Metabolism of Zearalenone in the Rumen of Dairy Cows with and without Application of a Zearalenone-Degrading Enzyme.

The mycotoxin zearalenone (ZEN) is a frequent contaminant of animal feed and is well known for its estrogenic effects in animals. Cattle are considered less sensitive to ZEN than pigs. However, ZEN has previously been shown to be converted to the highly estrogenic metabolite α-zearalenol (α-ZEL) in rumen fluid in vitro. Here, we investigate the metabolism of ZEN in the reticulorumen of dairy cows. To this end, rumen-fistulated non-lactating Holstein Friesian cows (n = 4) received a one-time oral dose of ZEN (5 mg ZEN in 500 g concentrate feed) and the concentrations of ZEN and ZEN metabolites were measured in free rumen liquid from three reticulorumen locations (reticulum, ventral sac and dorsal mat layer) during a 34-h period. In all three locations, α-ZEL was the predominant ZEN metabolite and ß-zearalenol (ß-ZEL) was detected in lower concentrations. ZEN, α-ZEL and ß-ZEL were eliminated from the ventral sac and reticulum within 34 h, yet low concentrations of ZEN and α-ZEL were still detected in the dorsal mat 34 h after ZEN administration. In a second step, we investigated the efficacy of the enzyme zearalenone hydrolase ZenA (EC 3.1.1.-, commercial name ZENzyme®, BIOMIN Holding GmbH, Getzersdorf, Austria) to degrade ZEN to the non-estrogenic metabolite hydrolyzed zearalenone (HZEN) in the reticulorumen in vitro and in vivo. ZenA showed a high ZEN-degrading activity in rumen fluid in vitro. When ZenA was added to ZEN-contaminated concentrate fed to rumen-fistulated cows (n = 4), concentrations of ZEN, α-ZEL and ß-ZEL were significantly reduced in all three reticulorumen compartments compared to administration of ZEN-contaminated concentrate without ZenA. Upon ZenA administration, degradation products HZEN and decarboxylated HZEN were detected in the reticulorumen. In conclusion, endogenous metabolization of ZEN in the reticulorumen increases its estrogenic potency due to the formation of α-ZEL. Our results suggest that application of zearalenone hydrolase ZenA as a feed additive may be a promising strategy to counteract estrogenic effects of ZEN in cattle.

Effects of soybean oil and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls.

The study evaluated the effects of soybean oil (SO) and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls. Eight Holstein rumen-cannulated bulls (14 ± 0.2 months of age and 326 ± 8.9 kg of body weight) were allocated into a replicated 4 × 4 Latin square design in a 2 × 2 factorial arrangement with factors being 0 or 40 g/kg dietary dry matter (DM) of SO and 0 or 7.68 mg/kg DM of Cu from copper sulphate (CS). The basal diet contained per kg DM 500 g of corn silage, 500 g of concentrate, 28 g of ether extract (EE) and 7.5 mg of Cu. The SO × CS interaction was significant (p < 0.05) for ruminal propionate proportion and acetate to propionate ratio. Dietary SO addition increased (p < 0.05) intake and total tract digestibility of EE but did not affect average daily gain (ADG) of bulls. Dietary CS addition did not affect nutrient intake but increased (p < 0.05) ADG and total tract digestibility of DM, organic matter, crude protein and neutral detergent fibre. Ruminal pH was not affected by treatments. Dietary SO addition did not affect ruminal total volatile fatty acids (VFA) concentration, decreased (p < 0.05) acetate proportion and ammonia N and increased (p < 0.05) propionate proportion. Dietary CS addition did not affect ammonia N, increased (p < 0.05) total VFA concentration and acetate proportion and decreased (p < 0.05) propionate proportion. Acetate to propionate ratio decreased (p < 0.05) with SO addition and increased (p < 0.05) with CS addition. Dietary SO addition decreased (p < 0.05) activity of carboxymethyl cellulase, cellobiase and xylanase as well as population of fungi, protozoa, methanogens, Ruminococcus albus and R. flavefaciens but increased (p < 0.05) α-amylase activity and population of Prevotella ruminicola and Ruminobacter amylophilus. Dietary CS addition increased (p < 0.05) activity of cellulolytic enzyme and protease as well as population of total bacteria, fungi, protozoa, methanogens, primary cellulolytic and proteolytic bacteria. Microbial protein synthesis was unchanged with SO addition but increased (p < 0.05) with CS addition. The results indicated that the addition of CS promoted nutrient digestion and ruminal fermentation by stimulating microbial growth and enzyme activity but did not relieve the negative effects of SO addition on ruminal fermentation in dairy bulls.

Metagenomic Mining and Functional Characterization of a Novel KG51 Bifunctional Cellulase/Hemicellulase from Black Goat Rumen.

A novel KG51 gene was isolated from a metagenomic library of Korean black goat rumen and its recombinant protein was characterized as a bifunctional enzyme (cellulase/hemicellulase). In silico sequence and domain analyses revealed that the KG51 gene encodes a novel carbohydrate-active enzyme that possesses a salad-bowl-like shaped glycosyl hydrolase family 5 (GH5) catalytic domain but, at best, 41% sequence identity with other homologous GH5 proteins. Enzymatic profiles (optimum pH values and temperatures, as well as pH and thermal stabilities) of the recombinant KG51 bifunctional enzyme were also determined. On the basis of the substrate specificity data, the KG51 enzyme exhibited relatively strong cellulase (endo-ß-1,4-glucanase [EC 3.2.1.4]) and hemicellulase (mannan endo-ß-1,4-mannosidase [EC 3.2.1.78] and endo-ß-1,4-xylanase [EC 3.2.1.8]) activities, but no exo-ß-1,4-glucanase (EC 3.2.1.74), exo-ß-1,4-glucan cellobiohydrolase (EC 3.2.1.91), and exo-1,4-ß-xylosidase (EC 3.2.1.37) activities. Finally, the potential industrial applicability of the KG51 enzyme was tested in the preparation of prebiotic konjac glucomannan hydrolysates.

Effects of tributyrin supplementation on ruminal microbial protein yield, fermentation characteristics and nutrients degradability in adult Small Tail ewes.

Two trials were conducted to assess the effects of tributyrin (TB) supplementation on ruminal microbial protein yield and fermentation characteristics in adult sheep. In an in vitro trial, substrate was made to offer TB at 0, 2, 4, 6, and 8 g/kg on a dry matter (DM) basis and incubated for 48 hr. In an in vivo trial, 45 adult ewes were randomly assigned by initial body weight (55 ± 5 kg) to five treatments of nine animals over an 18-day period. Total mixed ration was made to offer TB to ewes at 0, 2, 4, 6, and 8 g/kg on a DM basis. The in vitro trial showed that TB enhanced apparent degradation of DM (p = .009), crude protein (p < .001), neutral detergent fiber (p = .007) and acid detergent fiber (p = .010) and increased methanogenesis (p < .001), respectively. The in vivo trial showed that TB decreased DM intake (p < .001) and enhanced rumen microbial N synthesis (p < .001), respectively. Both in vitro and in vivo trials showed that TB increased total volatile fatty acid concentration and enhanced fibrolytic enzyme activity. The results indicated that TB might exert positive effects on microbial protein yield and fermentation in the rumen.

Effects of traditional Chinese medicine formula on ruminal fermentation, enzyme activities and nutrient digestibility of beef cattle.

This study was conducted to evaluate effects of traditional Chinese medicine formula (TCMF) combined with several herbs on ruminal fermentation, enzyme activities and nutrient digestibility. Twenty finishing bulls were assigned to control or different TCMFs (Yufeisan-1, -2, -3; 2.5% dry matter (DM) in concentrate). Results showed that DM intake was higher (P < 0.05) in the Yufeisan-3 group than others. Compared to control, apparent digestibility of crude protein and neutral detergent fiber were increased (P < 0.05) by Yufeisan-3. No changes were observed in ruminal pH, concentrations of ammonia-N, microbial crude protein and total volatile fatty acid, whereas ratio of acetate to propionate was lower (P < 0.05) and propionate proportion tended to be higher (P < 0.1) in three TCMFs than control. Ruminal xylanase (P = 0.061) and carboxymethylcellulase (P < 0.05) activities were higher in Yufeisan-3 than control. No changes were observed in abundance of total bacteria, fungi and protozoa, whereas Fibrobacter succinogenes (P = 0.062) and Ruminococcus flavefaciens (P < 0.05) were increased and total methanogens was reduced (P = 0.069) by Yufeisan-3 compared to control. Yufeisan-3 improved nutrient digestibility and ruminal enzyme activity, and modified fermentation and microbial community, maybe due to the presence of Herba agastaches, Cortex phellodendri and Gypsum fibrosum.

Effects of dietary Greek oregano (Origanum vulgare ssp. hirtum) supplementation on rumen fermentation, enzyme profile and microbial communities in goats.

This study was conducted to examine in vivo long-term effects of dietary dried oregano (Origanum vulgare ssp. hirtum) whole plant on rumen fermentation, enzyme profile and microbial communities. For this purpose, eight healthy, adult, non-lactating Alpine goats were kept in tie stalls equipped for individual feeding and randomly divided into two homogeneous groups: one fed 0.6 kg of a concentrate mixture and 0.6 kg of wheat straw without any supplementation and served as control group (CON) while the other group (OR) fed the same diet of CON but supplemented with 20 g of dried oregano plants (OPs) to provide daily dosage of 1 ml of essential oil (EO) per animal. The experimental period lasted 69 days and individual rumen fluid samples were obtained every 2 weeks at 0 and 4 hr after feeding. The results showed that dietary supplementation with OPs increased the protease activity (p < .001) and ammonia concentration (p < .05) in the rumen. Among the studied microbial populations, Peptostreptococcus anaerobius (p = .028) and Clostridium sticklandii (p < .001) were found to be the most sensitive to oregano at the current dosage. Furthermore, the total methanogen population significantly decreased (p < .05). It is concluded that a long-term dietary administration of OPs can suppress specific rumen micro-organisms and modify rumen fermentation favourably at least by means of suppressing methanogens.

Modulation of in vitro rumen biohydrogenation by Cistus ladanifer tannins compared with other tannin sources.

BACKGROUND: Tannins are polyphenolic compounds able to modify the ruminal biohydrogenation (BH) of unsaturated fatty acids, but their activity may vary among different tannin sources. The effect of rockrose (Cistus ladanifer) on BH has never been compared with other more common tannin sources. Tannin extracts (100 g kg-1 substrate dry matter) from chestnut (CH), quebracho (QB), grape seed (GS) and rockrose (CL) were incubated in vitro for 6 h with ruminal fluid using as substrate a feed containing 60 g kg-1 of sunflower oil. A control treatment with no added tannins was also included. RESULTS: Compared with the control, GS and CL, but not CH and QB, increased (P < 0.05) the disappearance of c9,c12-18:2 with a consequent larger production of c9,t11-18:2 and t11-18:1. However, no differences among treatments (P > 0.05) were observed for the disappearance of c9-18:1 and c9,c12,c15-18:3. The production of 18:0 was not different (P > 0.05) among treatments, although its proportion in the total BH products was lower (P < 0.05) for GS than for the other treatments. CONCLUSION: Condensed tannins from GS and, to a lesser extent, from CL stimulate the first steps of BH, without a clear inhibition of 18:0 production. © 2016 Society of Chemical Industry.

Effects of isobutyrate supplementation on ruminal microflora, rumen enzyme activities and methane emissions in Simmental steers.

The objective of this study was to evaluate the effects of isobutyrate supplementation on rumen microflora, enzyme activities and methane emissions in Simmental steers consuming a corn stover-based diet. Eight ruminally cannulated Simmental steers were used in a replicated 4 × 4 Latin square experiment. The treatments were control (without isobutyrate), low isobutyrate (LIB), moderate isobutyrate (MIB) and high isobutyrate (HIB) with 8.4, 16.8 and 25.2 g isobutyrate per steer per day respectively. Isobutyrate was hand-mixed into the concentrate portion. Diet consisted of 60% corn stover and 40% concentrate [dry matter (DM) basis]. Dry matter intake (averaged 9 kg/day) was restricted to a maximum of 90% of ad libitum intake. Population of total bacteria, cellulolytic bacteria and anaerobic fungi were linearly increased, whereas that of protozoa and total methanogens was linearly reduced with increasing isobutyrate supplementation. Real-time PCR quantification of population of Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio fibrisolvens and Fibrobacter succinogenes was linearly increased with increasing isobutyrate supplementation. Activities of carboxymethyl cellulase, xylanase and ß-glucosidase were linearly increased, whereas that of protease was linearly reduced. Methane production was linearly decreased with increasing isobutyrate supplementation. Effective degradabilities of cellulose and hemicellulose of corn stover were linearly increased, whereas that of crude protein in diet was linearly decreased with increasing isobutyrate supplementation. The present results indicate that isobutyrate supplemented improved microflora, rumen enzyme activities and methane emissions in steers. It was suggested that the isobutyrate stimulated the digestive micro-organisms or enzymes in a dose-dependent manner. In the experimental conditions of this trial, the optimum isobutyrate dose was approximately 16.8 g isobutyrate per steer per day.

Establishment of ruminal bacterial community in dairy calves from birth to weaning is sequential.

AIM: Establishment of ruminal bacterial community in dairy calves. METHODS AND RESULTS: Rumen bacterial community was analysed on 6 calves bred according to commercial practices from day one to weaning at day 83 of age, using 454 16S rRNA-based pyrosequencing. Samples taken at day 1 did not produce amplicons. Analysis of data revealed a three-stage implantation process with a progressive but important shift of composition. At day 2, the bacterial community was mainly composed of Proteobacteria (70%) and Bacteroidetes (14%), and Pasteurellaceae was the dominant family (58%). The bacterial community abruptly changed between days 2 and 3, and until day 12, dominant genera were Bacteroides (21%), Prevotella (11%), Fusobacterium (5%) and Streptococcus (4%). From 15 to 83 days, when solid food intake rapidly increased, Prevotella became dominant (42%) and many genera strongly decreased or were no longer detected. A limited number of bacteria genera correlated with feed intake, rumen volatile fatty acids and enzymatic activities. CONCLUSION: The ruminal bacterial community is established before intake of solid food, but solid food arrival in turn shapes this community. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insight into the establishment of calves' rumen bacterial community and suggests a strong effect of diet.

Time course of changes in antioxidant activity of milk from dairy cows fed a trehalose-supplemented diet.

This study was to investigate the time course of changes to the antioxidant activity of milk from cows fed a trehalose-supplemented diet, and to determine possible underlying mechanisms for observed changes. Six Holstein cows were used, and subjected to two experimental feeding periods consisting of a 1% trehalose-supplemented diet for 10 days, followed by a basal diet only (no trehalose) for 10 days. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities in milk were gradually increased during the trehalose supplementation period and were highest at the end of the second period. However, trehalose was not detected in the milk and plasma of dairy cows fed a diet supplemented with trehalose for 10 days, indicating that the increased antioxidant activity in the milk of trehalose-fed cows is not due to the direct transfer of trehalose to the milk. Plasma DPPH activities exhibited a similar time course to that seen for milk. Relative superoxide dismutase (SOD) activities in the rumen were higher 3 days after the end of trehalose supplementation than at any other time during the experimental periods. These results suggested that the improved antioxidant activity in milk and plasma of cows fed a trehalose-supplemented diet was due to improved ruminal relative SOD activity.

Effects of dietary supplementation with alkyl polyglycoside, a nonionic surfactant, on nutrient digestion and ruminal fermentation in goats.

The effects of dietary alkyl polyglycoside [APG, a nonionic surfactant (NIS), derived from a reaction of corn starch glucose and a natural fatty alcohol] inclusion on digestion of nutrients and ruminal fermentation in goats were examined in a 4 × 4 Latin square design using 4 ruminally and duodenally cannulated wethers (mean BW: 19.5 ± 0.8 kg). The animals were assigned to 4 dietary treatments of APG supplementation at 0, 3, 6, and 12 g/kg of DM diets and were designated as control, APG3, APG6, and APG12, respectively. The results showed that dietary APG inclusion tended to increase the intestinal digestibility of OM (linear, P = 0.09) and NDF (linear, P = 0.1), and quadratically increased (P ≤ 0.02) total tract digestibility of OM and NDF, the duodenal microbial N flow, and efficiency of microbial protein synthesis. The true ruminal digestibility and apparent total tract digestibility of N quadratically increased (P < 0.01) with increasing dietary APG. The ruminal pH values were not affected by dietary APG inclusion (P > 0.05), but the concentration of NH(3)-N (P < 0.01) and total VFA (linear and quadratic, P < 0.01) increased in the rumen fluid. Dietary APG inclusion also increased the activities of ruminal carboxymethyl cellulase (quadratic, P < 0.01) and xylanase (linear and quadratic, P < 0.01). It is concluded that APG is a potential feed additive that can be used in ruminant production; 6 g/kg in the total mixed rations for goats is recommended. It is necessary to validate the effectiveness of dietary APG inclusion in ruminant diets with more animals in further studies.

The effect of steam-flaked or dry ground corn and supplemental phytic acid on phosphorus partitioning and ruminal phytase activity in lactating cows.

The effect of starch source and phytic acid (PA) supplementation on phosphorus (P) partitioning and ruminal phytase activity was evaluated in eight midlactation cows (four ruminally cannulated). Cows were randomly assigned to treatments in replicated 4 x 4 Latin squares with four 18-d periods. Diets included dry ground corn (DG) or steam-flaked corn (SF), with no supplemental P (L; 0.33% P) or supplemental purified PA (0.44% P) to provide additional P from a nonmineral source. Total collection of milk, urine, and feces was conducted on d 16 to 18 of each period. Ruminal fluid was sampled and ruminal pH measured every 8 h on d 17 and 18. Milk yield was unaffected by starch source, despite lower DMI by cows fed SF. Cows fed SF had increased DM digestibility compared with those fed DG, and tended to have higher efficiency of milk yield (1.40 vs. 1.35 kg of milk/kg of DMI). Intake and fecal excretion of P was lower in cows fed SF than in cows fed DG. In cows fed SF, milk P as a percentage of P intake increased compared with cows fed DG. Ruminal pH was unaffected by diet, but milk fat content was lower for cows fed SF. Milk yield, DMI, and feed to milk ratio were not affected by supplementation with PA. Although cows fed PA had increased P intake compared with cows fed low P diet, increased P excretion resulted in no differences in apparent P digestibility. Phosphorus balance tended to be higher in cows fed PA, but milk P as a percentage of intake was reduced. The interaction of starch source and PA affected ruminal phytase activity. Altering starch source to improve efficiency of milk yield in lactating dairy cows may help reduce P losses from dairy farms.

Degradability of forage proteins by in situ and in vitro enzymatic methods.

The overall objective of these two studies was to evaluate the efficacy of using the proteolytic enzyme from Streptomyces griseus to estimate concentrations of ruminally degradable protein (RDP) in a wide array of forages. In the first study, alfalfa and prairie hays that previously had been evaluated in vivo for RDP were incubated in a replicated 3 x 3 factorial combination of enzyme concentrations (6.6, 0.66, and 0.066 activity units/ml of incubation medium) and incubation times (2, 4, and 48 h). Two treatment combinations (6.6 activity units for 4 h and 0.066 activity units for 48 h) yielded respective RDP estimates for alfalfa and prairie hay that were close to the known in vivo values. In the second study, 20 diverse forages were evaluated for RDP by using the in situ technique. These forages also were evaluated for RDP with the two enzyme concentrations identified in the first study, but incubation times were expanded to include 1, 2, 3, 4, and 5 h at the high concentration and 24, 30, 36, 42, 48, and 54 h at the low concentration. At the high enzyme concentration, r2 statistics from linear regressions of enzymatic estimates of RDP on corresponding estimates obtained by the in situ procedure were high (r2 > or = 0.898) at all incubation times; in addition, slopes (range = 0.88 to 1.00) and intercepts (range = -9.4 to 3.5%) approached unity and 0, respectively. At the lower enzyme concentration, r2 statistics were still good (> 0.81), but slopes (0.59 to 0.67) and intercepts (18.5 to 21.9%) for all incubation times did not meet the respective goals of unity and 0.

Effect of dietary or abomasal supplementation of exogenous polysaccharide-degrading enzymes on rumen fermentation and nutrient digestibility.

The effect of site of supplementation of a mixture of two crude preparations (Enzyme C and Enzyme X) of exogenous polysaccharide-degrading enzymes (EPDE) was studied in vivo using four ruminally and duodenally cannulated heifers (Exp. 1). The treatments were as follows: control (no EPDE), EPDE supplied through the diet (EF, 47.0 g/d), and EPDE infused continuously into the abomasum (EA, 41.6 g/d). Enzyme treatment increased the concentration of soluble reducing sugars (P < .05) and decreased NDF content (P < .05) in the treated feed, but this did not increase the rate or extent of in sacco disappearance of DM from the feed. Compared with control, ruminal fermentation was not affected by EF, but abomasal infusion increased (P < .05) rumen ammonia levels and shifted ruminal VFA patterns. Ruminal carboxymethylcellulase (CMCase) and xylanase activities were not affected by treatment. Abomasal infusion increased (P < .05) duodenal xylanase activity as compared with control and EF, but apparent digestion of DM, NDF, and CP were not affected by treatment. Negligible levels of CMCase and amylase reached the duodenum. During an in vitro experiment (Exp. 2), abomasal stability of the two EPDE was studied over a range of pH from 3.39 to .85, with or without pepsin. Carboxymethylcellulase activity (in Enzymes C and X) and beta-glucanase activity (in Enzyme C) were largely unstable against pepsin proteolysis (P < .001) and low pH (P < .001). Xylanase and amylase activities were resistant to pepsin but irreversibly inactivated at low pH. These two experiments showed that abomasal supplementation of EPDE did not successfully supply cellulases and amylases to the intestine, due partially to their limited resistance to low pH and pepsin proteolysis. Although EPDE significantly increased the level of xylanase activity at the duodenum, this did not significantly improve total tract digestion.

Effects of forage species and particle size on bacterial cellulolytic activity and colonization in situ.

Two ruminally cannulated Holstein heifers were used to evaluate effects of three forage species and two particle sizes in a 3 x 2 factorial arrangement on in situ fiber disappearance and carboxymethylcellulase activity (CMCase). A second objective was to compare CMCase and 15N as relative bacterial colonization markers. At time zero, 15N-enriched ammonium sulfate was dosed into the rumen. Residues in nylon bags incubated for 3, 6, 9, 12, 18, 24, 30, 36, 42, 48, 60, and 72 h were analyzed for NDF, ADF, CMCase, and 15N. Rates of NDF and ADF disappearance were faster (P < .05) for red clover (RC) than for gamagrass (GG) or orchardgrass (OG) and for forages ground to pass a 2-mm screen (P < .05) than for those ground to pass a 5-mm screen. Cumulative area under the curve (AUC) of CMCase activity was related to both NDF disappearance (r2 = .93) and 15N AUC (r2 = .99), indicating that CMCase can be used to estimate relative colonization of cellulolytic particle-associated bacteria (PAB). During the first 18 h of incubation, RC had greater (P < .05) CMCase and 15N enrichment of in situ residues than did OG or GG. The RC residues also had a greater (P < .05) ratio of CMCase activity:15N enrichment; this was presumed to indicate that, during this time, RC supported a greater total PAB population and had a greater proportion of cellulolytic PAB than did either GG or OG. Differences in the colonization rate of forage particles by cellulolytic PAB relative to total PAB may account for differences in rate of fiber digestion.

Substitution of DL-methionine for soybean meal as a winter supplement for gestating cows grazing native range.

A winter grazing study was conducted to determine whether DL-methionine could replace soybean meal as a N supplement for gestating beef cows. During two winters (Trial 1, n = 51; Trial 2, n = 60), crossbred beef cows grazed native foothill range. Three treatment groups were supplemented with either none (CON), DL-methionine (7.5 g Trial 1 and 9 g Trial 2) in .5 kg beet pulp carrier (BPM) or .4 kg soybean meal (SBM). Cows were supplemented individually every other day. Small differences were noted in cow BW, condition score and blood metabolites. Unsupplemented cows lost the greatest amount of BW (P less than .01) in both trials and lost more (P less than .05) condition during Trial 1 than cows fed BPM or SBM supplements. Blood samples were obtained on two consecutive days during each trial (45 d and 25 d prepartum) and analyzed for blood urea N, total bilirubin, creatinine, albumin, total protein and cholesterol. A treatment x day preparatum interaction (P less than .05) was noted for blood urea. Blood urea nitrogen declined as gestation length increased for CON and SBM cows, but blood urea of BPM-supplemented cows remained low and unchanged. In situ forage digestion was measured in 12 ruminally cannulated cows (four/treatment). In both trials, in situ rate of NDF disappearance was greater (P less than .05) for SBM than for BPM. In Trial 2, a treatment x sampling hour interaction was detected for purine concentration of whole ruminal contents; SBM maintained greater purine concentrations throughout the 48-h supplementation cycle than BPM did. Principal component analysis suggested that ruminal ammonia limited the microbial growth response to DL-methionine. Therefore, alternate-day supplementation of DL-methionine plus beet pulp did not effectively substitute for soybean meal in these trials.

Effect of tannin-rich leaves of oak (Quercus incana) on various microbial enzyme activities of the bovine rumen.

1. The objective of the present experiment was to study the effects of oak (Quercus incana) leaves rich in tannins on various enzyme activities of the bovine rumen. 2. The procedure employed was incubation of tannin-rich, very-low-tannin or virtually tannin-free leaves in nylon-gauze bags in the rumen, and determination of enzyme activities in microbes tightly bound to the solid matrix and in microbes loosely plus tightly attached to the solid matrix. 3. The activities of urease (EC 3.5.1.5), carboxymethylcellulose, glutamate dehydrogenase (EC 1.4.1.2) and alanine aminotransferase (glutamic-pyruvic transaminase) (EC 2.6.1.2) were significantly lower in the tannin-rich group, whereas the activities of glutamate ammonia ligase (glutamine synthetase) (EC 6.3.1.2; both gamma-glutamyltransferase (EC 2.3.2.2) and the forward reaction) were higher in the tannin-rich group. These changes were more marked in micro-organisms tightly bound to the solid matrix than in the more complex microbial compartment. 4. The protein, DNA and RNA contents, and protein: RNA ratio, were significantly lower in the tannin-rich group, whereas no difference was observed for protein: DNA between the groups. 5. Effects of tannin-containing extracts of oak leaves on various rumen enzymes in vitro showed a trend similar to that observed in nylon-gauze bags, suggesting that the changes observed in various compartments were due to the tannins of oak leaves.

Thiamin and niacin in the rumen.

Thiamin analogs, produced in the rumen by thiaminase I, in the presence of a cosubstrate appear to be responsible for the central nervous system disorder, polioencephalomalacia (PEM). For PEM to occur, an analog must be produced that inhibits an essential thiamin-requiring reaction, and results from a cosubstrate present in the rumen. In high concentrate diets, thiaminase I is produced by rumen microbes. However, PEM can also be caused by thiaminase I of plant origin. Based on physical characteristics and cosubstrate specificity, the thiaminase I enzymes produced by Bacillus thiaminolyticus and Clostridium sporogenes appear to be different from the enzyme produced by the rumen. Because niacin and certain antihelmentics are thiaminase I cosubstrates, they should be used cautiously. Supplementary niacin increased microbial protein synthesis in vitro and in vivo, and was more effective with urea than soybean meal. Supplementary niacin (5 to 6 g X cow-1 X d-1) increased milk production in postpartum cows but not in those in mid-lactation, and in cows fed soybean meal but not in those fed urea. We believe the heating of soybean meal during commercial processing decreased the availability of niacin for rumen protozoa. Supplementary niacin for postpartum cows increased blood glucose, decreased blood ketones and reduced the incidence of ketosis. Niacin flow to the small intestine and its absorption from the small intestine increased with niacin supplementation. Supplemental niacin prevented the postpartum decrease in red blood cell niacin observed in control cows.

Nickel as a "newer trace element" in the nutrition of domestic animals.

Evidence is reviewed that indicates Ni is an essential element for the chick, rat, pig, sheep and goat. Although a number of possible functions for Ni have been proposed based on in vitro and in vivo studies, the physiological role of Ni in the mammalian or avian system is presently unknown. Rumen bacterial urease has been shown to be a Ni-dependent enzyme and Ni is a component of factor F430 present in methanogenic bacteria. Nickel can interact or influence the metabolism of a number of minerals. Interactions of Ni and Fe, Zn and Cu are discussed. The requirement for Ni is low (50 to 60 ppb) in chicks fed semipurified diets. Insufficient data are available to estimate the Ni requirement of swine. In ruminants, the Ni requirement appears to be higher than that for other animal species. Nickel supplementation to practical diets has increased gain, feed efficiency and ruminal urease activity in ruminants, but performance results have been inconsistent. Level of crude protein and urea are two factors that influence ruminant responses to dietary Ni. The greatest responses have been observed in ruminants fed low protein diets. Little is known concerning levels, forms and bioavailability of Ni in different feedstuffs. Nickel is homeostatically controlled in the animal's body and high levels of Ni are required to cause toxicity.