• Asian-Australasian Journal of Animal Sciences
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• 제14권6호
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• pp.880-884
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• 2001

Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.

• Asian-Australasian Journal of Animal Sciences
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• 제28권3호
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• pp.351-359
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• 2015

Twelve multiparous Holstein dairy cows in mid-lactation were selected for a replicated $4{\times}4$ Latin square design with a $2{\times}2$ factorial arrangement to investigate the effects of corn and soybean meal (SBM) types on rumen fermentation, N metabolism and lactation performance in dairy cows. Two types of corn (dry ground [DGC] and steam-flaked corn [SFC]) and two types of SBM (solvent-extracted and heat-treated SBM) with different ruminal degradation rates and extents were used to formulate four diets with the same basal ingredients. Each period lasted for 21 days, including 14 d for adaptation and 7 d for sample collection. Cows receiving SFC had a lower dry matter (DM) and total N intake than those fed DGC. However, the milk yield and milk protein yield were not influenced by the corn type, resulting in higher feed and N utilization efficiency in SFC-fed cows than those receiving DGC. Ruminal acetate concentrations was greater and total volatile fatty acids concentrations tended to be greater for cows receiving DGC relative to cows fed SFC, but milk fat content was not influenced by corn type. The SFC-fed cows had lower ruminal ammonia-N, less urea N in their blood and milk, and lower fecal N excretion than those on DGC. Compared with solvent-extracted SBM-fed cows, cows receiving heat-treated SBM had lower microbial protein yield in the rumen, but similar total tract apparent nutrient digestibility, N metabolism measurements, and productivity. Excessive supply of metabolizable protein in all diets may have caused the lack of difference in lactation performance between SBM types. Results of the present study indicated that increasing the energy degradability in the rumen could improve feed efficiency, and reduce environmental pollution.

• Asian-Australasian Journal of Animal Sciences
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• 제29권2호
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• pp.230-240
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• 2016

Information on the effects of different yeast species on ruminal fermentation is limited. This experiment was conducted in a $3{\times}4$ factorial arrangement to explore and compare the effects of addition of three different live yeast species (Candida utilis 1314, Saccharomyces cerevisiae 1355, and Candida tropicalis 1254) at four doses (0, $0.25{\times}10^7$, $0.50{\times}10^7$, and $0.75{\times}10^7$ colony-forming unit [cfu]) on in vitro gas production kinetics, fiber degradation, methane production and ruminal fermentation characteristics of maize stover, and rice straw by mixed rumen microorganisms in dairy cows. The maximum gas production (Vf), dry matter disappearance (IVDMD), neutral detergent fiber disappearance (IVNDFD), and methane production in C. utilis group were less (p<0.01) than other two live yeast supplemented groups. The inclusion of S. cerevisiae reduced (p<0.01) the concentrations of ammonia nitrogen ($NH_3$-N), isobutyrate, and isovalerate compared to the other two yeast groups. C. tropicalis addition generally enhanced (p<0.05) IVDMD and IVNDFD. The $NH_3$-N concentration and $CH_4$ production were increased (p<0.05) by the addition of S. cerevisiae and C. tropicalis compared with the control. Supplementation of three yeast species decreased (p<0.05) or numerically decreased the ratio of acetate to propionate. The current results indicate that C. tropicalis is more preferred as yeast culture supplements, and its optimal dose should be $0.25{\times}10^7$ cfu/500 mg substrates in vitro.

• Asian-Australasian Journal of Animal Sciences
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• 제17권3호
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• pp.355-365
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• 2004

Two experiments were conducted to determine the effect of alkaline hydrogen peroxide (AHP) treatment (1% NaOH+1.5% $H_2O_2$; 1 AHPMS, 2% NaOH+1.5% $H_2O_2$; 2AHPMS) on rate and extent of degradation of mustard straw (MS) in sacco in sheep, and its in vivo digestion and ruminal fermentation characteristics when fed to sheep with concentrate (200 g per sheep daily). The treatment of straw with 1 and 2% AHP increased its sodium content by 148 and 296% to that of untreated straw (UMS). There was significant decrease in NDF and hemicellulose contents of AHP treated straw and increase in cellulose and lignin contents. Phenolic acids like ferrulic, $\rho$-coumaric and o-coumaric significantly (p<0.001) reduced by AHP treatment of mustard straw. In first experiment the in sacco degradation of DM, OM and NDF was significantly (p<0.01) greater for 2 AHPMS than for UMS at all incubation periods. The disappearance of nutrient from 1 AHPMS and 2 AHPMS treated straws continue to increase up to 96 h whereas in UMS the peak disappearance was found at 48 h. By using the equation {(y=a+b) ($1-e^{-ct}$)} the degradation rates (c) for DM, OM, and NDF were significantly higher for UMS than AHP treated straws. Level of alkali (1 and 2%) had significant effect on degradation characteristics (a, b, c and $P_{0.05}$) of DM and NDF fraction of MS. However, the effect was not pronounced on OM fraction of MS. In feeding experiment, the intake of nutrients for DM, OM, cell wall constituents and energy was higher on 2 AHPMS, whereas no effect on the digestibility of these nutrients was observed. The apparent nitrogen retention was higher (p<0.05) both in 1 and 2 AHPMS groups. Water intake by animals was significantly increased due to AHP treated mustard straw feeding. Rumen liquor pH was higher in 2 AHPMS fed animals. The $NH_3-N$ of rumen liquor was not affected by feeding of AHP treated MS based diets. Total VFA concentration was significantly (p<0.01) higher in UMS fed group. The fractional out flow rate of DM was higher (p<0.05) in animals fed on 2 AHPMS diets compared to UMS and lAHPMS fed groups. The population of large holotrichs was higher (p<0.05) on AHP treated MS fed diets compared to UMS. The study indicated that treatment of mustard straw with AHP changed its chemical composition towards a better feed. The nutritive value of 2% AHP treated mustard straw was better in terms of dry matter intake and apparent nitrogen retention. The higher in sacco DM, OM and NDF disappearance however, was not confirmed by in vivo data in this study.

• Asian-Australasian Journal of Animal Sciences
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• 제13권6호
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• pp.785-790
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• 2000

The effect of formaldehyde treatment of solvent and mechanical extracted cottonseed meal on the performance, digestibility and nitrogen balance was assessed in lambs. Four total mixed rations viz., A, B, C and D containing 40% untreated and treated solvent and mechanical extracted meal were prepared. Sixteen male lambs with average BW of 20-22 kg were randomly allocated to experimental rations and were fed individually during ninety days growth trial. The treatment of solvent extracted cottonseed meal resulted in a linear decrease in ruminal protein degradation. Maximum decrease (64%) in protein degradation was observed at 4 h incubation time with 0.3% formaldehyde treatment. Highest daily BW gain was observed in lambs fed on rations Band D compared to lambs fed on rations A and C. Daily BW gain was higher on rations having 0.3% for fromaldehyde treated cottonseed meals. Higher DM digestibility was observed on ration D compared to other rations. Higher (p<0.05) CP and CF digestibility was observed on rations Band D compared to rations A and C. Nitrogen retention as % age of nitrogen intake was (p<0.05) higher for lambs fed rations B and D compared to rations A and C. Similar pattern was observed for nitrogen retention as percent of nitrogen absorbed. The present study suggested that oil extraction methods of cottonseed did not alter their meal utilization in lambs, however, formaldehyde (0.3%) treatment of meals enhanced its efficiency for growth, digestibility and nitrogen balance in lambs.

• Asian-Australasian Journal of Animal Sciences
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• 제25권10호
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• pp.1381-1388
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• 2012

An in vitro experiment was conducted to examine the effects of defaunation (removal of protozoa) on ruminal fermentation characteristics, $CH_4$ production and degradation by rumen microbes when incubated with cereal grains (corn, wheat and rye). Sodium lauryl sulfate as a defaunation reagent was added into the culture solution at a concentration of 0.000375 g/ml, and incubated anaerobically for up to 12 h at $39^{\circ}C$. Following defaunation, live protozoa in the culture solution were rarely observed by microscopic examination. A difference in pH was found among grains regardless of defaunation at all incubation times (p<0.01 to 0.001). Defaunation significantly decreased pH at 12 h (p<0.05) when rumen fluid was incubated with grains. Ammonia-N concentration was increased by defaunation for all grains at 6 h (p<0.05) and 12 h (p<0.05) incubation times. Total VFA concentration was increased by defaunation at 6 h (p<0.05) and 12 h (p<0.01) for all grains. Meanwhile, defaunation decreased acetate and butyrate proportions at 6 h (p<0.05, p<0.01) and 12 h (p<0.01, p<0.001), but increased the propionate proportion at 3 h, 6 h and 12 h incubation (p<0.01 to 0.001) for all grains. Defaunation increased in vitro effective degradability of DM (p<0.05). Production of total gas and $CO_2$ was decreased by defaunation for all grains at 1 h (p<0.05, p<0.05) and then increased at 6 h (p<0.05, p<0.05) and 12 h (p<0.05, p<0.05). $CH_4$ production was higher from faunation than from defaunation at all incubation times (p<0.05).

• Asian-Australasian Journal of Animal Sciences
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• 제21권11호
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• pp.1604-1609
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• 2008

A series of in vitro and in vivo experiments were conducted to investigate the effects of the mixture of Tween 80 and cellulolytic enzymes (xylanase and cellulase) on total tract nutrient digestibility and rumen cellulolytic bacterial adhesion rates in Holstein steers. Ground timothy hay sprayed with various levels of Tween 80 and cellulolytic enzymes was used as substrates in an in vitro experiment to find out the best combinations for DM degradation. The application level of 2.5% (v/w) Tween 80 and the combination of 5 U xylanase and 2.5 U cellulase per gram of ground timothy hay (DM basis) resulted in the highest in vitro dry matter degradation rate (p<0.05). Feeding the same timothy hay to Holstein steers also improved in vivo nutrient (DM, CP, CF, NDF and ADF) digesibilities compared to non-treated hay (p<0.05). Moreover, Tween 80 and enzyme combination treatment increased total ruminal VFA and concentrations of propionic acid and isovaleric acid with decreased acetate to propionate ratio (p<0.001). However, adhesion rates of Fibrobacter succinogenes and Ruminococcus flavefaciens determined by Real Time PCR were not influenced by the treatment while that of Ruminococcus albus was decreased (p<0.05). The present results indicate that a mixture of Tween 80 and cellulolytic enzymes can improve rumen environment and feed digestibility with variable influence on cellulolytic bacterial adhesion on feed.

• Asian-Australasian Journal of Animal Sciences
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• 제32권3호
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• pp.357-365
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• 2019

Objective: The experiment was conducted to evaluate the effects of four fungal pretreatments on the nutritional value of Camellia seed residues, and to evaluate the feeding value of pretreated Camellia seed residues for ruminants. Methods: Camellia seed residues were firstly fermented by four lignin degrading fungi, namely, Phanerochaete chrysosporium (P. chrysosporium)-30942, Trichoderma koningiopsis (T. koningiopsis)-2660, Trichoderma aspellum (T. aspellum)-2527, or T. aspellum-2627, under solid-state fermentation (SSF) conditions at six different incubation times. The nutritional value of each fermented Camellia seed residues was then analyzed. The fermentation profiles, organic matter degradability and metabolizable energy of each pre-treated Camellia seed residue were further evaluated using an in vitro rumen fermentation system. Results: After 5 days of fermentation, P. chrysosporium-30942 had higher degradation of lignin (20.51%), consumed less hemicellulose (4.02%), and the SSF efficiency reached 83.43%. T. koningiopsis-2660 degraded more lignin (21.54%) and consumed less cellulose (20.94%) and hemicellulose (2.51%), the SSF efficiency reached 127.93%. The maximum SSF efficiency was 58.18% for T. aspellum-2527 and 47.61% for T. aspellum-2627, appeared at 30 and 15 days respectively. All the fungal pretreatments significantly improved the crude protein content (p<0.05). The Camellia seed residues pretreated for 5 days were found to possess significantly increased organic matter degradability, volatile fatty acid production and metabolizable energy (p<0.05) after the treatment of either P. chrysosporium-30942, T. koningiopsis-2660 or T. aspellum-2527. The fungal pretreatments did not significantly change the rumen fermentation pattern of Camellia seed residues, with an unchanged ratio of acetate to propionate. Conclusion: The fungi showed excellent potential for the solid-state bioconversion of Camellia seed residues into digestible ruminant energy feed, and their shorter lignin degradation characteristics could reduce loss of the other available carbohydrates during SSF.

• Animal Bioscience
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• 제35권9호
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• pp.1367-1378
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• 2022

Objective: The present study was conducted to examine the gas production, fermentation characteristics, nutrient degradation, and methanogenic community composition of a rumen fluid culture with Broussonetia papyrifera (B. papyrifera) subjected to ensiling or steam explosion (SE) pretreatment. Methods: Fresh B. papyrifera was collected and pretreated by ensiling or SE, which was then fermented with ruminal fluids as ensiled B. papyrifera group, steam-exploded B. papyrifera group, and untreated B. papyrifera group. The gas and methane production, fermentation characteristics, nutrient degradation, and methanogenic community were determined during the fermentation. Results: Cumulative methane production was significantly improved with SE pretreatment compared with ensiled or untreated biomass accompanied with more volatile fatty acids production. After 72 h incubation, SE and ensiling pretreatments decreased the acid detergent fiber contents by 39.4% and 22.9%, and neutral detergent fiber contents by 10.6% and 47.2%, respectively. Changes of methanogenic diversity and abundance of methanogenic archaea corresponded to the variations in fermentation pattern and methane production. Conclusion: Compared with ensiling pretreatment, SE can be a promising technique for the efficient utilization of B. papyrifera, which would contribute to sustainable livestock production systems.

• Asian-Australasian Journal of Animal Sciences
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• 제26권5호
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• pp.668-674
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• 2013

An in vitro fermentation was conducted to determine the effects of hainanmycin on protein degradation and populations of ammonia-producing bacteria. The substrates (DM basis) for in vitro fermentation consisted of alfalfa hay (31.7%), Chinese wild rye grass hay (28.3%), ground corn grain (24.5%), soybean meal (15.5%) with a forage: concentrate of 60:40. Treatments were the control (no additive) and hainanmycin supplemented at 0.1 (H0.1), 1 (H1), 10 (H10), and 100 mg/kg (H100) of the substrates. After 24 h of fermentation, the highest addition level of hainanmycin decreased total VFA concentration and increased the final pH. The high addition level of hainanmycin (H1, H10, and H100) reduced (p<0.05) branched-chain VFA concentration, the molar proportion of acetate and butyrate, and ratio of acetate to propionate; and increased the molar proportion of propionate, except that for H1 the in molar proportion of acetate and isobutyrate was not changed (p>0.05). After 24 h of fermentation, H10 and H100 increased (p<0.05) concentrations of peptide nitrogen and AA nitrogen and proteinase activity, and decreased (p<0.05) $NH_3$-N concentration and deaminase activity compared with control. Peptidase activitives were not affected by hainanmycin. Hainanmycin supplementation only inhibited the growth of Butyrivibrio fibrisolvens, which is one of the species of low deaminative activity. Hainanmycin supplementation also decreased (p<0.05) relative population sizes of hyper-ammonia-producing species, except for H0.1 on Clostridium aminophilum. It was concluded that dietary supplementation with hainanmycin could improve ruminal fermentation and modify protein degradation by changing population size of ammonia-producing bacteria in vitro; and the addition level of 10 mg/kg appeared to achieve the best results.