• Asian-Australasian Journal of Animal Sciences
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• v.30 no.3
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• pp.370-378
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• 2017

Objective: An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO), containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods: Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control), 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results: Ruminal pH, total volatile fatty acids (VFA) concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05) compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05) in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP) bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion:The results of the present study suggested that supplementation of MEO may have limited effects on apparent nutrient digestibility, ruminal fermentation and protozoa and HAP bacteria count, blood cells and metabolites.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1458-1468
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• 2002

There is increasing interest in exploiting natural products as feed additives to solve problems in animal nutrition and livestock production. Essential oils and saponins are two types of plant secondary compounds that hold promise as natural feed additives for ruminants. This paper describes recent advances in research into these additives. The research has generally concentrated on protein metabolism. Dietary essential oils caused rates of NH$_3$ production from amino acids in ruminal fluid taken from sheep and cattle receiving the oils to decrease, yet proteinase and peptidase activities were unchanged. Hyper-ammonia-producing (HAP) bacteria were the most sensitive of ruminal bacteria to essential oils in pure culture. Essential oils also slowed colonisation and digestion of some feedstuffs. Ruminobacter amylophilus may be a key organism in mediating these effects. Saponin-containing plants and their extracts appear to be useful as a means of suppressing the bacteriolytic activity of rumen ciliate protozoa and thereby enhancing total microbial protein flow from the rumen. The effects of some saponins seems to be transient, which may stem from the hydrolysis of saponins to their corresponding sapogenin aglycones, which are much less toxic to protozoa. Saponins also have selective antibacterial effects which may prove useful in, for example, controlling starch digestion. These studies illustrate that plant secondary compounds, of which essential oils and saponins comprise a small proportion, have great potential as 'natural' manipulators of rumen fermentation, to the potential benefit of the farmer and the environment.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.918-921
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• 2000

A comparative study on rumen bacterial and protozoal population and fungal zoospores in cattle (Brahman$\times$Native) and swamp buffalo (Bubalus bubalis) was conducted. Forty animals, twenty of each, with same sex and similar age which were raised under similar condition in the Northeast of Thailand, were used. Rumen digesta were sampled immediately post slaughtering for total microscopic counts of bacteria, protozoa and fungal zoospores. It was found that total bacterial population were higher in swamp buffalo that those in cattle (1.6 vs $1.36{\times}10^{8}cells/ml$) having more population of cocci, rods and ovals. Lower rumen protozoal pupulation in swamp buffalo with lower numbers of Holotrichs and Entodiniomorphs were found as compared to those in cattle. Significant higher fungal zoospore counts were in swamp buffalo than those in cattle being 7.30 and $3.78{\times}10^6$, respectively. Study under electron microscope, revealed Anaeromyces sp. with acuminate apex were more predominant in the rumen of swamp buffalo. With these findings, cattle and swamp buffaloes showing differences in rumen bacterial, protozoal population and fungal zoospore counts, offer new additional information as why swamp buffaloes exhibit conditionally better than cattle especially during long dry season without green grass.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.957-961
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• 2000

A series of experiments was conducted to evaluate phytin phosphohydrolysis actlVlty in the rumen and to isolate phytase positive rumen bacteria. Endogenous phytase activity of wheat bran was estimated and compared with that of bacterial phytin phosphohydrolysis. Substantial phytase activity was detected in wheat bran during in vitro rumen incubation. Bacterial phytase activity was suggested not to be high. Only two facultative anaerobes, Klebsiella sp. and Corynebacterium sp. were isolated as phytase producing organisms. These belonged to a minor microbial group in the rumen population. Protozoal fraction showed an initial velocity of phytin phosphohydrolysis 7 times higher than the bacterial fraction.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.2
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• pp.285-294
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• 2011

The abatement of methane emission from ruminants is an important global issue due to its contribution to greenhouse gas with carbon dioxide. Methane is generated in the rumen by methanogens (archaea) that utilize metabolic hydrogen ($H_2$) to reduce carbon dioxide, and is a significant electron sink in the rumen ecosystem. Therefore, the competition for hydrogen used for methanogenesis with alternative reductions of rumen microbes should be an effective option to reduce rumen methanogenesis. Some methanogens parasitically survive on the surface of ciliate protozoa, so that defaunation or decrease in protozoa number might contribute to abate methanogenesis. The most important issue for mitigation of rumen methanogenesis with manipulators is to secure safety for animals and their products and the environment. In this respect, prophylactic effects of probiotics, prebiotics and miscellaneous compounds to mitigate rumen methanogenesis have been developed instead of antibiotics, ionophores such as monensin, and lasalocid in Japan. Nitrate suppresses rumen methanogenesis by its reducing reaction in the rumen. However, excess intake of nitrate causes intoxication due to nitrite accumulation, which induces methemoglobinemia. The nitrite accumulation is attributed to a relatively higher rate of nitrate reduction to nitrite than nitrite to ammonia via nitroxyl and hydroxylamine. The in vitro and in vivo trials have been conducted to clarify the prophylactic effects of L-cysteine, some strains of lactic acid bacteria and yeast and/or ${\beta}$1-4 galactooligosaccharide on nitrate-nitrite intoxication and methanogenesis. The administration of nitrate with ${\beta}$1-4 galacto-oligosaccharide, Candida kefyr, and Lactococcus lactis subsp. lactis were suggested to possibly control rumen methanogenesis and prevent nitrite formation in the rumen. For prebiotics, nisin which is a bacteriocin produced by Lactococcus lactis subsp. lactis has been demonstrated to abate rumen methanogenesis in the same manner as monensin. A protein resistant anti-microbe (PRA) has been isolated from Lactobacillus plantarum as a manipulator to mitigate rumen methanogenesis. Recently, hydrogen peroxide was identified as a part of the manipulating effect of PRA on rumen methanogenesis. The suppressing effects of secondary metabolites from plants such as saponin and tannin on rumen methanogenesis have been examined. Especially, yucca schidigera extract, sarsaponin (steroidal glycosides), can suppress rumen methanogenesis thereby improving protein utilization efficiency. The cashew nutshell liquid (CNSL), or cashew shell oil, which is a natural resin found in the honeycomb structure of the cashew nutshell has been found to mitigate rumen methanogenesis. In an attempt to seek manipulators in the section on methane belching from ruminants, the arrangement of an inventory of mitigation technologies available for the Clean Development Mechanism (CDM) and Joint Implementation (JI) in the Kyoto mechanism has been advancing to target ruminant livestock in Asian and Pacific regions.

• Asian-Australasian Journal of Animal Sciences
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• v.2 no.3
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• pp.493-496
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• 1989

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.1
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• pp.96-114
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• 2000

Genetically selected beef cattle are fed high-energy diets in intensive production systems developed in industrial countries. This type of feeding can induce rumen dysfunctions that have to be corrected by farmers to optimise cost-effectiveness. The risk of rumen acidosis can be reduced by using slowly degradable starch, which partly escapes rumen fermentation and goes on to be digested in the small intestine. Additives are proposed to stabilise the rumen pH and restrict lactate accumulation, thus favouring the growth of cellulolytic bacteria and stimulating the digestion of the dietary plant cell wall fraction. This enhances the energy value of feeds when animals are fed maize silage for example. Supplementation of lipids to increase energy intake is known to influence the population of rumen protozoa and some associated rumen functions such as cellulolysis and proteolysis. The end products of rumen fermentation are also changed. Lipolysis and hydrogenation by rumen microbes alter the form of fatty acids supplied to animals. This effect is discussed in relation with the quality of lipids in beef and the implications for human health. Conditions for optimising the amount of amino acids from microbial proteins and dietary by-pass proteins flowing to the duodenum of ruminants, and their impact on beef production, are also examined.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.8
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• pp.1088-1097
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• 2014

The study aimed to investigate the effects of gynosaponin on in vitro methanogenesis under different forage-concentrate ratios (F:C ratios). Experiment was conducted with two kinds of F:C ratios (F:C = 7:3 and F:C = 3:7) and gynosaponin addition (0 mg and 16 mg) in a $2{\times}2$ double factorial design. In the presence of gynosaponin, methane production and acetate concentration were significantly decreased, whereas concentration of propionate tended to be increased resulting in a significant reduction (p<0.05) of acetate:propionate ratio (A:P ratio), in high-forage substrate. Gynosaponin treatment increased (p<0.05) the butyrate concentration in both F:C ratios. Denaturing gradient gel electrophoresis (DGGE) analysis showed there was no apparent shift in the composition of total bacteria, protozoa and methanogens after treated by gynosaponin under both F:C ratios. The real-time polymerase chain reaction (PCR) analysis indicated that variable F:C ratios significantly affected the abundances of Fibrobacter succinogenes, Rumninococcus flavefaciens, total fungi and counts of protozoa (p<0.05), but did not affect the mcrA gene copies of methanogens and abundance of total bacteria. Counts of protozoa and abundance of F.succinogenes were decreased significantly (p<0.05), whereas mcrA gene copies of methanogens were decreased slightly (p<0.10) in high-forage substrate after treated by gynosaponin. However, gynosaponin treatment under high-concentrate level did not affect the methanogenesis, fermentation characteristics and tested microbes. Accordingly, overall results suggested that gynosaponin supplementation reduced the in vitro methanogenesis and improved rumen fermentation under highforage condition by changing the abundances of related rumen microbes.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.11
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• pp.1583-1591
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• 2013

The objective of this study was to investigate microbial population in the rumen of dairy steers as influenced by supplementing with dietary condensed tannins and saponins and different roughage to concentrate ratios. Four, rumen fistulated dairy steers (Bos indicus) were used in a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. The main factors were two roughage to concentrate ratios (R:C, 60:40 and 40:60) and two supplementations of rain tree pod meal (RPM) (0 and 60 g/kg of total DM intake). Chopped 30 g/kg urea treated rice straw was used as a roughage source. All animals received feed according to respective R:C ratios at 25 g/kg body weight. The RPM contained crude tannins and saponins at 84 and 143 g/kg of DM, respectively. It was found that ruminal pH decreased while ruminal temperature increased by a higher concentrate ratio (R:C 40:60) (p<0.05). In contrast, total bacterial, Ruminococus albus and viable proteolytic bacteria were not affected by dietary supplementation. Numbers of fungi, cellulolytic bacteria, Fibrobactor succinogenes and Ruminococus flavefaciens were higher while amylolytic bacteria was lower when steers were fed at 400 g/kg of concentrate. The population of Fibrobactor succinogenes, was found to be higher with RPM supplementation. In addition, the use of real-time PCR technique indicated that the population of protozoa and methanogens were decreased (p<0.05) with supplementation of RPM and with an increasing concentrate ratio. Supplementation of RPM and feeding different concentrate ratios resulted in changing the rumen microbes especially, when the animals were fed at 600 g/kg of concentrate and supplemented with RPM which significantly reduced the protozoa and methanogens population.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.9
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• pp.1285-1292
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• 2014

An in vitro experiment was conducted to evaluate the effects of Aspergillus oryzae culture (AOC) and 2-hydroxy-4-(methylthio)-butanoic acid (HMB) on rumen fermentation and microbial populations between different roughage sources. Two roughage sources (Chinese wild rye [CWR] vs corn silage [CS]) were assigned in a $2{\times}3$ factorial arrangement with HMB (0 or 15 mg) and AOC (0, 3, or 6 mg). Gas production (GP), microbial protein (MCP) and total volatile fatty acid (VFA) were increased in response to addition of HMB and AOC (p<0.01) for the two roughages. The HMB and AOC showed inconsistent effects on ammonia-N with different substrates. For CWR, neither HMB nor AOC had significant effect on molar proportion of individual VFA. For CS, acetate was increased (p = 0.02) and butyrate was decreased (p<0.01) by adding HMB and AOC. Increase of propionate was only occurred with AOC (p<0.01). Populations of protozoa ($p{\leq}0.03$) and fungi ($p{\leq}0.02$) of CWR were differently influenced by HMB and AOC. Percentages of F. succinogenes, R. albus, and R. flavefaciens (p<0.01) increased when AOC was added to CWR. For CS, HMB decreased the protozoa population (p = 0.01) and increased the populations of F. succinogenes and R. albus ($p{\leq}0.03$). Populations of fungi, F. succinogenes (p = 0.02) and R. flavefacien (p = 0.03) were increased by adding AOC. The HMB${\times}$AOC interactions were noted in MCP, fungi and R. flavefacien for CWR and GP, ammonia-N, MCP, total VFA, propionate, acetate/propionate (A/P) and R. albus for CS. It is inferred that addition of HMB and AOC could influence rumen fermentation of forages by increasing the number of rumen microbes.