Red seaweed (Asparagopsis taxiformis) supplementation reduces enteric methane by over 80 percent in beef steers.

The red macroalgae (seaweed) Asparagopsis spp. has shown to reduce ruminant enteric methane (CH4) production up to 99% in vitro. The objective of this study was to determine the effect of Asparagopsis taxiformis on CH4 production (g/day per animal), yield (g CH4/kg dry matter intake (DMI)), and intensity (g CH4/kg ADG); average daily gain (ADG; kg gain/day), feed conversion efficiency (FCE; kg ADG/kg DMI), and carcass and meat quality in growing beef steers. Twenty-one Angus-Hereford beef steers were randomly allocated to one of three treatment groups: 0% (Control), 0.25% (Low), and 0.5% (High) A. taxiformis inclusion based on organic matter intake. Steers were fed 3 diets: high, medium, and low forage total mixed ration (TMR) representing life-stage diets of growing beef steers. The Low and High treatments over 147 days reduced enteric CH4 yield 45 and 68%, respectively. However, there was an interaction between TMR type and the magnitude of CH4 yield reduction. Supplementing low forage TMR reduced CH4 yield 69.8% (P <0.01) for Low and 80% (P <0.01) for High treatments. Hydrogen (H2) yield (g H2/DMI) increased (P <0.01) 336 and 590% compared to Control for the Low and High treatments, respectively. Carbon dioxide (CO2) yield (g CO2/DMI) increased 13.7% between Control and High treatments (P = 0.03). No differences were found in ADG, carcass quality, strip loin proximate analysis and shear force, or consumer taste preferences. DMI tended to decrease 8% (P = 0.08) in the Low treatment and DMI decreased 14% (P <0.01) in the High treatment. Conversely, FCE tended to increase 7% in Low (P = 0.06) and increased 14% in High (P <0.01) treatment compared to Control. The persistent reduction of CH4 by A. taxiformis supplementation suggests that this is a viable feed additive to significantly decrease the carbon footprint of ruminant livestock and potentially increase production efficiency.

Effects of transport stress on pathological injury and expression of main heat shock proteins in the caprine stomach.

BACKGROUND: Transportation is necessary to introduce new breeds of goats to the farm and move the adult meat goat from the farm to the slaughterhouse. However, these actions may give rise to transport stress. Heat shock proteins (HSPs) are playing some important regulate roles during transport stress. The aim of this study was to evaluate the effects of transport stress on the pathological injury and HSPs expression in the stomach of goats. A total of three batches of Ganxi goats from western Jiangxi province were enrolled in this study. For each batch, twelve healthy adult male goats were randomly divided into three groups (four goats per batch and per group): Control group, stress group transported during 2 h and stress group transported during 6 h. RESULTS: Our results showed that the different degrees of stomach walls damage, with the change of expression levels of heat shock protein 27 (HSP27), heat shock protein 70 (HSP70) and heat shock protein 90 (HSP90), occurred after goats transportation. In rumen, the mRNA and protein expressions of HSP27 and HSP70 were increased after transport stress, but not HSP90. In reticulum, all three HSPs mRNA and protein levels were upregulated after 2 h transport, but decreased after 6 h transport. In omasum, HSP27 and HSP70 mRNA and protein were increased after transport stress, however, HSP90 mRNA level only had a slightly enhancement after transport stress. In abomasum, HSP70 and HSP90 mRNA and protein levels were increased after transport stress, but HSP27 was decreased after transport stress. CONCLUSIONS: Taken together, these results revealed that the pathological changes in the gastric tissues and the stomach HSPs expression in goats are related to transport stress and duration. Moreover, this study also provides some new data to advocate reducing transport stress of goats and improving animal welfare.

Enteric methane emission from Jersey cows during the spring transition from indoor feeding to grazing.

Organic dairy cows in Denmark are often kept indoors during the winter and outside at least part time in the summer. Consequently, their diet changes by the season. We hypothesized that grazing might affect enteric CH4 emissions due to changes in the nutrition, maintenance, and activity of the cows, and they might differentially respond to these factors. This study assessed the repeatability of enteric CH4 emission measurements for Jersey cattle in a commercial organic dairy herd in Denmark. It also evaluated the effects of a gradual transition from indoor winter feeding to outdoor spring grazing. Further, it assessed the individual-level correlations between measurements during the consecutive feeding periods (phenotype × environment, P × E) as neither pedigrees nor genotypes were available to estimate a genotype by environment effect. Ninety-six mixed-parity lactating Jersey cows were monitored for 30 d before grazing and for 24 d while grazing. The cows spent 8 to 11 h grazing each day and had free access to an in-barn automatic milking system (AMS). For each visit to the AMS, milk yield was recorded and logged along with date and time. Monitoring equipment installed in the AMS feed bins continuously measured enteric CH4 and CO2 concentrations (ppm) using a noninvasive "sniffer" method. Raw enteric CH4 and CO2 concentrations and their ratio (CH4:CO2) were derived from average concentrations measured during milking and per day for each cow. We used mixed models equations to estimate variance components and adjust for the fixed and random effects influencing the analyzed gas concentrations. Univariate models were used to precorrect the gas measurements for diurnal variation and to estimate the direct effect of grazing on the analyzed concentrations. A bivariate model was used to assess the correlation between the 2 periods (in-barn vs. grazing) for each gas concentration. Grazing had a weak P × E interaction for daily average CH4 and CO2 gas concentrations. Bivariate repeatability estimates for average CH4 and CO2 concentrations and CH4:CO2 were 0.77 to 0.78, 0.73 to 0.80, and 0.26, respectively. Repeatability for CH4:CO2 was low (0.26) but indicated some between-animal variation. In conclusion, grazing does not create significant shifts compared with indoor feeding in how animals rank for average CH4 and CO2 concentrations and CH4:CO2. We found no evidence that separate evaluation is needed to quantify enteric CH4 and CO2 emissions from Jersey cows during in-barn and grazing periods.

Bioactive polysaccharides and oligosaccharides as possible feed additives to manipulate rumen fermentation in Rusitec fermenters.

Bioactive polysaccharides and oligosaccharides (POSs) have been used as safe non-antibiotic alternatives in animal diets; however, only limited data are available on their effects in rumen manipulation. This study was undertaken to systematically evaluate and compare the efficacies of six different bioactive POSs (10g/kg diet) with respect to ruminal fermentation, feed digestibility, biohydrogenation and bacterial population using a rumen simulation technique. The experiment consisted of 4 independent 12-d incubation periods (each including 8 d for acclimation, followed with 4 d for sample collection). The results demonstrated that Lycium barbarum polysaccharide had no effect and Astragalus polysaccharide only increased the total VFA production. Chitooligosaccharide decreased the propionate proportion with a corresponding increase of acetate to propionate ratio. Fructooligosaccharide increased fatty acid biohydrogenation and the abundances of R. albus, B. fibrisolvens and S. ruminantium. Both Lentinan polysaccharide and xylooligosaccharide increased the total VFA production, fatty acid biohydrogenation and the abundance of B. fibrisolvens. In addition, Lentinan polysaccharide increased 48-h dietary protein degradation and the abundance of R. flavefaciens, while xylooligosaccharide increased 24-h dietary fiber degradation and the abundance of R. albus. In conclusion, bioactive POSs, especially Lentinan polysaccharide and xylooligosaccharide, can serve as potential feed additives to manipulate rumen fermentation.

Abomasal dysfunction and cellular and mucin changes during infection of sheep with larval or adult Teladorsagia circumcincta.

This is the first integrated study of the effects on gastric secretion, inflammation and fundic mucins after infection with L3 T. circumcincta and in the very early period following transplantation of adult worms. At 3 months-of-age, 20 Coopworth lambs were infected intraruminally with 35,000 L3; infected animals were killed on Days 5, 10, 15, 20 and 30 post-infection and 6 controls on either Day 0 or 30 post-infection. Another 15 Romney cross lambs received 10,000 adult worms at 4-5 months-of-age though surgically-implanted abomasal cannulae and were killed after 6, 12, 24 and 72 hours; uninfected controls were also killed at 72 hours. Blood was collected at regular intervals from all animals for measurement of serum gastrin and pepsinogen and abomasal fluid for pH measurement from cannulated sheep. Tissues collected at necropsy were fixed in Bouin's fluid for light microscopy, immunocytochemistry and mucin staining and in Karnovsky's fluid for electron microscopy. Nodules around glands containing developing larvae were seen on Day 5 p.i., but generalised effects on secretion occurred only after parasite emergence and within hours after transplantation of adult worms. After L3 infection, there were maximum worm burdens on Days 10-15 post-infection, together with peak tissue eosinophilia, inhibition of gastric acid secretion, hypergastrinaemia, hyperpepsinogenaemia, loss of parietal cells, enlarged gastric pits containing less mucin and increased numbers of mucous neck cells. After adult transplantation, serum pepsinogen was significantly increased after 9 hours and serum gastrin after 18 hours. Parallel changes in host tissues and the numbers of parasites in the abomasal lumen suggest that luminal parasites, but not those in the tissues, are key drivers of the pathophysiology and inflammatory response in animals exposed to parasites for the first time. These results are consistent with initiation of the host response by parasite chemicals diffusing across the surface epithelium, possibly aided by components of ES products which increased permeability. Parietal cells appear to be a key target, resulting in secondary increases in serum gastrin, pit elongation, loss of surface mucins and inhibition of chief cell maturation. Inflammation occurs in parallel, and could either cause the pathology or exacerbate the direct effects of ES products.

[The Patterns of Behavior of Radioactive Particles in the Food Chain of Cattle and Transport in the Gastrointestinal Tract of Animals].

The analysis of the patterns of behavior of polydisperse radioactive silicate particles in the components of the food chain of cattle is presented. It is shown that the composition of the size distribution of radioactive particles taken into animal organisms differs from the original composition of the particles deposited on the surface of pasture vegetation, and from dispersion of the particles in the aboveground biomass of vegetation at the time of grazing. The intake of particles into animal organisms is reduced with the increase of their size, and for the particle fraction of 400-800 microns it is about 10 times less than for the fine fraction (< 100 microns). The mathematical compartment model ofthe transport of polydisperse radioactive particles in the digestive tract of cattle has been developed. It is found that the elimination rate of radioactive particles from the animal organism depends on their sizes. Deposition of particles on the fundic surface of the wall ventral sac of rumen and reticulum as well as their long stay in comparison with the chyme in abomasum was noted. The maximum levels of irradiation are formed in these parts of the digestive tract of cattle.

Anaerobic digestion of corn stovers for methane production in a novel bionic reactor.

To improve the biogas production from corn stovers, a new bionic reactor was designed and constructed. The bionic reactor simulated the rumen digestion of ruminants. The liquid was separated from corn stovers and refluxed into corn stovers again, which simulated the undigested particles separated from completely digested materials and fed back again for further degradation in ruminant stomach. Results showed that the bionic reactor was effective for anaerobic digestion of corn stovers. The liquid amount and its reflux showed an obvious positive correlation with biogas production. The highest biogas production rate was 21.6 ml/gVS-addedd, and the total cumulative biogas production was 256.5 ml/gVS-added. The methane content in biogas ranged from 52.2% to 63.3%. The degradation of corn stovers were greatly enhanced through simulating the animal digestion mechanisms in this bionic reactor.

Intravenous lipopolysaccharide challenge alters ruminal bacterial microbiota and disrupts ruminal metabolism in dairy cattle.

In the present study, three primiparous lactating Holstein cows (260-285 d in lactation) were used in a 3 × 3 Latin square design to assess the effects of three doses (0.0, 0.4 and 0.8 µg/kg body weight) of lipopolysaccharide (LPS, Escherichia coli 0111:B4) on changes in ruminal microbiota and ruminal fermentation. Ruminal pH was linearly decreased (P< 0.001) by LPS challenge, and the concentrations of acetate, propionate, butyrate, total volatile fatty acids and amino N increased linearly (P< 0.001) according to the LPS dose. LPS infusion linearly decreased (P< 0.001) the organic matter degradability of alfalfa hay and soyabean meal in the rumen, but did not affect (P>0.10) the gene expression of Na⁺/K⁺-ATPase and monocarboxylic acid transporter-1, -2 and -4. A plot of principal coordinate analysis based on unweighted UniFrac values and analysis of molecular variance revealed that the structure of ruminal bacterial communities in the control was distinct from that of the ruminal microbiota in the cattle exposed to LPS. At the phylum level, when compared with the control group, LPS infusion in the tested cows linearly increased (P< 0.05) the abundance of Firmicutes, and linearly decreased (P< 0.05) the percentage of Bacteroidetes, Tenericutes, Spirochaetes, Chlorobi and Lentisphaerae. To our knowledge, this is the first study to report that intravenously LPS challenge altered the ruminal bacterial microbiota and fermentation profiles. The present data suggest that systemic LPS could alter ruminal environment and ruminal microbiota composition, leading to a general decrease in fermentative activity.

Effect of pH on conjugated linoleic acid (CLA) formation of linolenic acid biohydrogenation by ruminal microorganisms.

Conventional beliefs surrounding the linolenic acid (LNA; cis-9 cis-12 cis-15 C18:3) biohydrogenation (BH) pathway propose that it converts to stearic acid (SA) without the formation of conjugated linoleic acid (CLA) as intermediate isomers. However, an advanced study (Lee and Jenkins, 2011) verified that LNA BH yields multiple CLAs. This study utilized the stable isotope tracer to investigate the BH intermediates of (13)C-LNA with different pH conditions (5.5 and 6.5). The (13)C enrichment was calculated as a (13)C/(12)C ratio of labeled minus unlabeled. After 24 h, eight CLA isomers were significantly enriched on both pH treatment, this result verifies that these CLAs originated from (13)C-LNA BH which supports the results of Lee and Jenkins (2011). The enrichment of cis-cis double bond CLAs (cis-9 cis-11 and cis-10 cis-12 CLA) were significantly higher at low pH conditions. Furthermore, the concentration of cis-10 cis-12 CLA at low pH was four times higher than at high pH conditions after a 3 h incubation. These differences support the LNA BH pathways partial switch under different pH conditions, with a strong influence on the cis-cis CLA at low pH. Several mono-, di-, and tri-enoic fatty acid isomers were enriched during 24 h of incubation, but the enrichment was decreased or restricted at low pH treatment. Based on these results, it is proposed that low pH conditions may cause a changed or limited capacity of the isomerization and reduction steps in BH.

Net transfer of nutrients to the duodenum and disappearance of n-alkanes in the reticulo-rumen and the hindgut of sheep fed grass/legume combinations.

An experiment was carried out to examine the effect of increasing the proportion of Wimmera ryegrass hay in a lucerne hay-based diet on net transfer of nutrients to the intestine, and on the disappearance of n-alkanes in the reticulo-rumen and the hindgut of sheep. Following a latin square design, four adult ewes were fed 1:0, 0·33:0·67, 0·67:0·33 and 0:1 proportions of legume and grass. Increasing the proportion of ryegrass in the diet linearly decreased the intake of DM (P= 0·017), organic matter (P= 0·021) and N (P= 0·001). However, neutral-detergent fibre intake was not affected (P= 0·148), nor was its digestibility coefficient (P>0·10). Diet had no effect on duodenal flows of nutrients (P>0·10), although the proportion of N intake (NI) recovered at the duodenum as non-NH3 N (NAN) increased linearly with Lolium rigidum in the diet (P= 0·002). Full recovery of NI as NAN was achieved at NH3 concentrations in the rumen below 110 g/l. Microbial N contribution to NAN varied in a quadratic manner (P< 0·05) with the proportion of grass in the diet, although efficiency of microbial synthesis was not affected (P>0·10). Duodenal recovery of consumed n-alkanes was not affected by diet and was complete for those present in higher concentrations in the forages. Isolated rumen bacteria contained significant amounts of n-alkanes, contributing to the duodenal flow of these hydrocarbons in variable proportions depending on the diet consumed.

Ruminal degradation of soybean, canola and cottonseed meal using In sacco procedure in sheep.

This research was conducted in order to investigate rumen degradability of some factors includes; Dray Matter (DM), Organic Matter (OM), Crude Protein (CP), Acid Detergent Fiber (ADF) in three different plant protein supplements includes; soybean, canola and cottonseed meal. The experiment was carried out using in three castrated and fistulated male Zel sheep. Each feedstuff was weighed into duplicate nylon bags and incubated in each of the three rumen fistulated sheep for 0, 2, 4, 8, 16, 24 and 48 h. Results revealed that effective degradability of DM of soybean, canola and cottonseed meal were 55.8, 73.8 and 48.5%, respectively. Effective degradabilities of the CP in feedstuffs were 55.8, 62 and 48.3% for the respective feedstuffs. Effective degradabilities of the OM were 55.7, 56.4 and 47.4%, respectively. Results also showed that effective degradabilities of the ADF were 55, 56.4 and 37.6, respectively. According to the results the researchers concluded that canola and soybean were more degradable in the rumen of the sheep while cottonseed meal were less degradable and, hence resulted in higher rumen undegradable protein.

Sulfated cholecystokinin-8 increases ghrelin secretion but does not affect oxyntomodulin in Holstein steers.

The effect of appetite regulatory hormone cholecystokinin (CCK) on the secretions of oxyntomodulin (OXM) and ghrelin, and the effect of ghrelin on the secretions of CCK and OXM were studied in ruminants. Eight Holstein steers, 7 months old, 243 ± 7 kg body weight (BW), were arranged in an incomplete Latin square design (8 animals × 4 treatments × 4 days of sampling). Steers were intravenously injected with 10 µg of sulfated CCK-8/kg BW, 20 µg of acyl ghrelin/kg BW, 100 µg of des-acyl ghrelin/kg BW or vehicle. Blood samples were collected from -60 min to 120 min relative to time of injection. Plasma concentrations of ghrelin, sulfated CCK and OXM were measured by double-antibody radioimmunoassay. Plasma acyl ghrelin was increased to peak level (428.3 ± 6 pg/mL) at 60 min after injection of CCK compared with pre-injected levels (203.3 ± 1 pg/mL). These results showed for the first time, that intravenous bolus injection of CCK increased ghrelin secretion in ruminants. In contrast, injection of ghrelin did not change CCK secretion. Administration of ghrelin or CCK has no effect on plasma OXM concentrations. In conclusion, our results show that administration of CCK increased ghrelin secretion but did not affect OXM release in ruminants. Ghrelin did not affect the secretions of CCK and OXM.

Effect of replacing corn with brown rice in a total, mixed, ration silage on milk production, ruminal fermentation and nitrogen balance in lactating dairy cows.

Nine multiparous Holstein cows were used in a replicated 3 × 3 Latin square design to determine the effects of substituting corn grain with brown rice (BR) grain in total mixed ration (TMR) silage on milk yield, ruminal fermentation and nitrogen (N) balance. The TMR silages were made from the ensiling of TMR containing (dry matter basis) 50.1% forage in rice silage and corn silage combination, and 49.9% concentrate. The grain portion of the diets contained 31.2% steam-flaked corn, 31.2% steam-flaked BR or an equal mixture of corn and BR. Dietary treatments did not affect dry matter intake, milk yield and milk fat, protein and lactose yields. The ruminal pH and total volatile fatty acid concentrations were not affected by dietary treatment. The urinary N excretion decreased linearly (P < 0.01) in response to increased levels of BR, with no dietary effect on N intake, N secretion in milk and fecal N excretion. Our results indicate that steam-flaked BR is a suitable replacement for steam-flaked corn in dairy cow diets, and that it can be included in rations to a level of at least 31.2% of dry matter without adverse effects on milk production, when cows were fed rice silage and corn silage-based diets.

Distribution of acute phase proteins in the bovine forestomachs and abomasum.

Acute phase proteins (APPs) are produced mainly by the liver and their concentration is increased during the systemic inflammatory response. Expression of haptoglobin (Hp), serum amyloid A (SAA), lipopolysaccharide-binding protein (LBP) and α-1 acid glycoprotein (AGP) was determined in the mucosa of the normal bovine forestomachs and abomasum by qualitative and quantitative reverse transcriptase-PCR for mRNA and by Western blot analysis and immunohistochemistry for proteins. Although expression of SAA mRNA was evident in the forestomachs and abomasum, SAA protein was identified only in the abomasum. Expression of Hp protein was high in the forestomachs and abomasum, even though expression of Hp mRNA was negligible. The main site of expression of LBP mRNA was the omasum, whereas the highest protein expression was evident in the abomasum. AGP was expressed at low levels in the bovine forestomachs. Western blot analysis revealed a heterogeneous electrophoretic pattern for AGP, LBP and Hp, indicating that different stomach compartments produce isoforms that are different to those expressed by the liver. Expression of APPs by the bovine forestomachs and abomasum may contribute to regulation of the innate immune response against pathogens.

Identification of differentially expressed proteins in ruminal epithelium in response to a concentrate-supplemented diet.

Ruminal epithelium adapts to dietary change with well-coordinated alterations in metabolism, proliferation, and permeability. To further understand the molecular events controlling diet effects, the aim of this study was to evaluate protein expression patterns of ruminal epithelium in response to various feeding regimes. Sheep were fed with a concentrate-supplemented diet for up to 6 wk. The control group received hay only. Proteome analysis with differential in gel electrophoresis technology revealed that, after 2 days, 60 proteins were significantly modulated in ruminal epithelium in a comparison between hay-fed and concentrate-fed sheep (P < 0.05). Forty proteins were upregulated and 20 proteins were downregulated in response to concentrate diet. After 6 wk of this diet, only 14 proteins were differentially expressed. Among these, 11 proteins were upregulated and 3 downregulated. To identify proteins that were modulated by dietary change, two-dimensional electrophoresis was coupled with liquid chromatography electrospray ionization mass spectrometry. The differential expression of selected proteins, such as esterase D, annexin 5, peroxiredoxin 6, carbonic anhydrase I, and actin-related protein 3, was verified by immunoblotting and/or mRNA analysis. The identified proteins were mainly associated with functions related to cellular stress, metabolism, and differentiation. These results suggest new candidate proteins that may contribute to a better understanding of the signaling pathways and mechanisms that mediate rumen epithelial adaptation to high-concentrate diet.

Characterization of glucagon-like peptide 2 pathway member expression in bovine gastrointestinal tract.

Glucagon-like peptide 2 (GLP-2), secreted by enteroendocrine cells, has several physiological effects on the intestine of monogastric species, including promotion of growth of intestinal epithelium, reduction of epithelial cell apoptosis, and enhancement of intestinal blood flow, nutrient absorption, and epithelial barrier function. The regulatory functions of GLP-2 in the ruminant gastrointestinal tract (GIT) have not been well studied. The objectives of this investigation were to characterize the mRNA expression of 4 members of the GLP-2 pathway throughout the bovine GIT, including (1) proglucagon (GCG), the parent peptide from which GLP-2 is derived through cleavage by prohormone convertase; (2) prohormone convertase (PCSK1); (3) GLP-2 receptor (GLP2R); and (4) dipeptidyl peptidase IV (DPP4), the enzyme that inactivates GLP-2. Gene expression was evaluated in rumen, reticulum, omasum, abomasum, duodenum, jejunum, ileum, cecum, and rectum collected at slaughter from prepubertal heifers, mature cows in early, mid, and late lactation, and nonlactating cows (n=3 per stage) by a gene expression profiling assay. In addition, mRNA expression of 14 genes involved in nutrient transport, enzyme activity, blood flow, apoptosis, and proliferation were evaluated in the 9 GIT tissues for their association with GCG and GLP2R mRNA expression. Immunohistochemistry was used to localize GLP2R protein in tissues of the lower GIT. Results indicated that mRNA expression of GCG, PCSK1, GLP2R, and DPP4 varies across the 9 GIT tissues, with greatest expression in small and large intestines, and generally nondetectable levels in forestomachs. Expression of DPP4 and GLP2R mRNA varied by developmental stage or lactational state in intestinal tissues. Expression of GCG or GLP2R mRNA was correlated with molecular markers of proliferation, apoptosis, blood flow, enzyme activity, and urea transport, depending on the tissue examined, which suggests a potential for involvement of GLP-2 in these physiological processes in the ruminant GIT. The GLP2R protein was expressed in intestinal crypts of the bovine GIT, which is consistent with the distribution in monogastric species. Our findings support a functional role of the GLP-2 pathway in bovine GIT and the potential for use of GLP-2 as a therapy to improve intestinal function and nutrient absorption in ruminants.

Hepatic and extra-hepatic metabolic pathways involved in flubendazole biotransformation in sheep.

Flubendazole (FLBZ) is a broad-spectrum benzimidazole anthelmintic compound used in pigs, poultry and humans. Its potential for parasite control in ruminant species is under investigation. The objective of the work described here was to identify the main enzymatic pathways involved in the hepatic and extra-hepatic biotransformation of FLBZ in sheep. Microsomal and cytosolic fractions obtained from sheep liver and duodenal mucosa metabolised FLBZ into a reduced FLBZ metabolite (red-FLBZ). The keto-reduction of FLBZ led to the prevalent (approximately 98%) stereospecific formation of one enantiomeric form of red-FLBZ. The amounts of red-FLBZ formed in liver subcellular fractions were 3-4-fold higher (P<0.05) compared to those observed in duodenal subcellular fractions. This observation correlates with the higher (P<0.05) carbonyl reductase (CBR) activities measured in the liver compared to the duodenal mucosa. No metabolic conversion was observed following FLBZ or red-FLBZ incubation with sheep ruminal fluid. Sheep liver microsomes failed to convert red-FLBZ into FLBZ. However, this metabolic reaction occurred in liver microsomes prepared from phenobarbital-induced rats, which may indicate a cytochrome P450-mediated oxidation of red-FLBZ. A NADPH-dependent CBR is proposed as the main enzymatic system involved in the keto-reduction of FLBZ in sheep. CBR substrates such as menadione and mebendazole (a non-fluoride analogue of FLBZ), inhibited this liver microsomal enzymatic reaction, which may confirm the involvement of a CBR enzyme in FLBZ metabolism in sheep. This research is a further contribution to the understanding of the metabolic fate of a promissory alternative compound for antiparasitic control in ruminant species.

In vitro fermentation vessel type and method alter fiber digestibility estimates.

Selected vessel types and conditions used for in vitro fermentation were compared to evaluate their effects on determinations of neutral detergent fiber (NDF) digestibility (NDFD) in 2 replicate 48-h fermentations. Treatments included 50-mL polyethylene centrifuge tubes with gas-release valves (treatment 1); 50-mL polyethylene centrifuge tubes with continuous gassing with CO2 (treatment 2); 50-mL polyethylene centrifuge tubes sealed, oriented horizontally, and shaken continuously parallel to the long axis of the tube, with manual gas release (treatment 3); 125-mL Erlenmeyer flasks with continuous gassing with CO2 (treatment 4); and 125-mL serum vials sealed with stoppers and crimp seals with (treatment 5) or without (treatment 6) manual gas release. Goering and Van Soest medium and blended ruminal inoculum from 4 lactating cows were used. Substrates were alfalfa hay, corn silage, ryegrass hay, and soyhulls. Gas was released and measured in treatments 3 and 6 at 3.0, 5.5, 9.0, 11.5, 23.5, 29.5, and 47.5 h by using a syringe with a hypodermic needle. Vessels from each treatment were harvested at 0, 6, 12, 24, 30, and 48 h for NDF analysis, with NDFD calculated as 1 - [(residual NDF, g - residual NDF in fermentation blank, g)/sample NDF, g]. Medium pH did not decline below 6.3 for any treatment. Average values for NDFD for 24 through 48 h were 0.576, 0.639, 0.688, 0.668, 0.679, and 0.681 for treatments 1 through 6, respectively (standard error of the difference = 0.008). The lowest NDFD was noted for treatment 1, which differed from all other treatments; treatments 3, 4, 5, and 6 did not differ by treatment or by the interaction of treatment and substrate. Treatments 1 and 2 gave lower NDFD values than the other treatments, but these differences were not consistent and differed by substrate, with alfalfa showing the fewest differences among treatments and soyhulls the most. Net energy of lactation values for substrates, as predicted from differences in 48-h NDFD, were 7 to 15% lower for treatment 1 than for the average of all other treatments. Slopes of the gas production per gram of substrate dry matter curves differed between treatments 3 and 5. In conclusion, measured NDFD was altered by fermentation treatment, with polyethylene tubes + gas-release valves giving the lowest values. Consequently, NDFD values may not be comparable across fermentation methods, but the effect will vary among feedstuffs. The combination of methods used for sealing, gassing, or agitating vessels may have a greater impact on NDFD than does vessel type.

[The regulation of fatty acid metabolism and other lipid compounds in ruminant animals].

The review deals with peculiarities of molecular mechanisms of metabolism regulation of fatty acids and other lipid components in preruminant and ruminant animals (i.e. before and after formation of the functioning of the mixed microbial population in the rumen). A characteristic of possible biosynthesis regulation processes, transformation, transport, utilization and catabolism of fatty acids with different molecular masses and peculiarities and of triacylglycerols and other lipid substances in the liver compartments, skeletal and heart muscle cells has been shown. Peculiarities of intracellular changes in metabolism of fatty acids and apolipoprotein B in the liver of neonatal calves under development of alimentary enteropathology have been considered. Main factors in the mechanism of regulation of intracellular metabolism of fatty acids and lipid substances have been defined.

Assessment of the main metabolism pathways for the flukicidal compound triclabendazole in sheep.

Triclabendazole (TCBZ) is an halogenated benzimidazole (BZD) compound worldwide used to control immature and adult stages of the liver fluke Fasciola hepatica. The purpose of this investigation was to characterize in vitro the patterns of hepatic and ruminal biotransformation of TCBZ and its metabolites in sheep. TCBZ parent drug was metabolized into its sulphoxide (TCBZSO), sulphone (TCBZSO2) and hydroxy derivatives by sheep liver microsomes. The same microsomal fraction was also able to oxidize TCBZSO into TCBZSO2 and hydroxy-TCBZSO (HO-TCBZSO). TCBZ sulphoxidation was significantly (P < 0.001) inhibited after inactivation of the flavin-monooxygenase (FMO) system (77% inhibition) as well as in the presence of the FMO substrate methimazole (MTZ) (71% inhibition). TCBZ sulphoxidative metabolism was also reduced (24% inhibition, P < 0.05) by the cytochrome P450 inhibitor piperonyl butoxide (PB). The rate of TCBZSO conversion into TCBZSO2 was also significantly inhibited by PB (55% inhibition), MTZ (52% inhibition) and also following FMO inactivation (58% inhibition). The data reported here indicate that the FMO is the main enzymatic pathway involved in TCBZ sulphoxidation (ratio FMO/P450 = 3.83 +/- 1.63), although both enzymatic systems participate in a similar proportion in the sulphonation of TCBZSO to form the sulphone metabolite (ratio FMO/P450 = 1.31 +/- 0.23). Additionally, ketoconazole (KTZ) did not affect TCBZ sulphoxidation but decreased (66% inhibition, P < 0.05) the formation of TCBZSO2. Similarly, inhibition of TCBZSO2 production was observed after incubation of TCBZSO in the presence of KTZ and erythromycin (ETM). Conversely, thiabendazole (TBZ) and fenbendazole (FBZ) did not affect the oxidative metabolism of both incubated substrates. The sheep ruminal microflora was able to reduce the sulphoxide (TCBZSO) into the parent thioether (TCBZ). The ruminal sulphoreduction of the HO-TCBZSO derivative into HO-TCBZ was also demonstrated. The rate of sulphoreduction of HO-TCBZSO was significantly (P < 0.05) higher than that observed for TCBZSO. The metabolic approach tested here contributes to the identification of the different pathways involved in drug biotransformation in ruminant species. These findings on the pattern of hepatic and ruminal biotransformation of TCBZ and its main metabolites are a further contribution to the understanding of the pharmacological properties of widely used anthelmintics in ruminants. Comprehension of TCBZ metabolism is critical to optimize its flukicidal activity.