scholarly journals Ruminal microbes of adult sheep do not degrade extracellular l-citrulline

2020 ◽  
Vol 98 (6) ◽  
Author(s):  
Kyler R Gilbreath ◽  
Fuller W Bazer ◽  
M Carey Satterfield ◽  
Jason J Cleere ◽  
Guoyao Wu
Keyword(s):  
Jugular Vein ◽  
Rumen Fluid ◽  
Feeding Time ◽  
Ruminal Bacteria ◽  
Animal Blood ◽  
Extrahepatic Tissues ◽  
Baseline Values ◽  
Hydrolysis Of ◽  
Predetermined Time ◽  
Suffolk Sheep

Abstract This study determined whether extracellular citrulline is degraded by ruminal bacteria of sheep. In the first experiment, whole rumen fluid (3 mL) from six adult Suffolk sheep was incubated at 37 °C with 5 mM l-glutamine (Gln), l-glutamate (Glu), l-arginine (Arg), or l-citrulline (Cit) for 0, 0.5, 1, and 2 h or with 0, 0.5, 2, or 5 mM Gln, Glu, Arg, or Cit for 2 h. An aliquot (50 µL) of the incubation solution was collected at the predetermined time points for amino acids (AA) analyses. Results showed extensive hydrolysis of Gln into Glu and ammonia, of Arg into l-ornithine and l-proline, but little or no degradation of extracellular Cit or Glu by ruminal microbes. In the second experiment, six adult Suffolk sheep were individually fed each of three separate supplements (8 g Gln , Cit, or urea) on three separate days along with regular feed (800 g/animal). Blood (2 mL) was sampled from the jugular vein prior to feeding (time 0) and at 0.5, 1, 2, and 4 h after consuming the supplement. Plasma was analyzed for AA, glucose, ammonia, and urea. The concentrations of Cit in the plasma of sheep consuming this AA increased (P < 0.001) by 117% at 4 h and those of Arg increased by 23% at 4 h, compared with the baseline values. Urea or Gln feeding did not affect (P > 0.05) the concentrations of Cit or Arg in plasma. These results indicate that Cit is not metabolized by ruminal microbes of sheep and is, therefore, absorbed as such by the small intestine and used for the synthesis of Arg by extrahepatic tissues.

scholarly journals Identification of Catalposide Metabolites in Human Liver and Intestinal Preparations and Characterization of the Relevant Sulfotransferase, UDP-glucuronosyltransferase, and Carboxylesterase Enzymes

Pharmaceutics ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 355 ◽  
Author(s):  
Deok-Kyu Hwang ◽  
Ju-Hyun Kim ◽  
Yongho Shin ◽  
Won-Gu Choi ◽  
Sunjoo Kim ◽  
...  
Keyword(s):  
Human Liver ◽  
Individual Variability ◽  
Hydroxybenzoic Acid ◽  
Catalpa Ovata ◽  
Anti Oxidant ◽  
Extrahepatic Tissues ◽  
Udp Glucuronosyltransferase ◽  
Hydrolysis Of

Catalposide, an active component of Veronica species such as Catalpa ovata and Pseudolysimachion lingifolium, exhibits anti-inflammatory, antinociceptic, anti-oxidant, hepatoprotective, and cytostatic activities. We characterized the in vitro metabolic pathways of catalposide to predict its pharmacokinetics. Catalposide was metabolized to catalposide sulfate (M1), 4-hydroxybenzoic acid (M2), 4-hydroxybenzoic acid glucuronide (M3), and catalposide glucuronide (M4) by human hepatocytes, liver S9 fractions, and intestinal microsomes. M1 formation from catalposide was catalyzed by sulfotransferases (SULTs) 1C4, SULT1A1*1, SULT1A1*2, and SULT1E1. Catalposide glucuronidation to M4 was catalyzed by gastrointestine-specific UDP-glucuronosyltransferases (UGTs) 1A8 and UGT1A10; M4 was not detected after incubation of catalposide with human liver preparations. Hydrolysis of catalposide to M2 was catalyzed by carboxylesterases (CESs) 1 and 2, and M2 was further metabolized to M3 by UGT1A6 and UGT1A9 enzymes. Catalposide was also metabolized in extrahepatic tissues; genetic polymorphisms of the carboxylesterase (CES), UDP-glucuronosyltransferase (UGT), and sulfotransferase (SULT) enzymes responsible for catalposide metabolism may cause inter-individual variability in terms of catalposide pharmacokinetics.

scholarly journals Hydrolysis of14C-labelled proteins by rumen micro-organisms and by proteolytic enzymes prepared from rumen bacteria

1983 ◽  
Vol 50 (2) ◽  
pp. 345-355 ◽  
Author(s):  
R. J. Wallace
Keyword(s):  
Proteolytic Enzymes ◽  
Rumen Fluid ◽  
Performic Acid ◽  
Cross Linking ◽  
Acid Oxidation ◽  
Rumen Bacteria ◽  
Rate Of Hydrolysis ◽  
Micro Organisms ◽  
Hydrolysis Of

1. Proteins were labelled with14C in a limited reductive methylation using [14C]formaldehyde and sodium borohydride.2. The rate of hydrolysis of purified proteins was little (< 10%) affected by methylation and the14C-labelled digestion products were not incorporated into microbial protein during a 5 h incubation with rumen fluid in vitro. It was therefore concluded that proteins labelled with14C in this way are valid substrates for study with rumen micro-organisms.3. The patterns of digestion of14C-labelled fish meal, linseed meal and groundnut-protein meal by rumen micro-organisms in vitro were similar to those found in vivo.4. The rates of hydrolysis of a number of14C-labelled proteins, including glycoprotein II and lectin from kidney beans (Phaseolus vulgaris), were determined with mixed rumen micro-organisms and with proteases extracted from rumen bacteria. Different soluble proteins were digested at quite different rates, with casein being most readily hydrolysed.5. Proteins modified by performic acid oxidation, by cross-linking using 1,6-di-iso-cyanatohexane or by diazotization were labelled with14C. Performic acid treatment generally increased the susceptibility of proteins to digestion, so that the rates of hydrolysis of performic acid-treated proteins were more comparable than those of the unmodified proteins. Cross-linking resulted in a decreased rate of hydrolysis except with the insoluble proteins, hide powder azure and elastin congo red. Diazotization had little effect on the rate of hydrolysis of lactoglobulin and albumin, but inhibited casein hydrolysis and stimulated the breakdown of γ-globulin.

scholarly journals Evaluation of the microbial population in ruminal fluid using real time PCR in steers treated with virginiamycin

2010 ◽  
Vol 55 (No. 7) ◽  
pp. 276-285 ◽  
Author(s):  
T.J. Guo ◽  
J.Q. Wang ◽  
D.P. Bu ◽  
K.L. Liu ◽  
J.P. Wang ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Rumen Fluid ◽  
Control Diet ◽  
Basal Diet ◽  
Ammonia Nitrogen ◽  
Control Treatment ◽  
Ruminal Fermentation ◽  
Ruminal Bacteria

The objective of the present study was to investigate the effects of virginiamycin (VM ) supplementation on ruminal fermentation and microbial populations in steers. Four ruminally cannulated Chinese Luxi steers (BW 559.4 &plusmn; 30.1 kg) were used in a crossover design experiment with an experimental period of 28 days. The forage to concentrate ratio of the basal diet was 35:65 on dry matter basis. The experiment consisted of control treatment and treatment with control diet plus VM at a dose of 30 mg/kg concentrate (DM basis). Rumen fluid was collected at 07:30 prefeeding, at 11:30 and 17:30 postfeeding on day 27 and 28. A part of the pooled sample from rumen fluid was transferred to anaerobic culture by a roll-tube technique and analysed for species-specific real-time PCR quantification. The remaining pooled rumen fluid sample was analyzed for pH, VFA, ammonia N and l-lactic acid. The results showed that VM increased the ruminal pH (6.70 vs. 6.63; P &lt; 0.05), but it decreased ammonia nitrogen (4.94 vs. 6.19 mg/100 ml; P &lt; 0.01) and mean counts of amylolytic bacteria and proteolytic bacteria (P &lt; 0.01) as compared to the control. The additive VM did not affect the l-lactic acid concentration (1.39 vs. 1.26 mmol/l) in rumen fluid. Compared to the control, the steers receiving VM have altered a trend of quantification of Selenomonas ruminantium, Anaerovibrio lipolytica, Ruminococcus albus and Streptococcus bovis in rumen fluid (0.05&lt;p&lt;0.1) as compared to the control. However, VM had no significant effect on Lactobacillus spp. (P = 0.41), Butyrivibrio fibrisolvens (P = 0.35), on the genus Ruminococcus (P = 0.25), Ruminococcus flavefaciens (P = 0.52), Prevotella ruminicola (P = 0.54), on the genus Prevotella (P = 0.67) and Megasphaera elsdenii (P = 0.97). In this study, we found that VM had selective effects on ruminal bacteria and influenced ruminal fermentation by changing a part of the specific ruminal bacteria populations.

scholarly journals Metatranscriptomic Analysis of Sub-Acute Ruminal Acidosis in Beef Cattle

Animals ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 232 ◽  
Author(s):  
Ibukun Ogunade ◽  
Andres Pech-Cervantes ◽  
Hank Schweickart
Keyword(s):  
Beef Cattle ◽  
Rumen Fluid ◽  
Control Diet ◽  
Bacterial Biofilm ◽  
Ruminal Bacteria ◽  
Subacute Ruminal Acidosis ◽  
Rumen Microbiota ◽  
Ruminal Acidosis ◽  
Rrna Sequencing ◽  
Illumina Hiseq4000

Subacute ruminal acidosis (SARA) is a metabolic disease of ruminants characterized by low pH, with significant impacts on rumen microbial activity, and animal productivity and health. Microbial changes during subacute ruminal acidosis have previously been analyzed using quantitative PCR and 16S rRNA sequencing, which do not reveal the actual activity of the rumen microbial population. Here, we report the functional activity of the rumen microbiota during subacute ruminal acidosis. Eight rumen-cannulated Holstein steers were assigned randomly to acidosis-inducing or control diet. Rumen fluid samples were taken at 0, 3, 6, and 9 h relative to feeding from both treatments on the challenge day. A metatranscriptome library was prepared from RNA extracted from the samples and the sequencing of the metatranscriptome library was performed on Illumina HiSeq4000 following a 2 × 150 bp index run. Cellulolytic ruminal bacteria including Fibrobacter succinogenes, Ruminococcus albus, and R. bicirculans were reduced by an induced acidotic challenge. Up to 68 functional genes were differentially expressed between the two treatments. Genes mapped to carbohydrate, amino acid, energy, vitamin and co-factor metabolism pathways, and bacterial biofilm formation pathways were enriched in beef cattle challenged with sub-acute acidosis. This study reveals transcriptionally active taxa and metabolic pathways of rumen microbiota during induced acidotic challenge.

On the role of higher plant and microbial lipases in the ruminal hydrolysis of grass lipids

1977 ◽  
Vol 38 (2) ◽  
pp. 225-232 ◽  
Author(s):  
R. M. C. Dawson ◽  
N. Hemington ◽  
G. P. Hazlewood
Keyword(s):  
Rumen Fluid ◽  
Higher Plant ◽  
Microbial Enzymes ◽  
Heat Treated ◽  
Ruminal Degradation ◽  
Complex Lipids ◽  
Microbial Lipases ◽  
Micro Organisms ◽  
Hydrolysis Of

1. The galactolipids of heat-treated, 14C-labelled rye grass S24 administered intraruminally to a sheep fed on an autoclaved diet were rapidly catabolized.2. When grass was homogenized with rumen contents devoid of higher plant lipases the grass galactolipids were rapidly metabolized, but were not metabolized when the rumen contents were boiled to destroy microbial galactolipases.3. 14C-labelled monogalactosyldiglyceride, digalactosyldiglyceride and triolein were metabolized, with the release of 14C-labelled fatty acids when incubated with a homogenate (100 g/l) of grass or clover in rumen fluid from a starved sheep, but not when the rumen fluid was heat-treated to destroy microbial enzymes.4. It is concluded that in the sheep the lipases of rumen micro-organisms play a major part in the ruminal degradation of ingested complex lipids of pasture.

scholarly journals Howardella ureilytica gen. nov., sp. nov., a Gram-positive, coccoid-shaped bacterium from a sheep rumen

2007 ◽  
Vol 57 (12) ◽  
pp. 2940-2945 ◽  
Author(s):  
Anthony R. Cook ◽  
Paul W. Riley ◽  
Heather Murdoch ◽  
Paul N. Evans ◽  
Ian R. McDonald
Keyword(s):  
New Genus ◽  
Gene Sequence ◽  
Rumen Fluid ◽  
Sequence Divergence ◽  
Major Fatty Acid ◽  
Rrna Gene ◽  
Gram Positive ◽  
Gene Sequence Analysis ◽  
Sheep Rumen ◽  
Hydrolysis Of

An unidentified obligately anaerobic, fastidious, Gram-positive, non-motile, non-spore-forming, non-fermentative coccoid-shaped bacterium (designated strain GPC 589T) was isolated from the rumen fluid of a sheep. The major fatty acid constituents (>5 %) were C16 : 0 (29.2 %), C18 : 0 (40.7 %) and an unidentified compound (19.7 %) with an equivalent chain-length of 13.523. The G+C content of the DNA was 34 mol%. The organism was strongly ureolytic and generated ATP through the hydrolysis of urea. Comparative 16S rRNA gene sequence analysis demonstrated that strain GPC 589T was far removed, phylogenetically, from the ruminococci and related Gram-positive anaerobic cocci but exhibited a phylogenetic association with Clostridium rRNA cluster XIVa [as defined by Collins, M. D., Lawson, P. A., Willems, A., Cordoba, J. J., Fernandez-Garayzabal, J., Garcia, P., Cai, J., Hippe, H. & Farrow, J. A. E. (1994). Int J Syst Bacteriol 44, 812–826]. Sequence divergence values of 12.5 % or more were observed between strain GPC 589T and all other recognized species within this and related rRNA clostridial clusters. Phylogenetic analysis showed that strain GPC 589T represents a new genus within cluster XIVa. On the basis of both phylogenetic and phenotypic evidence, it is proposed that strain GPC 589T should be classified as representing a new genus and novel species, Howardella ureilytica gen. nov., sp. nov. The type strain is strain GPC 589T (=DSM 15118T=JCM 13267T).

scholarly journals Impact of Cadmium Intoxication on Health Status, Rumen and Blood Constituents in Egyptian Ossimi Sheep

2021 ◽  
Vol 10 (2) ◽  
pp. 102-106
Keyword(s):  
Drinking Water ◽  
Health Status ◽  
Rumen Fluid ◽  
Animal Health ◽  
Ammonia Nitrogen ◽  
Strong Positive Correlation ◽  
Toxic Heavy Metals ◽  
Animal Blood ◽  
Antioxidant Markers ◽  
The Impact

Toxic heavy metals particularly Cadmium (Cd) have a hazardous impact on animal health and productivity because of their ill-degradability and bio-accumulation for long periods. This study carried out on 52 Ossimi sheep belonging to Giza Governorate, including 12 sheep considered as control (kept in a private farm) and 40 sheep grazed on an area polluted with cadmium. Drinking water, animal blood and rumen fluid samples were collected from all sheep. Complete blood, rumen and serum constituents were analyzed. Iron, copper, zinc, oxidant, and antioxidant markers were evaluated and Cd levels in water, rumen fluid and serum were investigated to show the impact of Cd on those parameters. Physical examination revealed significant disturbance in health status of Cd-exposed sheep. Rumen fluid examination showed significant increase in rumen pH, significant decrease in rumen ammonia-nitrogen, TVFAs, AST, ALT, GGT, Ca and Ph. Blood constituent revealed significant alteration as significant decrease in RBCs count, Hb, PCV, MCHC and TLC as well as impaired hepatic and renal function and significant decrease of antioxidant markers. These alterations associated with strong positive correlation between these altered parameters and cadmium level in drinking water, rumen fluid and blood samples which markedly increased more than permissible limits; these results should be put in consideration in interpretation of affected animals' status and during treatment and control of cadmium exposed sheep cases as well as consumption of such animals' meat and offal is not recommended.

scholarly journals Diet dominates age in shaping the rumen bacteria community and function in dairy cattle

2021 ◽  
Author(s):  
Yangyi Hao ◽  
Yue Gong ◽  
Shuai Huang ◽  
Shoukun Ji ◽  
Wei Wang ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Rumen Fluid ◽  
Variation Partitioning ◽  
Rumen Bacteria ◽  
Ruminal Bacteria ◽  
Linear Discriminant ◽  
High Fiber ◽  
Fiber Diet ◽  
Α Diversity ◽  
And Function

Abstract Background To understand the effects of diet and age on the rumen bacterial community and function, forty-eight dairy cattle at 1.5 (M1.5), 6 (M6), 9 (M9), 18 (M18), 23 (M23), and 27 (M27) months old were selected. The M1.5, M6, and M27 had the high protein and starch dietary, while the M9, M18, and M23 had the high fiber dietary. Fermentation profile, enzyme activity, and bacteria community in rumen fluid were measured. Results The acetate to propionate ratio (A/P) at M9, M18, and M23 (high fiber diet) was higher than other ages, and M6 was the lowest (P < 0.05). The total volatile fatty acid (TVFA) at M23 and M27 was higher than other ages (P < 0.05). The urease at M18 was lower than M1.5, M6, and M9, and the xylanase at M18 was higher than M1.5, M23, and M27 (P < 0.05). The α-diversity indexes (Ace and Chao1) of ruminal bacteria increased from M1.5 to M23, while they decreased from M23 to M27 (P < 0.05). Thirty-three bacteria were identified as biomarkers of the different groups based on the linear discriminant analysis (LDA) when the LDA score > 4. The variation partitioning approach analysis showed that the age and diet had a 7.98% and 32.49% contribution to the rumen bacteria community variation, respectively. The richness of Succinivibrionaceae_UCG-002 and Fibrobacter were positive correlated with age (r > 0.60, P < 0.01) and also positively correlated with TVFA and acetate (r > 0.50, P < 0.01). The Lachnospiraceae_AC2044_group, Pseudobutyrivibrio, and Saccharofermentans has a positive correlation (r > 0.80, P < 0.05) with diet NDF and negative correlation (r < -0.80, P < 0.05) with diet CP and starch, which were also positively correlated with the acetate and A/P (r > 0.50, P < 0.01). Conclusion These findings indicated that the quantitative effect of diet and age on the rumen bacteria were 7.98% and 32.49%, respectively. The genera of Lachnospiraceae_AC2044_group, Pseudobutyrivibrio, and Saccharofermentans could be worked as the target bacteria to modulate the rumen fermentation by diet; meanwhile, the high age-correlated bacteria such as Succinivibrionaceae_UCG-002 and Fibrobacter also should be considered when shaping the rumen function.

scholarly journals Diversity and Composition of Rumen Bacteria, Fungi, and Protozoa in Goats and Sheep Living in the Same High-Altitude Pasture

Animals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 186 ◽  
Author(s):  
Suo Langda ◽  
Chenguang Zhang ◽  
Ke Zhang ◽  
Ba Gui ◽  
De Ji ◽  
...  
Keyword(s):  
High Altitude ◽  
Volatile Fatty Acid ◽  
Cellulose Degradation ◽  
Rumen Fluid ◽  
Gene Sequencing ◽  
Alpha Diversity ◽  
Total Volatile ◽  
Ruminal Bacteria ◽  
The Family ◽  
Bacteria And Fungi

Environmental adaptation of ruminants was highly related to microbiota in the rumen. To investigate the diversity and composition of bacteria, fungi, and protozoa in the rumen of high-altitude animals, amplicon gene sequencing was performed using rumen fluid samples derived from both Tibetan goats and sheep at the same pasture in a highland (altitude > 4800 m). Between these two species, the ruminal bacteria and fungi were significantly different at multiple taxonomic levels. The alpha diversity of bacteria was significantly high in goats (p < 0.05). One hundred and sixty-four and 29 Operational Taxonomy Units (OTUs) with significant differences were detected in bacteria and fungi, respectively. The abundance of bacteria, fungi, and protozoa in the rumen was characterized at multiple taxonomic levels, and we determined that Firmicutes, Bacteroidetes, Neocallimastigomycota, and Ciliophora were the most abundant bacteria, fungi, and protozoa. The family Neocallimastigaceae and the genus Metadinium had cellulose degradation capacity in the rumen with high abundance, thereby, suggesting that fungi and protozoa played an essential role in rumen fermentation. In addition, by comparing microbiota in the rumen of goats and sheep it was found, that the fiber-degrading fungi genus (Cyllamyces) was increased in the rumen of sheep (p < 0.05) whereas VFA-producing bacteria (Saccharofermentans and Lachnospiraceae_XPB1014) were increased in the rumen of goats (p < 0.05). Interestingly, in the rumen, no differences in protozoa were observed between goats and sheep (p > 0.05). Furthermore, when compared to sheep, level of acetic acid, propionic acid, and total volatile fatty acid (TVFA) were significantly increased in the rumen of goats (p < 0.05). Taken together, these results suggested microbiota in the rumen drive goats to better adapt to high-altitude grazing conditions.

METABOLISM OF SELENOMETHIONINE IN THE RUMEN

1974 ◽  
Vol 54 (3) ◽  
pp. 325-330 ◽  
Author(s):  
K. J. JENKINS ◽  
M. HIDIROGLOU ◽  
J. E. KNIPFEL
Keyword(s):  
Ion Exchange ◽  
Rumen Fluid ◽  
In Vitro Study ◽  
Exchange Chromatography ◽  
Elemental Selenium ◽  
Bacterial Protein ◽  
Time Intervals ◽  
Ruminal Bacteria ◽  
Enzymatic Hydrolysate

Five rumen-fistulated wethers were dosed intraruminally with a single dose of 75Se-selenomethionine, and the rumen liquor was collected after various time intervals and separated into bacterial, cell-free, and protozoa plus plant material fractions. Maximum 75Se activity in rumen fluid was observed 2 h after labelled selenomethionine was administered. At 6 h post-dosing, 50% of the rumen liquor label was in the bacterial fraction, decreasing to 20% at 96 h. The majority of bacterial 75Se activity was found to be protein-bound. In a second experiment, five wethers were given 75Se-selenomethionine intraruminally, and 2 h later ruminal bacteria were isolated for identification of selenocompounds in the bacterial protein. Paper chromatographic separation of protein enzymatic hydrolysate showed the presence of selenocystine, selenomethionine, elemental selenium, and 40–50% 75Se activity as unidentifiable components. In an in vitro study, 75Se-selenomethionine was incubated with a rumen bacterial fraction, and ion-exchange and paper chromatography were used to identify selenocompounds formed. The results indicated that 75Se-selenomethionine was metabolized by the bacteria to 75Se-selenocystine, with both selenoamino acids incorporated into bacterial protein. With ion-exchange chromatography, unless carrier selenomethionine was added, 75Se-selenomethionine was degraded and a high proportion of 75Se activity remained on the column.

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