• Animal Bioscience
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• v.34 no.2
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• pp.213-222
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• 2021

Objective: The metabolites that constitute the rumen fluid and milk in dairy cattle were analyzed using proton nuclear magnetic resonance (1H-NMR) spectroscopy and compared with the results obtain for other dairy cattle herds worldwide. The aim was to provide basic dataset for facilitating research on metabolites in rumen fluid and milk. Methods: Six dairy cattle were used in this study. Rumen fluid was collected using a stomach tube, and milk was collected using a pipeline milking system. The metabolites were determined by 1H-NMR spectroscopy, and the obtained data were statistically analyzed by principal component analysis, partial least squares discriminant analysis, variable importance in projection scores, and metabolic pathway data using Metaboanalyst 4.0. Results: The total numbers of metabolites in rumen fluid and milk were measured to be 186 and 184, and quantified as 72 and 109, respectively. Organic acid and carbohydrate metabolites exhibited the highest concentrations in rumen fluid and milk, respectively. Some metabolites that have been associated with metabolic diseases (acidosis and ketosis) in cows were identified in rumen fluid, and metabolites associated with ketosis, somatic cell production, and coagulation properties were identified in milk. Conclusion: The metabolites measured in rumen fluid and milk could potentially be used to detect metabolic diseases and evaluate milk quality. The results could also be useful for metabolomic research on the biofluids of ruminants in Korea, while facilitating their metabolic research.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.2
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• pp.198-205
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• 2011

This study was conducted to investigate the effect of feed types on Ochratoxin A (OTA) degradation by Korean native goats. Rumen fluid from canulated goats fed whole roughage or 50% roughage served as a source of micro-organisms. Experiments were undertaken i) to investigate OTA degradation ability in a $2{\times}4$ factorial arrangement with different feed types (100% roughage vs. 50% roughage) and rumen fluid fractions (whole rumen fluid, cells, autoclaved rumen fluid and supernatant) supplemented with OTA ii) to evaluate OTA degradation by the rumen fluid of goats fed two different diets at different time points (0, 3, 6, 9 and 12 h) of feeding iii) to isolate potential rumen microorganisms and iv) to identify elements responsible for OTA degradation. Rumen fluid from goats fed 100% roughage had higher (p<0.05) OTA degradability than 50% roughage diets. OTA degradation based on rumen fluid collection times showed that rumen fluid at 0 h showed significantly higher (p<0.05) degradability. Carboxypeptidase A (CPA) enzyme has been reported to be responsible for OTA degradation. Thus, using real time PCR, primers designed to target the CPA gene from Bacillus licheniformis could be amplified using genomic DNA from rumen fluid of goats and sequenced, thus enabling evaluation of the Bacillus population under different feeding condition and times. Our findings showed that the Bacillus population was significantly higher (p<0.05) before feeding (0 h) in animals which were fed a whole roughage diet, giving indirect evidence of OTA degradation being influenced by Bacillus sps. Thus, it can be concluded that OTA degradability is influenced by feed, feeding time and Bacillus licheniformis population.

• Animal Bioscience
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• v.36 no.1
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• pp.53-62
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• 2023

Objective: In this study, metabolites that changed in the rumen fluid, urine and feces of dairy cows fed different feed ratios were investigated. Methods: Eight Holstein cows were used in this study. Rumen fluid, urine, and feces were collected from the normal concentrate diet (NCD) (Italian ryegrass 80%: concentrate 20% in the total feed) and high concentrate diet (HCD) groups (20%: 80%) of dairy cows. Metabolite analysis was performed using proton nuclear magnetic resonance (NMR) identification, and statistical analysis was performed using Chenomx NMR software 8.4 and Metaboanalyst 4.0. Results: The two groups of rumen fluid and urine samples were separated, and samples from the same group were aggregated together. On the other hand, the feces samples were not separated and showed similar tendencies between the two groups. In total, 160, 177, and 188 metabolites were identified in the rumen fluid, urine, and feces, respectively. The differential metabolites with low and high concentrations were 15 and 49, 14 and 16, and 2 and 2 in the rumen fluid, urine, and feces samples, in the NCD group. Conclusion: As HCD is related to rumen microbial changes, research on different metabolites such as glucuronate, acetylsalicylate, histidine, and O-Acetylcarnitine, which are related to bacterial degradation and metabolism, will need to be carried out in future studies along with microbial analysis. In urine, the identified metabolites, such as gallate, syringate, and vanillate can provide insight into microbial, metabolic, and feed parameters that cause changes depending on the feed rate. Additionally, it is thought that they can be used as potential biomarkers for further research on subacute ruminal acidosis.

• Korean Journal of Veterinary Service
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• v.41 no.4
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• pp.221-228
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• 2018

Rumen cannulation is used for nutritional and microbiological research, clinical diagnosis, and rumen component transfaunation. However, the cannulation procedure can affect parameters such as complete blood count findings, serum chemistry, and rumen fluid pH. The objective of this study was to evaluate the health risks related to the rumen cannulation procedure over a 1-month period. We did not identify significant differences in red blood cell numbers or morphologies between pre- and postoperative timepoints. Moreover, no inflammation or infection was detected. Despite the absence of apparent clinical signs after surgery, serum chemistry results revealed changes in blood urea nitrogen levels and the activities of liver enzymes, including aspartate transaminase, lactate dehydrogenase, and creatinine kinase, from postoperative days 1 to 14. Rumen fluid pH, as measured from samples collected via an orogastric tube, was slightly increased after a preoperative fasting period and on postoperative day 1 but decreased thereafter from postoperative day 4, indicating a minor influence of cannulation surgery on ruminal fluid pH. This is the first study to evaluate hematological parameters and rumen pH before and after rumen cannulation surgery in Hanwoo cattle. Further research is required to better elucidate the potential effects of rumen cannulation surgery on animal health.

• Asian-Australasian Journal of Animal Sciences
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• v.8 no.3
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• pp.249-254
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• 1995

The effects of bentonite (B) on rumen protozoal population and rumen fluid characteristics of sheep fed palm kernel cake (PKC) were studied for a period of 21 days. Two groups, each comprising two sheep were fed either PKC or PKC + B ad libitum A third group was left at pasture. Rumen fluid was sampled through a rumen cannula three times daily from all animals. Palm kernel cake contained 16% crude protein, 1 % crude fat and high amounts of copper, zinc, iron and manganese. Protozoal population in the rumen fluid decreased significantly (p < 0.05) after the onset of feeding PKC or PKC + B. However, sheep given bentonite supplementation at 2% of the dietary dry matter, maintained higher protozoal densities ($15{\times}10^4/ml$) when compared to animals fed only PKC ($8{\times}10^4/ml$). With both diets, the protozoa were mainly of the small entodinia species. Animals at pasture had higher protozoal population ($47{\times}10^4/ml$) with varying species of entodiniomorphids and holotrichs. Rumen fluid pH and ammonia concentration was significantly (p < 0.05) higher in animals at pasture compared to animals fed PKC or PKC + B. Volatile fatty acid concentration was significantly (p < 0.05) lower in animals fed PKC when compared to animals at pasture. There was a shift in fermentation pattern in animals fed PKC or PKC + B towards a lower acetate; and higher propionate, isovalerate and valerate. Studies in vitro also showed the positive effect of bentonite on protozoal numbers.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.8
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• pp.1233-1241
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• 2020

Objective: The present investigation was aimed to explore the potential of lactobacilli for conjugated linoleic acid (CLA) production, isolated from rumen fluid samples of lactating goats. Methods: A total of 64 isolates of lactobacilli were obtained using deMan-Rogosa-Sharpe (MRS) agar from rumen fluid of goats and further subjected to morphological and biochemical characterizations. Isolates found as gram-positive, catalase negative rods were presumptively identified as Lactobacillus species and further confirmed by genus specific polymerase chain reaction (PCR). The phylogenetic tree was constructed from the nucleotide sequences using MEGA6. Results: Out of the 64 isolates, 23 isolates were observed positive for CLA production by linoleate isomerase gene-based amplification and quantitatively by UV-spectrophotometric assay for the conversion of linoleic acid to CLA as well as gas chromatography-based assay. In all Lactobacillus species cis9, trans11 isomer was observed as the most predominant CLA isomer. These positive isolates were identified by 16S rRNA gene-based PCR sequencing and identified to be different species of L. ingluviei (2), L.salivarius (2), L. curvatus (15), and L. sakei (4). Conclusion: The findings of the present study concluded that lactic acid bacteria isolated from ruminal fluid samples of goat have the potential to produce bioactive CLA and may be applied as a direct fed microbial to enhance the nutraceutical value of animal food products.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.6
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• pp.776-782
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• 2019

Objective: Fasting may lead to changes in the microbiota and activity in the rumen. In the present study, the effects of fasting on rumen microbiota and the impact of fasting on in vitro rumen fermentation were evaluated using molecular culture-independent methods. Methods: Three ruminally cannulated Holstein steers were fed rice straw and concentrates. The ruminal fluids were obtained from the same steers 2 h after the morning feeding (control) and 24 h after fasting (fasting). The ruminal fluid was filtrated through four layers of muslin, collected for a culture-independent microbial analysis, and used to determine the in vitro rumen fermentation characteristics. Total DNA was extracted from both control and fasting ruminal fluids. The rumen microbiota was assessed using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction. Microbial activity was evaluated in control and fasting steers at various intervals using in vitro batch culture with rice straw and concentrate at a ratio of 60:40. Results: Fasting for 24 h slightly affected the microbiota structure in the rumen as determined by DGGE. Additionally, several microorganisms, including Anaerovibrio lipolytica, Eubacterium ruminantium, Prevotella albensis, Prevotella ruminicola, and Ruminobacter amylophilus, decreased in number after fasting. In addition, using the ruminal fluid as the inoculum after 24 h of fasting, the fermentation characteristics differed from those obtained using non-fasted ruminal fluid. Compared with the control, the fasting showed higher total gas production, ammonia, and microbial protein production (p<0.05). No significant differences, however, was observed in pH and dry matter digestibility. Conclusion: When in vitro techniques are used to evaluate feed, the use of the ruminal fluid from fasted animals should be used with caution.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1439-1444
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• 2002

An objective of this study was to determine the effects of increasing contents of rumen undegradable protein (RUP) from formalin treated soy bean (FSBM) on rumen functions. Four rumen canulated non-lactating cows were randomly allocated to total mixed rations (TMR) containing different proportions of soy bean meal (SBM) and FSBM. Of rumen fermentation characteristics, concentrations of ruminal fluid ammonia and molar proportions of isoacids decreased with increasing contents of RUP in diets (p<0.01). The animals on TMR containing only SBM gained less weight and had smaller rumen volume than those on TMR containing RUP from FSBM (p<0.05). Organic matter and neutral detergent fiber digestibility in sacco were not different (p>0.05). The density of protozoa particularly small Entodinium sp. in ruminal fluid was higher in animal fed TMR containing SBM:FSBM (34:66) and FSBM than those fed TMR containing SBM:FSBM (66:34) and SBM (p<0.01). Total viable count, and net microbial protein synthesis as indicated by purine derivatives in urine increased with increasing contents of RUP from FSBM (p<0.01). It can be concluded that a reduction in net microbial protein synthesis in the rumen with increasing contents of RUP in the diet can be due to the reduction of preformed protein available for microbial growth as well as an increased turnover rate of microbial cells by predatory activity of protozoa.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.4 no.2
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• pp.137-141
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• 1991

Rumen liquor characteristics and disappearance rate of dry matter were studied in Kedah-Kelantan cattle and swamp buffaloes fed grass of rice straw-based diet. Cobalt-EDTA and chromium mordented fibres prepared from the faecal material were used to determine the liquid and solid particles movement in both animal species fed with rice straw. Swamp buffaloes showed a more intense rumen fermentation activity than Kedah-Kelantan cattle when both species were fed straw-based diet. The buffaloes also demonstrated faster rates of grass and straw degradation in situ. The fluid outflow rate from the rumen of buffalo ($1.06{\pm}0.19l/h$) was observed to be slower than that of cattle ($1.55{\pm}0.01l/h$). No significant differences between cattle and buffaloes were observed in rumen fluid volume and passage rate of small particles from the rumen.