• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.10
• /
• pp.1411-1416
• /
• 2004

The objective of this work was to isolate a microorganism, able to produce high lactate dehydrogenase (LDH) activity, for use as a microbial feed additive. The LDH is an important enzyme for lactate conversion in the rumen, thereby possibly overcoming lactic acidosis owing to sudden increases of cereal in the diets of ruminants. In the present study, various bacterial strains were screened from a variety of environments. Among the isolated microorganisms, strain FFy 111-1 isolated from a Korean traditional fermented vegetable food called Kimchi showed the highest enzyme activity, along with retaining strong enzyme activity even in rumen fluid in vitro. Based on morphological and biochemical characteristics as well as compositions of cellular fatty acids plus API analyses, this strain was identified as Lactobacillus sp. The optimum temperature and pH for growth were found to be 30$^{\circ}C$ and pH 6.5, respectively. A maximum cell growth of 2.2 at $A_{650}$ together with LDH activity of 2.08 U per mL was achieved after 24 h of incubation. Initial characterization of FFy 111-1 suggested that it could be a potential candidate for use as a direct-fed microbial in the ruminant animals.

• Asian-Australasian Journal of Animal Sciences
• /
• v.9 no.2
• /
• pp.189-193
• /
• 1996

A study was conducted to estimate nutritive value of forage sources used in deer diets. Bags containing feedstuffs were incubated four times for periods up to 72 hours in two chemostats filled with ruminal fluid from deer or cattle. Parameters estimated were water-soluble and $65{\mu}m$ filterable plus insoluble digestible fractions(No. = 4) and extent of disappearance(No. = 8) of feed neutral detergent fiber(NDF). Among tested feeds, the ranking of values of these parameters were soybean hulls > alfalfa pellets > corn cobs or rice straw > cottonseed hulls or rice hulls > sawdust, indicating that soybean hulls and alfalfa pellets were more fermentable than other agricultural residues. It is possible to utilize variability among tested feeds of fraction, disappearance rate and (or) extent of DM and NDF when these feeds are used as roughage sources in deer diets.

• Journal of Life Science
• /
• v.22 no.10
• /
• pp.1324-1329
• /
• 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
• /
• v.12 no.4
• /
• pp.544-547
• /
• 1999

The object of this study was to determine the influence of monensin and virginiamycin (VM) on in vitro ruminal fermentation of rice straw or ammoniated rice straw. Rumen fluid was collected from 4 wethers fed 200 g of concentrate supplement with 400 g of untreated (U) or ammoniated (A) rice straw once daily for 28 days. Mixed ruminal microorganisms were incubated in anaerobic media that contained 20% (vol/vol) ruminal fluid and 0.3 g of either U or A rice straw. Monensin and/or VM, dissolved in ethanol, were added in centrifuge tubes at final concentrations of 0, 15, 30, 15+15 and 30+30 ppm of culture fluid. The addition of monensin and VM combination to A rice straw fermentation decreased (p<0.05) the acetate to propionate ratio, total VFA and lactate production, but increased (p<0.05) pH. Total gas production tended to be decreased by the addition of monensin plus VM. Antimicrobial agents decreased $NH_3$ N concentration and dry matter digestibility.

• Journal of Animal Science and Technology
• /
• v.58 no.1
• /
• pp.4.1-4.7
• /
• 2016

Background: This study was conducted to evaluate the effects of adding Korean rice wine residue (RWR) in total mixed ration (TMR) on in vitro ruminal fermentation and growth performance of growing Hanwoo steers. Methods: For in vitro fermentation, the experimental treatments were Control (Con: 0 % RWR + TMR), Treatment 1 (T1: 10 % RWR + TMR), and Treatment 2 (T2: 15 % RWR + TMR). The rumen fluid was collected from three Hanwoo steers and mixed with buffer solution, after which buffered rumen fluid was transferred into serum bottles containing 2 g dry matter (DM) of TMR added with or without RWR. The samples were then incubated for 0 h, 12 h, 24 h, or 48 h at $39^{\circ}C$ and 100 rpm. For the in vivo experiment, 27 Hanwoo steers (6 months old) with an average weight of $196{\pm}8.66kg$ were subjected to a 24-week feeding trial. The animals were randomly selected and equally distributed into three groups. After which the body weight, feed intake and blood characteristics of each group were investigated. Results: The pH of the treatments decreased significantly relative to the control during the 12 h of incubation. Total gas production and ammonia nitrogen ($NH_3-N$) was not affected by RWR addition. The total volatile fatty acid (VFA) was lower after 24 h of incubation but at other incubation times, the concentration was not affected by treatments. Feed cost was 8 % and 15 % lower in T1 and T2 compared to control. Blood alcohol was not detected and a significant increase in total weight gain and average daily gain were observed in Hanwoo steers fed with RWR. Conclusion: Overall, the results of this study suggest that TMR amended with 15 % RWR can be used as an alternative feed resource for ruminants to reduce feed cost.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.4
• /
• pp.479-485
• /
• 2017

Objective: The effects of particle size of processed barley grain, enzyme addition and microwave treatment on in vitro dry matter (DM) disappearance (DMD), gas production and fermentation pH were investigated for feedlot cattle. Methods: Rumen fluid from four fistulated feedlot cattle fed a diet of 860 dry-rolled barley grain, 90 maize silage and 50 supplement g/kg DM was used as inoculum in 3 batch culture in vitro studies. In Experiment 1, dry-rolled barley and barley ground through a 1-, 2-, or 4-mm screen were used to obtain four substrates differing in particle size. In Experiment 2, cellulase enzyme (ENZ) from Acremonium cellulolyticus Y-94 was added to dry-rolled and ground barley (2-mm) at 0, 0.1, 0.5, 1, and 2 mg/g, while Experiment 3 examined the interactions between microwaving (0, 30, and 60 s microwaving) and ENZ addition (0, 1, and 2 mg/g) using dry-rolled barley and 2-mm ground barley. Results: In Experiment 1, decreasing particle size increased DMD and gas production, and decreased fermentation pH (p<0.01). The DMD (g/kg DM) of the dry-rolled barley after 24 h incubation was considerably lower (p<0.05) than that of the ground barley (119.1 dry-rolled barley versus 284.8 for 4-mm, 341.7 for 2-mm; and 358.6 for 1-mm). In Experiment 2, addition of ENZ to dry-rolled barley increased DMD (p<0.01) and tended to increase (p = 0.09) gas production and decreased (p<0.01) fermentation pH, but these variables were not affected by ENZ addition to ground barley. In Experiment 3, there were no interactions between microwaving and ENZ addition after microwaving for any of the variables. Microwaving had minimal effects (except decreased fermentation pH), but consistent with Experiment 2, ENZ addition increased (p<0.01) DMD and gas production, and decreased (p<0.05) fermentation pH of dry-rolled barley, but not ground barley. Conclusion: We conclude that cellulase enzymes can be used to increase the rumen disappearance of barley grain when it is coarsely processed as in the case of dry-rolled barley. However, microwaving of barley grain offered no further improvements in ruminal fermentation of barley grain.

• Journal of Animal Science and Technology
• /
• v.60 no.11
• /
• pp.27.1-27.8
• /
• 2018

Background: Enteric methane ($CH_4$) accounts for about 70% of total $CH_4$ emissions from the ruminant animals. Researchers are exploring ways to mitigate enteric $CH_4$ emissions from ruminants. Recently, nano zinc oxide (nZnO) has shown potential in reducing $CH_4$ and hydrogen sulfide ($H_2S$) production from the liquid manure under anaerobic storage conditions. Four different levels of nZnO and two types of feed were mixed with rumen fluid to investigate the efficacy of nZnO in mitigating gaseous production. Methods: All experiments with four replicates were conducted in batches in 250 mL glass bottles paired with the ANKOM$^{RF}$ wireless gas production monitoring system. Gas production was monitored continuously for 72 h at a constant temperature of $39{\pm}1^{\circ}C$ in a water bath. Headspace gas samples were collected using gas-tight syringes from the Tedlar bags connected to the glass bottles and analyzed for greenhouse gases ($CH_4$ and carbon dioxide-$CO_2$) and $H_2S$ concentrations. $CH_4$ and $CO_2$ gas concentrations were analyzed using an SRI-8610 Gas Chromatograph and $H_2S$ concentrations were measured using a Jerome 631X meter. At the same time, substrate (i.e. mixed rumen fluid+ NP treatment+ feed composite) samples were collected from the glass bottles at the beginning and at the end of an experiment for bacterial counts, and volatile fatty acids (VFAs) analysis. Results: Compared to the control treatment the $H_2S$ and GHGs concentration reduction after 72 h of the tested nZnO levels varied between 4.89 to 53.65%. Additionally, 0.47 to 22.21% microbial population reduction was observed from the applied nZnO treatments. Application of nZnO at a rate of $1000{\mu}g\;g^{-1}$ have exhibited the highest amount of concentration reductions for all three gases and microbial population. Conclusion: Results suggest that both 500 and $1000{\mu}g\;g^{-1}$ nZnO application levels have the potential to reduce GHG and $H_2S$ concentrations.

• Asian-Australasian Journal of Animal Sciences
• /
• v.7 no.1
• /
• pp.83-89
• /
• 1994

The effects of inorganic selenium (Se), selenate and selenite on Se balance levels in the different ruminal fluid fractions were studied using Japanese Corriedale wethers with an average body weight of 47 kg. A $3{\times}3$ Latin square design was used with three animal, three periods and three treatments. In each period, there was 7 d dietary adjustment followed by 5 d total collection of urine and feces. Ruminal fluid samples were obtained at 0, 1, 3, 5 and 7 h postprandially on the final day of the collection period. The three dietary treatments were: (1) without Se supplementation (control); (2) with Se supplement as sodium selenate; and (3) sodium selenite at a rate of 0.2 mg Se/kg dietary DM. The basal diet was timothy hay (Phleum pratense L.) fed 2% of body weight/d. Results indicated that Se balance were higher (p < 0.05) for those animals under supplementation than those animals under control. Overall data gathered showed a similar digestion balance of selenate and selenite in sheep. Inorganic Se, both selenate and selenite produced positive Se contents of the ruminal feed particles and protozoa. Bacterial Se increased (p < 0.05) on the first three hours post-prandially in Se supplemented diets. Gross ruminal fluid fraction, although there was improvement on their Se content under the supplemented diets, the changes were insignificant over the control. free inorganic Se and Se in soluble protein of the ruminal fluid were not significantly different for selenate and selenite. Most of the Se in the ruminal fluids of the animals under supplementation were insoluble, indicating the influence of rumen environments on Se bioavaliability.

• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.12
• /
• pp.1738-1746
• /
• 2002

Research was carried out to ascertain whether or not the volume of saliva flowing into the rumen regulates dry forage intake in ruminants. Goats with a parotid fistula were fed roughly crushed alfalfa hay cubes, concentrated beef cattle feed and $NaHCO_3$ wice daily (10:00-12:00, 16:00-18:00). Except for the days on which experiments were conducted, the animals were free access to drinking water. The animals were intraruminally infused every day prior to the morning feeding period with parotid saliva collected from the parotid fistula over a 24 h period. The present experiment consisted of three treatments, non-infusion (NI), intraruminal infusion of parotid saliva (RSI), and intraruminal infusion of warm water (RWI). In the RSI treatment, approximately 4-5 kg of parotid saliva (280-290 mOsm/l) collected over a 24 h period was intraruminally infused 1 h prior to the commencement of morning feeding. In the RWI treatment, parotid saliva was substituted for warm water ($36^{\circ}C$). After infusions, the animals were fed on roughly crushed alfalfa hay cubes for 2 h. During feeding, eating and saliva secretion rates were measured. Blood samples were also periodically collected from the jugular vein. After 2 h feeding, water intake was measured for 30 min. These measurements were used to define thirst levels. On the day of the experiment, the animals were not access to drinking water during the morning feeding. It is thought that rumen fill in RSI and RWI treatments was higher than the NI treatment. In comparison with the NI treatment however, cumulative feed intake increased by 39.3% with RSI treatment and by 45.9% with RWI treatment after completion of the 2 h feeding period. After 2 h feeding, thirst level in the RSI treatment showed only a 10% decrease compared to the NI treatment, but thirst level in the RWI treatment decreased 49.8%. Despite the significant differences in thirst levels between RSI and RWI treatments, the cumulative feed intake in both treatments was similar. When comparing accumulated saliva secretion volumes 2 h after feeding, volumes in the RSI treatment were significantly 35.9% lower than the NI treatment while volumes in the RWI treatment were unchanged. However, the volumes of saliva and fluid flowing into the rumen were greater in both RSI and RWI treatments when compared to the NI treatment. The results indicate that the amount of saliva flowing into the rumen is a factor regulating feed intake in ruminants fed on dry forage.

• Asian-Australasian Journal of Animal Sciences
• /
• v.26 no.4
• /
• pp.517-522
• /
• 2013

The effect on methanogens attached to the surface of rumen ciliate protozoa by the addition of plant extracts (pine needles and ginkgo leaves) was studied with particular reference to their effectiveness for decreasing methane emission. The plant extracts (pine needles and ginkgo leaves) were added to an in vitro fermentation incubated with rumen fluid. The microbial population including bacteria, ciliated-associated methanogen, four different groups of methanogens and Fibrobacter succinogenes were quantified by using the real-time PCR. Gas profiles including methane, carbon dioxide and hydrogen, and runinal fermentation characteristics were observed in vitro. The methane emission from samples with an addition of individual juices from pine needles, ginkgo leaves and 70% ethanol extract from ginko leaves was significantly lower (p<0.05, 27.1, 28.1 and 28.1 vs 34.0 ml/g DM) than that of the control, respectively. Total VFAs in samples with an addition of any of the plant extracts were significantly lower than that of the control (p<0.05) as well. The order Methanococcales and the order Methanosarcinales were not detected by using PCR in any incubated mixtures. The ciliate-associated methanogens population decreased from 25% to 49% in the plant extacts as compared to control. We speculate that the supplementation of juice from pine needles and ginkgo leaves extract (70% ethanol extract) decreased the protozoa population resulting in a reduction of methane emission in the rumen and thus inhibiting methanogenesis. The order Methanobacteriales community was affected by addition of all plant extracts and decreased to less than the control, while the order Methanomicrobiales population showed an increase to more than that of the control. The F. succinogenes, the major fibrolytic microorganism, population in all added plant extracts was increased to greater than that of the control. In conclusion, pine needles and ginkgo leaves extracts appear to have properties that decrease methanogenesis by inhibiting protozoa species and may have a potential for use as additives for ruminants.