The stimulatory effects of cattle rumen liquor and digested sludge supernatant on the methane fermentation /

Thesis (M.S.)--University of Illinois at Urbana-Champaign, 1965.

The effectiveness of adapted rumen fluid versus PEG to ferment tannin-containing substrates in vitro

This study investigated the potential of the goat's ruminal adaptation to reduce the negative effect of tannins on in vitro fermentation. Rumen fluid was obtained from goats fed a mixture of tannin-containing tree fruits (adapted rumen fluid) or tannin-free commercial protein supplements (unadapted rumen fluid) for 85 days. Dry, mature fruits of Acacia nilotica, Acacia erubescens, Acacia erioloba, Dichrostachys cinerea and Piliostigma thonningii were used as substrates for the in vitro fermentation. The effectiveness of adapted rumen fluid to ferment tannin-containing substrates was compared to the extent of fermentation when tannins were inactivated with polyethylene glycol (PEG), a known tannin-binding agent. Adapted rumen fluid (P < 0.05) increased gas production from all five substrates between 15.8% and 73.7%. In A. nilotica, D. cinerea and P thonningii, this increase was less than that obtained through PEG treatment. When PEG was added to adapted rumen fluid a further improvement in extent of fermentation was observed in four out of the five fruit samples. The largest PEG effect when incubated with adapted rumen fluid was observed in A. nilotica (43.1%) and D. cinerea (42.9%) fruits. It is concluded that some tannin-rich feedstuffs may still benefit from treatment even when these are offered to adapted animals. (c) 2006 Elsevier B.V. All rights reserved.

Relationship between condensed tannin structures and their ability to precipitate feed proteins in the rumen

Abstract BACKGROUND Tannins can bind to and precipitate protein by forming insoluble complexes resistant to fermentation and with a positive effect on protein utilisation by ruminants. Three protein types, Rubisco, rapeseed protein and bovine serum albumin (a single high-molecular weight protein), were used to test the effects of increasing concentrations of structurally different condensed tannins on protein solubility/precipitation. RESULTS Protein type (PT) influenced solubility after addition of condensed tannins (P < 0.001) in the order: Rubisco < rapeseed < BSA (P < 0.05). The type of condensed tannin (CT) affected protein solubility (P = 0.001) with a CT × PT interaction (P = 0.001). Mean degree of polymerisation, proportions of cis- versus trans-flavanol subunits or prodelphinidins versus procyanidins among CTs could not explain precipitation capacities. Increasing tannin concentration decreased protein solubility (P < 0.001) with a PT × CT concentration interaction. The proportion of low-molecular weight rapeseed proteins remaining in solution increased with CT concentration but not with Rubisco. CONCLUSIONS Results of this study suggest that PT and CT type are both of importance for protein precipitation but that the CT structures investigated did not allow identification of parameters that contribute most to precipitation. It is possible that the three-dimensional structures of tannins and proteins may be more important factors in tannin–protein interactions. © 2013 Society of Chemical Industry

The effect of adding stinging nettle (Urtica dioica) haylage to a total mixed ration on performance and rumen function of lactating dairy cows

In vitro studies found that inclusion of dried stinging nettle (Urtica dioica) at 100 mg/g dry matter (DM) increased the pH of a rumen fluid inoculated fermentation buffer by 30% and the effect was persistent for 7 days. Our objective was to evaluate the effects of adding stinging nettle haylage to a total mixed ration on feed intake, eating and rumination activity, rumen pH, milk yield, and milk composition of lactating dairy cows. Six lactating Holstein-Friesian cows were used in a replicated 3 × 3 Latin Square design experiment with 3 treatments and 3 week periods. Treatments were a control (C) high-starch (311 g/kg DM) total mixed ration diet and two treatment diets containing 50 (N5) and 100 (N10) g nettle haylage (DM/kg) as a replacement for ryegrass silage (Lolium perenne). There was an increase (linear, P < 0.010) in the proportion of large particles and a reduction in medium (linear, P = 0.045) and fine particles (linear, P = 0.026) in the diet offered with increasing nettle inclusion. A numerical decrease (linear, P = 0.106) in DM intake (DMI) was observed as nettle inclusion in the diet increased. Milk yield averaged 20.3 kg/day and was not affected by diet. There was a decrease (quadratic, P = 0.01) in the time animals spent ruminating as nettle inclusion in the diet increased, in spite of an increase in the number of boli produced daily for the N5 diet (quadratic, P = 0.031). Animals fed the N10 diet spent less time with a rumen pH below 5.5 (P < 0.05) than cows fed the N5 diet. Averaged over an 8.5 h sampling period, there were no changes in the concentration or proportions of acetate or propionate in the rumen, but feeding nettle haylage reduced the concentrations of n-butyrate (quadratic, P < 0.001), i-butyrate (linear, P < 0.009) and n-caproate (linear, P < 0.003). Milk and fat and protein corrected milk yield were not affected when nettles replaced ryegrass silage in the diet of lactating dairy cows, despite a numerical reduction in feed intake. Rumination activity was reduced by the addition of nettle haylage to the diet, which may reflect differences in fibre structure between the nettle haylage and ryegrass silage fed. Changes observed in rumen pH suggest potential benefits of feeding nettle haylage for reducing rumen acidosis. However, the extent to which these effects were due to the fermentability and structure of the nettle haylage compared to the ryegrass silage fed, or a bioactive component of the nettles, is not certain

Diversity of condensed tannin structures affects rumen in vitro methane production in sainfoin (Onobrychis viciifolia) accessions

Sainfoin is a non-bloating temperate forage legume with a moderate-to-high condensed tannin (CT) content. This study investigated whether the diversity of sainfoin accessions in terms of CT structures and contents could be related to rumen in vitro gas and methane (CH4) production and fermentation characteristics. The aim was to identify promising accessions for future investigations. Accessions differed (P < 0·0001) in terms of total gas and CH4 productions. Fermentation kinetics (i.e. parameters describing the shape of the gas production curve and half-time gas production) for CH4 production were influenced by accession (P ≤ 0·038), but not by PEG. Accession, PEG and time affected (P < 0·001) CH4 production, but accession and PEG interaction showed only a tendency (P = 0·08). Increase in CH4 due to PEG addition was not related to CT content. Further analysis of the relationships among multiple traits (nutritional composition, CT structure and CH4 production) using principal component analysis (PCA) based on optimally weighted variables revealed differences among accessions. The first two principal component axes, PC1 (57·6%) and PC2 (18·4%), explained 76·0% of the total variation among accessions. Loading of biplots derived from both PCAs made it possible to establish a relationship between the ratio of prodelphinidin:procyanidin (PD:PC) tannins and CH4 production in some accessions. The PD:PC ratio seems to be an important source of variation that is negatively related to CH4 production. These results suggested that sainfoin accessions collected from across the world exhibited substantial variation in terms of their effects on rumen in vitro CH4 production, revealing some promising accessions for future investigations.

Impact of variation in structure of condensed tannins from sainfoin (Onobrychis viciifolia) on in vitro ruminal methan production and fermentation characteristics

Our study investigated the effects of condensed tannins (CT) on rumen in vitro methane (CH4) production and fermentation characteristics by incubating lucerne in buffered rumen fluid in combination with different CT extracts at 0 (control), 40, 80 and 120 g CT/kg of substrate DM. Condensed tannins were extracted from four sainfoin accessions: Rees ‘A’, CPI63763, Cotswold Common and CPI63767. Gas production (GP) was measured using a fully automated GP apparatus with CH4 measured at distinct time points. Condensed tannins differed substantially in terms of polymer size and varied from 13 (Rees ‘A’) to 73 (CPI63767) mean degree of polymerization, but had relatively similar characteristics in terms of CT content, procyanidin: prodelphinidin (PC: PD) and cis:trans ratios. Compared to control, addition of CT from CPI63767 and CPI63763 at 80 and 120 g CT/kg of substrate DM reduced CH4 by 43% and 65%, and by 23% and 57%, respectively, after 24-h incubation. Similarly, CT from Rees ‘A’ and Cotswold Common reduced CH4 by 26% and 46%, and by 28% and 46% respectively. Addition of increasing level of CT linearly reduced the maximum rates of GP and CH4 production, and the estimated in vitro organic matter digestibility. There was a negative linear and quadratic (p < 0.01) relation between CT concentration and total volatile fatty acid (VFA) production. Inclusion of 80 and 120 g CT/kg of substrate DM reduced (p < 0.001) branched-chain VFA production and acetate: propionate ratio and was lowest for CPI63767. A decrease in proteolytic activity as indirectly shown by a change in VFA composition favouring a shift towards propionate and reduction in branched-chain VFA production varied with type of CT and was highest for CPI63767. In conclusion, these results suggest that tannin polymer size is an important factor affecting in vitro CH4 production which may be linked to the CT interaction with dietary substrate or microbial cells.

Structural features of condensed tannins affect in vitro ruminal methane production and fermentation characteristics

An in vitro study was conducted to investigate the effects of condensed tannins (CT) structural properties, i.e. average polymer size (or mean degree of polymerization); percentage of cis flavan-3-ols and percentage of prodelphinidins in CT extracts on methane production (CH4) and fermentation characteristics. CT were extracted from eight plants in order to obtain different CT types: black currant leaves, goat willow leaves, goat willow twigs, pine bark, red currant leaves, sainfoin plants, weeping willow catkins and white clover flowers. They were analysed for CT content and CT composition by thiolytic degradation, followed by HPLC analysis. Grass silage was used as a control substrate. Condensed tannins were added to the substrate at a concentration of 40 g/kg, with or without polyethylene glycol (+ or −PEG 6000 treatment) to inactivate tannins, and then incubated for 72 h in mixed buffered rumen fluid from three different lactating dairy cows per run. Total cumulative gas production (GP) was measured by an automated gas production system. During the incubation, 12 gas samples (10 μl) were collected from each bottle headspace at 0, 2, 4, 6, 8, 12, 24, 30, 36, 48, 56 and 72 h of incubation and analyzed for CH4. A modified Michaelis–Menten model was fitted to the CH4 concentration patterns and model estimates were used to calculate total cumulative CH4 production (GPCH4). Total cumulative gas production and GPCH4 curves were fitted using biphasic and monophasic modified Michaelis-Menten models, respectively. Addition of PEG increased GP, GPCH4, and CH4 concentration compared to the −PEG treatment. All CT types reduced GPCH4 and CH4 concentration. All CT increased the half time of GP and GPCH4. Moreover, all CT decreased the maximum rate of fermentation for GPCH4 and rate of substrate degradation. The correlation between CT structure and GPCH4 and fermentation characteristics showed that the proportion of prodelphinidins within CT had the largest effect on fermentation characteristics, followed by average 27 polymer size and percentage of cis-flavan-3-ols.

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.

Effect of pelleting and pellet size of a concentrate for intensively reared beef cattle on in vitro fermentation by two different approaches

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of Aspergillus spp. exogenous fibrolytic enzymes on in vitro fermentation of tropical forages

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.

In vitro fermentation parameters and biohydrogenation of vegetable oils without glycerol

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Bioconversion of Cellulose to Hydrogen by dark fermentation

Hydrogen is known as a clean energy resource. The biological production of hydrogen has been attracting attention as an environmentally friendly processs that does not consume fossil fuels. Cellulosic plant and waste materials are potential resources for fermentative hydrogen production. Cellulose is a linear biopolymer of glucose molecules, connected by β-1,4-glycosidic bonds. Enzymatic hydrolysis of cellulose requires the presence of cellulase. The present study aimed to investigate the efficiency of acid pretreatment on ruminal fluid in order to enrich H2 producing bacteria consortia to enhance biohydrogen rate and substrate removal efficiency. In this study, fermentative hydrogen producers were enriched on cellulose (2g/L) in a modificated Del Nery medium (DNM) at 37ºC and initial pH 7.0 using rumen fluid (10% v/v) as inoculum. To increase the hydrogen production it was added cellulose (10mL) to the medium. The gas products (mainly H2 and CO2) was analyzed by gas chromatography (Shimadzu GC 2010) using a thermal conductivity detector. The volatile fatty acids and ethanol were also detected by GC using a flame ionization detector. Cellulose degradation was quantified by using the phenolsulfuric acid method. Analysis showed that the biogas produced from the anaerobic fermentation contained only hydrogen and carbon dioxide, without detectable methane after acid pretreatment test. On DNM the hydrogen production started with 4 h (5,3 x 105 mmol H2/L) of incubation, and the maximum H2 concentration was observed with 34 h (7,1 x 106 mmol H2/L) of incubation. During the process, it was observed a predominance of acetic acid and butyric acid as well as a low production of acetone, ethanol and nbutanol in all experimental phases. Butyrate accounted for more than 77% of total. As a result of the accumulation of volatile fatty acids (VFAs), the pH value in anaerobic digestion system was reduced to 4,0. On microscopy analyses there were observed rods with endospores. The batch anaerobic fermentation assays performed on anaerobic mixed inoculum from rumen fluid demonstrated the feasibility of H2 generation utilizing cellulose as substrate. Based on the results, it can be concluded that the acid treatment was efficient to inhibit the methanogenic archaea cells present in rumen fluid. The rumen fluid cells present a potential route in converting renewable biomass such as cellulose into hydrogen energy.

Biohidrogenação microbiana e o fluxo duodenal de ácidos graxos do rumen em novilhos de corte suplementados com fontes lipídicas em dietas contendo glicerina

Pós-graduação em Zootecnia - FCAV

Effects of Aspergillus spp. exogenous fibrolytic enzymes on in vitro fermentation of tropical forages

BACKGROUND: Cellulose and hemicellulose are quantitatively the most important structural carbohydrates present in ruminant diets. Rumen micro-organisms produce enzymes that catalyse their hydrolysis, but the complex network formed by structural carbohydrates and lignin reduces their digestibility and restricts efficient utilisation of feeds by ruminants. This study aimed to produce two enzymatic extracts, apply them in ruminant diets to determine the best levels for ruminal digestibility and evaluate their effects on in vitro digestibility. RESULTS: In experiment 1 a two-stage in vitro technique was used to examine the effects of different enzymatic levels of Aspergillus japonicus and Aspergillus terricola on tropical forages. Enzyme addition had minor effects on corn silage at the highest enzymatic level. In experiment 2 an in vitro gas production (GP) technique was applied to determine apparent in vitro organic matter digestibility and metabolisable energy. The addition of enzymes in GP showed interesting results. Good data were obtained using sugar cane and Tifton-85 hay supplemented with extracts of A. japonicus and A. terricola respectively. CONCLUSION: Overall, the study suggests that addition of crude extracts containing exogenous fibrolytic enzymes to ruminant diets enhances the effective utilisation of ruminant feedstuffs such as forages. Copyright (c) 2012 Society of Chemical Industry