Influence of exogenous fibrolytic enzyme level and incubation pH on the in vitro ruminal fermentation of alfalfa stems

A series of in vitro experiments was carried out to examine the impact of enzyme application rate and incubation medium pH on the rate and extent of fermentation of alfalfa stems. In Experiment 1, a commercial enzyme product (Liquicell 2500, Specialty Enzyme and Biochemicals, Fresno, CA, USA) was added to alfalfa stems at six levels: 0, 0.51, 1.02, 2.55, 5.1, and 25.5 mu l/g (control and L1-L5, respectively) to forage DM in a completely randomized design, with a factorial arrangement of treatments. Rate and extent of fermentation and apparent organic matter degradation (OMD) were determined in vitro, using a gas production technique. Addition of enzyme linearly increased (P < 0.01) gas production for up to 12 h (68.9, 70.9, 67.6, 67.9, 71.9, and 74.9 ml/g OM for control, L1-L5, respectively) and OMD for up to 19 h incubation (0.425, 0.444, 0.433, 0.446, 0.443, and 0.451 for control, L1-L5, respectively), but no increases (P > 0.05) were detected thereafter. In Experiment 2, the effect of the same enzyme as used previously (added at 0.51 mu l/g forage DM, directly into the incubation medium), and buffer pH were examined using the ANKOM system, in a completely randomized design. Incubation medium pH was altered using 1 M citric acid, in order to obtain target initial pH values of 6.8 (control, no citric acid added), 6.2, 5.8, and 5.4. Actual initial pH values achieved were 6.72, 6.50, 6.20, and 5.72. Lowering the pH decreased (P < 0.01) dry matter disappearance (DMD) at 18 h incubation (0.339, 0.341, 0.314, and 0.291 for 6.72, 6.50, 6.20, and 5.72, respectively), whereas enzyme addition increased (P < 0.05) DMD at 24 h (0.363 versus 0.387 for control and enzyme-treated, respectively). Addition of enzyme increased (P < 0.05) neutral detergent fibre (NDF), acid detergent fibre (ADF), and hemicellulose (HC) degradation at pH 6.50 (0.077 versus 0.117; 0.020 versus 0.051; 0.217 versus 0.270 for control and enzyme-treated NDF, ADF and hemicellulose degradation, respectively) and 6.72 (0.091 versus 0.134; 0.041 versus 0.079; 0.205 versus 0.261 for control and enzyme-treated NDF, ADF and HC degradation, respectively). It is concluded that the positive effects of this enzyme product were independent of the pre-treatment period, but pH influenced the responses to enzyme supplementation. Under the conditions of this experiment, exogenous fibrolytic enzymes seemed to work better at close to neutrality ruminal pH conditions. (C) 2006 Elsevier B.V. All rights reserved.

Chemical composition and in vitro ruminal fermentation of common tree forages in the semi-arid rangelands of Swaziland

Browse plants play an important role in providing feed for livestock in semi-arid rangelands of Africa. Chemical composition and in vitro ruminal fermentation of leaves collected from Acacia burkei, Acacia tortilis, Acacia nilotica, Dichrostachys cinerea and Ehretia obtusifolia in communal grazing lands in the lowveld of Swaziland is presented. Leaves were collected from trees located on two soil types (i.e., lithosol and vertisol) in the communal land but it had no effect on the chemical composition of tree leaves. The NDFom and ADFom content were highest in D. cinerea and A. burkei and lowest in E. obtusifolia and A. nilotica. Crude protein (CP) contents ranged between 108 g/kg and 122 g/kg DM. D. cinerea had the highest Ca and Mg content, while A. tortilis had the lowest. There were marked variations in K level amongst browse species, with A. tortilis (9.1 g/kg DM) having the highest value. The P, Zn and Fe did not differ between browse species. Soil type and tree species interaction impacted in vitro fermentation parameters. Extent of fermentation, as measured by 48 h cumulative gas production, and organic matter degradability was highest in E. obtusifolia leaves and lowest in D. cinerea leaves within soil type. Fermentation efficiency, as measured by partitioning factors, was highest in A. nilotica leaves. Leaves of E. obtusifolia could be a valuable supplementary feedstuff for ruminant livestock due to its in vitro fermentation characteristics as well as low fibre and moderate CP levels. (c) 2007 Elsevier B.V. All rights reserved.

Intake, water consumption, ruminal fermentation, and stress response of beef heifers fed after different lengths of delays in the daily feed delivery time

Four rumen-fistulated Holstein heifers (134 +/- 1 kg initial BW) were used in a 4 x 4 Latin square design to determine the effects of delaying daily feed delivery time on intake, ruminal fermentation, behavior, and stress response. Each 3-wk experimental period was preceded by 1 wk in which all animals were fed at 0800 h. Feed bunks were cleaned at 0745 h and feed offered at 0800 h (T0, no delay), 0900 (T1), 1000 (T2), and 1100 (T3) from d1 to 21 with measurements taken during wk 1 and 3. Heifers were able to see each other at all times. Concentrate and barley straw were offered in separate compartments of the feed bunks, once daily and for ad libitum intake. Ruminal pH and saliva cortisol concentrations were measured at 0, 4, 8, and 12 h postfeeding on d 3 and 17 of each experimental period. Fecal glucocorticoid metabolites were measured on d 17. Increasing length of delay in daily feed delivery time resulted in a quadratic response in concentrate DMI (low in T1 and T2; P = 0.002), whereas straw DMI was greatest in T1 and T3 (cubic P = 0.03). Treatments affected the distribution of DMI within the day with a linear decrease observed between 0800 and 1200 h but a linear increase during nighttimes (2000 to 0800 h), whereas T1 and T2 had reduced DMI between 1200 and 1600 h (quadratic P = 0.04). Water consumption (L/d) was not affected but decreased linearly when expressed as liters per kilogram of DMI (P = 0.01). Meal length was greatest and eating rate slowest in T1 and T2 (quadratic P <= 0.001). Size of the first meal after feed delivery was reduced in T1 on d 1 (cubic P = 0.05) and decreased linearly on d 2 (P = 0.01) after change. Concentrate eating and drinking time (shortest in T1) and straw eating time (longest in T1) followed a cubic trend (P = 0.02). Time spent lying down was shortest and ruminating in standing position longest in T1 and T2. Delay of feeding time resulted in greater daily maximum salivary cortisol concentration (quadratic P = 0.04), which was greatest at 0 h in T1 and at 12 h after feeding in T2 (P < 0.05). Daily mean fecal glucocorticoid metabolites were greatest in T1 and T3 (cubic P = 0.04). Ruminal pH showed a treatment effect at wk 1 because of increased values in T1 and T3 (cubic P = 0.01). Delaying feed delivery time was not detrimental for rumen function because a stress response was triggered, which led to reduced concentrate intake, eating rate, and size of first meal, and increased straw intake. Increased salivary cortisol suggests that animal welfare is compromised.

Effects of adding a spray-dried polyclonal antibody preparation on ruminal fermentation patterns and digestibility of cows fed high concentrate diets

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of adding polyclonal antibody preparations on ruminal fermentation patterns and digestibility of cows fed different energy sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Replacement of dicalcium phosphate by rock phosphate in cattle diets: nutrients digestibility, plasma parameters, ruminal fermentation and microbial synthesis efficiency

Este estudo foi conduzido para avaliar os efeitos da substituição do fosfato bicálcico pelo fosfato de rocha na dieta de bovinos em crescimento. Foram determinados a digestibilidade aparente das dietas, a absorção aparente do fósforo, cálcio e flúor, o pH ruminal, a concentração de amônia ruminal, a eficiência microbiana e o fósforo no plasma utilizando-se cinco bovinos da raça Holandesa Preto-e-Branco, fistulados, pesando entre 275 e 283 kg. O delineamento estatístico foi um quadrado latino 5 × 5 e as dietas consistiram de 0, 25, 50, 75 e 100% de substituição do fosfato bicálcico pelo fosfato de rocha no suplemento mineral. A adição de fosfato de rocha nas dietas ocasionou aumento linear na ingestão, no fluxo omasal, no fluxo fecal e no desaparecimento total do flúor. As dietas não diferiram quanto à absorção aparente do cálcio, assim como em relação à ingestão, excreção, digestão e digestibilidades aparentes parcial e total da matéria seca, matéria orgânica, proteína bruta, fibra em detergente neutro e carboidratos não-fibrosos. O fósforo no plasma não foi influenciado pelos tratamentos e a média foi de 5,93 mg/dL. Não houve diferença para o pH ruminal e concentração de amônia ruminal. A substituição do fosfato bicálcico não afetou a síntese microbiana aparente e verdadeira de proteína. A total substituição do fosfato bicálcico pelo fosfato de rocha em suplementos minerais em bovinos em crescimento não afetou o ambiente ruminal e a síntese de proteína no rúmen. Assim, a substituição do fosfato bicálcico em dietas para bovinos em crescimento diminui a absorção de fósforo e deveria ser vista com cuidado dependendo dos requerimentos.

Methane emissions and estimates of ruminal fermentation parameters in beef cattle fed different dietary concentrate levels

Using sorghum silage, the effect of roughage/concentrate ratios was evaluated on nutrient intake, digestibility, ruminal parameters and methane production by beef cattle. Three treatments (0, 30 and 60% of concentrate in DM of the diet) were distributed in three Latin squares, with nine animals and three periods. Dry matter intake increased as the grain concentration in diet increased; pH showed opposite behavior. Methane emissions were lower for animals fed the diet exclusively with sorghum silage as compared with those fed 30% of concentrate, but was similar to that of animals receiving 60% of concentrate. Losses of ingested gross energy as methane were reduced by 33% when grain concentration was increased in the diet. Concentrations of propionic and butyric acids were greater in diets with grain concentrate; acetic acid concentration was not affected. Concentrate in diet increases available energy for the metabolism, measured by lower losses of ingested gross energy as ruminal methane. © 2013 Sociedade Brasileira de Zootecnia.

Effects of feeding corn silage inoculated with microbial additives on the ruminal fermentation, microbial protein yield, and growth performance of lambs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of supplementation frequency on the ruminal fermentation and enteric methane production of beef cattle grazing in tropical pastures

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparative in vitro evaluation of forage legumes (prosopis, acacia, atriplex, and leucaena) on ruminal fermentation and methanogenesis

Two experiments in vitro were conducted to evaluate four Egyptian forage legume browses, i.e., leaves of prosopis (Prosopis juliflora), acacia (Acacia saligna), atriplex (A triplex halimus), and leucaena (Leucaena leucocephala), in comparison with Tifton (Cynodon sp.) grass hay for their gas production, methanogenic potential, and ruminal fermentation using a semi-automatic system for gas production (first experiment) and for ruminal and post ruminal protein degradability (second experiment). Acacia and leucaena showed pronounced methane inhibition compared with Tifton, while prosopis and leucaena decreased the acetate:propionate ratio (P<0.01). Acacia and leucaena presented a lower (P<0.01) ruminal NH3-N concentration associated with the decreasing (P<0.01) ruminal protein degradability. Leucaena, however, showed higher (P<0.01) intestinal protein digestibility than acacia. This study suggests that the potential methanogenic properties of leguminous browses may be related not only to tannin content, but also to other factors.

Effects of flavomycin on ruminal fermentation, In situ degradability and In vivo digestibility in bovine fed sugarcane diets

Problem statement: The aim of the present study was to characterize and differentiate the effects of addition of flavomycin or monensin on ruminal fermentation and degradability as well as on total digestibility in bovine. Approach: Twelve non-pregnant and non-lactating cows (736 kg of BW) were randomly assigned to three treatments: control, flavomycin (20 mg animal-1 day-1) and monensin (300 mg animal-1 day-1). The trial lasted 21 days. The last 10 days were used for external marker administration (15 g of chromic oxide animal-1 day-1). The last 5 days of the trial were used for feces collection and evaluation of corn grain, soybean meal or sugarcane ruminal degradability and the 21st day was used for ruminal fluid sampling. Results: Monensin increased 27.2%, on average, propionate molar proportion at 0, 4, 6, 8, 10 and 12 h after feeding, compared to control and flavomycin groups. When compared to control, flavomycin reduced the degradation rate of soybean meal CP in 31.0%, decreasing the effective degradability when passage rates of 5 and 8% h-1 were used. Dry matter intake, pH, total Short Chain Fatty Acids (tSCFA) or ammoniacal Nitrogen (NH3-N) concentration were not influenced by the addition of either antibiotics. Effective degradability of sugarcane NDF was not influenced by the use of either antibiotic; neither were the TDN nor the digestibility of DM, CP, EE, NFE, ADF, NDF, GE or starch of the diet. Conclusion/Recommendations: In the present study, it was possible to show the beneficial effects of monensin but not of flavomycin, on rumen fermentation

In vitro ruminal fermentation of diets containing wheat straw and date pits as forage.

The pH, VFA concentration, total gas and met hane production were determined in the rumen of four Sicilo- Sarde rams fitted with permanent canulas. Rams received a ration that included 1.5 kg DM of oat hay and were supplemented with one of four concentrates: CC (10% barley, 43.3% corn, 25% wheat bran, 17.7% soybean meal, 4% sheep Vitamin and Mineral Mixture (VMM)), SC (66% white sorghum, 30% faba, 4% sheep VMM); TC (71% triticale, 18% faba, 7%, soybean meal, 4% VMM) or BC (71.5% barley, 17.5% faba, 7% soybean meal and 4% VMM). 50 ml samples were taken before, 2, 5 and 8 hours after the morning meal. Total gas was determined on rumen content before the morning meal. The rumen pH was statistically different (P<0.05) before and 2 hours after the morning meal among concentrates feed. It was in favour of TC and BC (P<0.05) concentrates but was comparable at the end of the day. The concentration of VFA was significantly higher (P<0.05) for diets TC and BC following the meal and became comparable among concentrates thereafter. The proportion of acetate and butyrate acids evolved in the same way during the day regardless of the regimen. The total volu me of gas was different (P<0.05) among diets, the BC showed the highest value (87.00±17.29 ml) while the lowest value was found in the TC concentrate (56.58±13.06 ml). The CH4 production for the BC was significantly different (P<0.05) from that of TC. Quantities produced by the CC and SC were similar (22.08±4.18vs . 21.16±3.21).

The influence of diet on the effectiveness of garlic oil and cinnamaldehyde to manipulate in vitro ruminal fermentation and methane production.

The objective of this study was to evaluate the effects of increasing doses [0 (control: CON), 20, 60, 180 and 540 mg/L incubation medium] of garlic oil (GO) and cinnamaldehyde (CIN) on in vitro ruminal fermentation of two diets. Batch cultures of mixed ruminal microorganisms were inoculated with ruminal fluid from four sheep fed a medium-concentrate diet (MC; 50 : 50 alfalfa hay : concentrate) or four sheep fed a high-concentrate diet (HC; 15 : 85 barley straw : concentrate). Diets MC and HC were representative of those fed to dairy and fattening ruminants, respectively. Samples of each diet were used as incubation substrates for the corresponding inoculum, and the incubation was repeated on 4 different days (four replicates per experimental treatment). There were GO × diet-type and CIN × diet-type interactions (P < 0.001–0.05) for many of the parameters determined, indicating different effects of both oils depending on the diet type. In general, effects of GO were more pronounced for MC compared with HC diet. Supplementation of GO did not affect (P > 0.05) total volatile fatty acid (VFA) production at any dose. For MC diet, GO at 60, 180 and 540 mg/L decreased (P < 0.05) molar proportion of acetate (608, 569 and 547 mmol/mol total VFA, respectively), and increased (P < 0.05) propionate proportion (233, 256 and 268 mmol/mol total VFA, respectively), compared with CON values (629 and 215 mmol/mol total VFA for acetate and propionate, respectively). A minimum dose of 180 mg of GO/L was required to produce similar modifications in acetate and propionate proportions with HC diet, but no effects (P > 0.05) on butyrate proportion were detected. Methane/VFA ratio was reduced (P < 0.05) by GO at 60, 180 and 540 mg/L for MC diet (0.23, 0.16 and 0.10 mol/mol, respectively), and by GO at 20, 60, 180 and 540 mg/L for HC diet (0.19, 0.19, 0.16 and 0.08 mol/mol, respectively), compared with CON (0.26 and 0.21 mol/mol for MC and HC diets, respectively). No effects (P = 0.16–0.85) of GO on final pH and concentrations of NH3-N and lactate were detected. For both diet types, the highest CIN dose decreased (P < 0.05) production of total VFA, gas and methane, which would indicate an inhibition of fermentation. Compared with CON, CIN at 180 mg/L increased (P < 0.05) acetate proportion for the MC (629 and 644 mmol/mol total VFA for CON and CIN, respectively) and HC (525 and 540 mmol/mol total VFA, respectively) diets, without affecting the proportions of any other VFA or total VFA production. Whereas for MC diet CIN at 60 and 180 mg/L decreased (P < 0.05) NH3-N concentrations compared with CON, only a trend (P < 0.10) was observed for CIN at 180 mg/L with the HC diet. Supplementation of CIN up to 180 mg/L did not affect (P = 0.18–0.99) lactate concentrations and production of gas and methane for any diet. The results show that effectiveness of GO and CIN to modify ruminal fermentation may depend on diet type, which would have practical implications if they are confirmed in vivo.

Efeito de Diferentes Volumosos sobre a Degradabilidade in situ de Nutrientes e Variáveis da Fermentação Ruminal, Mediante a Aplicação da Somatotropina Bovina

The objective of this work was to evaluate the effect of diets with different forages (sugarcane, corn silage, hydrolyzed bagasse and sugarcane + corn silage) on the ruminal Fermentation and ruminal nutrients degradability, by the application of bovine somatotropin. Three bovines with ruminal cannulas were used in a factorial scheme (4 x 2). The pH, number of protozoa, ammonia and volatile fatty acids concentration were quantified. The potential and the effective degradability and degradation rate of dry matter and crude protein in each diet were evaluated, beyond gross energy disappearance. There were differences among diets on the effective degradability of dry matter and potential degradability, effective degradability and potentially degradable fraction of crude protein, being verified the same for the rBST application. There was effect of different forages on the variables of ruminal Fermentation. The treatments did not affect the gross energy disappearance. The rBST application did not affect the variables of ruminal fermentation.

Fermentação ruminal de ovinos alimentados com alto concentrado e grãos de girassol ou gordura protegida

Seis ovinos machos, não castrados, da raça Santa Inês, com média de peso de 30kg, fistulados no rúmen, foram distribuídos em delineamento de quadrado-latino duplo (3x3). Três períodos e três dietas, uma controle, sem inclusão de fonte de lipídio, e duas com inclusão de grãos de girassol ou gordura protegida, foram testados quanto aos parâmetros ruminais. Foram verificadas diferenças (P<0,05) entre as dietas quanto à concentração ruminal de amônia (18mg/dL), mas não houve efeito sobre o pH (6,1), a produção total de ácidos graxos de cadeia curta (98mM), a proporção de acetato (66,4%), de propionato (20%) e de butirato (13%) e sobre a razão acetato:propionato (3,2:1). As bactérias sólido-aderidas isoladas do conteúdo ruminal dos animais recebendo a dieta-controle apresentaram maior teor de nitrogênio (10,7%) que as das dietas com gordura protegida (9,8%) ou com grãos de girassol (9,1%). A produção de nitrogênio pelas bactérias sólido-aderidas da dieta-controle (170mg/g) não diferiu da dieta com grãos de girassol (153mg/kg) ou com gordura protegida (160mg/kg). A inclusão de grãos de girassol ou gordura protegida na dieta com alto concentrado para ovinos propiciou ambiente adequado para fermentação ruminal.