Protozoa Counts Research Articles

Effect of Saccharomyces cerevisiae fermentation product on ruminal fermentation and nutrient utilization in dairy cows

The goal of this experiment was to investigate the effect of yeast culture (Saccharomyces cerevisiae) on rumen fermentation, nutrient utilization, and ammonia and methane emission from manure in dairy cows. Eight ruminally cannulated Holstein cows were allocated to 2 dietary treatments in a crossover design. Treatments were control (no yeast culture) and XP (yeast culture, fed at 56 g/head per day; XP, Diamond V Mills Inc., Cedar Rapids, IA). Dry matter intake, milk yield, milk composition, and body weight were similar between treatments. Milk urea nitrogen concentration was also not affected by treatment. Rumen pH was similar between the control and XP treatments, but rumen ammonia concentration tended to be lower with XP than with the control. Treatment had no effect on concentrations of total or individual volatile fatty acids, protozoal counts, polysaccharide-degrading activities (except amylase activity that tended to be increased by XP), or methane production in the rumen. Urinary N losses did not differ significantly between treatments, but allantoin and total purine derivative excretions and the estimated microbial N outflow from the rumen tended to be increased by XP compared with the control treatment. Total-tract apparent digestibility of dietary nutrients was not affected by XP. Milk fatty acid composition was also not altered by XP supplementation. Cumulative (253 h) ammonia and methane emissions from manure, measured in a steady-state gas emission system, were slightly decreased by XP. Overall, the yeast culture tested had little effect on ruminal fermentation, digestibility, or N losses, but tended to reduce rumen ammonia concentration and increase microbial protein synthesis in the rumen, and decreased ammonia and methane emissions from manure.

Utilization of complete rumen modifier on sheep fed high fibrous forages.

A research to improve livestock productivity and lower enteric methane production on ruminant was conducted by manipulation approach on rumen system using a complete rumen modifier ( CRM ). An in vivo experiment was carried out using twenty four sheep ( mean weight 18 kg) which were distributed into 3 treatment groups of feed additive: I. control ( without treatment: K); II: K + CRM -LG; III: K + CRM -EL. Diet given consisted of fermented rice straw ( ad libitum ) + concentrate containing 16 % protein (400 g/head/day), and drinking water was given ad libitum . The experiment was conducted for 14 weeks based on completely randomized design. By the end of the experiment, animals were placed in the metabolism cages for 2 weeks (ie. 1 week for adaptation and 1 week for data collection). Rumen liquid of each treated animal was taken for the measurement of rumen characteristics. Parameter measuremed were: total gas production; gas composition of CO 2 and CH 4 ; in vitro DMD; NH 3 and VFA contents; pH; bacterial and protozoal counts; consumption/ DMI; in vivo DMD; ADG and FCR. The results showed that productivity of sheep was improved by CRM treatments followed by lowered enteric methane production. The ADG values of CRM treatments (71.4 to 73.5 g) were significantly higher (P < 0.05) than that of control (50 g). The improvement of average daily gain was followed by a better feed conversion (P < 0.05) (ie. 10.6 vs. 12.8). The CRM treatments lowered the percentage of CH 4 by 24% compared to Control (P < 0.05). The total and composition of VFA of CRM -treated rumen liquor were significantly different (P<0.05) compared to that of rumen liquor of Control (ie. the total VFA: 85.3 vs 73.5 mM and the percentage of acetic acid: 67.8 vs 60.3%). It is concluded that CRM treatment resulted in positive effects on growth of ruminant fed high fibrous forages such as rice straw and could lower enteric methane production. Key Words : Rumen Modifier, Productivity, Enteric Methane, Sheep, Rice Straw

Effects of supplemental humic acid on ruminal fermentation and blood variables in rams

In this study, we particularly aimed to research the effect of supplemental humic acid on ruminal fermentation and blood variables in rams. A trial was conducted to evaluate the effect of humic acid (HA) on protozoa count, percentages of different protozoa types and blood parameters. Three male Kivircik rams with ruminal cannula were used in a Latin square design, during 22 days periods (15 days for adaptation, 7 days for collection). They received 0 control group (CG), 5 g/day or 10 g/day of HA (HA5, HA10, assay groups). HA were added to the ration with grain diet. Ration was consisted of 5% grain diet and 95% alfalfa hay. Rumen contents collected before and, 3h and 6h after morning feeding on days 1 and 7 in each collection period were analyzed. Blood samples were also collected the same days. No significant difference in biochemical and hematological parameters (except eosinophils levels, P<0.05), variables of ruminal fluid (except sodium levels before feeding) and species of rumen protozoa organism (except the percentage Epidinium spp.) were evidenced with the addition of HA. In conclusion, we think that they might have been true the widely use in animal food of humates which don’t have any negative effect on biochemical and hematological parameters in particular.

Effect of eucalyptus (Eucalyptus citriodora) fresh or residue leaves on methane emission in vitro

Rumen fermentation and methane emission for eucalyptus (Eucalyptus citriodora) fresh leaves (FL) or residue leaves (RL), after essential oil extraction from eucalyptus leaves in comparison with alfalfa (Medicago sativa) hay, were investigated in vitro. Eucalyptus FL and RL were obtained from the Distillery Trees Barras Company, Torrinha City, Sao Paulo, Brazil. The semi-automatic system of gas production was used to measure gas production, methane emission and rumen fermentation after 24 h incubation in vitro.The results showed that the crude protein (CP) contents were 76.4, 78.1 and 181.9 g kg−1 DM for eucalyptus FL, RL and alfalfa hay, respectively. The neutral-detergent fibre (NDF) and acid-detergent fibre (ADF) were significantly lower in eucalyptus FL and RL than alfalfa hay. The Eucalyptus fresh and residue leaves were rich in total phenols (TP) and total tannins (TT) but had negligible content of condensed tannins (CT). There was significant reduction in cumulative gas production about 54 and 51% with eucalyptus FL and RL, respectively, compared with alfalfa hay. The methane emission (mL/g DM) was reduced (P<0.05) by 53 and 57% with eucalyptus FL and RL, respectively, but the reduction was 21 and 16% when expressed on truly digested organic matter basis. There were a decline (P<0.05) in true dry and organic matter degradation in vitro in eucalyptus FL and RL compared with alfalfa hay substrate. The partitioning factor values were higher (P<0.05) in eucalyptus FL and RL than alfalfa hay. There was no significant difference observed between eucalyptus FL, RL and alfalfa hay in protozoa count. It is concluded that the eucalyptus leaves have potential effect to mitigate CH4 production in vitro, which may be attributed to a decrease in fermentable substrate rather than to a direct effect on methanogenesis.

Effects of propolis on in vitro rumen microbial fermentation

Bu araştırmada, Rusitec tekniği (Rumen Simulation Technique) kullanılarak, farklı yoğunluklardaki etanolik propolis ekstraktlarının rumen mikrobiyal fermantasyonu üzerine etkilerinin belirlenmesi amaçlandı. Gerçek hacimleri 750 ml olan 6 fermenterin kullanıldığı araştırma 14 gün sürdü. Her bir fermenterde günlük olarak 6 g yonca otu peleti ve 4 g konsantre pelet yemden oluşan bir rasyon inkübe edildi. Araştırmadaki uygulama grupları kontrol (propolis içermeyen % 70’lik etanol çözeltisinden 0,5 ml/gün), propolisin % 20’lik etanolik ekstraktı (0,5 ml/gün) ve propolisin % 60’lık etanolik ekstraktı (0,5 ml/gün) şeklindeydi. Rusitec fermenterlerine ilave edilen propolisin her iki yoğunluktaki ekstraktı da ruminal pH, toplam kısa zincirli yağ asitleri ve asetat üretimleri, asetatın propiyonata oranı, toplam protozoon sayısı ve yem kuru maddesi sindirilebilirliklerinde istatistiksel bir değişime neden olmadı. Ancak % 60’lık propolis ekstraktı propiyonat üretimini belirgin bir şekilde azaltırken (p<0.05), hem % 60 hem de % 20’lik propolis ekstraktı ruminal bütirat üretimini belirgin bir şekilde artırdı (p<0.05). Ayrıca her iki yoğunluktaki propolis ektraktı da ruminal bakterilerin toplam sayısında azalmaya (p<0.05) neden oldu. NH3-N konsantrasyonunda hem düşük hem de yüksek yoğunluktaki propolis ekstraktı ilavesinden sonra sırasıyla % 24 ve % 39’luk bir azalma (p<0.05) belirlendi. Bu araştırmadan elde edilen sonuçlar propolisin ruminal amonyak üretimini azaltmada ve ruminal azot değerlendirilebilirliğini iyileştirmede kullanılabilecek bir katkı maddesi olabileceğine işaret etmektedir

Utility of buckwheat ( Fagopyrum esculentum Moench) as feed: Effects of forage and grain on in vitro ruminal fermentation and performance of dairy cows

Buckwheat is of high value in crop rotations and overall agricultural ecology because of strong rooting and intensive flowering properties, but it is rarely cultivated and information on its nutritional value to ruminants is scarce. The contents of net energy for lactation (NEL), as estimated with the Hohenheim Gas Test ( n = 3), were 4.3, 4.9 and 7.5 MJ NEL/kg dry matter (DM) for fresh and ensiled whole buckwheat plants and buckwheat grain, respectively. In two experiments with the Rumen Simulation Technique (Rusitec), ruminal fermentation characteristics of buckwheat forages and buckwheat grain ( n = 4/diet) were evaluated. In the first experiment, 0, 300 or 600 g/kg of a pure hay diet were replaced by either fresh or ensiled buckwheat to create five diets. Neither form of buckwheat forages had effects on in vitro ruminal degradability and short chain fatty acid (SCFA) concentrations and composition. The use of fresh buckwheat reduced ruminal ammonia concentrations and enhanced estimated microbial N growth efficiency. These differences did not occur with silage, indicating a change in nutritional value by ensiling. Fresh buckwheat reduced the number of bacteria in the incubated fluid, while ensiled buckwheat reduced that of holotrich protozoa. Methane formation was not influenced by the buckwheat forages. In the second experiment, wheat meal (400 g/kg dietary DM), was replaced stepwise (0.5 and 1.0) by buckwheat grain meal. This did not cause differences in parameters of nutrient degradability, relative N efficiency and total amount and composition of SCFA. Holotrich protozoa counts increased, but total gas formation decreased with increasing dietary level of buckwheat grain. In a final experiment, cows yielding about 40 kg milk/day were fed mixed silage-concentrate diets ( n = 4). A control diet contained no buckwheat. In a second diet, maize silage was partly substituted by buckwheat silage (98 g/kg dietary DM). In a third group, part of the energy concentrate was substituted by buckwheat grain meal (94 g/kg). There were no effects on feed intake, milk yield and milk composition. Buckwheat proved to be a plant that offers different feeds of a quality sufficient to be considered suitable in ruminant nutrition.

Difference in the nature of tannins on in vitro ruminal methane and volatile fatty acid production and on methanogenic archaea and protozoal populations

Six plant sources of hydrolyzable tannins (HT) or HT and condensed tannins (CT; designated as HT1, HT2, HT3, HT + CT1, HT + CT2, and HT + CT3) were evaluated to determine their effects in vitro on CH4 production and on ruminal archaeal and protozoa populations, and to assess potential differences in biological activities between sources containing HT only or HT and CT. Samples HT1, HT2, and HT3 contained only HT, whereas samples HT + CT1, HT + CT2, and HT + CT3 contained HT and CT. In experiment 1, in vitro incubations with samples containing HT or HT + CT resulted in a decrease in CH4 production of 0.6 and 5.5%, respectively, compared with that produced by incubations containing the added tannin binder polyethylene glycol-6000. Tannin also suppressed the population of methanogenic archaea in all incubations except those with HT2, with an average decrease of 11.6% in HT incubations (15.8, 7.09, and 12.0 in HT1, HT2, and HT3) and 28.6% in incubations containing HT + CT (35.0, 40.1, and 10.8 in HT + CT1, HT + CT2, and HT + CT3) when compared with incubations containing added polyethylene glycol-6000. The mean decrease in protozoal counts was 12.3% in HT and 36.2% in HT + CT incubations. Tannins increased in vitro pH, reduced total VFA concentrations, increased propionate concentrations, and decreased concentrations of iso-acids. In experiment 2, when a basal diet was incubated with graded levels of HT + CT1, HT + CT2, and HT + CT3, the total gas and CH4 production and archaeal and protozoal populations decreased as the concentration of tannins increased. Our results confirm that tannins suppress methanogenesis by reducing methanogenic populations in the rumen either directly or by reducing the protozoal population, thereby reducing methanogens symbiotically associated with the protozoal population. In addition, tannin sources containing both HT and CT were more potent in suppressing methanogenesis than those containing only HT.

Effect of lauric acid and coconut oil on ruminal fermentation, digestion, ammonia losses from manure, and milk fatty acid composition in lactating cows

This experiment (replicated 3×3 Latin square design) was conducted to investigate the effects of lauric acid (LA) or coconut oil (CO) on ruminal fermentation, nutrient digestibility, ammonia losses from manure, and milk fatty acid (FA) composition in lactating cows. Treatments consisted of intraruminal doses of 240g of stearic acid/d (SA; control), 240g of LA/d, or 530g of CO/d administered once daily, before feeding. Between periods, cows were inoculated with ruminal contents from donor cows and allowed a 7-d recovery period. Treatment did not affect dry matter intake, milk yield, or milk composition. Ruminal pH was slightly increased by CO compared with the other treatments, whereas LA and CO decreased ruminal ammonia concentration compared with SA. Both LA and CO decreased protozoal counts by 80% or more compared with SA. Methane production rate in the rumen was reduced by CO compared with LA and SA, with no differences between LA and SA. Treatments had no effect on total tract apparent dry matter, organic matter, N, and neutral detergent fiber digestibility coefficients or on cumulative (15d) in vitro ammonia losses from manure. Compared with SA, LA and CO increased milk fat 12:0, cis-9 12:1, and trans-9 12:1 content and decreased 6:0, 8:0, 10:0, cis-9 10:1, 16:0, 18:0, cis 18:1, total 18:2, 18:3 n-3 and total polyunsaturated FA concentrations. Administration of LA and 14:0 (as CO) in the rumen were apparently transferred into milk fat with a mean efficiency of 18 and 15%, respectively. In conclusion, current data confirmed that LA and CO exhibit strong antiprotozoal activity when dosed intraruminally, an effect that is accompanied by decreases in ammonia concentration and, for CO, lowered methane production. Administration of LA and CO in the rumen also altered milk FA composition.

Parâmetros fermentativos, produção de proteína microbiana, concentrações de ureia no leite e no plasma e balanço de nitrogênio de vacas alimentadas com silagem de milho ou cana-de-açúcar com caroço de algodão

Doze vacas da raça Holandesa foram distribuídas em três quadrados latinos 4 × 4 com o objetivo de avaliar o pH, o N-amoniacal, a produção de proteína microbiana, o número de protozoários no rúmen, a concentração de N-ureia no plasma e no leite e balanço de nitrogênio. Os tratamentos consistiram de silagem de milho ou cana-de-açúcar contendo 0, 7 ou 14% de caroço de algodão. Não houve diferença no pH quando fornecida cana-de-açúcar, apesar da diminuição dos valores após o início da alimentação. Os valores de N-amoniacal mantiveram-se acima de 15 mg/dL, após 6 horas de alimentação, apenas quando fornecida silagem de milho e cana-de-açúcar com 7% de caroço de algodão. O maior consumo de MS e nutrientes obtido com a silagem de milho implicou em menor eficiência de uso do nitrogênio e maiores excreções de N-ureia na urina em relação às demais dietas (150,6 vs 96,0 mg/kg PV). O fornecimento de cana-de-açúcar com 7% de caroço de algodão promoveu maiores quantidades de purinas totais (PT), purinas absorvidas (PA), nitrogênio microbiano (207, 6 vs 185,2 g/dia) e melhor balanço de compostos nitrogenados (31,5 vs 15,5) em relação às demais dietas com cana-de-açúcar. O menor consumo de MS e extrato etéreo quando fornecida cana-de-açúcar sem caroço de algodão resultou em maior número de protozoários (129,4 vs 34,0 × 10³/mL), maior excreção urinária de nitrogênio e menor síntese de proteína microbiana em relação às demais dietas, o que indica falta de sincronização na degradação da energia e proteína na relação volumoso:concentrado proposta neste experimento.

Investigating unsaturated fat, monensin, or bromoethanesulfonate in continuous cultures retaining ruminal protozoa. I. Fermentation, biohydrogenation, and microbial protein synthesis

Methane is an end product of ruminal fermentation that is energetically wasteful and contributes to global climate change. Bromoethanesulfonate, animal-vegetable fat, and monensin were compared with a control treatment to suppress different functional groups of ruminal prokaryotes in the presence or absence of protozoa to evaluate changes in fermentation, digestibility, and microbial N outflow. Four dual-flow continuous culture fermenter systems were used in 4 periods in a 4×4 Latin square design split into 2 subperiods. In subperiod 1, a multistage filter system (50-μm smallest pore size) retained most protozoa. At the start of subperiod 2, conventional filters (300-μm pore size) were substituted to efflux protozoa via filtrate pumps over 3 d; after a further 7 d of adaptation, the fermenters were sampled for 3 d. Treatments were retained during both subperiods. Flow of total N and digestibilities of NDF and OM were 18, 16, and 9% higher, respectively, for the defaunated subperiod but were not different among treatments. Ammonia concentration was 33% higher in the faunated fermenters but was not affected by treatment. Defaunation increased the flow of nonammonia N and bacterial N from the fermenters. Protozoal counts were not different among treatments, but bromoethanesulfonate increased the generation time from 43.2 to 55.6h. Methanogenesis was unaffected by defaunation but tended to be increased by unsaturated fat. Defaunation did not affect total volatile fatty acid production but decreased the acetate:propionate ratio; monensin increased production of isovalerate and valerate. Biohydrogenation of unsaturated fatty acids was impaired in the defaunated fermenters because effluent flows of oleic, linoleic, and linolenic acids were 60, 77, and 69% higher, and the ratio of vaccenic acid:unsaturated FA ratio was decreased by 34% in the effluent. This ratio was increased in both subperiods with the added fat diet, indicating an accumulation of intermediates of biohydrogenation. However, the flow of 18:2 conjugated linoleic acid was unaffected by defaunation or by treatments other than added fat. The flows of trans-10, trans-11, and total trans-18:1 fatty acids were not affected by monensin or faunation status.

Efeitos da fonte de enxofre sobre a população de protozoários e degradabilidade no rúmen

Estudaram-se os efeitos de fontes de enxofre na dieta sobre os parâmetros ruminais de bovinos Nelore, utilizando-se oito novilhos canulados no rúmen e submetidos a quatro tratamentos, segundo a fonte de enxofre. Os animais do grupo-controle não receberam suplementação de enxofre. Os dos outros três grupos receberam flor de enxofre ou metionina ou carboquelatado, como suplementação. Os animais foram arraçoados com dieta total, utilizando cana-de-açúcar picada como volumoso. O período experimental foi de 76 dias, dividido em quatro subperíodos de 19 dias, dos quais 14 eram para adaptação à dieta e cinco para as colheitas. Não foram encontradas diferenças entre as fontes de enxofre na degradabilidade da matéria seca, proteína bruta e fibras em detergente ácido e neutro e no pH ruminal. O tratamento com carboquelatado resultou em maior número de protozoários ciliados e o tratamento metionina em menor contagem. O carboquelatado pode ser uma boa alternativa para a suplementação de enxofre para bovinos em confinamento.

Efficiency of monolaurin in mitigating ruminal methanogenesis and modifying C-isotope fractionation when incubating diets composed of either C3or C4plants in a rumen simulation technique (Rusitec) system

Mitigation of methanogenesis in ruminants has been an important goal for several decades. Free lauric acid, known to suppress ruminal methanogenesis, has a low palatability; therefore, in the present study the aim was to evaluate the mitigation efficacy of its esterified form (monolaurin). Further, 13C-isotope abundance (delta13C) and 13C-12C fractionation during methanogenesis and fermentation were determined to evaluate possible microbial C-isotope preferences. Using the rumen simulation technique, four basal diets, characterised either by the C3 plants grass (hay) and wheat (straw and grain), or the C4 plant (13C excess compared with C3 plants) maize (straw and grain), and a mixture of the latter two, were incubated with and without monolaurin (50 g/kg dietary DM). Added to hay, monolaurin did not significantly affect methanogenesis. When added to the other diets (P < 0.05 for the wheat-based diet) methane formation was lowered. Monolaurin decreased fibre disappearance (least effect with the hay diet), acetate:propionate ratio, and protozoal counts. Feed residues and SCFA showed the same delta13C as the diets. Methane was depleted in 13C while CO2 was enriched in 13C compared with the diets. Monolaurin addition resulted in 13C depletion of CO2 and enrichment in CH4 (the latter only in the hay diet). In conclusion, monolaurin proved to effectively decrease methanogenesis in the straw-grain diets although this effect might partly be explained by the concomitantly reduced fibre disappearance. The influence on 13C-isotope abundance and fractionation supports the hypothesis that ruminal microbes seem to differentiate to some extent between C-isotopes during methanogenesis and fermentation.

Crushed sunflower, flax, or canola seeds in lactating dairy cow diets: Effects on methane production, rumen fermentation, and milk production

The objective of this study was to investigate the potential of reducing enteric methane production from dairy cows by incorporating into the diet various sources of long-chain FA varying in their degree of saturation and ruminal availability. The experiment was conducted as a crossover design with 16 lactating dairy cows maintained in 2 groups and fed 4 dietary treatments in four 28-d periods. Eight ruminally cannulated primiparous cows (96±18 d in milk) were assigned to group 1 and 8 multiparous cows (130±31 d in milk) were assigned to group 2. The dietary treatments were: 1) a commercial source of calcium salts of long-chain fatty acids (CTL), 2) crushed sunflower seeds (SS), 3) crushed flaxseed (FS), and 4) crushed canola seed (CS). The oilseeds added 3.1 to 4.2% fat to the diet (DM basis). All 3 oilseed treatments decreased methane production (g/d) by an average of 13%. When corrected for differences in dry matter intake (DMI), compared with CTL, methane production (g/kg of DM intake) was decreased by feeding FS (−18%) or CS (−16%) and was only numerically decreased (−10%) by feeding SS. However, compared with the CTL, feeding SS or FS lowered digestible DMI by 16 and 9%, respectively, because of lowered digestibility. Thus, only CS lowered methane per unit of digestible DM intake. Feeding SS and CS decreased rumen protozoal counts, but there were no treatment effects on mean ruminal pH or total volatile fatty acid concentration. Milk efficiency (3.5% fat corrected milk/DMI), milk yield, and component yield and concentrations were not affected by oilseed treatments. The study shows that adding sources of long-chain fatty acids to the diet in the form of processed oilseeds can be an effective means of reducing methane emissions. However, for some oilseeds such as SS or FS, the reduction in methane can be at the expense of diet digestibility. The use of crushed CS offers a means of mitigating methane without negatively affecting diet digestibility, and hence, milk production.

Effects of supplementary inosine on nutrient digestibility, ruminal fermentation and nitrogen balance in goats fed high amount of concentrate

A study was conducted to compare responses of nutrient intake, digestibility, ruminal fermentation, and N balance in goats fed high amounts of concentrate supplemented with inosine or urea. Four Japanese Saneen goats (49.2 ± 5 kg), housed in individual metabolic cages, were blocked by weight into two pairs, and assigned inosine or urea as non-protein N (NPN) supplements to their basal diet at 0.15 g per kg body weight on a urea basis, in a 2 × 2 switch back experimental design. The basal diet consisted of 0.4 timothy hay, and 0.6 concentrates, and was fed to the goats as a total mixed ration (TMR). The experimental period was 13 days, including 7 days of adaptation, 5 days of sampling of feces, urine and orts, and 1 day of sampling of rumen fluid. Intake of dry matter (DM), organic matter (OM), neutral detergent fiber (NDF) and crude protein (CP) and digestibility in the total tract, as well as N balance, were assessed by total fecal collection. Rumen fluid was sampled at 0, 1, 4 and 7 h after feeding, and analyzed for pH, ammonia and volatile fatty acids (VFA). Additionally, samples collected at 0 and 4 h after feeding were assessed for protozoal counts. Daily intakes of DM, OM, NDF and CP did not differ among treatments. Digestibility in the total tract tended (P=0.07) to be lower for NDF and was numerically lower for DM (P=0.11), OM (P=0.11), and NDF (P=0.12) in goats supplemented with inosine versus urea. Ammonia concentration and ruminal pH were similar among treatments, and protozoal counts 4 h after feeding was lower (P=0.02) in goats supplemented with inosine compared to those on urea. No differences between treatments occurred in concentrations of rumen VFA (i.e., acetate, propionate, valerate, iso-acids, and total VFA) except for butyrate, which was higher (P=0.03) in goats supplemented with inosine versus urea 4 h after feeding. There were no differences in intake, absorption and retention of N between treatments. Thus, responses in nutrient intake, digestibility, ruminal fermentation and N balance in goats fed high amounts of concentrate with supplementary inosine and urea were similar.

Physiological changes in rumen fermentation during acidosis induction and its control using a multivalent polyclonal antibody preparation in heifers

Physiological changes in rumen fermentation during acidosis induction and its control using a multivalent polyclonal antibody preparation (PAP) were studied in a completely randomized experiment using 12 crossbred heifers (452 +/- 20 kg of BW). Treatments were control (CTR) or PAP. The acidosis induction protocol consisted of 3 periods: 3 mo of 100% fescue hay fed for ad libitum intake, 10 d (from d 1 to 10 of the experiment) of adaptation to the treatment (100% forage feeding + 10 mL/d of PAP top-dressed to the treatment group), and 5 d (from d 11 to 15 of the experiment) of transition, which consisted of increasing the concentrate (16.5% CP) 2.5 kg/d up to 12.5 kg/d while maintaining ad libitum intake of fescue and providing 10 mL/d of PAP to the treated heifers. Concentrate feeding of 12.5 kg/d was maintained until heifers developed acidosis (from d 16 to 22 of the experiment). When an animal was considered acidotic, it was changed to a 50:50 forage:concentrate diet, monitored for 4 d, and removed from the experiment. Samples of ruminal fluid were collected before and 6 h after feeding to determine pH, VFA, lactate, protozoa counts, and DNA extraction for quantitative real-time PCR and denaturing gradient gel electrophoresis analyses. Only samples collected during adaptation to the treatment, at 3 and 1 d before acidosis, on the acidosis day, and at 1 and 4 d after acidosis were analyzed. Differences were declared at P < 0.05. Heifers (83% for CTR, and 50% for PAP) entered into acidosis 5.25 +/- 0.17 d after the beginning of the transition. The fermentation profile of animals with acidosis was similar between treatments. From 3 d before acidosis to acidosis day, decreases in pH and in acetate-to-propionate ratio and increases in total VFA, butyrate, and entodiniomorph counts were observed. However, the greatest concentrations of Streptococcus bovis and Megasphaera elsdenii (79 +/- 54 and 104 +/- 73 ng of DNA/mL of ruminal fluid, respectively) and a decrease in DMI (10.6 vs. 6.46 kg, respectively) were recorded 1 d after acidosis. Compared with CTR heifers, heifers fed PAP had greater pH before feeding on d 6 (6.70 vs. 6.11), 8 (6.54 vs. 5.95), and 9 (7.26 vs. 6.59) after the beginning of the feeding challenge. Heifers fed PAP tended to have greater total VFA concentrations than CTR (124 and 114 +/- 4.0 mM, respectively). These results indicate that PAP may be effective in controlling acidosis of heifers during a rapid transition to a high-concentrate diet.

Soybean oil and linseed oil supplementation affect profiles of ruminal microorganisms in dairy cows

The objective of this study was to evaluate changes in ruminal microorganisms and fermentation parameters due to dietary supplementation of soybean and linseed oil alone or in combination. Four dietary treatments were tested in a Latin square designed experiment using four primiparous rumen-cannulated dairy cows. Treatments were control (C, 60 : 40 forage to concentrate) or C with 4% soybean oil (S), 4% linseed oil (L) or 2% soybean oil plus 2% linseed oil (SL) in a 4 × 4 Latin square with four periods of 21 days. Forage and concentrate mixtures were fed at 0800 and 2000 h daily. Ruminal fluid was collected every 2 h over a 12-h period on day 19 of each experimental period and pH was measured immediately. Samples were prepared for analyses of concentrations of volatile fatty acids (VFA) by GLC and ammonia. Counts of total and individual bacterial groups (cellulolytic, proteolytic, amylolytic bacteria and total viable bacteria) were performed using the roll-tube technique, and protozoa counts were measured via microscopy in ruminal fluid collected at 0, 4 and 8 h after the morning feeding. Content of ruminal digesta was obtained via the rumen cannula before the morning feeding and used immediately for DNA extraction and quantity of specific bacterial species was obtained using real- time PCR. Ruminal pH did not differ but total VFA (110 v. 105 mmol/l) were lower (P < 0.05) with oil supplementation compared with C. Concentration of ruminal NH3-N (4.4 v. 5.6 mmol/l) was greater (P < 0.05) due to oil compared with C. Compared with C, oil supplementation resulted in lower (P < 0.05) cellulolytic bacteria (3.25 × 108v. 4.66 × 108 colony-forming units (CFU)/ml) and protozoa (9.04 × 104v. 12.92 × 104 cell/ml) colony counts. Proteolytic bacteria (7.01 × 108v. 6.08 × 108 CFU/ml) counts, however, were greater in response to oil compared with C (P < 0.05). Among oil treatments, the amount of Butyrivibrio fibrisolvens, Fibrobacter succinogenes and Ruminococcus flavefaciens in ruminal fluid was substantially lower (P < 0.05) when L was included. Compared to C, the amount of Ruminococcus albus decreased by an average of 40% regardless of oil level or type. Overall, the results indicate that some ruminal microorganisms, except proteolytic bacteria, are highly susceptible to dietary unsaturated fatty acids supplementation, particularly when linolenic acid rich oils were fed. Dietary oil effects on ruminal fermentation parameters seemed associated with the profile of ruminal microorganisms.

Evidence for the inhibition of the terminal step of ruminal α-linolenic acid biohydrogenation by condensed tannins

Effects of condensed tannins (CT), either via extract or plant-bound, and saponin extract on ruminal biohydrogenation of α-linolenic acid (ALA) were investigated in vitro. Grass-clover hay served as basal diet (control). The control hay was supplemented with extracts contributing either CT from Acacia mearnsii [7.9% of dietary dry matter (DM)] or saponins from Yucca schidigera (1.1% of DM). The fourth treatment consisted of dried sainfoin (Onobrychis viciifolia), a CT-containing forage legume, in an amount also providing 7.9% CT in dietary DM. All diets were supplemented with linseed oil at a level contributing 60% of total dietary ALA in all treatments. Diets were incubated for 10 d (n=4) in the rumen simulation technique system, using the last 5 d for statistical evaluation. Fatty acids were analyzed in feed, feed residues, incubation fluid, and its effluent. Data were subjected to ANOVA considering diet and experimental run as main effects. Both CT treatments reduced ruminal fiber and crude protein degradation, and lowered incubation fluid ammonia concentration. Only the CT extract suppressed methane formation and shifted microbial populations toward bacteria at cost of protozoa. The saponin extract remained without clear effects on fermentation characteristics except for increased protozoal counts. The extent of ALA biohydrogenation was 20% less with the CT plant, but this probably resulted from reduced organic matter degradability rather than from an inhibition of biohydrogenation. After incubation analysis of incubation fluid effluent and feed residues showed a considerable proportion of the 3 biohydrogenation intermediates, cis-9, trans-11, cis-15 C18:3, trans-11, cis-15 C18:2, and trans-11 C18:1, which did not occur in the initial feeds. Only the CT-extract diet led to a different profile in the effluent compared with the control diet with trans-11 C18:1 being considerably increased at cost of C18:0. This could have been achieved by suppressing protozoa and enhancing the bacterial population, thus removing potential microbes involved in biohydrogenation and increasing competition between bacteria involved in biohydrogenation and others. The elevation of trans-11 C18:1 as the precursor of cis-9, trans-11 conjugated linoleic acid formed in body tissue and mammary gland is probably favorable from a human health point of view.

Evaluation of a stratified continuous rumen incubation system

We present a continuous culture system based on one developed by [Teather, R.M., Sauer, F.D., 1988. A naturally compartmented rumen simulation system for the continuous culture of rumen bacteria and protozoa. J. Dairy Sci. 71, 666–673]. In which programmable stirring speeds allow a raft mat to form similar to that found in the rumen to produce a de facto dual flow system with different liquid and solid turnover rates that helps to maintain the protozoal population. Novel features include a non-clogging outflow, and a computer controlled, compact feeding system able to deliver ground substrates or pellets. The system comprises 12 thermostat controlled fermenter vessels with a working volume of 1 L each. The fermenters were primed with rumen fluid from two cows and continuously provided with the same diet as the donor animals (15 g/day for 3 days then 10 g/day) and artificial saliva (1 L/day). The concentration of short chain fatty acids (SCFA), microbial ribosomal RNA (rRNA) concentrations and total enzymatic activities were all lower in the fermenter than in the donor animals, but the proportions of SCFA remained the same. There was no difference in specific cellulase activity, but specific xylanase activity and total protozoal counts were significantly higher in the fermenters. The proportions of Bacteria, and Eukaryota (i.e., protozoa and fungi) were similar in the fermenters compared to the donor animal, but the proportion of the Archaea (methanogens) was increased. Total rRNA from cellulolytic organisms ( Chytridiomycetes, Fibrobacter sp. Ruminococcus albus and R. flavefaciens) was similar to that in the donor animals. However, the composition of the cellulolytic community changed with the proportion of R. albus and the Chytridiomycetes increasing while the proportion of R. flavefaciens decreased, and the genus Fibrobacter was unchanged. Despite these differences, this in vitro system has potential to study effects of feed components on fermentation parameters. Whether the system will prove useful for modelling changes in the composition of the rumen microbial community needs to be evaluated further.

Effect of DL-malic acid supplementation on feed intake, methane emission, and rumen fermentation in beef cattle

The objective of this study was to determine the effect of dietary concentration of dl-malic acid (MA) on DMI, CH(4) emission, and rumen fermentation in beef cattle. Two Latin square experiments were conducted. In Exp. 1, six beef heifers (19 +/- 1 mo old) were assigned in a duplicated Latin square to 1 of 3 dietary concentrations of MA on a DMI basis (0%, MA-0; 3.75%, MA-3.75; or 7.5%, MA-7.5) over 3 periods. In Exp. 2, four rumen-fistulated steers (48 +/- 1 mo old) were assigned to 1 of 4 dietary concentrations of MA (0%, MA-0; 2.5%, MA-2.5; 5.0%, MA-5.0; or 7.5%, MA-7.5) on a DMI basis, over 4 periods. Both experimental diets consisted of grass silage and pelleted concentrate (containing MA). Silage was fed ad libitum once daily (a.m.), whereas concentrate was fed twice daily (a.m. and p.m.) with the aim of achieving a total DMI of 40:60 silage:concentrate. In both Exp. 1 and 2, experimental periods consisted of 28 d, incorporating a 13-d acclimatization, a 5-d measurement period, and a 10-d washout period. In Exp. 1, enteric CH(4), feed apparent digestibility, and feed intake were measured over the 5-d measurement period. In Exp. 2, rumen fluid was collected on d 16 to 18, immediately before (a.m.) feeding and 2, 4, 6, and 8 h thereafter. Rumen pH was determined and samples were taken for protozoa count, VFA, and ammonia analysis. Enteric CH(4) emissions were estimated by using the sulfur hexafluoride tracer technique and feed apparent digestibility was estimated by using chromic oxide as an external marker for fecal output. In Exp. 1, increasing dietary MA led to a linear decrease in total DMI (P < 0.001) and total daily CH(4) emissions (P < 0.001). Compared with the control diet, the greatest concentration of MA decreased total daily CH(4) emissions by 16%, which corresponded to a 9% reduction per unit of DMI. Similarly, in Exp. 2, inclusion of MA reduced DMI in a linear (P = 0.002) and quadratic (P < 0.001) fashion. Increasing dietary MA led to a linear decrease in molar proportion of acetic (P = 0.004) and butyric acids (P < 0.001) and an increase in propionic acid (P < 0.001). Ruminal pH tended to increase (P = 0.10) with increasing dietary MA. Dietary inclusion of MA led to a linear (P = 0.01) decrease in protozoa numbers. Increasing supplementation with MA decreased CH(4) emissions, but DMI was also decreased, which could have potentially negative effects on animal performance.

Effect of different liquid cultures of live yeast strains on performance, ruminal fermentation and microbial protein synthesis in lambs

Three yeast strains, Kluyveromyces marximanus NRRL-3234 (KM), Saccharomyces cerevisiae NCDC-42 (SC) and Saccharomyces uvarum ATCC-9080 (SU), and a mixed culture (1:1:1 ratio) were evaluated for their value as probiotics in lamb feeding in two experiment. In experiment I and II, 20 and 30 pre-weaner lambs were fed for 63 and 60 days in two and three equal groups respectively. All lambs were offered ad libitum a creep mixture and Zizyphus nummularia leaves, and yeasts were dosed orally. In experiment I, one group received no yeast, the other of the mixed culture (1.5-2 x 10(10) live cells/ml). In experiment II, yeast cultivation was modified yielding 1.5-2 x 10(13) live cells/ml. Lambs of the three experimental groups received 1 ml/kg live weight of one of the individual yeasts. Feed intake did not differ among groups of both experiments with the exception of SC-supplemented lambs in experiment II which showed a trend to higher intakes per kg metabolic body weight and in percentage of body weight when compared with KM- and SU-supplemented lambs. Supplementation of the mixed yeast culture had no effect on intakes of digestible crude protein and metabolisable energy, nutrient digestibility, nitrogen balance and rumen fermentation characteristics (pH, ammonia, volatile fatty acid concentration, protozoa count) and urinary allantoin as an indicator of microbial protein synthesis. The same was true for comparisons in experiment II except ciliate protozoa counts, which showed a trend to be the highest with SU and the lowest with SC. The results of present study show that the response of lambs to supplemented live yeast cultures is inconsistent, as it lacked to have an effect in the present study, and that differences among strains were small, even when supplemented at a much higher live cell count.