Liquid-phase denaturant gradient gel electrophoresis profiles of rumen bacteria from Brahman cross steers selected into two groups on the basis of post-weaning liveweight gain on low crude protein pasture

2012 ◽  
Vol 52 (7) ◽  
pp. 647 ◽  
Author(s):  
E. D. Martínez ◽  
K. E. Turnbull ◽  
S. P. Quigley ◽  
S. J. Streeter ◽  
A. Swain ◽  
...  
Keyword(s):  
Liquid Phase ◽  
Gel Electrophoresis ◽  
Crude Protein ◽  
Rumen Fluid ◽  
Rumen Bacteria ◽  
Ruminal Ph ◽  
Gradient Gel Electrophoresis ◽  
Denaturant Gradient Gel Electrophoresis ◽  
The Relationship ◽  
Protein Diets

Two experiments were conducted to evaluate the relationship between rumen liquid-associated bacterial community structures and post-weaning liveweight gain (LWG) of Brahman crossbred steers. Bacterial diversity was assessed using denaturant gradient gel electrophoresis (DGGE). In Experiment 1, 16 steers were selected from a group of 100 steers by pairing steers with the same weaning weight, but different LWG 90 days after weaning (n = 8 highest growth, 0.21 ± 0.01 kg/day; and n = 8 lowest growth –0.07 ± 0.01 kg/day). Thereafter, steers were allocated to a 28-day pen study and fed Mitchell grass (Astrebla spp.) hay to examine DM intake and digestibility, rumen parameters and rumen microbial community in these two groups. Rumen fluid samples were taken by stomach tube 3 h after feeding on the last day of the pen phase. In Experiment 2, 12 pairs of weaned steers were selected from a group of 203 steers on the same basis as Experiment 1. The post-weaning LWG were 0.20 ± 0.03 and 0.02 ± 0.03 kg/day for the 12 highest and 12 lowest growth animals selected, respectively. Steers then grazed dry season Sabi grass (Urochloa mosambicensis) dominant pasture for 21 days, before rumen sampling 3 h after morning grazing by stomach tubing on the last day. In Experiment 1, there were no significant differences between the two groups in DM intake, digestibility, ruminal pH, total volatile fatty acid (VFA) concentration or the VFA proportion. In Experiments 1 and 2, rumen ammonia-N concentration was similar between LWG groups and there was no evidence of a relationship between liquid-phase DGGE profiles of rumen bacteria and high or low post-weaning LWG using multivariate analyses. Furthermore, the number of detected DGGE bands, the Shannon–Wiener and evenness indexes were not different between LWG groups. This DGGE analysis of the most abundant groups of rumen fluid-associated bacteria suggests that microbial populations were not related with the differences observed in post-weaning LWG within a group of weaners fed low crude protein diets.

Segregation of partly melted molecules: isolation of CpG islands by polyacrylamide gel electrophoresis

2004 ◽  
Vol 385 (10) ◽  
pp. 967-973 ◽  
Author(s):  
Masahiko Shiraishi ◽  
Adam J. Oates ◽  
Xu Li ◽  
Ying H. Chuu ◽  
Takao Sekiya
Keyword(s):  
Efficient Method ◽  
Gel Electrophoresis ◽  
Human Cancer ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cpg Islands ◽  
Theoretical Background ◽  
Dna Fragments ◽  
Practical Application ◽  
Gradient Gel Electrophoresis ◽  
Denaturant Gradient Gel Electrophoresis

Abstract The technique of segregation of partly melted molecules (SPM) is a convenient and efficient method to isolate DNA fragments associated with CpG islands. The approach is conceptually simple and uses denaturant gradient gel electrophoresis to separate DNA molecules digested with restriction endonucleases. The SPM methodology has successfully been applied to the identification of genes from anonymous, unsequenced DNA fragments and CpG islands methylated in human cancer. In this article the theoretical background and practical application of the SPM method is reviewed.

scholarly journals Mango (Mangifera indica) seed kernels as untraditional source of energy in Rahmani sheep rations

2019 ◽  
Vol 43 (1) ◽  
Author(s):  
Hamed A. A. Omer ◽  
Mohamed A. Tawila ◽  
Sawsan M. Gad ◽  
Soha S. Abdel-Magid
Keyword(s):  
Crude Protein ◽  
Rumen Fluid ◽  
Nutrient Digestibility ◽  
Sampling Time ◽  
Water Metabolism ◽  
Ruminal Ph ◽  
Yellow Corn ◽  
Live Body Weight ◽  
Mango Seed ◽  
Dietary Treatments

Abstract Background The present work aimed to investigate the impact of partial replacement yellow corn by mango seed kernels (MSK) at different levels of (0, 25, and 50%) on feed and water consumption, water metabolism, nutrient digestibility, nitrogen balance, and rumen fluid parameters. Methods Twelve mature male Rahmani sheep of about 58.39 ± 1.22 kg live body weight on average were used to investigate the influence of replacing yellow corn contents (45% in the control ration) by mango seed kernels (MSK) at different levels (0, 25, and 50%) that equal (0, 11.25, and 22.5% of total ration contents) on feed and water intakes, water metabolism, nutrient digestibility coefficients, nitrogen utilization, and some rumen fluid parameters. The animals were randomly assigned to three experimental groups (four animals in each treatment). Sheep fed rations are composed of 50% concentrate feed mixture (CFM) and 50% peanut vein hay (PVH) at portion 3% dry matter (DM) of live body weight (LBW). Results The results showed that values of crude protein (CP), crude fiber (CF), and nitrogen-free extract (NFE) of MSK and yellow corn were similar. Meanwhile, MSK has superior ether extract content compared to yellow corn; meanwhile, ash content of yellow corn was lower than ash content of MSK. All tested CFM were isonitrogenous (16.17% CP), but it differs in their contents of gross and digestible energy. On the other hand, all experimental total mixed rations (TMR) were isonitrogenous (14.52% CP) and isocaloric (4177 kcal/kg DM in average). Dietary treatments significantly (P < 0.05) decreased total dry matter intake (DMI) and crude protein intake (CPI) that expressed as (g/h/day, g/kgW0.75, and kg/100 kg LBW) or gross energy intake (GEI) and digestible energy intake (DEI) that expressed as (Mcal/h/day, kcal/kgW0.75, and Mcal/100 kg LBW). Inclusion MSK in sheep ration insignificantly (P > 0.05) increased drinking water by 20% and 6.89% for experimental groups fed TMR2 and TMR3, respectively, compared to the control one (TMR1). Incorporation MSK in the rations significantly (P < 0.05) increased insensible losses of water. Dietary treatments significantly (P < 0.05) increased nutrient digestibility coefficients of DM, organic matter (OM), CP, CF, and NFE. Meanwhile, it significantly (P < 0.05) decreased EE digestibility. On the other hand, both values of total digestible nutrient (TDN) and digestible crude protein (DCP) were significantly (P < 0.005) improved. Nitrogen retention (NR) (NR % of nitrogen intake (NI) and NR % of digested nitrogen (DN)) values were significantly (P < 0.05) increased with increasing level of MSK in the sheep ration. Inclusion MSK significantly (P < 0.05) increased ruminal pH; however, it had no significant (P > 0.05) on both ammonia nitrogen (NH3-N) and total volatile fatty acid (TFVA) concentrations. Sampling time at 3 h post feeding significantly (P < 0.05) decreased ruminal pH in comparison with before and 6 h post feeding; meanwhile, sampling time had no significant effect (P > 0.05) on NH3-N concentration. Dietary treatment significantly (P < 0.05) increased the interaction between treatments and sampling time for ruminal pH. However, no interaction between treatments and sampling time was observed for both NH3-N and TVFA concentrations. Conclusion The present results revealed that replacement up to 50% of yellow corn by MSK as alternative source of energy could be useful in feeding sheep without any adverse effect on digestion and fermentation processing. Also, it can be used to depress the rations cost.

DIGESTION IN THE RUMEN AND AMINO ACID SUPPLY TO THE DUODENUM OF SHEEP FED ENSILED AND FRESH ALFALFA

1986 ◽  
Vol 66 (4) ◽  
pp. 1019-1027 ◽  
Author(s):  
D. A. FLORES ◽  
L. E. PHILLIP ◽  
D. M. VEIRA ◽  
M. IVAN
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Rumen Fluid ◽  
Essential Amino Acids ◽  
Ruminal Ph ◽  
Sheep Rumen ◽  
Branched Chain ◽  
High Degree ◽  
The Relationship ◽  
Adaptation Phase

A study was conducted with six sheep equipped with rumen and duodenal re-entrant cannulae to investigate differences in duodenal amino acid supply with fresh and ensiled alfalfa. The experiment was conducted as a cross-over design with two 12-d periods consisting of an adaptation phase (5 d) followed by feces collection (5 d), and finally on day 12, 24 h continuous collection of duodenal digesta and sampling of rumen fluid. During the experiment the sheep were restricted on feed to 52 g DM d−1 kg−0.75. Analyses of the forages revealed extensive breakdown of threonine, lysine and other basic amino acids, and high degree of preservation of branched chain amino acids. Relative to the amount consumed, there was a net loss of amino acids at the duodenum with fresh alfalfa but a net gain with silage. The relationship between intake of individual essential amino acids and duodenal recovery was inconsistent. Duodenal recovery of total essential amino acids was 28% higher with fresh than with ensiled alfalfa, a difference which was most pronounced with threonine and lysine. Digestion of organic matter and fiber was unaffected (P > 0.05) by ensiling of alfalfa. Estimates of ruminal pH were higher (P < 0.05) for silage than fresh forage (6.9 vs. 6.7) but VFA concentrations were lower (P < 0.05) for silage (52.1 vs. 64.1 mmol L−1). Rumen ammonia levels were similar for the two forages. The deficit in duodenal supply of essential amino acids with silage is discussed in relation to the problem of silage intake in ruminants. Key words: Alfalfa silage, duodenal amino acids, sheep, rumen digestion

EFFECT OF TANNIN CONTENT IN TWO ISOSYNTHETIC STRAINS OF BIRDSFOOT TREFOIL (Lotus corniculatus L.) ON FEED DIGESTIBILITY AND RUMEN FLUID COMPOSITION IN SHEEP

1989 ◽  
Vol 69 (4) ◽  
pp. 1031-1039 ◽  
Author(s):  
J. CHIQUETTE ◽  
K.-J. CHENG ◽  
L. M. RODE ◽  
L. P. MILLIGAN
Keyword(s):  
Crude Protein ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Lotus Corniculatus ◽  
Ammonia Nitrogen ◽  
Birdsfoot Trefoil ◽  
Fluid Composition ◽  
Protein Nitrogen ◽  
Ruminal Ph ◽  
Feed Digestibility

Low tannin (LT) and high tannin (HT) isosynthetic strains of birdsfoot trefoil containing 1 and 3% condensed tannins on a dry matter (DM) basis were cut fresh daily and fed to eight ruminally cannulated Suffolk wethers. Rumen fluid samples were collected before feeding and 3 and 6 h after feeding for pH, chlorophyll, supernatant protein nitrogen (N), ammonia nitrogen (NH3-N) and volatile fatty acids (VFA) determinations, and protozoa counts. Daily samples of the forage fed to the animals, and of feces, were analyzed for crude protein (CP) content, acid detergent fiber (ADF) and organic matter (OM). Results from rumen fluid analyses indicated that sheep receiving the HT strain had lower (P < 0.01) supernatant protein N and NH3-N concentrations, and consistently but not significantly lower VFA and chlorophyll concentrations. Animals fed HT showed higher (P < 0.05) protozoa counts than LT-fed animals, but only in the samples taken 3 h postfeeding. Ruminal pH was similar on LT and HT diets. Apparent digestibility of CP was 9% less (P < 0.05) in sheep fed the HT strain. However, apparent digestibilities of DM, OM and ADF were not affected by the tannin level. Key words: Lotus corniculatus L., birdsfoot trefoil, tannin, digestibility, rumen, sheep

THE DISTRIBUTION OF CHLOROPHYLL IN RUMEN CONTENTS AND THE ONSET OF BLOAT IN CATTLE

1986 ◽  
Vol 66 (1) ◽  
pp. 97-102 ◽  
Author(s):  
W. MAJAK ◽  
J. W. HALL ◽  
R. E. HOWARTH
Keyword(s):  
Particulate Matter ◽  
Liquid Phase ◽  
Key Words ◽  
Rumen Fluid ◽  
Gas Production ◽  
Solid Phases ◽  
Relationship Of ◽  
The Relationship ◽  
Liquid And Solid Phases ◽  
Chlorophyll Concentrations

Chlorophyll levels in rumen contents from the dorsal and ventral sac were examined before alfalfa ingestion to determine the relationship of chloroplast particles to the onset of pasture bloat in cattle. Chlorophyll concentrations were significantly higher in cattle that subsequently bloated than in those that did not. The higher levels were observed in both the liquid and solid phases of rumen contents from cattle predisposed to bloat but the liquid phase showed a greater increase. Chlorophyll levels were uniformly distributed in the rumen with little or no difference between samples obtained from the ventral and dorsal sacs. In agreement with previous results, animals predisposed to bloat showed higher rates of gas production and greater buoyancy of particulate matter in prefeeding rumen fluid. The results suggest that chloroplast particles in the rumen have a slower rate of clearance in bloating animals than in nonbloating ones. Key words: Bloat, alfalfa, cattle, rumen, chlorophyll, chloroplast particles

scholarly journals 413 Effect of chitosan inclusion and dietary crude protein level on rumen fermentation in beef heifers fed a total mixed ration

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 166-167
Author(s):  
Stuart Kirwan ◽  
Tommy Boland ◽  
Eleonora Serra ◽  
Gaurav Rajauria ◽  
Karina Pierce
Keyword(s):  
Crude Protein ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Rumen Fermentation ◽  
Ruminal Fermentation ◽  
Beef Heifers ◽  
Total Mixed Ration ◽  
Ruminal Ph ◽  
Dietary Crude Protein ◽  
Dietary Treatments

Abstract Chitosan (N-acetyl-D-glucosamine polymer) is a natural biopolymer, which is the second most abundant organic compound on earth, found in lower plants and some arthropods and crustaceans. Studies with ruminants have shown that supplementing with chitosan can alter ruminal fermentation, including higher propionate concentration and lower acetate to propionate ratio, improving the energy efficiency within the rumen though data on nitrogen metabolism is lacking. Therefore, the objective of this study was to evaluate the effects of chitosan with different dietary crude protein (CP) inclusion on rumen fermentation in beef heifers. Eight ruminally cannulated Belgian Blue x Holstein Friesian heifers (752 ± 52Kg BW) were allocated one of four dietary treatments in a 2 x 2 factorial Latin Square design (n = 8). Diets were offered as total mixed ration (TMR) consisting of grass silage plus concentrate. Diets were offered once daily ad libitum. Dietary treatments were as follows: high CP (16%) +/- chitosan (10g kg-1 DMI) supplementation (HP+) or (HP-) and low CP (12%) +/- chitosan supplementation (LP+) or (LP-). Experimental periods were 25d in duration (14 d dietary adaptation and 10 d sampling). Rumen fluid was collected via cannula every 2 h post feeding for analysis of ruminal pH, ammonia (NH3) and volatile fatty acids (VFA) over a 48-hour period. Data was analysed using Proc Mixed procedure in SAS. Significant differences were considered when P &lt; 0.05. Dietary treatment had no effect on ruminal pH (P &gt; 0.05). Chitosan supplementation had no effect on ruminal NH3 and VFA concentrations (P &gt; 0.05), whereas, animals offered 16% CP had higher concentrations of ruminal NH3 (P &lt; 0.001). In conclusion, supplementing a TMR diet fed to beef heifers with chitosan had no effect on rumen fermentation. However, reducing dietary CP concentration fed to beef animals resulted in lower rumen NH3 concentrations.

scholarly journals Effects of dietary forage and calf starter on ruminal pH and transcriptomic adaptation of the rumen epithelium in Holstein calves during the weaning transition

2016 ◽  
Vol 48 (11) ◽  
pp. 803-809 ◽  
Author(s):  
Yo-Han Kim ◽  
Noriyuki Toji ◽  
Keiichiro Kizaki ◽  
Shiro Kushibiki ◽  
Toshihiro Ichijo ◽  
...  
Keyword(s):  
Growth Factor ◽  
Rumen Fluid ◽  
Expression Profiles ◽  
Ruminal Ph ◽  
Ruminal Acidosis ◽  
Rumen Epithelium ◽  
Transcription Regulators ◽  
Holstein Calves ◽  
The Relationship ◽  
Upstream Regulators

We investigated the relationship between ruminal pH and transcriptomic adaptation of the rumen epithelium (RE) of calves fed calf starter with and without forage during the weaning transition. Holstein calves were assigned to groups fed calf starter either with forage (HAY group, n = 3) or without forage (CON group, n = 4). Ruminal pH was measured continuously, and rumen fluid and epithelium were collected 3 wk after weaning. mRNA expression profiles of the RE were examined by one-color microarray. Differentially expressed genes (DEGs) were investigated using the Ingenuity Pathway Analysis (IPA). Mean and maximum ruminal pH were significantly ( P < 0.05) higher, and the duration of pH < 5.8 during 1 day was significantly ( P < 0.05) shorter, in the HAY group. The proportion of ruminal acetate and the acetate-to-propionate ratio were significantly ( P < 0.05) lower in the CON group. DEGs encoding transcription regulators (SREBP1), insulin-like growth factor binding proteins (IGFBP7 and CTGF), ketogenic enzymes (HMGCL, BDH1, and BDH2), and a transporter (SLC16A3) were identified ( P < 0.05) between the two groups. A growth factor (TGFB1) and signaling pathway (EGF and EGFR) were activated as upstream regulators. These results suggest that dietary forage alleviates ruminal acidosis, and the decrease in ruminal pH may damage the RE, leading to changes in gene expression to repair the damage. Furthermore, rumen development may be regulated by growth factor (TGFB1) and signaling pathways (EGF and IGFBP) for adaptation to feeding on calf starter with and without forage during the weaning transition.

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