An in vitro trial was conducted to examine the effects of total mixed rations (TMR) on fermentation characteristics and effective degradability (ED) by rumen microbes. Three TMR diets were growing period TMR (GR-TMR, 67% TDN), early fattening period TMR (EF-TMR, 75.4% TDN) and late fattening TMR (LF-TMR, 80% TDN). Three TMR diets (3 g of TMRs in each incubation bottles) was added to the mixed culture solution of stained rumen fluid with artificial saliva (1:1, v/v) and incubated anaerobically for 48 hours at $39^{\circ}C$. The pH in all incubation solutions tended to decrease up to 48h, but the opposite results were found in concentration of total gas production, ammonia-N and total VFA in all incubations.The total gas production (p<0.05) in LF-TMR was highest compared with those of other diets. Also, concentration of total VFA was tended to increase in LF-TMR compared with other TMR diets in all incubations. The EDDM in both EF-TMR and LF-TMR was tended to high compared with GR-TMR (p=0.100). In this in vitro trials, concentration of propionate in all incubation solution was not affected by increased concentration of TDN. The results of the present in vitro study indicate that TMR may provide more favorable condition for nutrient digestion both in the rumen.
Bureenok, Smerjai;Yuangklang, Chalermpon;Vasupen, Kraisit;Schonewille, J. Thomas;Kawamoto, Yasuhiro
Asian-Australasian Journal of Animal Sciences
/
v.25
no.9
/
pp.1248-1254
/
2012
The effect of silage additives on ensiling characteristics and nutritive value of Napier grass (Pennisetum purpureum) silages was studied. Napier grass silages were made with no additive, fermented juice of epiphytic lactic acid bacteria (FJLB), molasses or cassava meal. The ensiling characteristics were determined by ensiling Napier grass silages in airtight plastic pouches for 2, 4, 7, 14, 21 and 45 d. The effect of Napier grass silages treated with these additives on voluntary feed intake, digestibility, rumen fermentation and microbial rumen fermentation was determined in 4 fistulated cows using $4{\times}4$ Latin square design. The pH value of the treated silages rapidly decreased, and reached to the lowest value within 7 d of the start of fermentation, as compared to the control. Lactic acid content of silages treated with FJLB was stable at 14 d of fermentation and constant until 45 d of ensiling. At 45 d of ensiling, neutral detergent fiber (NDF) and acid detergent fiber (ADF) of silage treated with cassava meal were significantly lower (p<0.05) than the others. In the feeding trial, the intake of silage increased (p<0.05) in the cow fed with the treated silage. Among the treatments, dry matter intake was the lowest in the silage treated with cassava meal. The organic matter, crude protein and NDF digestibility of the silage treated with molasses was higher than the silage without additive and the silage treated with FJLB. The rumen parameters: ruminal pH, ammonia-nitrogen ($NH_3$-N), volatile fatty acid (VFA), blood urea nitrogen (BUN) and bacterial populations were not significantly different among the treatments. In conclusion, these studies confirmed that the applying of molasses improved fermentative quality, feed intake and digestibility of Napier grass.
Kim, J.H.;Yokota, H.;Ko, Y.D.;Okajima, T.;Ohshima, M.
Asian-Australasian Journal of Animal Sciences
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v.6
no.1
/
pp.53-59
/
1993
In situ degradability and fermentation characteristics in the rumen of goats fed whole crop corn forage ensiled with (MS silage) or without (CS silage) 30% of cage layer manure (CLM) were investigated. The two silages were well preserved. To adjust nitrogen intake of CS silage to that of MS silage, the 3rd group of goats was given urea with CS silage at feeding time (US silage). Each goat was given a diet of 2% of the body weight (dry matter basis) daily. In situ degradability of dry matter (DM) and crude protein (CP) of MS silage in the rumen were higher than those of CS and US silages. Total potentially degradable portions of DM and CP in MS silage were also higher than those in CS and US silages. Blood urea nitrogen and rumen ammonia nitrogen concentration of goats fed US and MS silages were significantly (p<0.05) higher than those of goats fed CS silage. Acetic, propionic and butyric acids in ruminal fluids of goats fed MS silage were significantly (p<0.05) higher than those of goats fed CS and US silages.
Alam, Mohammad Khairul;Ogata, Yasumichi;Sato, Yukari;Sano, Hiroaki
Asian-Australasian Journal of Animal Sciences
/
v.29
no.4
/
pp.523-529
/
2016
An isotope dilution method using $[U-^{13}C]glucose$ and $[1-^{13}C]leucine$ (Leu) was conducted to evaluate the effects of rice straw supplemented with urea and molasses (RSUM-diet) on plasma glucose and Leu turnover rates in sheep. Nitrogen (N) balance, rumen fermentation characteristics and blood metabolite concentrations were also determined. Four sheep were fed either mixed hay (MH-diet), or a RSUM-diet with a crossover design for two 21 days period. Feed allowance was computed on the basis of metabolizable energy at maintenance level. The isotope dilution method was performed as the primed-continuous infusion on day 21 of each dietary period. Nitrogen intake was lower (p = 0.01) for the RSUM-diet and N digestibility did not differ (p = 0.57) between diets. Concentrations of rumen total volatile fatty acids tended to be higher (p = 0.09) for the RSUM-diet than the MH-diet. Acetate concentration in the rumen did not differ (p = 0.38) between diets, whereas propionate concentration was higher (p = 0.01) for the RSUM-diet compared to the MH-diet. Turnover rates as well as concentrations of plasma glucose and Leu did not differ between diets. It can be concluded that kinetics of plasma glucose and Leu metabolism were comparable between the RSUM-diet and the MH-diet, and rumen fermentation characteristics were improved in sheep fed the RSUM-diet compared to the MH-diet.
Objective: To isolate and identify new methanogens from the rumen of Holstein steers in Korea. Methods: Representative rumen contents were obtained from three ruminally cannulated Holstein steers ($793{\pm}8kg$). Pre-reduced media were used for the growth and isolation of methanogens. Optimum growth temperature, pH, and sodium chloride (NaCl) concentration as well as substrate utilization and antibiotic tolerance were investigated to determine the physiological characteristics of the isolated strain. Furthermore, the isolate was microscopically studied for its morphology. Polymerase chain reaction of 16S rRNA and mcrA gene-based amplicons was used for identification. Results: One strain designated as KOR-2 was isolated and found to be a non-motile irregular coccus with a diameter of 0.2 to $0.5{\mu}m$. KOR-2 utilized $H_2+CO_2$ and formate but was unable to metabolize acetate, methanol, trimethylamine, 2-propanol, and isobutanol for growth and methane production. The optimum temperature and pH for the growth of KOR-2 were $38^{\circ}C$ and 6.8 to 7.0, respectively, while the optimum NaCl concentration essential for KOR-2 growth was 1.0% (w/v). KOR-2 tolerated ampicillin, penicillin G, kanamycin, spectromycin, and tetracycline. In contrast, the cell growth was inhibited by chloramphenicol. Phylogenetic analysis of 16S rRNA and mcrA genes revealed the relatedness between KOR-2 and Methanoculleus bourgensis. Conclusion: Based on the physiological and phylogenetic characteristics, KOR-2 was thought to be a new strain within the genus Methanoculleus and named Methanoculleus bourgensis KOR-2.
Darlis, N. Abdullah;Halim, R.A.;Jalaludin, S.;Ho, Y.W.
Asian-Australasian Journal of Animal Sciences
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v.13
no.7
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pp.922-928
/
2000
The effects of animal species and supplements on rumen fluid characteristics, plasma urea-N (PUN) concentration, plasma urea-N pool size, urea-N degradation in the gut and urea-N net flux (urea-N synthesis rate) were studied in goats and sheep, with some minor differences detected. The animals were fed either chopped rice straw ad libitum+200 g soybean meal (SBM), or chopped rice straw ad libitum+190 g soybean meal+300 g sago meal (SBM+SM) for 14 days. The supplements were isonitrogenous (80 g crude protein/animal/d). [$^{14}C$]-urea was used as the marker for urea metabolism studies. Two animals from each species were fed either supplement in a cross-over design in two periods. The results showed that rumen pH was significantly (p<0.001) lower in animals fed SBM+SM than those fed SBM supplement. The ammonia concentrations of rumen fluid were significantly (p<0.01) higher in sheep (382.9 mg N/L) than goats (363.1 mg N/L) when fed SBM supplement but lower (282.5 mg N/L) than that of goats (311.0 mg N/L) when fed SBM+SM supplement. Total VFA concentrations were significantly (p<0.05) higher in animals fed SBM+SM supplement than those fed SBM supplement. Goats had significantly (p<0.01) higher molar proportions of acetate (79.1, 77.7%, respectively) than sheep (75.8, 74.0%, respectively) in both supplements. The molar proportion of acetate was significantly (p<0.05) higher, while that of butyrate lower in animals fed SBM supplement than those fed SBM+SM supplement. In animals fed SBM supplement, the molar proportion of propionate was significantly (p<0.01) higher in sheep (18.0%) than in goats (15.6%), but in animals fed SBM+SM, the molar proportion of butyrate was significantly (p<0.01) higher (9.6%) in sheep than in goats (7.2%). Plasma urea-N concentration, plasma urea-N pool size, urea-N degradation in the gut, urea-N net flux and the fraction of urea-C from the blood entering the rumen were not significantly different between goats and sheep fed either supplement. However, PUN concentration was significantly (p<0.05) lower in animals fed SBM+SM supplement (average of 13.8 mg N/100 ml) than in those fed SBM supplement (average of 16.5 mg N/100 ml). The urea net flux was significantly (p<0.05) higher in goats (average of 14.5 g N/d) than sheep (average of 12.9 g N/d), and animals fed SBM supplement showed higher (average of 14.9 g N/d) urea net flux than animals fed SBM+SM supplement (average of 12.9 g N/d). A significant (p<0.05) positive correlation was observed between urea-N net flux and urea-N degradation; urea-N net flux and pool size; urea-N net flux and urea excretion in the urine; and PUN and rumen ammonia in goats. While in sheep, significant (p<0.05) positive correlation was observed between urea-N net flux and urea excretion in the urine; and PUN and rumen ammonia.
The objectives were to compare the ability of various rumen microbial fractions to reduce nitrate and to assess the effect of nitrate on in vitro fermentation characteristics. Physical and chemical methods were used to differentiate the rumen microbial population into the following fractions: whole rumen fluid (WRF), protozoa (Pr), bacteria (Ba), and fungi (Fu). The three nitrogen substrate treatments were as follows: no supplemental nitrogen source, nitrate or urea, with the latter two being isonitrogenous additions. The results showed that during 24 h incubation, WRF, Pr and Ba fractions had an ability to reduce nitrate, and the rate of nitrate disappearance for the Pr fraction was similar to the WRF fraction, while the Ba fraction needed an adaptation period of 12 h before rapid nitrate disappearance. The WRF fraction had the greatest methane ($CH_4$) production and the Pr fraction had the greatest prevailing $H_2$ concentration (p<0.05). Compared to the urea treatment, nitrate diminished net gas and $CH_4$ production during incubation (p<0.05), and ammonia-N ($NH_3$-N) concentration (p<0.01). Nitrate also increased acetate, decreased propionate and decreased butyrate molar proportions (p<0.05). The Pr fraction had the highest acetate to propionate ratio (p<0.05). The Pr fraction as well as the Ba fraction appears to have an important role in nitrate reduction. Nitrate did not consistently alter total VFA concentration, but it did shift the VFA profile to higher acetate, lower propionate and lower butyrate molar proportions, consistent with less $CH_4$ production by all microbial fractions.
Lee, Shin Ja;Shin, Nyeon Hak;Jeong, Jin Suk;Kim, Eun Tae;Lee, Su Kyoung;Lee, Sung Sill
Asian-Australasian Journal of Animal Sciences
/
v.31
no.1
/
pp.54-62
/
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.
Total mixed ration (TMR) including concentrate diet and roughage together have been used for the ruminant animal. Relatively high concentrations of moisture and water soluble carbohydrate are representative feature of TMR. Those moisture and water can also provide a niche for bacterial growth. Therefore, a possible fermentation of TMR induced by micro-organism is generally accepted. The present study hypothesized that different lactic acid bacteria could alter fermentation of TMR and subsequently rumen fermentation. Three lactic acid bacteria, Lactobacillus paracasei (A), L. plantarum (B) and L. parabuchneri (C), were employed and 7 treatments under full factorial design were compared with control without inoculation. TMR for dairy cow was used. Significant alterations by treatments were detected at lactic acid and butyric acid contents in TMR (p<0.05). Treatment AC (mixture of A and C) and BC (mixture of B and C) showed great lactate production. Great butyrate production was found at treatment C. At in vitro rumen fermentation, treatments B, C and AB (mixture of A and B) showed significantly great total gas production (p<0.05). All treatments except treatments B and AB, showed less dry matter digestibility, significantly (p<0.05). Total volatile fatty acid production at treatment AC was significantly greater than others (p<0.05). In individual volatile fatty acid production, treatment AB and AC showed great acetate and propionate productions, significantly (p<0.05). This study investigated correlation between organic acid production in TMR and rumen volatile fatty acid production. And it was found that butyric acid in TMR had significant negative correlation with acetate, propionate, total volatile fatty acid, AP ratio and dry matter digestibility.
Yang, Sung Jae;Kim, Han Been;Moon, Joon Beom;Kim, Na Eun;Park, Joong Kook;Park, Byung Ki;Lee, Se Young;Seo, Jakyeom
Korean Journal of Agricultural Science
/
v.45
no.4
/
pp.741-748
/
2018
This study was conducted to evaluate the nutritional value of total mixed rations (TMR) containing rice grain in an in vitro rumen fermentation system. Three types of grains (corn, wheat, and rice), timothy, and soybean meal (SBM) were used to prepare the experimental TMR: Corn TMR, Wheat TMR, and Rice TMR. The rumen fermentation characteristics of all the experimental TMRs were evaluated by an in vitro anaerobic system using rumen fluid for 24 and 48 h. The digestibility of the nutrients (dry matter [DM], crude protein [CP], and neutral detergent fiber [NDF]), pH, ammonia ($NH_3-N$), and volatile fatty acids (VFA) were determined. Rice TMR showed a higher DM digestibility than that of the Corn TMR at 48 h (p < 0.05). In all treatments, the CP digestibility was more than 80% at 48 h, but no significant differences were observed among the treatments. The NDF digestibility tended to be the lowest in the Wheat TMR (p = 0.06), and the pH tended to be the lowest in the Rice TMR (p = 0.09) among the treatments for the 48 h incubation. The Wheat TMR had the highest $NH_3-N$ concentration among the treatments (p < 0.01). Rice TMR had a lowest total VFA concentration among the treatments (p = 0.05) at 24 h, but no significant differences were observed at 48 h. Based on this in vitro result, it was considered that a rice grain has the potential to replace conventional grain ingredients when the TMR was formulated.
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