• Asian-Australasian Journal of Animal Sciences
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• v.5 no.3
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• pp.571-582
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• 1992

Soybeans were dry extruded at three different temperatures (125, 135 and $145^{\circ}C$) for 30 s. Four lambs fitted with cannulae in the rumen and abomasums were used in a balanced $4{\times}4$ Latin square design. Lambs were fed at 2 h intervals for 12 times a day with automatic feeder to maintain steady state conditions in digestive tract. A dual-phase marker system was used to estivate ruminal flow rate of both liquid and solid digesta. Objectives of this study were to determine the effect of extrusion temperature of raw soybean on the ruminal liquid and solid dilution rate, nitrogen digestion and flow at the abomasum and availability of amino acid in lambs. There were no significant effects of extrusion on liquid and solid dilution rate, and liquid volume. Ruminal liquid flow rate was not influenced by extrusion and ranged from 389 to 435 ml/hr. Extrusion had no influence on ruminal OM digestion and flow rate to the abomasums. Dietary N flow to the abomasums increased (p < 0.05) as extruding temperature increased. Extruding temperature had a significant effect (p < 0.05) on flow of N escaping ruminal degradation and ranged from 34.91 to 57.38%. Microbial N synthesized/kg OMTDR ranged from 27 to 37 g and highest with $145^{\circ}C$ ESB diet. Extrusion decreased the amount of degradable amino acid in the rumen and increased the supply of amino acid to the lower gut, especially with 135 and $145^{\circ}C$ ESB diets.

• Journal of the Korea Organic Resources Recycling Association
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• v.1 no.1
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• pp.103-113
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• 1993

Every year, over $3.37{\times}10^7$ ton of municipal solid waste is generated in Korea, of which about 28% is organic food waste from restaurant, dining halls and households etc. Methane conversion of the food waste by anaerobic digestion could be a viable approach for energy recovery as well as safe disposal of the waste. However, as food waste is composed of highmolecular complex polymers such as cellulose, lignin and protein, anaerobic digestion of food waste has not been efficient in terms of volumetric loading rate, solid retention time and extent of anaerobic degradation. In this research, the improved anaerobic degradation of food waste was attemped by applying rumen microorganisms to anaerobic digestion. Acidification efficiency of food waste by rumen microorganisms was compared with that of conventional acidogenesis. And optimum acidification conditions by rumen microorganisms were also determined. For the experiments, anaerobic batch reactors of 600 mL was fed with the processed (dried and milled) food waste obtained from a restaurant. Ultimate volatile fatty acid (VFA) yield produced by rumen microorganisms was about 8.4 meq VFA/g volatile solid (VS) that is 95% of the theoretical value. This yield was not much different from that of conventional acidogenesis, but hydrolysis rate was about twice faster. Cumulative VFA concentration increased from 66 meq/L to 480 meq/L, when the initial TS was increased from 1% to 15%. But VFA yield at 15% TS was half of that at 1% TS. This inhibition on the acidification might be caused by the rapid drop of pH and higher concentration of nonionized VFA. Optimal pH and temperature range for the acidification were about 6.0~7.5 and $35{\sim}45^{\circ}C$, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.6
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• pp.802-806
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• 2005

The objective of this study was to determine the effects of processing method and rapeseed variety on ruminal and intestinal protein digestibility of rapeseed meal in steers. Intestinal amino acid digestibility was assessed with an in situ ruminal incubation and precision-fed rooster bioassay. In this experiment one traditional rapeseed meal sample (sample A, prepress extraction) and three double low rapeseed meal samples (sample B, prepress extraction, sample C, screw press and sample D, low temperature press) were placed in polyester bags(8 cm${\times}$12 cm) and suspended in the ventral rumen of steers for 16 h. The residues of in situ incubations were intubated to roosters. Total excreta were collected for 48 h after incubation and then desiccated and amino acid concentrations were determined. Results showed that in ruminal incubation the degradation rate of amino acid and crude protein was higher for traditional rapeseed meal sample A than for double low rapeseed meal sample B, but was much lower than for double low sample C and D. In the group of double low rapeseed meal samples, sample D processed by low temperature press had the highest degradation rate of amino acids in the rumen. For all amino acids, the digestibility of the residual protein as measured by the precision-fed rooster bioassay tended to be lower for sample B than for sample A, which had the same processing method with sample B, and in the group of double low rapeseed meals, sample B had similar digestibility of amino acid in residual protein to sample D and higher than that of sample C. However, although the total amino acid availability involving the digestibility of amino acids in the rumen and rooster bioassay of double low rapeseed meal sample D (low temperature press) was higher than those of the other three samples by 7 to 9 percent, there were no significant differences. Results indicated that processing method markedly affected ruminal and post ruminal amino acid digestibility of rapeseed meal when the temperature exceeded 110$^{\circ}C$. Rapeseed meal that had a high content of fiber was not suitable for dry heat treatment at higher temperatures or the amino acids digestibility in rumen and total availability of amino acids could be reduced. Results also suggested the variety of rapeseed meal had no significant effect on the digestibility and availability of amino acids.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.7
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• pp.974-981
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• 2002

Faba beans (vicia faba) (FB) and lupin seeds (Lupinus Albus) (LS) were dry roasted at three temperatures (110, 130, $150^{\circ}C$) for 15, 30 or 45 min to determine the effects of dry roasting on rumen degradation of crude protein and starch free organic matter ($^{PSF}OM$). Rumen degradation characteristics of $^{PSF}OM$ were determined by the nylon bag incubation technique in dairy cows fed 60% hay and 40% concentrate. Measured characteristics of $^{PSF}OM$ were undegradable fraction (U), degradable fraction (D), soluble fraction (S), lag time (T0), and the rate of degradation (Kd). Based on the measured characteristics, rumen availability ($RA^{PSF}OM$) and bypass $^{PSF}OM$ ($B^{PSF}OM$) were calculated. Dry roasting did not have a greater impact on rumen degradation characteristics of $^{PSF}OM$ (p>0.05). S varied from 32.1 (raw) to 30.0, 27.8, 30.8% (LS) and 15.4 (raw) to 14.4, 20.8, 20.9% (FB); D varied from 65.4 (raw) to 66.3, 66.9, 55.9% (LS) and 54.9 (raw) to 55.0, 51.0, 64.7% (FB); U varied from 2.6 (raw) to 7.3, 7.0, 7.7% (LS) and 29.7 (raw) to 30.6, 28.2, 14.4% (FB); Kd varied from 6.0 (raw) to 7.3, 7.0, 7.7% (LS) and 22.4 (raw) to 24.4, 21.1, 7.9% (FB); $B^{PSF}OM$ varied from 35.5 (raw) to 33.8, 36.6, 38.2% (LS) and 41.3 (raw) to 41.5, 39.7, 47.6% (FB) at 110, 130 and $150^{\circ}C$, respectively. Therefore dry roasting did not significantly affect $RA^{PSF}OM$, which were 353.7, 367.9, 349.6, 336.9 (g/kg DM) (LS) and 12.82, 127.0, 133.7, 117.1 (g/kg DM) (FB) at 110, 130 and $150^{\circ}C$, respectively. These results alone with our previously published reports indicate dry roasting had the differently affected pattern of rumen degradation characteristics of various components in LS and FB. It strongly increased bypass crude protein (BCP) and moderately increased starch (BST) with increasing temperature and time but least affected $^{PSF}OM$. Such desirable degradation patterns in dry roasted LS and FB might be beneficial to the high yielding cows which could use more dry roasted $^{PSF}OM$ as an energy source for microbial protein synthesized in the rumen and absorb more amino acids and glucose in the small intestine.

• Journal of agriculture & life science
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• v.46 no.2
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• pp.129-137
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• 2012

A carboxymethyl cellulase gene, cel5B, was cloned, sequenced, and expressed in Escherichia coli. pRCS20 in E. coli was identified from metagenomic cosmid library of cow rumen for cellulase activity on a carboxymethyl cellulose agar plates. Cosmid clone (RCS20) was partially digested with Sau3AI, ligated into BamHI site of pBluescript II SK+ vector, and transformed into E. coli $DH5{\alpha}$. The insert DNA of 1.3 kb was obtained, designated cel5B, which has the activity of hydrolyzation of CMC. The cel5B gene had an open reading frame (ORF) of 1,059 bp encoding 352 amino acids with a signal peptide of 48 amino acids and the conserved region, VIYEIYNEPL, belongs to the glycosyl hydrolase family 5. The molecular mass of Cel5B protein expressed from E. coli $DH5{\alpha}$ exhibited to be about 34 kDa by CMC-SDS-PAGE. The optimal pH was 8.0, and the optimal temperature was about $50^{\circ}C$ for its enzymatic activity.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.3
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• pp.376-380
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• 2006

An experiment was conducted to study the effect of temperature and pH on in vitro nutrient degradability, volatile fatty acid profile and methane production. The fermenter used was the semi-continuous system, known as the rumen simulation technique (RUSITEC). Sixteen cylinders were used at one time with a volume of 800 ml, the dilution rate was set at 3.5%/hour, the infused buffer being McDougall's artificial saliva. Basal diet (9.6 g DM) used in RUSITEC consisted of (DM) 6.40 g Timothy hay, 1.86 g crushed corn and 1.34 g soybean meal. The food for the fermentation vessel was provided in nylon bags, which were gently agitated in the liquid phase. The experiment lasted for 17 d with all the samples taken during the last 5 d. Treatments were allocated at random to four vessels each and were (1) two temperature levels of $39^{\circ}C$ and $41^{\circ}C$ (2) two pH levels of 6.0 and 7.0. The total diet contained ($g\;kg^{-1}$ DM) 957 OM, 115 CP and $167MJ\;kg^{-1}$ (DM) GE. Although increase in temperature from $39^{\circ}C$ to $41^{\circ}C$ reduced degradation of major nutrients in vitro, it was non-significant. Interaction effect of temperature with pH also reflected a similar trend. However, pH showed a significant (p<0.05) negative effect on the degradability of all the nutrients in vitro. Altering the in vitro pH from 7 to 6 caused marked reduction in DMD from 60.2 to 41.8, CPD from 76.3 to 55.3 and GED from 55.3 to 35.1, respectively. Low pH (6) depressed total VFA production (61.9 vs. 34.9 mM) as well as acetate to propionate ratio in vitro (from 2.0 to 1.5) when compared to pH 7. Compared to pH 7, total gas production decreased from 1,841 ml to 1,148 ml at pH 6, $CO_2$ and $CH_4$ production also reduced from 639 to 260 ml and 138 to 45 ml, respectively. This study supported the premise that pH is one of the principal factors affecting the microbial production of volatile fatty acids and gas. Regulating the ruminal pH to increase bacterial activity may be one of the methods to optimize VFA production, reduce methane and, possibly, improve animal performance.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1617-1622
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• 2006

Long-term storage of feeds or feedstuffs in high temperature and humid conditions can be difficult because of microbial contamination. Essential oil isolated from industrial waste citrus peel could be used as a preservative because it is likely to have anti-bacterial and anti-fungal activity. Our objective was to determine whether different levels (0.028, 0.056 and 0.112 g/kg) of citrus essential oil (CEO) would provide anti-microbial activity and enhance preservation of animal feed without influencing rumen fermentation. At 0.112 g/kg, CEO inhibited growth of Escherichia coli (ATCC 25922) and Salmonela enteritidis (IFO 3313). Growth of E. coli recovered after 24 h of incubation, but S. enteritidis continued to be inhibited for 72 h. Preservation of antibiotic-free diets for swine was assessed by observing anti-aflatoxin activity. Aflatoxin was detected in control feed samples on days 16 (8 ppb) and 21 (8 ppb) and in anti-fungal agent (AA) treated samples on days 16 (2 ppb) and 21 (4 ppb). However, aflatoxin was not detected in feed samples treated with CEO. Treatment with CEO and AA did not influence ruminal pH, dry matter digestibility (DMD) or organic matter digestibility (OMD) over 48 h of incubation in rumen fluid. Acetate and propionate were slightly higher with CEO treatment (p<0.05), but total concentration of volatile fatty acid (VFA) was not significantly affected by treatment. Ammonia-N concentration was slightly higher for the control treatment (p<0.05). This study showed that treating feed with CEO enhances preservation of animal feed without influencing in vitro rumen fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.4
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• pp.452-461
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• 2012

Four, ruminally fistulated crossbred (Brahman${\times}$native) beef cattle with initial body weight of $420{\pm}15kg$ were randomly assigned according to a $4{\times}4$ Latin square design. The dietary treatments were mulberry leaf pellet (MUP) supplementation at 0, 200, 400 and 600 g/hd/d with rice straw fed to allow ad libitum intake. All steers were kept in individual pens and supplemented with concentrate at 5 g/kg of body weight daily. The experiment was 4 periods, and each lasted 21 d. During the first 14 d, all steers were fed their respective diets ad libitum and during the last 7 d, they were moved to metabolism crates for total urine and fecal collection. It was found that increasing MUP levels resulted in linearly increasing rice straw and total intakes (p<0.05). Ruminal temperature and pH were not significantly affected by MUP supplementation while $NH_3$-N concentration was increased (p<0.05) and maintained at a high level (18.5 mg/dl) with supplementation of MUP at 600 g/hd/d. Similarly, viable total bacteria in the rumen and cellulolytic bacteria were enriched by MUP supplementation at 600 g/hd/d. However, the rumen microbial diversity determined with a PCR-DGGE technique showed similar methanogenic diversity between treatments and sampling times and were similar at a 69% genetic relationship as determined by a UPGMA method. Based on this study, it could be concluded that supplementation of MUP at 600 g/hd/d improved DM intake, ruminal $NH_3$-N, and cellulolytic bacteria thus iimproving rumen ecology in beef cattle fed with rice straw.

• Animal Bioscience
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• v.36 no.8
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• pp.1285-1292
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• 2023

Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn²⁺ or Zn²⁺ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.625-634
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• 2004

The objective of this experiment is to study the possibility of lactate dehydrogenase(LDH) enzyme to prevent lactate accumulation in the rumen, For understanding capacity of bacterial LDH in rumen environments, this study was conducted to explore the effects of temperature, pH, VFAs and metal ions on Lactobacillus sp. FFy111-1's LDH activity, and the LDH activation in rumen fluid accumulated lactate. The optimum pH and temperature of LDH were pH 7.5 and 40$^{\circ}C$, respectively. The LDH activity had a good thennostability at range from 30 to 50$^{\circ}C$. The highest pH stability of the enzyme was at ranges from pH 7.0 to 8.0 and the enzyme activities showed above 64% level of non-treated one at pH 6.0 and 6.5. The LDH was inactivated by VFAs treatments but was enhanced by metal ion treatments without NaCl and $CuSO_4$ Especially, the LDH activity was increased to 127% and 124% of its original activity by 2 mM of $BaCl_2$ and $MnSO_4$, addition, respectively. When the acidic rumen fluid was treated by LDH enzyme of Lactobacillus sp. FFy111-1, the lactate concentration in the rumen fluid was lower compared with non-treated rumen fluid(P<0.05). This lactate reduction was resulted from an action of LDH. It was proved by result of purified D,L-LDH addition that showed the lowest lactate concentration among the treatments(P<0.05). Although further investigation of microbial LDH and ruminal lactate is needed, these findings suggest that the bacterial LDH has the potential capability to decrease the lactate accumulated in an acidic rumen fluid. Also, screening of super LDH producing bacteria and technical development for improving enzyme activity in rumen environment are essential keys for practical application.