Effects of defaunation and various nitrogen supplementation regimens on microbial numbers and activity in the rumen of sheep

Five sheep (average BW 62 kg) were fed 65% roughage: 35% concentrate diets (CP = 15%) in a 5 x 5 Latin square design to study the effects of combinations of defaunation and N supplements (soybean meal [SBM], corn gluten meal [CGM], blood meal [BM], urea, and casein) differing in ruminal degradation on ruminal microbial numbers and activity. Diets were fed twice daily (DM intake 1,759 g/d). Defaunation was accomplished with doses of 30 ml of alkanate 3SL3.sheep-1.d-1 for 3 d with 2 d of fasting. Treatment 1 (control) involved feeding faunated sheep a diet in which the supplemental N was 67% SBM N and 33% urea N. Treatment 2 involved feeding defaunated sheep the same diet as the control. Treatments 3, 4, and 5 involved feeding defaunated sheep diets in which the supplemental N source was either 67% CGM-BM N (CGM and BM combined on a 1:1 N ratio): 33% urea N, or 33% CGM-BM N:67% urea N or 33% CGM-BM N:33% urea N:33% casein N, respectively. Compared with the faunated control, defaunation (Treatments 2, 3, 4, and 5) increased (P less than .05) total direct counts of ruminal bacteria (2.7 vs 1.3 x 10(11)/ml), fungal zoospores (2.8 vs 1.4 x 10(5)/ml), and ruminal microbial protease activity (1.4 vs 1.0 mg azocasein/[ml ruminal fluid.h]). Defaunation did not have a consistent effect on ruminal microbial deaminase activity. Compared with the control, defaunation resulted in lower (P less than .05) total perchloric acid-soluble amino N in ruminal fluid at 4 and 10 h after the morning feeding. Defaunation did not decrease (P greater than .05) total free amino acid concentrations in ruminal fluid, but it altered the profile of free amino acids. Although defaunation increased (P less than .05) ruminal bacterial numbers, no increases in total microbial CP or OM concentrations in ruminal contents were observed.     

Manipulation of nitrogen digestion by sheep using defaunation and various nitrogen supplementation regimens

Five ruminally, duodenally, and ileally cannulated sheep (average BW 62 kg) were fed 65% roughage: 35% concentrate diets (CP = 15%) in a 5 x 5 Latin square design to study the applicability of using a combination of defaunation with N supplements (soybean meal [SBM], corn gluten meal [CGM], blood meal [BM], urea, and casein) with different extents of ruminal degradation to manipulate microbial protein synthesis and amount of ruminal escape protein. Diets were fed twice daily (1,759 g DM/d). Defaunation was accomplished with 30-ml doses of alkanate 3SL3 (active ingredient: sodium lauryl diethoxy sulfate)/sheep daily for 3 d with 2 d of fasting. Treatment 1 (control) involved feeding faunated sheep a diet in which the supplemental N (45% of total dietary N) was 67% SBM N and 33% urea N. Treatment 2 involved feeding defaunated sheep the same diet as the control. Treatments 3, 4, and 5 involved feeding defaunated sheep diets in which the supplemental N source was either 67% CGM-BM (1:1 N ratio) N:33% urea N, or 33% CGM-BM N:67% urea N or 33% CGM-BM N:33% urea N:33% casein N, respectively. Compared with the faunated control, defaunation decreased (P less than .05) ruminal ammonia concentration (19 vs 26 mg/dl) and increased (P less than .05) CP flow to the duodenum (253 vs 214 g/d) due to a trend for increases in both bacterial (BCP) and nonbacterial (NBCP) CP flows.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of urea and sodium bicarbonate supplementation of a high-fiber diet on nutrient digestion and ruminal characteristics of defaunated sheep.

Five sheep (average BW 48 kg) with ruminal, duodenal, and ileal cannulas were fed 63% roughage: 37% concentrate diets (CP = 14.5%) in a 5 x 5 Latin square design to study effects of urea and sodium bicarbonate supplementation on nutrient digestion and ruminal characteristics of defaunated sheep. Diets were fed twice daily (DMI = 1,076 g/d). Defaunation was accomplished with 25-ml doses of alkanate 3SL3/sheep daily for 3 d. Control sheep were faunated (Treatment 1) and fed soybean meal as the major N supplement. Remaining sheep were maintained defaunated and fed either the same diet as Treatment 1 (Treatment 2), Treatment 1 with urea replacing 30% of the soybean meal N (Treatment 3), or Treatment 1 with 2% sodium bicarbonate in the diet (Treatment 4). Treatment 5 was a combination of Treatments 3 and 4. Compared with the faunated control, defaunation decreased (P less than .05) total tract DM, OM, NDF, ADF, and CP digestibilities (71.5 vs 69.4, 73.8 vs 71.7, 64.6 vs 61.4, 58.7 vs 55.8, and 74.2 vs 70.6%, respectively) and average (2 to 12 h postfeeding) ruminal fluid ammonia (23.5 vs 13.7 mg/dl) and isobutyrate (.9 vs .7 mM) concentrations. However, defaunation increased (P less than .05) linoleic and linolenic acid flows (.58 vs .45 g C18:2/d; .17 vs .14 g C18:3/d) to and disappearance (.50 vs .39 g C18:2/d; .14 vs .11 g C18:3/d) from the small intestine. Urea supplementation increased (P less than .05) total tract DM (70.2 vs 68.6%) and OM (72.3 vs 71.0%) digestibilities of defaunated sheep but lowered (P less than .05) ruminal fluid isobutyrate concentration (.6 vs .8 mM). Sodium bicarbonate supplementation increased (P less than .05) ruminal fluid pH (6.4 vs 6.2), isobutyrate concentration (.75 vs .60 mM), total tract ADF digestibility (57.6 vs 54.2%), and ruminal NDF (41.6 vs 28.5%), ADF (36.6 vs 22.8%), and CP (-5.5 vs -26.8%) digestibilities in defaunated sheep. Dietary supplementation of urea or sodium bicarbonate increased nutrient digestion by defaunated sheep.     

Effects of defaunation on in situ dry matter and nitrogen disappearance in steers and growth of lambs.

Trials were conducted to determine effects of defaunation procedures on protozoal concentrations and in situ nutrient disappearance in steers and to determine effects of defaunation and supplemental protein source on performance of lambs. Four ruminally cannulated steers were isolated from other ruminants and fed a dehydrated alfalfa-cracked corn diet for three periods with four replicates (steers) per period. Treatments were as follows: 1) control (no defaunation), 2) dosing fasted steers for two consecutive days with 40 g dioctyl sulfosuccinate (DSS) and 3) daily feeding of 40 g DSS to defaunated, nonfasted steers. Ten days post-dosing with DSS (treatment 2), three steers were free of protozoa but one steer still had a ruminal concentration of .6 x 10(4) protozoa/ml. Compared to steers prior to defaunation, treating steers for 2 d with DSS decreased (P less than .05) both in situ soybean meal (SBM) N disappearance at 3, 6 and 9 h of incubation and in situ orchardgrass DM disappearance at 24 h of incubation. Feeding 40 g of DSS daily for 10 d was not successful in maintaining the rumen free of protozoa. Thirty crossbred Targhee lambs (avg wt, 25 kg) were defaunated with DSS and allotted by BW and sex to five treatments: 1) defaunated, fish meal supplemented at 9.5% dietary CP (FM-9.5% CP), 2) defaunated, SBM-9.5% CP, 3) refaunated, FM-9.5% CP, 4) refaunated, SBM-9.5% CP and 5) refaunated SBM-12% CP. Defaunated lambs remained free of protozoa during the 56-d performance trial that was initiated 24 d after the defaunation procedure.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of presence of rumen protozoa on degradation of cell wall constituents in gastrointestinal tract of cattle

The cell wall constituents of feces from three faunated and three defaunated (without ruminal ciliate protozoa) cattle fed on a Sudangrass hay and concentrate mixture (8:5) were analyzed. There was little difference in digestibility of dry matter between the faunated and defaunated cattle. Analysis of the fecal sugar residues revealed that the digestibilities of arabinose and galactose, derived from pectic and hemicellulosic substances located within the compound middle lamella, were higher in the defaunated cattle than the faunated cattle (P < 0.05), whereas the digestibilities of glucose and xylose, derived mainly from cellulose and xylan, were unchanged by the removal of protozoa. The digestibility of lignin was not different between the faunated and defaunated cattle, but those of mannose and p‐coumaric acid were lower in the defaunated than in the faunated animals (P < 0.05). The ratio of primary cell wall to secondary cell wall in fecal plant materials was lower for the defaunated than for the faunated cattle. The results in this study suggested that the defaunation enhanced the microbial degradation of the thin cell walls, but depressed the degradation of developed cell walls.     

Effects of protozoa on the antioxidant activity in the ruminal fluid and blood plasma of cattle

The objective of this study was to investigate the antioxidant status in ruminal fluid and blood plasma among three faunated and two defaunated (protozoa‐free) cattle (average bodyweight of 225 kg), fed hay plus concentrate. The extra cellular antioxidant activity of the mixed protozoa and bacteria suspensions were also studied in vitro. The antioxidant activity was detected by means of the free radical scavenging ability. The activity (units/microbial nitrogen) of the protozoal suspension increased from 59 (0 h) to 318 (18 h), and decreased to 40 (24 h) during incubation. The activity of the bacterial suspension also increased from 111 (0 h) to 644 (18 h), and decreased to 533 (24 h). The antioxidant activity (units/mL, U/mL) in the ruminal fluid of faunated (ranging from 116 to 254) and defaunated (ranging from 66 to 110) cattle was increased after 2 h and decreased after 5 h of feeding, being significantly higher in the faunated cattle. The antioxidant activity of blood plasma (U/mL) ranged from 248 to 316 in the faunated and 121–170 in the defaunated cattle during 0–5 h after feeding, being significantly higher in the faunated cattle. Therefore, defaunation possibly causes a decrease in the antioxidant level in the ruminal fluid, and may impair the health and performance of ruminants through an oxidant–antioxidant imbalance.     

Occurrence of 2-aminoethylphosphonic acid in feeds, ruminal bacteria and duodenal digesta from defaunated sheep

A quantitative method of analysis for 2-aminoethylphosphonic acid (AEP) was developed using reverse-phase HPLC. The detection limit for AEP was 15 nM, and the detector response (peak area) was linear from AEP levels up to 100 microM (R = .99). Mean recovery of AEP added to strained ruminal fluid from faunated sheep was 98.2%. When AEP was added to a fermentation mixture at a concentration of 22.6 micrograms/ml, 78% disappeared during a 24-h incubation. 2-Aminoethylphosphonic acid was readily detected in preparations of mixed ruminal ciliate protozoa as well as in mixed and pure strains of ruminal bacteria, feedstuffs, and ruminal fluid and duodenal digesta from defaunated sheep. The occurrence of AEP in feed and bacterial hydrolysates was confirmed by organic phosphorus analyses. The concentration of AEP in mixed ruminal protozoa was three times greater than its concentration in mixed ruminal bacteria (4,304 vs 1,383 micrograms/g DM, respectively). The AEP values for pure ruminal bacterial cultures ranged from 733 micrograms/g DM in Bacteroides succinogenes B21a to 1,166 micrograms/g DM in Butyrivibrio fibrisolvens H17c. Ruminal fluid and duodenal digesta from defaunated sheep contained AEP concentrations of 30 micrograms/ml and 90 micrograms/g DM, respectively. The concentration of AEP in feedstuffs ranged from 25 micrograms/g DM in wheat straw to 263 micrograms/g DM in oats. Because AEP occurrence is not limited to ruminal ciliate protozoa, it is of little value as a marker for protozoal presence in or passage out of the rumen.     

The effects of urea supplementation on production and parasitological responses of sheep infected with Haemonchus contortus and Trichostrongylus colubriformis.

Merino wether lambs were individually confined and fed a basal diet of oaten chaff containing essential minerals which was untreated or contained 3% urea. Within each dietary group animals were orally infected with either 200 H. contortus (H), 1000 T. colubriformis (T) or both species (H + T) thrice weekly or remained uninfected (C). Weight gain, wool production, and parasite burden were measured over a 19-week period. Sheep on the diet containing urea gained more weight, consumed more feed and grew more wool of higher fibre diameter than their counterparts given no urea. On both diets uninfected sheep consumed more feed than infected sheep and the sheep given no urea and infected with both H and T worm species consumed the least feed. Parasitised sheep gained less weight than uninfected control sheep. Sheep with urea in their diet had lower faecal egg counts when infected with H alone or with H and T but there was no effect of urea on egg count of sheep infected with T alone. In contrast, T numbers after slaughter were reduced in sheep fed diets containing urea whereas H numbers were not affected by diet. It was concluded that supplementation with urea can increase resilience to parasitism thereby improving production and also enhance resistance mechanisms against worms in young sheep on low quality roughage diets. These responses can be partly attributed to stimulation of feed intake, presumably due to enhanced ruminal digestion, but also to elevated rumen NH3-N levels which would be expected to have increased rumen microbial protein synthesis and availability to the intestines.     

Effects of defaunation and dietary coconut oil distillate on fermentation,digesta kinetics and methane production of Brahman heifers

A 2 × 2 factorial experiment was conducted to assess the effects of presence or absence of rumen protozoa and of dietary coconut oil distillate (COD) supplementation on rumen fermentation characteristics, digesta kinetics and methane production in Brahman heifers. Twelve Brahman heifers were selected to defaunate, with 6 being subsequently refaunated. After defaunation and refaunation, heifers were randomly allocated to COD supplement or no supplement treatments while fed an oaten chaff‐based diet. Methane production (MP; 94.17 v 104.72 g CH₄/d) and methane yield [MY; 19.45 v 21.64 g CH₄/kg dry matter intake (DMI)] were reduced in defaunated heifers compared with refaunated heifers when measured at 5 weeks after refaunation treatment (p < 0.01). Supplement of COD similarly reduced MP and MY (89.36 v 109.53 g/d and 18.46 v 22.63 g/kg DMI, respectively; p < 0.01), and there were no significant interactions of defaunation and COD effects on rumen fermentation or methane emissions. Concentration of total volatile fatty acid (VFA) and molar proportions of acetate, propionate and butyrate was not affected by defaunation or by COD. Microbial crude protein (MCP; g/d) outflow was increased by defaunation (p < 0.01) in the absence of COD but was unaffected by defaunation in COD‐supplemented heifers. There was a tendency towards a greater average daily gain (ADG) in defaunated heifers (p = 0.09), but COD did not increase ADG (p > 0.05). The results confirmed that defaunation and COD independently reduced enteric MP even though the reduced emissions were achieved without altering rumen fermentation VFA levels or gut digesta kinetics.     

Effect of dietary protein level on nitrogen metabolism in the growing bovine: II. Diffusion into and utilization of endogenous urea nitrogen in the rumen

Six Angus heifer calves (234 kg) were assigned to either a high (HP; 126.1 g N/d) or low (LP; 66.5 g N/d) protein intake to evaluate ruminal criteria associated with movement of blood urea-N (BUN)-derived NH3-N from the rumen wall into interior ruminal digesta. Calves received 4.8 kg DM/d of diets containing 30% cottonseed hulls and 70% cornsoybean meal in equal portions at 4-h intervals. Following single i.v. injections of 15N-urea, ruminal fluid was collected serially for 4 h postinjection from digesta located adjacent to the rumen wall (wall-proximate digesta; WPD) and from the center of the rumen digesta mass after manual agitation (center mixed digesta; CMD). Mean ruminal NH3-N (RAN) concentrations were higher (P less than .05) for HP than for LP, but were not affected (P greater than .05) by digesta sampling site. Ruminal urease activity was higher (P less than .05) for LP than for HP and tended (P = .14) to be higher for WPD than for CMD. Area under the 15N enrichment curve (AUC) ratios between sampling sites (WPD/CMD x 100) for RAN were greater (P less than .05) for LP than for HP. However, AUC ratios for bacterial N were not affected (P greater than .05) by protein level. Whereas BUN-derived 15NH3 appeared to thoroughly equilibrate with RAN in interior ruminal digesta with HP, there appeared to be a declining enrichment gradient for RAN from the rumen wall to the interior ruminal digesta with LP. Data are interpreted to suggest that bacteria at or near the rumen wall may preferentially utilize some BUN-derived NH3-N entering through the rumen wall in calves fed LP diets.     

Microbial protein production determined by urinary allantoin and renal urea sparing in normal and low protein fed Corriedale sheep

The aim of the present study was to compare the amount of microbial N entering the duodenum and the efficiency of N utilisation for microbial protein synthesis in normal (NP, 17.4 g N/d) and low protein (LP, 7.5 g N/d) fed Corriedale sheep. Renal functional tests for urea handling studies, and determination of urinary allantoin as an indirect method to estimate the microbial protein production were used. Although the N intake was 57% lower in LP sheep, the microbial N production was not very different between both diets (NP: 3.99 +/- 0.01 vs. LP: 3.79 +/- 0.02 g/d, P < 0.05). The efficiency of the microbial protein synthesis in the rumen, expressed as grams of microbial N per kg of digestible organic matter apparently digested in the rumen, was not statistically different for both diets. The urinary elimination of urea was reduced by 84% in LP sheep, essentially due to an important decrease in both renal plasma flow (-63%) and glomerular filtration rate (-71%). These haemodynamic changes would also reduce the filtered load and the urinary elimination of allantoin, thereby leading to an underestimation of the amount of microbial protein entering in the duodenum. Since the renal urea spared by the kidneys remains in the blood, it limits the drop ofthe available urea for ruminal recycling consecutive to a low nitrogen diet.     

Effect of dietary protein level on nitrogen metabolism in the growing bovine: I. Nitrogen recycling and intestinal protein supply in calves

Eight Angus heifer calves (234 kg) were assigned to either a high (HP; 126 g N/d) or low (LP; 66.5 g N/d) protein intake. Calves received 4.8 kg DM/d consisting of 30% cottonseed hulls and 70% corn-soybean meal in equal portions at 4-h intervals. Single doses of 14C- and 15N-urea and 15N-ammonium sulfate were injected into the blood urea-N (BUN) and ruminal NH3-N (RAN) pools, respectively, to measure rate of flux through, and transfer of N between, these and bacterial N. Nitrogen balance was greater (P less than .05) for HP than for LP (56.9 vs 25.1 g N/d), but abomasal N flow as a percentage of N intake was greater (P less than .05) for LP than for HP (124 vs 71.1%). Pool size and net synthesis rate for both RAN and BUN pools were greater (P less than .05) for HP than for LP. Calves fed HP degraded more (P less than .05) BUN in the gastrointestinal tract than calves fed LP (37.4 vs 14.0 g N/d). Quantities of RAN absorbed from the rumen also were greater (P less than .05) for HP than for LP (14.2 vs 2.8 g N/d). The proportion of total gastrointestinal BUN degradation occurring in the rumen averaged 53 and 26% for LP and HP. Data are interpreted to suggest that net incorporation of BUN into bacterial protein (urea recycling) is inversely related to level of protein intake.     

Quantification of intraruminal recycling of microbial nitrogen using nitrogen-15.

Studies were done to derive a technique to quantify intraruminal recycling of microbial nonammonia nitrogen (NAN). After dosing 15NH4+ into the rumen, ruminal NH3N and NAN pools were sampled over time for analyses of 15N enrichment. Compartmental analysis of both pools was used to quantify the percentage of microbial NAN that recycles in the rumen. Microbial NAN does not need to be fractionated from total NAN, assuming that only microbial NAN becomes labeled with 15N and that the ratio of microbial NAN:nonmicrobial NAN remains constant over time. Based on the data obtained from eight nonlactating cattle fed 85% corn silage diets at 1.5 to 1.0% BW, percentage of intraruminal recycling of microbial NAN was related to efficiency of bacterial protein synthesis (grams of CP/100 g of OM truly digested) in a curvilinear manner (y = 100 - .073x2). From this function, recycling was predicted to be approximately 75% for dairy cows consuming 50:50 forage:concentrate diets at 3.5% of BW. However, more data are needed to evaluate intraruminal recycling of microbial NAN using this technique under different dietary conditions.     

Effect of dietary protein level on nitrogen metabolism in lambs: studies using 15N-nitrogen

Six wether lambs (31 kg) were randomly assigned to two treatments (three lambs/treatment): a high protein intake (HP; 21 g N/d) or a low protein intake (LP; 12 g N/d). Each lamb received 860 g/d dry matter (DM) of a pelleted diet (75% corn-soybean meal, 25% cottonseed hulls) offered hourly in 24 equal portions. Single injections of 15N-labelled compounds were made into the ruminal NH3-N and blood urea-N pools to measure the rate of flux through, and transfer of N between, these and the bacterial N pool. Total tract digestibilities of DM and N were lower (P less than .05) for the LP than the HP treatment. Abomasal flows of total, feed or bacterial N tended to be greater (P greater than .05) in lambs fed HP than LP. Lambs fed HP excreted more (P less than .01) urinary N, yet retained a greater (P less than .01) amount of N than lambs fed LP (6.2 vs 1.8 and 9.7 vs 4.1 g N/d, respectively). Pool size and production rate for both ruminal NH3-N and blood urea-N were greater (P less than .05) for the HP than LP treatment. Lambs consuming HP degraded more (P less than .05) blood urea-N in the gastro-intestinal tract (13.4 vs 6.9 g N/d); however, lambs fed LP degraded a greater (P less than .05) percentage of synthesized body urea-N (88.7 vs 71.8%). Ruminal NH3-N absorption was greater (P less than .01) for the HP than LP treatment (3.1 vs .5 g N/d). Although the percentage of bacterial N derived from ruminal NH3-N was similar (P greater than .05) between diets (51.1 vs 63.9), a greater (P less than .05) percentage of bacterial N was derived from blood urea-N in lambs fed LP than HP (77.1 vs 30.2%). Lambs fed LP incorporated a greater (P less than .10) amount of blood urea-N into bacterial N than lambs fed HP (5.5 vs 2.6 g N/d). These data are interpreted to suggest that blood urea-N may provide a substantial quantity of N for bacterial protein synthesis and, thus, may be an important source of protein in the deficient animal. In addition, urea recycling may play an important role in the recovery of ruminal NH3-N lost through absorption in animals fed a high level of protein.     

Nutritive defaunation of the rumen in steers with subsequent refaunation using a cryopreserved monoculture of Entodinium caudatum

This study evaluated a technique for the nutritive defaunation of the rumen of cattle with subsequent single species refaunation using a cryopreserved monoculture of Entodinium caudatum (family Ophryoscolecidae). Four mature steers were nutritionally defaunated in two periods using two steers in each period. A diet containing (dry matter basis) 68% ground wheat grain, 7% wheat bran, 8% soybean oil and 17% wheat straw was used to decrease the pH of ruminal contents and to eliminate rumen ciliate protozoa. Protozoa‐free rumens were observed on day 8 and 9 in the first and second period, respectively, after the start of defaunation. A monoculture of E. caudatum (34/89/94) was transported from the Institute of Animal Physiology, Slovak Academy of Sciences in Ko?ice to the University of Kiel (Germany) in liquid nitrogen in October 1996. The inoculation was accomplished on day 15 in the second period by applying 30 ml culture medium with a monoculture of E. caudatum (34/89/94; average concentration of protozoal cells 2 650/ml) into the rumen of a defaunated steer via the ruminal fistula. The mono‐faunated steer was successfully inoculated with an average concentration of E. caudatum cells at 4.1 × 10³/ml (SD = 0.2) on day 2 after the inoculation.     

Fatty acid composition of ruminal bacteria and protozoa,and effect of defaunation on fatty acid profile in the rumen with special reference to conjugated linoleic acid in cattle

Objectives of this study were to compare fatty acid (FA) composition of ruminal bacterial (B) and protozoal (P) cells, and to investigate effect of protozoa on FA profile in the rumen of cattle. Three cows were used to prepare ruminal B and P cells. Four faunated and three defaunated cattle (half‐siblings) were used to study effect of protozoa on ruminal FA profile. Proportions of C16:0 and C18:0 in total fatty acids in B cells were 20.7% and 37.4%, whereas those in P cells were 33.4% and 11.6%, respectively. Proportions of trans‐vaccenic acid (VA) and cis‐9, trans‐11 conjugated linoleic acid (CLA) in B cells were 3.9% and 1.0%, and those in P cells were 5.5% and 1.6%, respectively, being higher in P cells. Proportions of C18:1, C18:2 and C18:3 in P cells were two to three times higher than in B cells. Proportions of unsaturated fatty acids, VA and CLA in B cells of faunated cattle were higher than those of defaunated. VA and CLA in the ruminal fluid of faunated were also 1.6 to 2.5 times higher than those of defaunated. This tendency was similar for cell‐free fraction of ruminal fluid. These results indicate that protozoa contribute greatly in VA and CLA production in the rumen.     

Effect of various levels of imbalance between energy and nitrogen release in the rumen on microbial protein synthesis and nitrogen metabolism in growing double-muscled Belgian Blue bulls fed a corn silage-based diet

Seven double-muscled Belgian Blue bulls (initial BW: 341 +/- 21 kg) with cannulas in the rumen and proximal duodenum were used in an incomplete replicated Latin square. The study examined the effect of imbalance between energy and N in the rumen on microbial protein synthesis and N metabolism by giving the same diet according to 3 different feeding patterns. The feed ingredients of the diet were separated into 2 groups supplying the same amount of fermentable OM (FOM) but characterized by different levels of ruminally degradable N (RDN). The first group primarily provided energy for the ruminal microbes (12.5 g of RDN/kg of FOM), whereas the second provided greater N (33.3 g of RDN/kg of FOM). These 2 groups were fed to the bulls in different combinations with the aim of creating 3 levels of imbalance (0, 20, and 40 g/ kg of DM) between energy and N supplies in the rumen. Imbalance was measured by the variation of the degradable protein balance (OEB value in the Dutch system) of the diet between the 2 meals each a day. Diurnal variations in ruminal NH3-N concentrations and plasma urea concentrations were greatly influenced by the feeding patterns of the diet. Introduction of imbalance affected neither microbial N flow at the duodenum (P = 0.97) nor efficiency of growth (P = 0.54). The feeding patterns of the diet had no negative impact on NDF degradation in the rumen (P = 0.33). Nitrogen retention was not affected by imbalance (P = 0.74) and reached 49.7, 52.0, and 51.3 g/d, respectively for 0, 20, and 40 g of OEB/kg of DM imbalance. It seems that introduction of an imbalance between energy and N supplies for the ruminal microbes by altering the feeding pattern of the same diet does not negatively influence the microbial activity in the rumen nor N retention of the animal. Nitrogen recycling in the rumen plays a major role in regulating the amount of ruminally available N and allows a continuous synchronization of N and energy-yielding substrates for the microorganisms in the rumen. Therefore, imbalance between dietary energy and N created over a 24-h interval was not detrimental to rumen microbial growth for the animal as long as the level of imbalance did not exceed 40 g of OEB/kg of DM. Thus, these feeding patterns of the diet can be used under practical feeding conditions with minimal impact on the performance of ruminant animals for meat production.     

Effect of defaunating the rumen on growth and carcass composition of lambs

The effect of defaunating the rumen on growth performance and carcass composition of lambs fed a molasses-urea diet was investigated. Before the growth trial, all the animals were defaunated. Based on live weight and daily gain during a preliminar period, the animals were divided in two groups whereafter one group was refaunated. Defaunation caused a decrease in propionic acid percentage in the rumen. Daily gain and food conversion efficiency were better in the defaunated group, but only during the first five weeks. The response over the whole trial (0-9 weeks) remained positive however. There was a trend towards more meat and less fat in the carcass of defaunated lambs. The fact that two animals died during the defaunation procedure indicates the need for a completely harmless and effective defaunating agent.     

Long-term defaunation increases the abundance of cellulolytic ruminococci and methanogens but does not affect the bacterial and methanogen diversity in the rumen of sheep

Protozoa are commensal eukaryotes in the rumen of herbivores. Protozoa are large producers of hydrogen, which is utilized by methanogenic archaea to produce methane, a greenhouse gas. The removal of protozoa from the rumen (defaunation) decreases methanogenesis, but also negatively affects fiber digestion, which is the main function of the rumen. The aim of this study was to examine the effect of long-term defaunation on the structure of the microbiota and particularly methanogenic archaea and fibrolytic bacteria to better understand the microbial mechanisms responsible for the decrease in methanogenesis and fibrolysis. The trial was conducted in 5 adult sheep subjected successively to long-term defaunation (2 yr), refaunation (12 wk), and short-term defaunation (10 wk). Methanogens were enumerated by quantitative PCR targeting the rrs (16S ribosomal RNA subunit) and mcrA (methyl coenzyme-M reductase) genes. The rrs gene was used to quantify the 3 major culturable rumen cellulolytic bacterial species (i.e., Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens) and total bacteria. Bacterial and methanogen diversity was also examined by PCR-DGGE (PCR-denaturing gradient gel electrophoresis) analysis targeting the rrs and mcrA genes, respectively. Total rumen bacterial density estimated as rrs copies per gram of DM of rumen content increased in response to long- and short-term defaunation (+1 log, P < 0.001), but without noticeable shifts in diversity. Defaunation increased the rrs copies per gram of DM of rumen content of R. albus and R. flavefaciens (+2 log, P < 0 0.001), but did not affect that of F. succinogenes. Despite a 20% reduction in methane emission in the 2 defaunated periods, the mcrA and rrs copies of methanogens per gram of DM of rumen content increased (+1 log, P < 0.001) in the absence of protozoa, whereas the diversity of the dominant methanogenic community was not modified. This study shows no major difference between long- and short-term defaunation in abundance and diversity of bacteria and archaea. It also provides evidence that monitoring the abundance and diversity of methanogens is not sufficient to comprehend the microbial mechanisms leading to a reduction in methane emissions by ruminants. This study also reports for the first time in sheep a selective effect of defaunation on the abundance of cellulolytic bacterial species.     

Manipulation of rumen ecology by dietary lemongrass (Cymbopogon citratus Stapf.) powder supplementation

This experiment was conducted to investigate the effect of lemongrass [Cymbopogon citratus (DC.) Stapf.] powder (LGP) on rumen ecology, rumen microorganisms, and digestibility of nutrients. Four ruminally fistulated crossbred (Brahman native) beef cattle were randomly assigned according to a 4 x 4 Latin square design. The dietary treatments were LGP supplementation at 0, 100, 200, and 300 g/d with urea-treated rice straw (5%) fed to allow ad libitum intake. Digestibilities of DM, ether extract, and NDF were significantly different among treatments and were greatest at 100 g/d of supplementation. However, digestibility of CP was decreased with LGP supplementation (P < 0.05), whereas ruminal NH(3)-N and plasma urea N were decreased with incremental additions of LGP (P < 0.05). Ruminal VFA concentrations were similar among supplementation concentrations (P > 0.05). Total viable bacteria, amylolytic bacteria, and cellulolytic bacteria were significantly different among treatments and were greatest at 100 g/d of supplementation (4.7 x 10(9), 1.7 x 10(7), and 2.0 x 10(9) cfu/mL, respectively). Protozoal populations were significantly decreased by LGP supplementation. In addition, efficiency of rumen microbial N synthesis based on OM truly digested in the rumen was enriched by LGP supplementation, especially at 100 g/d (34.2 g of N/kg of OM truly digested in the rumen). Based on this study, it could be concluded that supplementation of LGP at 100 g/d improved digestibilities of nutrients, rumen microbial population, and microbial protein synthesis efficiency, thus improving rumen ecology in beef cattle.