 2012

The objective of this study was to evaluate the in vitro effects of organic acids on methane emission and ruminal fermentation characteristics. We expected our methodology to result in a decrease of methanogens attached to the surface of rumen ciliate protozoa by addition of organic acids and in particular a decrease in methane emission. A fistulated Holstein cow of 650 kg body weight was used as a donor of rumen fluid. Organic acids (aspartic acid, fumaric acid, lactic acid, malic acid, and succinic acid) known to be propionate enhancers were added to an in vitro fermentation system and incubated with rumen fluid. The microbial population, including bacteria, protozoa, and fungi, were enumerated, and gas production, including methane and fermentation characteristics, were observed in vitro. Organic acids appeared to affect the rumen protozoan community. The rumen protozoal popuation decreased with the addition of aspartic acid, fumaric acid, lactic acid, and malic acid. In particular, the methane emission was reduced by addition of lactic acid. The concentration of propionate with all organic acids that were added appeared to be higher than that of the control at 12 h incubation. Addition of organic acids significantly affected rumen bacteria and microbial growth. The bacteria in added fumaric acid and malic acid was significantly higher (p<0.05) and protozoa was significantly lower (p<0.05) than that of the control. Microbial growth with the addition of organic acids was greater than the control after 48 h incubation.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.1
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• pp.103-109
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• 2019

Objective: Ensiling of tannin-rich fruit byproducts (FB) involves quantitative and qualitative changes in the tannins, which would consequently change the rumen fermentation characteristics. This study aimed to evaluate whether ensiled FBs are effective in mitigating methane emission from ruminants by conducting in vitro assessments. Methods: Fruit byproducts (grape pomace, wild grape pomace, and persimmon skin) were collected and subjected to four-week ensiling by Lactobacillus buchneri inoculant. A defined feed component with or without FB samples (both fresh and ensiled material) were subjected to in vitro anaerobic culturing using rumen fluid sampled from beef cattle, and the fermentation parameters and microbial populations were monitored. Results: Reduced methane production and a proportional change in total volatile fatty acids (especially enhanced propionate proportion) was noted in bottles containing the FBs compared with that in the control (without FB). In addition, we found lower gene copy number of archaeal 16S rRNA and considerably higher levels of one of the major fibrolytic bacteria (Fibrobacter succinogenes) in the bottles containing FBs than in the control, particularly, when it was included in a forage-based feed. However, in the following cultivation experiment, we observed that FBs failed to exhibit a significant difference in methane production with or without polyethylene glycol, implying that tannins in the FBs may not be responsible for the mitigation of methane generation. Conclusion: The results of the in vitro cultivation experiments indicated that not only the composition but also ensiling of FBs affected rumen fermentation patterns and the degree of methane generation. This is primarily because of the compositional changes in the fibrous fraction during ensiling as well as the presence of readily fermented substrates, whereas tannins in these FBs seemed to have little effect on the ruminal fermentation kinetics.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.11
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• pp.1601-1607
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• 2016

This study was conducted to determine the effect of lysozyme addition on in vitro rumen fermentation and to identify the lysozyme inclusion rate for abating methane ($CH_4$) production. An in vitro ruminal fermentation technique was done using a commercial concentrate to rice straw ratio of 8:2 as substrate. The following treatments were applied wherein lysozyme was added into 1 mg dry matter substrate at different levels of inclusion: Without lysozyme, 2,000, 4,000, and 8,000 U lysozyme. Results revealed that, lysozyme addition had a significant effect on pH after 24 h of incubation, with the highest pH (p<0.01) observed in 8,000 U lysozyme, followed by the 4,000 U, 2,000 U, and without lysozyme. The highest amounts of acetic acid, propionic acid (p<0.01) and total volatile fatty acid (TVFA) (p<0.05) were found in 8,000 U after 24 h of incubation. The $CH_4$ concentration was the lowest in the 8,000 U and the highest in the without lysozyme addition after 24 h of incubation. There was no significant differences in general bacteria, methanogen, or protozoan DNA copy number. So far, addition of lysozyme increased the acetate, propionate, TVFA, and decreased $CH_4$ concentration. These results suggest that lysozyme supplementation may improve in vitro rumen fermentation and reduce $CH_4$ emission.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.54-62
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• 2018

Objective: Due to the threat of global warming, the livestock industry is increasingly interested in exploring how feed additives may reduce anthropogenic greenhouse gas emissions, especially from ruminants. This study investigated the effect of Rhodophyta supplemented bovine diets on in vitro rumen fermentation and rumen microbial diversity. Methods: Cannulated Holstein cows were used as rumen fluid donors. Rumen fluid:buffer (1:2; 15 mL) solution was incubated for up to 72 h in six treatments: a control (timothy hay only), along with substrates containing 5% extracts from five Rhodophyta species (Grateloupia lanceolata [Okamura] Kawaguchi, Hypnea japonica Tanaka, Pterocladia capillacea [Gmelin] Bornet, Chondria crassicaulis Harvey, or Gelidium amansii [Lam.] Lamouroux). Results: Compared with control, Rhodophyta extracts increased cumulative gas production after 24 and 72 h (p = 0.0297 and p = 0.0047). The extracts reduced methane emission at 12 and 24 h (p<0.05). In particular, real-time polymerase chain reaction analysis indicated that at 24 h, ciliate-associated methanogens, Ruminococcus albus and Ruminococcus flavefaciens decreased at 24 h (p = 0.0002, p<0.0001, and p<0.0001), while Fibrobacter succinogenes (F. succinogenes) increased (p = 0.0004). Additionally, Rhodophyta extracts improved acetate concentration at 12 and 24 h (p = 0.0766 and p = 0.0132), as well as acetate/propionate (A/P) ratio at 6 and 12 h (p = 0.0106 and p = 0.0278). Conclusion: Rhodophyta extracts are a viable additive that can improve ruminant growth performance (higher total gas production, lower A/P ratio) and methane abatement (less ciliateassociated methanogens, Ruminococcus albus and Ruminococcus flavefaciens and more F. succinogenes.

• Korean Journal of Organic Agriculture
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• v.26 no.4
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• pp.775-787
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• 2018

This study was conducted to investigate for the natural methane emission inhibitor as a feed additive no adversely effect on rumen fermentation. Five different Control (Wheat barn (0.05 g), MRA(Methane Reduction Additive)-1 (Allium fistulosum L. (0.05 g)), MRA-2 (Sodium Lauryl Sulfate (0.025 g) + Wheat barn (0.025 g) mixed), MRA-3 (Sodium Dodecyl Sulfate (0.025 g) + Wheat barn (0.025 g) mixed), and MRA-4 (Allium fistulosum L. (0.02 g) + Tannic acid (0.02 g) + Wheat barn (0.01 g) mixed) contents were used to perform 3, 6, 9, 12, 24 and 48 h incubation for in vitro fermentation. Ruminal pH values were ranged within normal ruminal microbial fermentation. Dry matter digestibility was not significantly different across the treatments during the whole fermentation time. Also, the result of microbial growth had no adversely effect on during the whole fermentation time. At 24 h, methane emission was significantly lower (P<0.05) than all treatments except to MRA-1. Especially, MRA-4 carbon dioxide emission was significantly lower (P<0.05) than control at 9, 24 and 48 h incubation. In addition MRA-4 propionate concentration was significantly higher (P<0.05) than control at 24 h incubation. The result of RT-PCR Ciliate-associated methanogens were significantly lower (P<0.05) at MRA-1, MRA-3 and MRA-4 than control at 24 h incubation. Based on the present results, MRA-4 could be suggestible methane emission inhibitor as a natural feed additive.

• Korean Journal of Organic Agriculture
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• v.25 no.4
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• pp.901-916
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• 2017

This study was conducted to evaluate natural plant extracts for methane gas reduction in ruminants. Rumen fluid was collected from cannulated Hanwoo cow ($450{\pm}30kg$) consuming 400 g/kg concentrate and 600 g/kg timothy. The 15 ml of mixture comparing McDougall's buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically into 50 ml serum bottles. Rumen fluid contents were collected and in vitro fermentation prepared control (timothy, 300 mg), ginseng, balloon flower, yucca plant, camellia, tea plant and ogapi extracts were added at the level of 5% against 300 mg of timothy as a substrate (v/w) and incubated for 3, 6, 9, 12, 24, 48, and 72 h. In vitro pH values range 6.55~7.41, this range include rumen titration. The dry matter digestibility was not differ between all treatments and control. Total gas emission was significantly higher (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Carbon dioxide emission was not differ all treatments on 9 h than in control and significantly higher (p<0.05) yucca plant, camellia and tea plant treatments on 12 h than control. Methane emission was not differ all treatments on 6 h than in control. The rumen microbial growth rate was significantly higher (p<0.05) in ginseng, balloon flower on 12 h and significantly higher (p<0.05) in ginseng, yucca plant, tea plant and ogapi treatments on 24 h than in control. Total VFA was significantly higher (p<0.05) in tea plant and ogapi treatments on 12 h than in control and significantly higher (p<0.05) in ginseng, balloon flower treatments on 48 h than in control. Acetic acid was significantly lower (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Propionic acid was significantly higher (p<0.05) in ginseng and balloon flower treatments on 48 h than in control. As a results, sixth natural plant extracts had no significant effect dry matter digestibility and negative on rumen fermentation, but not effect methane reduction.

• Korean Journal of Organic Agriculture
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• v.29 no.2
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• pp.241-256
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• 2021

The current study was to evaluate the antioxidant activity of Phyllostachys bambusoides (PHB) as a feed additives and investigate whether its antioxidant activity could be helpful for increasing rumen fermentation characteristics and methane reduction. The antioxidant activity results showed that total polyphenols and flavonoids contents were 43.54 ± 8.68 mg CE/g and 17.13 ± 0.45 mg QE/g, respectively, and the IC₅₀ values for 1,1-diphenyl-2-prcrylhydrazyl (DPPH) and 2,2'-azino-bis (3- ethylbenzthiazoline-6- sulphonic acid) (ABTS) radical scavenging activity were 163.13 ± 19.25 ㎍/mL and 97.07 ± 4.46 ㎍/mL, respectively. Two heads of cannulated Hanwoo (450 ± 30 kg), consuming timothy hay and a commercial concentrate (60:40, w/w) twice daily (at 09:00 and 17:30) at 2% of body weight, with free access to water and a mineral block, were used as rumen fluid donors. An in vitro incubation experiment was performed after 6, 12, 24, 48, and 72 hr with PHB added at concentration of 2, 4, and 6% of timothy hay basis. Total gas emission decreased as the amount of PHB addition increased at 6 and 24 hr of incubation. However, PHB addition did not affect total volatile fatty acid production, and methane and carbon dioxide emission also decreased as the amount of addition increased at 48 hr of incubation. Therefore, PHB was expected to be used as methane reducing additives in the ruminants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.3
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• pp.197-205
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• 2013

An in vitro study was conducted to determine the effects of defaunation (removal of protozoa) and forage sources (rice straw, ryegrass and tall fescue) on ruminal fermentation characteristics, methane ($CH_4$) production and degradation by rumen microbes. Sodium lauryl sulfate, as a defaunation reagent, was added into the mixed culture solution to remove ruminal protozoa at a concentration of 0.375 mg/ml. Pure cellulose (0.64 g, Sigma, C8002) and three forage sources were incubated in the bottle of culture solution of mixed rumen microbes (faunation) or defaunation for up to 24 h. The concentration of ammonia-N was high under condition of defaunation compared to that from faunation in all incubations (p<0.001). Total VFA concentration was increased at 3, 6 and 12 h (p<0.05~p<0.01) but was decreased at 24 h incubation (p<0.001) under condition of defaunation. Defaunation decreased acetate (p<0.001) and butyrate (p<0.001) proportions at 6, 12 and 24 h incubation times, but increased propionate (p<0.001) proportion at all incubation times for forages. Effective degradability of dry matter was decreased by defaunation (p<0.001). Defaunation not only decreased total gas (p<0.001) and $CO_2$ (p<0.01~0.001) production at 12 and 24 h incubations, but reduced $CH_4$ production (p<0.001) at all incubation times for all forages. The $CH_4$ production, regardless of defaunation, in order of forage sources were rice straw > tall fescue > ryegrass > cellulose (p<0.001) up to 24 h incubation.